Associazione Internazionale per la Ricerca Dental1 - Maurizio ...

UNIVERSITA’ DEGLI STUDI DI
FIRENZE
FACOLTA’ DI MEDICINA E
CHIRURGIA DIPARTIMENTO DI
ODONTOSTOMATOLOGIA
DIRETTORE: Professor ROMANO GRANDINI
ANNO ACCADEMICO 2006-2007
CORSO DI PERFEZIONAMENTO IN:
“Utilizzo delle nuove tecnologie in igiene dentale:
OZONO E LASER”
firenzelaser@aiola.it
AUTORI: Dottori C. CIGLIOLA, G. SCOZZI, M. VALENTI
RELATORE: Professor MAURIZIO MAGGIONI
dott Cosimo Cigliola
TESI N°
1

INDICE
1. OZONO, INTRODUZIONE: .…………………………………. 3-5
2. SCOPO DEL LAVORO:…………………………………………. 6
3. MATERIALI E METODI:……………………………………….. 6
4. ABSTRACT IN ITALIANO:……………………………………. 7-45
5. ENGLISCH ABSTRACT:……………………………………….. 46-147
6. LASER, INTRODUZIONE:……………………………………. 148-156
7. ENGLISCH ABSTRACT:……………………………….………. 157-204
8. ABSTRACT IN ITALIANO:……………………………………. 205-234
9. CONCLUSIONI:………………………………………….………. 235
dott Cosimo Cigliola
2

Ozono
Introduzione
L'ossigeno-ozono terapia è una metodica consolidata da anni di esperienze clinico-scientifiche e
diffusa in numerosi paesi.
Consente, in un’elevata percentuale di casi, di risolvere o migliorare significativamente, con
notevoli benefici personali e sociali, numerose patologie altrimenti fortemente invalidanti.
Storia
Nel 1781 M. Van Marum sospettò l'esistenza di un composto gassoso di odore pungente, nell'aria
attraversata da scintille elettriche. Nel 1840 Schönbein chiamò ozono (Dal Greco Odorare)
questo gas. Fu nel 1845 che la molecola di Ozono, venne prodotta per la prima volta dal
ricercatore Ginevrino Auguste De la Rive. Essa si comporta come un versatile pacchetto di
energia che produce molti effetti. Furono però J. C. Marignac, H. Becquerel ed E. Fremy a
stabilirne la natura di gas molto reattivo, mentre Soret ne determinò la formula.
Chimica
È un gas di colore azzurro, di odore forte e penetrante, pericoloso a respirarsi perché attacca le
mucose. Di densità 1,66, liquefa a 112 °C dando luogo a un liquido di colore azzurro indaco molto
instabile. Molto più solubile in acqua dell'ossigeno, viene disciolto in molti solventi organici, come il
cloroformio, l'etere, ecc. L'ozono è un ossidante: ossida a freddo lo iodio e quasi tutti i metalli, in
particolare il mercurio e l'argento; sposta il cloro, il bromo e
lo iodio dai loro composti con l'idrogeno e con i metalli; porta al
massimo stadio di ossidazione i composti dello zolfo,
del fosforo e dell'arsenico; trasforma a freddo l'ammoniaca in
nitrito e nitrato di ammonio; le sostanze organiche (sughero,
caucciù) vengono profondamente alterate. Per
scoprire le tracce di ozono, ci si serve di una speciale carta
ozonoscopica, detta di Houzeau: è una carta impregnata di ioduro di potassio, una metà della
quale è stata immersa nella salda d'amido e l'altra in una soluzione acida, e quindi rossa, di
tornasole; l'ozono, dando luogo con lo ioduro a iodio libero e idrossido di potassio, colora in
azzurro le due metà della carta; il cloro o il tetrossido di diazoto colorano invece solo la prima
metà, l'ammoniaca la seconda.
L'ozono è impiegato per il suo potere ossidante e battericida; serve in particolare per il rinnovo
dell'aria negli ambienti chiusi e per la sterilizzazione dell'acqua. Viene anche impiegato per la
sbianca dei tessuti, della cera, dell'amido, dell'avorio, per l'invecchiamento del vino e del legno.
L'ozono (O3) è la forma triatomica dell'ossigeno (O2).
In natura l'ozono si forma, sia mediante i raggi ultravioletti, sia mediante il lampo, che riesce a
dare l'energia necessaria affinché 3 molecole di ossigeno diventino 2 molecole di ozono:
dott Cosimo Cigliola
3

3O2 + = 2O3
Per uso medicale viene prodotto estemporaneamente al momento dell'uso
da appositi macchinari. L'O3 è un gas molto instabile e reattivo e deve
essere usato immediatamente dopo il prelievo. Si deve sempre evitare di
disperdere O3 nell'ambiente di lavoro perché è un gas irritante, dall'odore
pungente e caratteristico.
Il nostro olfatto avverte subito tracce di O3 anche quando la concentrazione
del gas è inferiore alla dose permessa, cioè di 0,2 mg/m 3 di aria per un
periodo lavorativo di 8 ore. Ciò per precisare che piccole fughe di O3 non
sono preoccupanti (ma sempre da evitare) perché l'olfatto già rileva concentrazioni di 0,02 mg/m 3
, che sono ben al di sotto della concentrazione tossica di 0,3 mg/m 3 come stabilito dalla CEE.
È quindi dannoso respirare ozono al di sopra di precise concentrazioni, ma è molto utile alla salute
per tutte le applicazioni descritte.
Azioni dell’ossigeno-ozono
L’ossigeno-ozono terapia è una tecnica dolce che sfrutta le potenzialità dell’ozono, combinato con
l’ossigeno, di stimolare e aumentare i meccanismi di protezione nei confronti della produzione di
radicali liberi (determinandone la riduzione) e di sostanze tossiche per le cellule. Il tutto
utilizzando una miscela composta per il 99.99997% da ossigeno e dal 0,00003% da ozono, gas
simile all’ossigeno ma molto più potente e attivo di quest’ultimo.
Attiva la circolazione in tutti i tessuti favorendo il rilascio dell’ossigeno agli stessi svolgendo così
diverse azioni: analgesica, antinfiammatoria, antivirale, antibatterica, antimicotica,
immunomodulante, stimolando la rigenerazione dei tessuti stessi.
Valori limite
La massima concentrazione di immissione per mezz'ora (valore MIK nel breve periodo) e pari ad un valore di
ozono di 120 µg / m 3 . Chi pero si espone non più di una volta la settimana e per non più di 30 minuti ad una
concentrazione di 400 µg / m 3 di ozono non arreca alcun danno alla propria salute. Non esiste alcun rischio per
la salute fino ad un valore di soglia di 110 µg/m 3 di ozono in media per otto ore. La Kommission der Deutschen
Forschungsgemein-schaft (commissione della comunità di ricerca tedesca) (DFG), dopo aver valutato la
letteratura specializzata, è giunta alla conclusione che: la protezione sul posto di lavoro potrebbe essere
rispettata "con un valore MAK di 200 µg/m 3 per 4 ore oppure di 100 µg /m 3 per 8 ore."(DFG, OZON
MAK/Classificazione, Supplemento del 1995, Consegna n. 21, 1995) Per la valutazione soggettiva e importante
che l'ozono venga percepito dall'olfatto gia ad un quinto (0,04 mg/m 3 ) del valore limite ammesso.
INDICAZIONI GENERALI SULL'USO DELL'OSSIGENO OZONO TERAPIA
Tale metodica, ormai consolidata da anni di esperienza clinico-scientifica e diffusa in numerosi
paesi, consente in una elevata percentuale di casi di risolvere o perlomeno di migliorare, con
notevoli benefici personali e sociali, numerose patologie, altrimenti fortemente invalidanti.
dott Cosimo Cigliola
4

Patologie in cui è indicata l’ossigeno-ozonoterapia
1. ortopedia, reumatologia e traumatologia: artrosi e artriti comprese le artriti reumatoide e
psoriasica, tendiniti (epicondiliti, epitrocleiti …), sindrome del tunnel carpale, discopatie sia
cervicali che lombari (cervicobrachialgie, lombosciatalgie )
2. disturbi della circolazione arteriosa, venosa e linfatica: vasculopatie periferiche di
origine venosa e arteriosa (postumi di flebite, insufficienza venosa periferica, microangiopatia
diabetica, ulcere periferiche su base vascolare e dismetabolica, linfedema).
3. turbe neurologiche: dai disturbi della memoria fino agli esiti di ischemia cerebrale, sclerosi a
placche, cefalee di varia natura (su base tensiva e vascolare).
4. senescenza con caduta della efficienza mentale e fisica, morbo di Alzheimer, morbo di
Parkinson, otosclerosi.
5. retinopatie: su base vascolare e dismetabolica, retinopatia maculare (degenerazione
maculare senile).
6. epatopatie: dalle semplici turbe della funzionalità epatica fino alle epatiti.
7. apparato gastrointestinale: gastriti, duodeniti, coliti (colon irritabile, morbo di Crohn, Colite
Ulcerosa).
8. apparato genito urinario : cistiti ricorrenti, vaginiti.
9. malattie dermatologiche: herpes simplex e zoster, eczemi acuti e cronici, dermatiti da
contatto, acne, psoriasi, micosi, afte.
10.patologie allergiche.
11. disordini metabolici : ipercolesterolemia e iperglicemia con le loro complicanze.
12. stati di deficienza immunitaria e patologie autoimmuni.
13. astenie su base funzionale e organica.
14. sostegno ai pazienti neoplastici con o senza metastasi azione palliativa.
Le applicazioni di ossigeno-ozono sono assolutamente innocue e prive di effetti collaterali né
allergiche, non presentano, in linea di massima, controindicazioni si tratta di medicina naturale.
Le modalità di applicazione sono di vario tipo : dalle infiltrazioni peri e intrarticolari, alla
piccola e grande autoemo, alle insufflazioni endo rettali -vescicali-vaginali, alle applicazioni di
sacchetti, alla utilizzazione di acqua ozonizzata (prodotta tramite appositi gorgogliatori), fino
all’applicazione con apposite cappette in silicone applicata ad un manipolo dal quale viene erogato
l’ozono (Unità di HealOzone, CurOzone, U.S.A.).
dott Cosimo Cigliola
5

Scopo del lavoro:
Questo studio è stato condotto per valutare, attraverso la revisione della letteratura corrente, gli
effetti benefici o meno dell’Ozono e del LASER.
Verificare l’efficacia dell’ozono nel trattamento delle patologie dentarie, carie radicolari, dei solchi
e delle cavità durante la fase iniziale e stabilire se esso è in grado di determinare la regressione
del processo carioso iniziale, efficacia antibatterica dell’ozono contro i ceppi più importanti di
batteri orali, decontaminazione dei condotti del riunito, ecc.. Congiuntamente agli effetti del Laser
nella rilevazione del calcolo sottogengivale, effetti sui batteri patogeni della malattia parodontale,
efficacia del laser congiuntamente allo scaling e root planing, effetto battericida nel trattamento
delle tasche parodontali, ecc.. E di conseguenza stabilire se, entrambi, possono essere decisivi
nella pratica dell’Igiene Dentale, nella prevenzione Primaria e Secondaria.
Materiali e metodi:
Le ricerche informatiche tramite internet, sono state condotte per identificare gli studi pubblicati e
non pubblicati. Sono state cercate le seguenti basi di dati: MEDLINE (dal 1966 al maggio 2007),
EMBASE (dal 1980 al maggio 2004), MEDLINE plus (dal 17 maggio 2004), Biosis (dal 1985 al
maggio 2004), AMED (dal 1985 al maggio 2004), registro nazionale di ricerca della biblioteca di
Cochrane (2 edizioni, 2004).
dott Cosimo Cigliola
6

Revisione della letteratura corrente sull'utilizzo del LASER, Ozono e
nuove tecnologie in Igiene Orale
ABSTRACT IN ITALIANO:
Estratti
RISULTATI
Associazione Internazionale per la Ricerca Dentale (IADR)
81 a sessione generale
2 o Meeting della Federazione pan-Europea
Göteborg, Svezia, dal 25 al 28 giugno 2003
N. JOHNSON1, J. JOHNSON1, H. DOMINGO2, and E. LYNCH2,
1 Private practice,Wales, United Kingdom, 2 Queen's University Belfast, United Kingdom
Il trattamento di una lesione cariosa e la ricostruzione tradizionale sono metodiche molto invasive.
Il trattamento di una lesione cariosa primaria con Ozono non richiede alcun trattamento invasivo,
semplicemente l'isolamento del dente e l'applicazione di Ozono attraverso le cappette in silicone.
Obiettivi: Questo studio è stato condotto per valutare le differenze tra il dispendio di tempo utile
per eseguire il trattamento con Ozono rispetto al metodo tradizionale del “drilling and
filling” (trapanazione e otturazione) misurando il tempo reale necessario per compiere entrambe le
modalità di trattamento.
Metodo: 40 pazienti adulti, con lesioni cariose primarie dei solchi, sono stati divisi a caso in due
gruppi uguali. Il primo gruppo è stato trattato con il metodo tradizionale del “drilling and filling”; il
secondo gruppo è stato trattato con Ozono. Entrambi i gruppi sono stati curati dagli stessi
Odontoiatri ed assistenti dentali. Il trattamento d’ogni paziente è stato cronometrato, in minuti,
dal momento in cui il paziente si è seduto alla poltrona fino al completamento della procedura.
Risultati: il tempo necessario per trattare ogni paziente è stato in media di 35min con una
deviazione standard di 10min, mentre il tempo necessario per trattare il gruppo terapeutico con
Ozono è stato in media di 8min con una deviazione standard di 2min. (P

Conclusioni: l'uso di Ozono per il trattamento delle carie occlusali primarie è stato provato
richiedere meno di un quarto del tempo richiesto per praticare le tecniche convenzionali del
“drilling and filling”.
1HealOzone unit, CurOzone USA and KaVo Germany.
Seq #277 - Miscellaneous Caries Clinical Studies
11:00 AM-12:15 PM, Saturday, 28 June 2003 Svenska Massan Exhibition Hall B
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Back to the 81st General Session of the International Association for Dental Research
(June 25-28, 2003)
2408 Applicazione di Ozono ad una carie radicolare valutazione dopo 12
mesi.
A. BAYSAN, Birmingham University, Dental School, United Kingdom, and E. LYNCH,
Queen's University, Belfast, United Kingdom
Obiettivi: L'obiettivo era valutare la sicurezza e l'efficacia di un sistema di erogazione di Ozono,
con o senza sigillante, per la terapia di una carie radicolare.
Metodi: sono stati registrati un totale di 79 pazienti con 220 PRCLs (lesioni cariose primarie della
radice). Il gruppo 1 è stato trattato solo con Ozono, il gruppo 2 non è stato trattato né con Ozono
né con sigillante radicolare, mentre il gruppo 3 è stato trattato sia con Ozono che con sigillante ed
il gruppo 4 è stato trattato solo con sigillante. Come linea di base sono stati impiegati la
misurazione della Resistenza Elettrica3 (ECM) e il DIAGNOdent4, a 1, 3, 6, 9 e 12 mesi. Le lesioni
sono state valutate clinicamente. Inoltre sono stati eseguiti i test di verifica di USPHS modificati.
Risultati: 74 pazienti hanno terminato il trattamento, dopo 12 mesi. Non sono stati osservati
eventi avversi. Dopo 12 mesi, il 47% di PRCLs ha avuto un’inversione dell’indice di severità tra 1 e
0 (hard) solo nel gruppo trattato con Ozono, mentre nel gruppo di controllo non si è verificata
nessun’inversione delle lesioni (p

Conclusioni: La carie superficiale della radice non attiva può essere trattata con Ozono. Questo
tipo di trattamento con Ozono è un'alternativa efficace al “drilling and filling”. Il sigillante è durato
più a lungo nulle lesioni trattate con Ozono.
Seq #247 - Oral Function, Caries and Health Education (Morita Junior Investigator Award)
9:00 AM-11:00 AM, Saturday, 28 June 2003 Svenska Massan F5
Back to the Geriatric Oral Research Program Back to the 81st General Session of the International
Association for Dental Research (June 25-28, 2003)
2008 Effetto dell’Ozono sulla durezza delle superfici dei materiali da
ricostruzione
D. CAMPBELL1, D.L. HUSSEY2, L. CUNNINGHAM2, and E. LYNCH2,
1 Royal Group of Hospitals, Belfast, United Kingdom, 2 Queen's University, Belfast, UK
Obiettivo: in odontoiatria di recente c’è stato molto interesse sull'uso di Ozono come trattamento
alternativo. Non è stato segnalato l'effetto residuo di tale trattamento sulla durezza di superficie
dei materiali da ricostruzione. Lo scopo di questo studio è stato quello di verificare l'effetto del
trattamento con Ozono sulla durezza della superficie su una gamma di materiali da restauro
comunemente usati.
Metodo: Sei dischi prefabbricati per test contenenti ciascuno 3 pozzetti sono stati riempiti di
materiali da restauro comunemente usati: un composito ibrido (Spectrum), un vetro ionomerico
(Chemflex), un compomero (Dyract AP), un'amalgama (Disperalloy), un composito fluido
(Revolution) e una resina modificata vetro ionomerica (Fuji II LC). I materiali sono stati
compattati su una lastra di vetro per produrre una superficie piana. 24 ore dopo i campioni (18 di
ciascuno) sono stati sottoposti al Test di Micro Durezza Vickers (Mitutoyo) usando un carico di 1Kg
per 10 secondi. L'Ozono è stato applicato per 10 secondi (Healozone, Curozone, USA e Kavo,
Germany) dopodiché è stata ripetuta la prova della durezza superficiale.
Risultati: I risultati dei test di durezza Vickers ottenuti sui campioni sono i seguenti:
Materiale Durezza media superficiale
± deviazione standard
Composito ibrido 46.82± 2.82 46.33 ± 3.07
Vetro-ionomerico 38.40 ± 10.85 37.02 ± 4.75
Compomero 27.32 ± 2.77 27.64 ± 2.93
dott Cosimo Cigliola
Durezza media superficiale
dopo applicazione d’ozono
± deviazione standard
Amalgama 127.13 ± 15.65 124.30 ± 14.02
Composito fluido 14.59 ± 2.11 12.61 ± 1.41
Resina modificata
G.I.
32.74 ± 3.18 30.06 ± 2.14
9

dott Cosimo Cigliola
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È stata utilizzata un‘analisi statistica bidirezionale ANOVA che non ha rivelato alcuna differenza di
durezza delle superfici dopo il trattamento con Ozono (p>0.15).
Conclusione: Si è giunti a conclusione che l'applicazione di Ozono per 10 secondi non ha effetto
nocivo sulla durezza delle superfici dei materiali esaminati e quindi non dovrebbe interessare le
prestazioni cliniche di tali materiali in vivo.
Seq #209 - Polymer Materials: Properties, Clinical Variables
3:45 PM-5:00 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
2034 Trattamento con Ozono in bambini ansiosi con carie dentale:
Atteggiamenti dei genitori dopo la prima sessione
J.E. DÄHNHARDT, T. JAEGGI, N. SCHEIDEGGER, N. KELLERHOFF, P.FRANCESCUT, and A.
LUSSI,
Università di Berna, Svizzera
In odontoiatria pediatrica la cura dei bambini ansiosi rimane una sfida. Molte volte le uniche
opzioni sono il trattamento con i sedativi o l'anestesia totale che include un’alto costo, determinati
rischi e i bambini rimangono ansiosi.
Obiettivo: L'obiettivo era di valutare gli atteggiamenti dei genitori di bambini ansiosi nei confronti
del trattamento della carie con Ozono (Heal Ozone Unit, CurOzone, U.S.A.).
Metodi: Dopo il primo trattamento di 20 bambini di età tra 2-10 anni (che include lo
spazzolamento dei denti, lo scavolino (Neos Vanadium Excavator No. 591/3, Hawe Neos Dental,
Switzerland), misurazioni con DIAGNOdent (KaVo, Germany) e l'applicazione di Ozono), al padre
e/o alla madre è stato chiesto, con un questionario, quale fosse il loro atteggiamento nei confronti
del trattamento con Ozono.
Risultati: Tutti i genitori hanno risposto al questionario, ma la maggior parte dei bambini sono
stati accompagnati solo dalle loro madri (90%). Il 75% dei bambini avevano paura di andare dal
dentista secondo i loro genitori prima del trattamento con Ozono, ma successivamente alcuni di
questo gruppo hanno perso il loro timore e tutti i bambini erano felici di ritornare
all'appuntamento seguente. Il 40% dei genitori avevano paura che il trattamento con Ozono fosse
pericoloso per i loro bambini ma tutti erano contenti di aver iniziato il trattamento con Ozono.
Dopo la prima sessione il 75% dei genitori suggerivano il trattamento con Ozono ai famigliari o
agli amici e acconsentivano ad un altro trattamento con Ozono. L’80% dei genitori era disposto a
pagare di più il trattamento con Ozono rispetto al trattamento tradizionale “drilling and filling”.
Conclusioni: Dopo la prima sessione, il trattamento con Ozono sembra essere stato accettato
bene dai bambini e dai loro genitori e in oltre fa diminuire l'ansia dei bambini.
Seq #211 - Fear, Anxiety, and Behavior
dott Cosimo Cigliola
11

3:45 PM-5:00 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
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(June 25-28, 2003)
0989 Efficacia dell'Ozono nel trattamento delle carie Occlusali dei denti
primari
O. ABU-SALEM, M. MARASHDEH, and E. LYNCH
Queen's University, Belfast, United Kingdom
Attualmente si sta studiando l'Ozono come farmaco per il trattamento delle carie occlusali dei
denti primari e permanenti.
Obiettivi: valutare e monitorare la progressione, la stabilizzazione o l'inversione delle lesioni
cariose delle superfici occlusali con l’utilizzo del DIAGNOdent, la Misurazione della Resistenza
Elettrica (ECM) i valori della scala standard, la classificazione clinica (Ekstrand, 1998), le macchie
dello smalto (mm), l'indice di colore, l'indice di struttura, lo smalto scheggiato e l'indice di bisogno
percepito di trattamento dopo il trattamento dei denti primari con Ozono.
Metodi: Lo studio in vivo eseguito su 16 pazienti con 42 lesioni cariose occlusali superficiali è
stato fatto in un periodo di 6 mesi. In linea di massima, a tre mesi ed a sei mesi tutti i denti sono
stati puliti con un sistema abrasivo ad aria denominato Prophyflex 2® (KaVo, Germany). Quindi
sono stati presi i valori ® del DIAGNOdent (D) (KaVo, Germany), i valori della scala standard del
ECM (LODE BV, Paesi Bassi) e la classificazione clinica (Ekstrand, 1998). La metà delle lesioni è
stata trattata con Ozono a ciascuna di queste è stata trattata per tre volte, per mezzo di una
cappetta siliconica sigillante (unità di HealOzone, CurOzone U.S.A. KaVo, Germany) per 10
secondi, mentre, l'altra metà delle lesioni è stata riservata come gruppo di controllo.
Risultati: A sei mesi i valori del ECM erano significativamente migliorati (p

(June 25-28, 2003)
1671 Efficacia Antimicrobica dell’Ozono sull’Enterococcus faecalis
H.H. CHANG, C. FULTON, and E. LYNCH,
Queen's University of Belfast, United Kingdom
Scopo: studiare l'efficacia battericida dell’Ozono su differenti concentrazioni di Enteroccocus
faecalis
Metodi: Enteroccocus faecalis è stato coltivato durante la notte sulle piastre di sangue di Agar.
Una sospensione dei microorganismi era composta da una concentrazione approssimativa di 10 8
usando McFarland Standards un tamponato salino di fosfati (PBS). Sono state fatte diluizioni di
serie a 10 7 , 10 6 , 10 5 . In ciascuno di questi concentrati sono stati pipettati bene 50 µl di ognuno su
di una piastra del microscopio. Ciascun concentrato a sua volta è stato trattato con Ozono
(HealOzone Unit, CurOzone, USA and KAVO, Germany) con tempi di esposizione differenti di 10s,
20s, 30s e 60s. I campioni sono stati prelevati dai pozzetti, placca a spirale, incubati ed effettuate
l'enumerazione delle unità di formazione delle colonie (cfu).
Risultati:
Tempo di
esposizione?
Concentrazione?
60s 30s 20s 10s Trattamento
10 8 cfu 2 x 103cfu 4.0 x
2.0 x
105cfu 107cfu
10 7 cfu 0 cfu 0 cfu 3.4 x
5.5 x
107cfu
4.4 x
in no-PBS
6.0 x 10 8 cfu
5.6 x 10 7 cfu
105cfu 105cfu
10 6 cfu 0 cfu 0 cfu 0 cfu 0 cfu 6.5 x 10 6 cfu
10 5 cfu 0 cfu 0 cfu 0 cfu 0 cfu 5.2 x 10 5 cfu
Tempo di esposizione = Tempo di esposizione all’Ozono.
PBS = tamponato salino di fosfati.
Concentrazione = concentrazione approssimata della sospensione batterica.
Conclusioni: L'Ozono ha ottenuto l’eradicazione totale degli Enterococcus Faecalis alla relativa
concentrazione della sospensione di 10 6 o meno, con un tempo d’esposizione di 10 secondi.
Seq #174 - Oral Microbiology & Immunology I
11:00 AM-12:15 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
Back to the Microbiology / Immunology and Infection Control Program
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(June 25-28, 2003)
dott Cosimo Cigliola
13

2753 Efficacia dell’Ozono nell’invertire la Carie Occlusale
R. MORRISON, Harbour Dental Practice, Donaghadee, United Kingdom, and E. LYNCH, Queen's
University Belfast, United Kingdom
L'Ozono è stato indicato in grado d’invertire clinicamente le lesioni cariose primarie della radice e
la carie iniziale occlusale dei solchi.
Obiettivi: valutare l'effetto dell'Ozono sulle lesioni cariose primarie dei solchi della cavità in un
periodo di 13 settimane durante un trattamento Odontoiatrico.
Metodi: i soggetti sono stati scelti a caso da 1 di 2 gruppi. Su 145 pazienti sono stati registrate
240 lesioni cariose mentre sono stati inseriti altri 60 soggetti di controllo con 60 lesioni, i quali non
hanno ricevuto alcun trattamento. Per ognuna delle lesione cariose è stato diagnosticato un
trattamento convenzionale. Ad ogni lesione della prova è stato applicato Ozono per 40 secondi.
Dopo 13 settimane, i pazienti sono stati richiamati e sono state rivalutate clinicamente le severità
delle lesioni.
Risultati: alla visita di controllo si sono presentati 89 soggetti del test con 141 lesioni della prova.
Non sono stati osservati eventi avversi. 123 lesioni cariose primarie del solco e della cavità
trattate con Ozono sono regredite clinicamente basandosi sulla misurazione clinica della severità
della lesione mentre le altre 18 lesioni sono rimaste stabili e nessuna si è aggravata (P

Obiettivi: Questo studio è servito a valutare gli effetti dell'Ozono (HealOzone unit, CurOzone USA
and KaVo Germany) sulle lesioni cariose primarie dei solchi occlusali per un periodo di 12 mesi.
Metodi: sono stati inseriti 376 pazienti che necessitavano di cure odontoiatriche con 2364 lesioni
cariose primarie del solco occlusale e in 1170 denti erano estese fino a 2 mm nella dentina. Dopo
sono state selezionate a caso, le lesioni che non dovevano ricevere il trattamento con Ozono e
altre che dovevano essere trattate con Ozono. Per misurare obiettivamente le lesioni cariose
primarie del solco occlusale è stato impiegato Il DIAGNOdent (KaVo, Germany). L'Ozono è stato
applicato ad ogni lesione della prova per 10, 20, 30 o 40 secondi a seconda della severità clinica e
ripetuto per 3 volte mensili se non si fosse ottenuta l'inversione della carie. I pazienti sono stati
richiamati dopo 12 mesi e rivalutati clinicamente. Per la registrazione della severità clinica di ogni
lesione, è stato impiegato di nuovo il DIAGNOdent per misurare obiettivamente le cavità occlusali
primarie e le lesioni cariose delle fessure.
Risultati: per il controllo sono stati richiamati 315 pazienti. Non sono stati osservati eventi
avversi. Il 99% delle lesioni cariose primarie dei solchi occlusali trattate con Ozono (1918 lesioni)
avevano avuto un’inversione del processo e questo era stato constatato clinicamente con il
miglioramento dei valori riferiti dal DIAGNOdent (P

Obbiettivo: stabilire l’accettazione dei pazienti al trattamento con Ozono effettuato in uno studio
dentistico privato.
Metodi: Hanno partecipato a questo studio pazienti (n=250) che presentavano una lesione
cariosa trattata nei sei mesi precedenti con una preparazione convenzionale della cavità mediante
frese ( preparazioni minimamente invasive e/o “air abrasion”), seguite da otturazione, e che
mostravano una lesione similare che richiedeva trattamento dallo stesso dentista. Accettazione,
ansia per le cure dentali e soddisfazione sono stati valutati un mese dopo il trattamento, mediante
un questionario con scala Likert a 5 punti. Il 100% dei pazienti ha partecipato e il 94% ha
completato il questionario entro tre mesi dopo il trattamento. Un questionario simile è stato
proposto ai pazienti (n=45) che avevano ricevuto un trattamento convenzionale minimamente
invasivo negli ultimi sei mesi ma che non avevano sperimentato il trattamento con Ozono.
Risultati: tutti i pazienti erano contenti o soddisfatti del trattamento con Ozono che avevano
ricevuto, ed erano contenti o soddisfatti del tempo richiesto dal trattamento.
L’85% ha considerato poco fastidiosi gli appuntamenti di controllo per monitorare la progressione
della reversione della lesione. Il 55% sono stati soddisfatti di scegliere questo trattamento anche
se più costoso del trattamento regolare, il 100% raccomanderebbe questo trattamento ad amici e
parenti, e il 100% vorrebbe ricevere ancora questo trattamento. L’80% dei pazienti ha riportato
una riduzione dell’ ansietà (p

L'Ozono è stato indicato clinicamente capace d'invertire le lesioni cariose primaria della radice e la
carie iniziale del solco occlusale.
Obiettivi: Questo studio ha valutato l'efficacia dell'Ozono (HealOzone unit, CurOzone USA) sulle
lesioni cariose dei solchi occlusali primari in un periodo di un mese.
Metodi: sono stati inseriti 105 pazienti con 300 lesioni cariose primarie del solco occlusale che
necessitavano di un trattamento Odontoiatrico generale. Ogni lesione cariosa aveva bisogno di un
trattamento convenzionale ed aveva riportato una lettura con il DIAGNOdent (KaVo, Germany)
(come linea base) fra 20 e 60. Dopo la randomizzazione, sono state assegnate le lesioni che non
dovevano ricevere il trattamento con Ozono e quelle che dovevano ricevere il trattamento con
Ozono su ogni soggetto con almeno una lesione di controllo. L'Ozono è stato applicato ad ogni
lesione della prova per 20 secondi. Dopo 1 mese, i pazienti sono stati richiamati e rivalutati
clinicamente per valutare la severità della lesione. E’ stato impiegato nuovamente il DIAGNOdent
per misurare obiettivamente le lesioni cariose primarie del solco occlusale.
Risultati: Dopo 1 mese, i pazienti sono stati richiamati per la rivalutazione. Non si sono osservati
eventi avversi. In base alla misurazione clinica della severità della lesioni, le lesioni trattate con
Ozono indicavano segni significativi d’inversione (P

La maggior parte della ricerca sull'uso di Ozono nella remineralizzazione delle lesioni cariose della
dentina è stata limitata al trattamento della carie occlusale. Comunque molte lesioni cariose si
presentano in altri luoghi, la carie interprossimale è una delle preoccupazioni.
Obiettivi: Questa ricerca ha mirato a stabilire se le altri lesioni, particolarmente interprossimale e
carie occlusale occulta potrebbero essere trattate usando l’airbrasion, per poter accedere alla
lesione, seguita dal trattamento con Ozono, e per valutare se questo metodo conservatore possa
essere più veloce delle tecniche convenzionali.
Metodi: sono state selezionate le lesioni che richiedevano un trattamento convenzionale. 37
pazienti con un totale di 48 denti sono stati trattati con airabrasion e Ozono mentre altri 48 denti
negli stessi pazienti sono stati trattati convenzionalmente, ed è stato registrato il tempo richiesto
per ristabilire l’integrità. L'accesso alle lesioni è stato ottenuto usando l’airbrasion a 80 PSI
utilizzzando come agente di taglio l'ossido d’alluminio a 27 micron (Abradent DV1). Una volta
stabilito l'accesso alla carie, è stato erogato Ozono per 40 secondi usando un'unità di HealOzone
(CurOzone U.S.A. e KaVo Germany). Nel trattamento della carie interprossimale, sono state
esplorate diverse tecniche per assicurare una efficace sigillatura. Il vetro ionomerico per restauri
(Fuji 9 (GC) or Diamond Carve (Kemdent)) è risultato efficace per otturare le cavità.
Risultati: Tutte le lesioni sono state esposte ed è stata stabilita con successo una sigillatura per
l’azione dell’Ozono. Intorno a tutti i ripristini sono state realizzate sigillature clinicamente
accettabili. La tecnica dell’airabrasion e Ozono è risultata significativamente più veloce delle
tecniche convenzionali (P

2756 Trattamento con Ozono della carie Occlusale in vivo: effetti a uno e
due mesi con Spettroscopia a fluorescenza (QLF) usato come metodo
diagnostico
G.D. MEGIGHIAN, Private Practice, Verona, Italy, and L. BERTOLINI, Private Practice, Verona,
Italy
E’ stato provato che l'Ozono inverte clinicamente la carie della radice e le lesioni cariose del solco
e delle fessure. Sono stati studiati più a fondo gli effetti di un trattamento farmaceutico delle
lesioni cariose del solco e delle cavità (PFCLs) nello sviluppo degli studi clinici controllati
longitudinali precedenti. Lo studio applica i protocolli sviluppati nella Dental Clinic, Queen’s
University, Belfast, in uno Studio privato Odontoiatrico a nord-est dell’Italia.
Obiettivi: Lo scopo di questo studio era controllare longitudinalmente le letture indotte dalla
Spettroscopia di fluorescenza (QLF) e la classificazione clinica delle PFCLs e quindi studiare
l'effetto a medio termine dell'applicazione di Ozono su queste letture.
Metodi: su 80 pazienti sono state registrate un totale di 300 PFCLs. Dopo la pulitura di ogni
lesione usando airabrasion con particelle di 27.5 micron, è stato registrato un indice clinico di
severità, letture della scala standard di QLF (DIAGNOdent®, KaVo, Germany). Due terzi delle
lesioni sono stati trattati con Ozono (utilizzando l'unità di HealOzone da CurOzone U.S.A.) per 20,
30 e 40s secondo la classificazione clinica e l'altro terzo è stato riservato come controllo in ogni
paziente che aveva almeno una lesione di controllo. Le letture sono state ripetute dopo un mese e
dopo due mesi.
Risultati: dopo un mese dal trattamento con Ozono è stata prodotta una riduzione generale
significativa delle letture di QLF. La percentuale dei denti che hanno prodotto questa riduzione era
più dell’80%. Dopo due mesi le letture di QLF che mostravano la riduzione erano più del 90%
(p

2754 Trattamento efficace dell’Ozono sulla carie del solco Occlusale
N. JOHNSON1, J. JOHNSON1, K. JOHNSON1, and E. LYNCH2,
1Private practice, Wales, United Kingdom, 2Queen's University Belfast, United Kingdom.
L'Ozono è stato indicato in grado di invertire clinicamente le lesioni cariose primaria della radice e
la carie del solco occlusale nella fase iniziale.
Obiettivi: Questo studio ha valutato l'effetto di un sistema di erogazione dell'Ozono (unità di
HealOzone, CurOzone U.S.A.) sulle lesioni cariose primarie dei solchi occlusali in un periodo di un
mese.
Metodi: allo studio hanno partecipato 105 pazienti che necessitavano di un trattamento dentale
generale con 300 lesioni cariose primarie dei solchi occlusali. Ogni lesione cariosa aveva necessità
di un trattamento tradizionale e ad una lettura di base con il DIAGNOdent (KaVo, Germania),
avevano registrato dei valori tra 20 e 60. Dopo, le lesioni, sono state divise, a caso, tra quelle che
non dovevano essere trattate e quelle che dovevano essere trattate con Ozono per ogni soggetto
che aveva almeno una lesione di controllo. Ad ogni lesione della prova l'Ozono è stato applicato
per 20 secondi. Dopo 1 mese, i pazienti sono stati richiamati e rivalutate clinicamente le severità
delle lesioni. È stato impiegato di nuovo il DIAGNOdent per misurare obiettivamente le lesioni
cariose primaria delle fessure occlusali.
Risultati: Dopo 1 mese, i pazienti sono stati richiamati per la rivalutazione. Non sono stati
osservati eventi avversi. Sulla base della misurazione clinica della severità della lesione, le lesioni
trattate con Ozono hanno indicato i segni significativi dell'inversione (P

2750 Trattamento con Ozono della carie primaria di cavità e del solco
Occlusale: Risultati a sei mesi
M.A. CRONSHAW, Private Practice, Isle of Wight, United Kingdom
L'Ozono è stato indicato essere efficace nell'arresto e nell'inversione delle lesioni cariose primarie
della cavità, del solco e della radice.
Obiettivi: Questo studio ha valutato l'effetto dell’Ozono sullo stato clinico e anche le letture del
DIAGNOdent delle carie delle cavità e del solco occlusale in un periodo di sei (6) mesi.
Metodi: sono stati inseriti 18 soggetti con 49 lesioni cariose che avevano bisogno di un
trattamento tradizionale. 31 denti sono stati trattati con Ozono per 30 secondi (HealOzone,
CurOzone e KaVo, U.S.A. e Germany) per la lesione della prova. Ciascun soggetto aveva una
lesione di controllo alla quale non è stato effettuato alcun trattamento. Tutte le lesioni sono state
rivalutate dopo sei mesi. La determinazione della severità delle lesioni è stata fatta tramite la
misurazione con un indice di severità clinica con le letture del DIAGNOdent (KaVo, Germania).
Risultati: Dei 31 denti trattati, 25 sono migliorati secondo le misurzioni del DIAGNOdent mentre
6 sono rimasti invariati (P

Obiettivi: scopo di questo studio era valutare l'utilizzo di Ozono generato da un dispositivo (unità
di HealOzone, CurOzone U.S.A. e KaVo, Grmany) per controllare la carie della cavità e del solco in
un periodo di 3 mesi durante un’attività dentale generale.
Metodi: i soggetti sono stati assegnati a caso ad uno di 2 gruppi. 98 pazienti sono stati inseriti
con 279 lesioni cariose della prova mentre altri 20 soggetti sono stati inseriti con 49 lesioni di
controllo, che non hanno ricevuto alcun trattamento. era stata ritenuto che ogni lesione cariosa
richiedesse un trattamento convenzionale. L'Ozono è stato applicato ad ogni lesione della prova
per 10 - 30 secondi secondo la loro severità clinica. Dopo 3 mesi, i pazienti sono stati richiamati
per rivalutare la severità clinica delle lesioni.
Risultati: Fin qui, 32 soggetti con 69 lesioni della prova hanno risposto alla visita di controllo.
Non si sono osservati eventi avversi. 58 delle lesione cariose primarie della cavità e del solco
trattate con Ozono avevano registrato un’inversione clinica, basandosi sulla misura clinica della
severità della lesione, mentre le altre 11 lesioni della prova sono rimaste stabili o addirittura
peggiorate (P

Obiettivi: verificare l’efficacia dell'Ozono(O3) nell’eradicazione da colonizzazione primaria di DUWL
(Dental Unit Water Lines) ed essere un rischio per gli adulti Immunodepressi ed i bambini con Fibrosi
Cistica. da P.A.
Metodi: Due nuove unità dentali (KaVo, Germany) sono state controllate settimanalmente per la
contaminazione di DUWL. La prima unità è stata trattata con erogazione continua di H2O2
(Oxygenal, KaVo, Germania). La seconda unità, di controllo, è stata lavata con acqua (H2O)
conformemente alle raccomandazioni del BDA. Dopo la comparsa di P.A. nell'unità di controllo, è
stata trattata con O3 (HealOzone, CurOzone U.S.A.; KaVo, Germany). La macchina di O3 è stata
collegata ad una bottiglia d'acqua e l’O3 è stato fatto gorgogliare nell'acqua per 5 minuti di seguito
irrigando i cavi dell’acqua con l'acqua ozonata per 10 minuti. Dopo una settimana l'acqua è stata
campionata e dopo un'altra settimana, è stato ripetuto il trattamento con O3. Sono stati raccolti
campioni di acqua da 20 ml, prima e dopo ogni trattamento con O3, e coltivati con Agar nutriente
e incubati per 3 giorni a 25°C.
Risultati: Il trattamento continuo di Ossigenazione (O3) dell'acqua ha prodotto un TVC (Total
Viable Count) di circa 100 CFU/ml. Il TVC dell’acqua nell’unità di controllo era di 2.3 x 10 4 e 3.4 x
10 4 CFU/ml dopo 1 e 2 settimane dall’installazione. Il colonizzatore primario è stato
identificato(API 20 NE kit) come P.A. puro. Dopo il primo trattamento con O3 il TVC si è ridotto a
60 CFU/mL ed è salito a 3.9 x 10 4 CFU/mL dopo una settimana con poche colonie di Psedomonas.
Dopo due settimane,il TVC era di 2.8 x 10 3 CFU/ml senza P.A. rilevato e si è trasformato in 0 CFU/
ml dopo il trattamento. Il campione ripetuto dell'unità per 9 settimane non ha mostrato nessuna
ricrescita di P.A.
Conclusione: l’Ossigenazione (O3) può mantenere la conta di DUWL a < 100 CFU/ml mentre non
potrebbe un lavaggio con acqua. L’P.A. può essere il colonizzatore primario di DUWL e può essere
eradicato con O3. Sostenuto da CurOzone, U.S.A.; KaVo, Germany.
Seq #222 - Biofilms II
3:45 PM-5:00 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
Back to the Microbiology / Immunology and Infection Control Program
Back to the 81st General Session of the International Association for Dental Research
(June 25-28, 2003)
Efficacia dell’ozono sulla sopravvivenza e permeabilità dei
microorganismi orali.
Microbiologia orale Immunol 2004:19: 240-246.
Parole chiave: attività antimicrobica; biofilm della placca batterica dentale; disinfettante;
microorganismi orali; ozono.
dott Cosimo Cigliola
23

Estratto: Nello studio presente abbiamo esaminato l'effetto dell'acqua ozonata sui microorganismi
orali e sulla placca batterica dentale. Quasi nessun microorganismo è stato rilevato dopo essere
stato trattato con acqua ozonata (4 mg/l) per 10s.
Valutare l’efficacia dell’acqua ozonata nei confronti dello Streptococcus Mutans, le cellule
batteriche sono state esposte con il corredo batterico di attuabilità di LIVE/DEAD BacLight TM.
L'analisi al microscopio a fluorescenza ha rivelato che le cellule dello S. Mutans erano
istantaneamente state eliminate dopo trattamento con acqua ozonata. Con l’ausilio del
microscopio elettronico è stata accertata una certa rottura della parete cellulare dello S. Mutans
trattato con acqua ozonata. Quando la placca batterica sperimentale è stata esposta all'acqua
ozonata, il numero di S. Mutans si è ridotto drasticamente. L'acqua Ozonata è un forte inibitore
della placca batterica in vitro. Dopo che i campioni della placca batterica dei soggetti umani sono
stati esposti all'acqua ozonata in vitro, non è stata rilevata quasi nessuna cellula batterica .
Questi risultati suggeriscono che l'acqua ozonata dovrebbe essere utile nella riduzione delle
infezioni causate dai microorganismi orali della placca batterica.
OZONO, GENIALE: questo gas discutibile può essere tossico o medicamentoso?
Bocci V.
L'ozono è un gas intrinsecamente tossico e la relativa pericolosità ha indotto ad una relativa
considerazione dell’ozono-terapia.
Lo scopo di questa revisione è di indicare che un dogma errato e molte idee sbagliate contrastano
il progresso: in realtà, l’ozono-terapia correttamente utilizzata, ed effettuata da medici esperti,
può essere molto utile quando la medicina convenzionale sembra essere inadeguata. L’incredibile
versatilità dell’ozono terapia è dovuta alla cascata degli ozono-derivati in grado di agire alla base
di uno stato patologico con un’azione multifattoriale. Durante la decade passata, al contrario di
tutte le aspettative, è stato dimostrato che l'applicazione giudiziosa di ozono nelle infezioni
croniche , vasculopatie, ortopediche e persino odontoiatriche ha raccolto risultati impressionanti
ed è deplorevole che l'istituzione medica continui ad ignorare l’ozono terapia.
PMID: 15203558 [PubMed - indexed for MEDLINE]
È vero che l'ozono è sempre tossico? La conclusione di un dogma.
Bocci V.
Department of Physiology, University of Siena, via A. Moro 2, Siena 53100, Italy. bocci@unisi.it
Ci sono un certo numero di buoni studi sperimentali che indicano che l'esposizione da inalazione
prolungata ad ozono troposferico danneggia l'apparato respiratorio e gli organi extrapolmonari. La
pelle, se esposta estesamente, può anche subire danni. Indubbiamente la forte reattività
dott Cosimo Cigliola
24

dell’ozono ha contribuito a stabilire il dogma che l'ozono è sempre un tossico e la relativa
applicazione medica deve essere prescritta. Anche se meno conosciuto, l’ozonoterapia
giudiziosamente eseguita sta diventando molto utile applicata da sola o congiuntamente alla
medicina tradizionale in una vasta gamma di patologie. Il giudizio degli oppositori
dell’ozonoterapia sulla chimica dell'ozono, e medicinale, senza alcuna conoscenza del problema,
spesso è scettico. Durante gli ultimi 15 anni, è stata acquisita una chiara comprensione del
meccanismo d’azione dell’ozono in biologia e in medicina, oggi viene messo in discussione se è
vero che l'ozono è sempre tossico.
I punti fondamentali che sono discussi in questa relazione sono: le caratteristiche anatomiche,
biochimiche e topografiche, degli organi esposti quotidianamente ad ozono contro la potente
capacità antiossidante del sangue esposta per pochi minuti ad una piccola dose calcolata di ozono.
È straordinario come l'apparato respiratorio sottoposto ad uno sforzo cronico ossidativo può
liberarsi lentamente, ma costantemente, di una quantità enorme di residui tossici in grado di
entrare nella circolazione e di causare seri danni. Lo scopo di questa relazione è valutare
obiettivamente questa controversa questione.
PMID: 16890971 [PubMed - indexed for MEDLINE]
Effetto dell’ozono sulle cellule orali paragonato agli antimicrobici tradizionali.
Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K.
Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-University, Munich,
Germany. khuth@dent.med.uni-muenchen.de
L'ozono, in odontoiatria, è stato prospettato come agente antisettico alternativo, basandosi sui
rapporti dei relativi effetti antimicrobici sia in forma gassosa che acquosa. Questo studio ha
stabilito se l’ozono gassoso (4 x 10 6 µg/ml(-3)) e acquoso (1.25 -20 µg/ml(-1)) esercita tutti gli
effetti citotossici sulle cellule epiteliali orali umane (BHY) e sui fibroblasti gengivali (HGF-1)
rispetto agli antisettici imposti [digluconato di clorexidina (CHX) 2%, 0.2%; ipoclorito di sodio
(NaOCl) 5.25%, 2.25%; perossido di idrogeno (H2O2) 3%], per oltre 1 minuto e paragonato
all'antibiotico, metronidazolo, per oltre 24h. Sono stati valutati, la conta delle cellule, l’attività
metabolica, l’Sp-1 binding, i livelli di actina e l’apoptosi. È stato provato che il gas Ozono ha effetti
tossici su entrambi i tipi di cellule. Essenzialmente non è stato osservato alcun segno citotossico
per l’ozono acquoso. CHX (2%, 0.2%) era altamente tossica per le cellule di BHY, lievemente(2%)
e non tossico (0.2%) per le cellule HGF-1. L’NaOCl e la H2O2 hanno provocato una marcata
riduzione della sopravvivenza cellulare di(BHY, HGF-1), mentre il metronidazolo ha dimostrato una
dott Cosimo Cigliola
25

lieve tossicità soltanto alle cellule BHY. Si desume che, l'ozono acquoso ha evidenziato un livello
elevato di biocompatibilità tra gli antisettici esaminati.
PMID: 17026511 [PubMed - indexed for MEDLINE]
Biofactors. 2006;27(1-4):5-18.
Indagini con RMN ad alta risoluzione 1H sul consumo ossidativo delle biomolecole
salivari trattate con ozono: attinenza con le applicazioni terapeutiche di questo agente
in odontoiatria clinica.
Grootveld M, Silwood CJ, Lynch E.
Department of Applied Science, London South Bank University, 103 Borough Road, London, UK.
grootvm@lsbu.ac.uk
La risonanza magnetica nucleare (RMN) protonica(1H) ad alta risoluzione spettroscopica è stata
impiegata per valutare simultaneamente gli effetti ossidanti dell’Ozono (O3) verso una vasta
gamma di biomolecole salivari in considerazione delle relative applicazioni nelle pratica dentale, il
quale può essere utilizzato come possibile e conveniente mezzo per il trattamento della carie
dentale. Il trattamento dei supernatants derivati dai campioni (n=12) di saliva umana stimolati
con O3 (4.48 mmol) hanno rivelato che questa specie reattiva di ossigeno ha causato l’aumento
del consumo ossidativo di piruvato (generando come prodotti acetato e CO2), del lattato (a
piruvato e acetato e CO2), dei carboidrati (un processo che genera formiato), della metionina (che
produce il relativo ossido di zolfo) e dell’urato. Tuttavia, le minori modifiche O3-indotte hanno
incluso l'ossidazione di trimetilammina e del 3-D-hydroxybutyrate, la frammentazione dei
glicosaminoglicani salivari in frammenti saccaridici RMN-rilevabili e la conversione degli acidi
grassi polinsaturi nei loro ozonidi. Inoltre, è stata ottenuta la prova della capacità di O3 di indurre
il rilascio delle biomolecole salivari selezionate della piccola massa molecolare dai siti di legame
macromolecolari. Da molti dei prodotti di ossidazione rilevabili nei campioni O3-trattati sono
identici a quelli che risultano dall'attacco del radicale - OH ai componenti del biofilm, sembra che
almeno alcune delle modifiche osservate qui siano attribuibili all'ossidante (derivato da O3 -
generato dalla singola riduzione dell'elettrone di O3).
PMID: 17012760 [PubMed - in process]
Braz Dent J. 2006;17(2):134-8
Potenziale antimicrobico dell’ozono in un sistema di pulizia ultrasonica contro lo
Stafilococco Aureo.
Estrela C, Estrela CR, Decurcio Dde A, Silva JA, Bammann LL.
dott Cosimo Cigliola
26

Faculty of Dentistry, Federal University of Goias, Goiania, GO, Brazil. estrela3@terra.com.br
Lo scopo di questo studio era di valutare il potenziale antimicrobico dell’ozono applicato in 3
soluzioni differenti in un sistema di pulizia ultrasonica contro lo Stafilococco Aureo. Sono stati
miscelati un totale di 120 ml di S.Aureus in 6 l di soluzione sperimentale (acqua distillata sterile,
aceto e acqua distillata sterile + Endozime AWpluz) introdotte in un sistema di pulizia ultrasonica
(UCS). L'ozono è stato prodotto tramite una scarica elettrica e fatto gorgogliare attraverso un
flusso di ossigeno ad una portata di 7 g/h di ozono(1.2%) nella sospensione microbica. Sono stati
raccolti 10 ml di ogni sospensione sperimentale ed effettuate 5 diluizioni alla volta in 9 ml di BHI
ed incubati a 37 0 C per 48 h. È stato valutato lo sviluppo batterico attraverso la torbidezza del
terreno di coltura. Allo stesso tempo, è stato raccolto 1 ml del campione batterico ed inoculato in
piastre di BHIA. Dopo incubazione a 37 0 C per 48 h, è stato contato il numero delle unità delle
colonie formate (cfu) per ml sulla superficie di BHIA. Nella prova di diluizione su tubi di BHI e su
piastre di BHIA (cfu/ml), non è stato osservato alcuna attività batterica in nessuna delle soluzioni
sperimentali trattate con ozono. Nelle circostanze esaminate, può essere concluso che l'aggiunta
di ozono in un sistema di pulizia ultrasonica contenenti differenti soluzioni sperimentali risulta l’
attività antibatterica contro S.Aureus.
PMID: 16924341 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2006 Aug;114(4):349-53
Effetto antibatterico di un dispositivo che produce ozono messo a confronto con due
sistemi di adesione della dentina.
Polydorou O, Pelz K, Hahn P.
Department of Operative Dentistry and Periodontology, Dental School and Hospital, Albert-
Ludwigs-University Freiburg, Germany. olga.polydorou@uniklinik-freiburg.de
I microorganismi rimasti sotto i restauri possono causare carie secondaria e danni della polpa. per
questo, il trattamento antimicrobico è stato di grande aiuto.
Lo scopo di questo studio era di valutare l'effetto antibatterico del dispositivo di HealOzone sullo
Streptococcus Mutans e paragonarlo all'attività già provata di due sistemi di dentin-bonding. 35
molari sono stati divisi in 5 gruppi. Allora i denti sono stati sezionati in cavità (n = 28 cavità per
gruppo). Dopo la sterilizzazione, i denti sono stati lasciati nelle colture di brodo con 10 6 /ml (-1)
Unità Formanti Colonie (CFU) di S. Mutans a 36 0 C per 48 h. Il trattamento adatto seguito (gruppo
A, controllo; gruppo B, Clearfil SE Bond; gruppo C, Clearfil Protect Bond; gruppo D, 40 s di
dott Cosimo Cigliola
27

trattamento con ozono; ed il gruppo E, 80 s di trattamento con ozono) dopo le cavità sono state
riempito di resina composita. Dopo 72 h, i restauri sono stati rimossi, sono stati raccolti frammenti
di dentina con un escavatore, ed è stato determinato il numero totale di microorganismi. Tutti i
trattamenti hanno ridotto significativamente il numero dello S. Mutans rispetto al gruppo di
controllo. L'effetto antimicrobico di entrambi i sistemi di bonding il trattamento con ozono per 80s
è stato significativamente superiore al trattamento con ozono per 40s. In conclusione, HealOzone
ed i sistemi di bonding mostrano gli effetti antimicrobici contro lo S. Mutans.
PMID: 16911107 [PubMed - indexed for MEDLINE]
Am J Dent. 2006 Feb;19(1):67-72
Effetti del trattamento con ozono sulle proprietà fisiche dello smalto.
Celiberti P, Pazera P, Lussi A.
Department of Operative, Preventive and Pediatric Dentistry, School of Dental Medicine, University
of Bern, Freiburgstrasse 7, 3010 Bern, Switzerland. paulaceliberti@hotmail.com
SCOPO: valutare gli effetti della molecola altamente reattiva di ozono sulle proprietà fisiche dello
smalto sano ed i relativi effetti sulla capacità sigillante.
METODI: effetti dell’ozono sulla adesività del sigillante, sono stati valutati le microfessure e le
aree non trattate dei solchi di molari sani. Sulle superfici liscie sane sono stati effettuati i tests di
microdurezza, l'angolo di contatto e di resistenza all'acido. I campioni sono stati trattati con ozono
per 40 secondi (HealOzone). I campioni di controllo sono stati trattati con aria (HealOzone
modificato) o appena trattati.
RISULTATI: non è stata osservata alcuna differenza statisticamente rilevante in nessuno dei
campioni di controllo e di ozono trattati in tutte le prove. I solchi preparati non hanno presentato
zone non riempite e una microfessure statisticamente significativa è stata confrontata con i solchi
intatti. L'ozono è in grado di disidratare lo smalto e conseguentemente di aumentarne la relativa
microdurezza, che è reversibile.
PMID: 16555661 [PubMed - indexed for MEDLINE]
Oral Microbiol Immunol. 2005 Aug;20(4):206-10
Efficacia microbicida dell'acqua ozonata verso la Candida Albicans che aderisce alle
superfici acriliche della dentiera.
dott Cosimo Cigliola
28

Arita M, Nagayoshi M, Fukuizumi T, Okinaga T, Masumi S, Morikawa M, Kakinoki Y,
Nishihara T.
First Department of Prosthetic Dentistry, Kyushu Dental College, Kitakyushu, Japan.
SCOPO: L'ozono è conosciuto come potente agente antimicrobico contro i batteri,funghi e virus.
Abbiamo esaminato gli effetti dell'acqua ozonata sulla Candida Albicans presente sulle superfici
delle dentiere acriliche.
METODI: Le resine acriliche a caldo sono state coltivate con C. Albicans. Dopo il lavaggio con
acqua ozonata, è stato contato il numero di C. Albicans fissati. In alcuni esperimenti, i campioni
della prova sono stati trattati con acqua ozonata congiuntamente ad ultrasuoni.
RISULTATI: Dopo l'esposizione al flusso di acqua ozonata (2 o 4 mg/l) per 1 minuto, le possibili
cellule di C. Albicans erano quasi inesistenti. La combinazione di acqua ozonata e ultrasuoni ha
avuto un forte effetto sulla sopravvivenza della C. Albicans che aderisce alle superfici acriliche
della resina. Non ci sono differenze significative nell'attività antimicrobica contro la C. Albicans tra
le piastre immerse dentro l'acqua ozonata con ultrasuoni e quelle trattate con i pulitori per
dentiera disponibili in commercio. In più, l'analisi al microscopio elettronico ha rivelato che i piccoli
aumenti di C. Albicans sono rimasti sulla piastra dopo l'esposizione a lavaggi con acqua ozonata o
l'immersione dentro l'acqua ozonata con ultrasuoni.
CONCLUSIONE: I nostri risultati indicano che l'applicazione di l'acqua ozonata può essere utile
nella riduzione del numero di C. Albicans sulle superfici della dentiera.
PMID: 15943763 [PubMed - indexed for MEDLINE]
Int J Prosthodont. 1999 Mar-Apr;12(2):179-83.
Influenza dell’ozono sull’ossidazione delle leghe dentali.
Suzuki T, Oizumi M, Furuya J, Okamoto Y, Rosenstiel SF.
Department of Geriatric Dentistry, Tokyo Medical and Dental University Faculty of Dentistry,
Japan. t.suzuki.gero@dent.tmd.ac.jp
SCOPO: Lo scopo di questo studio era esaminare l'influenza dell’ozono sulle superfici delle leghe
di protesi parziali amovibili(RPD) per determinare la relativa utilità come metodo di pulizia per
RPDs, poiché l'ozono ha forti proprietà di sterilizzazione e di deodorizzazione.
dott Cosimo Cigliola
29

MATERIALI E METODI: sono stati usati due tipi di pulizia con ozono. Le quantità di ozono
generate con entrambi i metodi erano le stesse (20 mg/h). Nel metodo A l'ozono è stato erogato
per 10 minuti ogni 12 ore, nel metodo B l'ozono è stato erogato per più di 24 ore al giorno. Gli
esemplari della prova di 3 tipi di leghe dentali (Co-Cr, Au-Ag-Pt e Au-Cu-Ag-Pd) sono stati
sottoposti ai differenti metodi di pulizia per 7 giorni e sono stati misurati in termini di riflessione,
rugosità di superficie e peso. Per il confronto con i trattamenti dell'ozono sono state usate cinque
soluzioni differenti di pulitori (tre pulitori commerciali per dentiera, acido-elettrolitico acquoso con
un pH di 2.4 ed acqua pura).
RISULTATI: Nessun cambiamento significativo è stato rilevato dopo il trattamento del Co-Cr e
delle leghe Au-AG-Pt con ozono. L'ozono ha causato un leggero cambiamento nella lega Au-Cu-
AG-Pd in termini di riflessione, ma i cambiamenti erano meno significativi di quelli causati da
acido-elettrolitico acquoso e da uno dei pulitori commerciali per dentiera.
CONCLUSIONE: L'ozono ha influenzato in maniera minima l'ossidazione delle leghe dentali.
PMID: 10371921 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2007 Feb;115(1):77-80.
Efficacia del gas ozono e terapia fotodinamica sul biofilm orale multispecie in vitro.
Muller P, Guggenheim B, Schmidlin PR.
Clinic of Preventive Dentistry, Periodontology and Cariology, University if Zurich, Zurich,
Switzerland.
Il gas ozono e la terapia fotodinamica (PDT) hanno entrambi effetti antimicrobici. Questo studio ha
valutato il loro potenziale antimicrobico in vitro. I biofilms orali di sei specie mature sono stati
trattati come segue (n=9 per gruppo):
a) applicazione per 60 s con ozono sottovuoto o solo sottovuoto (sui campioni del biofilm
bagnati o asciugati con aria);
b) PDT (cioè blu di metilene con o senza soft laser a diodo e solo un soft laser);
c) soluzioni antimicrobiche: immersione dei biofilms per 60 s in clorexidina al 2% e allo 0.2%
o in una soluzione di ipoclorito al 5% e allo 0.5%. I trattamenti con clorexidina o ipoclorito
sono serviti da controllo positivo, mentre i campioni non trattati sono serviti da comandi
negativi. Sono state contate le Unità Formanti Colonie (CFU) con sangue di Agar. Soltanto la
dott Cosimo Cigliola
30

soluzione con ipoclorito al 5% poteva completamente eliminare i microorganismi del biofilm. La
riduzione osservata dei conteggi possibili dall'applicazione di ozono sottovuoto e da PDT era
meno d'un ceppo (10) punti. Nei termini dello studio corrente, il gas ozono e il PDT hanno
avuto un’effetto minimo sulla riproducibilità dei microorganismi organizzati in un biofilm
cariogenico.
PMID: 17305720 [PubMed - in process]
J Endod. 2004 Nov;30(11):778-81.
Effetto antimicrobico dell'acqua ozonata sui batteri che invadono i tubuli dentinali.
Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M.
Department of Operative Dentistry and Endodontics, Kyushu Dental College, Kitakyushu, Japan.
L'ozono è conosciuto come un forte agente antimicrobico contro batteri, funghi e virus. Nel
presente studio, abbiamo esaminato l'effetto dell'acqua ozonata contro l'Enterococcus Faecalis e
infezioni da Streptcoccus Mutans in vitro su dentina di bovino. Dopo l'irrigazione con acqua
ozonata, l'attività dell’E. Fecalis e l'invasione dei tubuli dentinali dello S. Mutans è diminuita
significativamente. Notevole, quando le specie sono state irrigate con sonicazione, l'acqua ozonata
ha avuto quasi la stessa attività antimicrobica dell'ipoclorito di sodio al 2.5% (NaOCl). Inoltre
abbiamo confrontato la citotossicità contro i fibroblasti di topo L-929 tra acqua ozonata e NaOCl.
L'attività metabolica dei fibroblasti era alta quando le cellule sono state trattate con acqua
ozonata, mentre è diminuita significativamente quando le cellule sono state trattate con NaOCl al
2.5%. Questi risultati suggeriscono che l'applicazione di acqua ozonata può essere utile per la
terapia endodontica.
PMID: 15505509 [PubMed - indexed for MEDLINE]
J Med Dent Sci. 1998 Jun;45(2):135-9.
Metodo di pulizia di una dentiera con l’utilizzo di ozono. Prova in vitro.
Oizumi M, Suzuki T, Uchida M, Furuya J, Okamoto Y.
Department of Geriatric Dentistry, Faculty of Dentistry, Tokyo Medical and Dental University,
Japan. m.oizumi.gero@dent.tmd.ac.jp
Lo scopo di questo studio era paragonare l'effetto microbicida dell’ozono gassoso a quello
dell'acqua ozonata per determinare la relativa utilità come metodo per la disinfezione delle
dott Cosimo Cigliola
31

dentiere. Anche se, sull'effetto battericida dell’ozono, sono stati fatti tantissimi studi di ricerca
poco è conosciuto circa i relativi effetti microbicidi sui microorganismi orali. Di conseguenza,
abbiamo verificato l'effetto dell’ozono su tre studi standard dei microorganismi orali:
Streptococcus Mutans (studio IID 973), Stafilococco Aureo (studio 209-P) e Candida Albicans (LAM
14322 studio). Quando è stato usato il metodo per iniezione di ozono gassoso, i numeri di cellule
di tutti e tre gli studi sono diminuiti a 1/10 (5) in un 1 minuto ed entro 3 minuti erano sotto il
limite di segnalazione. Quindi, in poco tempo, è stato accertato l'effetto microbicida dell’ozono
gassoso. In opposizione, quando sono stati usati l'acqua ozonata a 1 PPM e 3 PPM, la C. Albicans
è diminuiti a 1/10. per preparare 1 PPM di acqua ozonata era necessario un livello di produzione di
ozono di 700 mg/h, mentre dal generatore dell'ozono gassoso sono stati richiesti 20 mg/h di
ozono. Questi risultati indicano che l'esposizione diretta a ozono gassoso sembra avere un’attività
microbicida più efficace rispetto all'acqua ozonata e che l'ozono gassoso può essere clinicamente
utile per la disinfezione delle dentiere.
PMID: 11186199 [PubMed - indexed for MEDLINE]
Trattamento con ozono di lesioni cariose aperte in bambini ansiosi. Studio clinico
controllato in prospettiva.
Dahnhardt JE, Jaeggi T, Lussi A.
Department of Operative, Preventive and Pediatric Dentistry, School of Dental Medicine, University
of Bern, Switzerland. daehnhardt@zmk.unibe.ch
SCOPO: Per determinare se il trattamento della carie dentale con ozono sia possibile in bambini
apprensivi ed accertare se l'ozono inverte la carie nelle lesioni aperte della singole superfici. In più
è stata studiata, l'influenza di ozono sulle letture del laser a fluorescenza.
METODI: sono state valutate 82 lesioni su 28 bambini con almeno due lesioni aperte per ogni
singola superficie. I bambini sono stati giudicati, dall’odontoiatra di riferimento, come ansiosi e
quindi non trattabili. Per ogni lesione del test, che è stata trattata con ozono, è stata lasciata una
lesione di controllo senza trattamento. Sono stati valutati i valori del laser a fluorescenza ed i
cambiamenti dei valori nelle lesioni durante i test sono stati confrontati con i valori delle lesioni di
controllo a distanza di 2, 4, 6 e 8 mesi.
RISULTATI: il 94% dei bambini sono risultati trattabili e il 93% ha perso l’ansia da dentista. I
valori di durezza sono migliorati significativamente nelle lesioni ozono-trattate della test dopo 4, 6
e 8 mesi (P0.05). L'uso di ozono ha
dott Cosimo Cigliola
32

provocato una riduzione media di 13% dei valori del laser a fluorescenza subito dopo il
trattamento con ozono.
PMID: 17073201 [PubMed - indexed for MEDLINE]
Am J Dent. 2005 Aug;18(4):223-8.
Effetto dell’ozono sulle lesioni dei solchi non profonde in un molare permanente. Studio
clinico controllato in Prospettiva.
Huth KC, Paschos E, Brand K, Hickel R.
Department of Restorative Dentistry and Periodontology, Dental School, Ludwig-Maximilians-
University, Goethe Street 70, D-80336 Munich, Germany. khuth@dent.med.uni-muenchen.de
SCOPO: valutare, con uno studio clinico controllato randomizzato, l'effetto dell’ozono sulle carie
superficiali del solco occlusale confrontate alle lesioni di controllo controlaterali non trattate (split
mouth) prendendo in considerazione pazienti a rischio di carie.
METODI: allo studio hanno partecipato 41 pazienti con 57 paia di lesioni (età media 7.7 +/- 2.2
anni; mascellare superiore n=29, mascellare inferiore n=28). L'ozono gassoso (HealOzone) è
stato applicato una volta per 40 secondi a caso al molare del test di ogni accoppiamento senza
l'uso delle soluzioni rimineralizanti.
La progressione o l'inversione della lesione è stata controllata dal sistema del laser a fluorescenza
DIAGNOdent per oltre 3 mesi ed il deterioramento o il miglioramento confrontato con le lesioni
ozono-trattate e le lesioni non trattate, di controllo (negli accoppiamenti). Questo studio è stato
condotto sull’intero campione di popolazione e un sottogruppo di pazienti con alto rischio di carie
(coppia di lesioni n=26).
RISULTATI: Dopo 3 mesi, l'analisi dei dati ha rivelato che le lesioni ozono-trattate hanno
mostrato una sensibile inversione della carie o una ridotta progressione della carie rispetto alle
lesioni non trattate di controllo nel gruppo di pazienti ad alto rischio di carie (Wilcoxon-Prova, P =
0.035). Non c’era importanza statistica che esamina la popolazione di tutto lo studio. Dai dati può
essere concluso che l'applicazione di ozono ha migliorato significativamente la carie iniziale
superficiale del solco in pazienti con alto rischio di carie in un periodo di tre mesi.
[PubMed - indexed for MEDLINE]
Cochrane Database Syst Rev. 2004;(3):CD004153.
Ozono terapia per il trattamento delle carie dentali
Old Sandstone Dental Practice, The Chine, Dorking, Surrey, UK, RH4 1QT.
dott Cosimo Cigliola
33

SCOPO: La carie dentale è una malattia batterico mediata caratterizzata dalla demineralizzazione
della superficie del dente, che può condurre alla cavitazione, al disagio, al dolore ed alla perdita
finale del dente. L'ozono in vitro è tossico per i batteri ed è stato ipotizzato che trattando con
ozono una lesione cariosa esso possa ridurre il numero dei batteri cariogeni. Questo possibilmente
potrebbe arrestare il progredire del processo carioso e, in presenza di fluoruro, permettere la
remineralizzazione dello smalto. Se ciò fosse vero potrebbe, a sua volta, far ritardare o impedire
l'esigenza di effettuare le metodiche di conservativa dentale tradizionali.
OBIETTIVI: valutare l’efficacia dell'ozono ad arrestare o invertire la progressione del processo
carioso.
STRATEGIA DI RICERCA: le ricerche sono state effettuate su Cochrane gruppo di salute orale (7
novembre 2003); Registro centrale delle prove controllate (CENTRALI) (edizione 3 della biblioteca
di Cochrane, 2003) di Cochrane; MEDLINE e PREMEDLINE (OVID, dal 1966 a novembre del 2003);
EMBASE (OVID, 1980 a novembre del 2003); CINAHL (OVID, 1982 a novembre del 2003); AMED
(OVID, 1985 a novembre del 2003). Quintessence ricerca manuale 2002 e contatto KaVo come
fornitori dell'apparecchiatura di HealOzone per le prove pubblicate o non pubblicate
supplementari.
CRITERI DI SELEZIONE: Lo studio è stato valutato almeno da due critici indipendenti. Le prove
erano incluse solo se rispondessero ai seguenti test di verifica: prova controllata randomizzata;
singola lesione cariosa in vivo di superficie accessibile all'applicazione con ozono; dissimulazione
libera di ripartizione; applicazione di ozono alle lesioni nel gruppo del test; nessuna applicazione di
ozono nel gruppo di controllo; risultati misurati dopo almeno 6 mesi.
RACCOLTA DEI DATI ED ANALISI: I critici hanno estratto indipendentemente le informazioni
due volte. Una scarsità dei dati paragonabili non ha permesso la riunione meta-analitica degli
studi inclusi.
RISULTATI PRINCIPALI: sono state incluse tre prove, con un totale di 432 lesioni ripartite con
scelta casuale (137 partecipanti). Sono stati esclusi gli atti del congresso, 42 estratti e manifesti
(da un numero sconosciuto di studi). Il rischio di polarizzazione in tutto lo studio è sembrato alto.
Le analisi di tutte e tre le prove sono state condotte alivello della lesione, che non è indipendente
dalla persona, per questo motivo la riunione dei dati non era adatta o adattabile. L'individuo studia
gli effetti contradditori indicati dall’ozono sulla carie, attraverso le misure differenti della
progressione o della regressione della carie. Sono stati segnalati pochi risultati secondari, e una
prova ha segnalato un'assenza di eventi avversi.
CRITERI DI CONCLUSIONE: Dato l'elevato rischio di polarizzazione negli studi disponibili e di
mancanza di consistenza fra risultato e differente misura, non c’è prova certa che l’applicazione
del gas ozono sulla superficie dei denti inverte il processo di deperimento. C’è un'esigenza
dott Cosimo Cigliola
34

fondamentale di una maggiore prova di rigore adatto e qualità prima dell'uso dell’ozono può
essere accettata nella cura dentale primaria tradizionale o può essere considerata un'alternativa
possibile ai metodi correnti per la gestione ed il trattamento della carie dentale.
dott Cosimo Cigliola
35

Ozono, inversione clinica della carie radicolare, prova Random di 18 mesi a Doppio-
Cieco
Holmes J.
UKSmiles Dental Practice, Wokingham, Berkshire, UK. julian@o3dc.co.uk
OBIETTIVO: valutare l'effetto di un sistema di erogazione dell’ozono, combinato con un kit
paziente di remineralizzazione per uso quotidiano, sulla severità clinica delle lesioni cariose
primarie radicolari superficiali (PRCL), in un gruppo di popolazione anziana.
DISEGNO: È stato reclutato un totale di 89 soggetti, (range d'età 60-82, Deviazione Standard,
70.8 +/- 6 anni), ciascuno con due PRCL per superficie. In uno studio Odontoiatrico privato le
lesioni di ogni soggetto sono state assegnate a caso per il trattamento con ozono o con aria, con il
metodo a Doppio-cieco. I soggetti sono stati richiamati a 3, 6, 12 e 18 mesi. Le lesioni sono state
registrate clinicamente durante ogni richiamo come consistenza: molle, media o dura; effettuato
con un indice convalidato di severità della carie radicolare.
RISULTATI: Non si sono osservati eventi avversi.
Dopo tre mesi, nel gruppo ozono-trattato, 61 PRCL (69%) erano regredite e nessuna era
peggiorata, mentre nel gruppo di controllo, quattro PRCL (4%) erano peggiorate (p

Pubblicato su American Journal of Dentistry
MANAGEMENT OF ROOT CARIES USING OZONE
Trattamento della carie della radice con l’utilizzo di OZONO
A. BAYSAN 1 and E. LYNCH 2
1Department of Adult Oral Health,Barts and the Royal London Queen Mary's School of Medicine
and Dentistry, London, UK
2Division of Restorative Dentistry and Gerodontology, Dental School, Royal Victoria Hospital,
Queen's University, Belfast, Northern Ireland
Corrispondenza a:
Aylin Baysan BDS MSc Barts and the Royal London Queen Mary's School of Medicine and
Dentistry, Department of Adult Oral Health, Turner Street, E1 2AD, London, UK Tel: +44 20 7377
7000 ext. 2186 - Fax +44 20 7377 7375 E- mail: abaysan@mds.qmw.ac.uk
INTRODUZIONE
Il trattamento conservativo della carie della radice è di particolare interesse alla luce delle
difficoltà di visibilità, controllo dell’umidità, accesso a lesioni cariate, prossimità della polpa,
prossimità al margine gengivale, e contenuto altamente organico della dentina [Titus, 1991]. Molti
materiali per la conservativa, come i vetroionomerici, resine-modificate, i cementi di
vetroionomero [Lynch and Tay, 1989a,b; Burgess, 1995], composti e amalgama, sono stati usati
per il restauro di lesioni cariose: tuttavia sono sorti svariati problemi, come microfratture e scarsi
adattamenti marginali con la necessità di frequenti sostituzioni dei materiali di otturazione [Taylor
and Lynch, 1992; Taylor and Lynch 1993]. Ad oggi è riconosciuto che il trattamento più
desiderabile per le carie radicolari è la remineralizzazione [Allen et al.,1999]. L’Ozono può essere
considerato come una strategia di trattamento alternativa della carie radicolare. Recentemente
Baysan et al.,[2000] hanno riportato che l’applicazione di Ozono per 10 o 20s risulta efficace ad
eliminare la grande maggioranza di microrganismi in lesioni primarie di carie della radice (PRCL)
in vitro e questa applicazione per una durata di 10s risulta anche in grado di ridurre i numeri di
Streptococcus Mutans e S. Sobrinus in vitro. Beighton et al., [1993] hanno convalidato un indice
di severità per le carie della radice e un criterio di diagnosi clinica per le PRCL. L’Electrical Caries
Monitor III (ECM III, Monitoraggio elettrico della carie) è altresì stato dimostrato essere correlato
con la severità delle PRCL [Baysan et al., 2002].
Lo scopo di questo studio longitudinale è di valutare la sicurezza e l’efficacia di un nuovo sistema
di distribuzione dell’Ozono con o senza un sigillante della radice, utilizzando il Monitoraggio
elettrico della carie (ECM), DIAGNOdent e un criterio di individuazione clinica per il trattamento
della carie della radice.
dott Cosimo Cigliola
37

MATERIALI E METODI
Pazienti considerati per lo studio
L’approvazione etica è stata ottenuta dal District Ethics Committee della Queen's University di
Belfast. I dati sono stati ottenuti da 220 PRCL di 79 pazienti. E’ stato selezionato un totale di 49
(62%) partecipanti di sesso maschile e 30 (38%) di sesso femminile con almeno una PRCL.
All’inizio del lavoro l’età media (± errore standard) dei soggetti era di 65 anni (± 14.76) con un
minimo di 30 e un massimo di 72 anni. In questo studio sono state scelte solo carie della radice
nella categoria media di severità (lesioni coriose con indice di severità 2) secondo l’indice di
necessità percepita di trattamento [Beighton et al., 1993], su almeno due superfici di due denti
diversi che erano accessibili per le procedure diagnostiche.
Attrezzatura utilizzata
o Sistema di distribuzione di Ozono
Il sistema di distribuzione di Ozono è un apparecchio portatile con un generatore di Ozono per il
trattamento della carie che distribuisce Ozono ad una concentrazione di 2,100 ppm ± 10% (Figura
1). Una pompa a vuoto spinge aria attraverso il generatore a 615 cc/min in modo da fornire
Ozono alla lesione e purificare il sistema dall’Ozono al termine del trattamento. E’ dotato anche di
una cappetta in silicone monouso removibile da applicare al
manipolo, applicabile sull’area del dente da trattare. La ventosa,
che si serra strettamente allo strumento, sigilla l’area scelta del
dente in modo da prevenire la fuga di Ozono (Figura 2). Un
sistema di aspirazione rimuove poi ogni possibile traccia di
Ozono mentre la ventosa è ancora applicata alla PRCL (il
sistema di aspirazione trasmette il gas dal sistema di
distribuzione attraverso ioni di Manganesio (II)). Questo
neutralizzatore di Ozono converte l’Ozono in ossigeno. Il
sistema, infine, emette un liquido che contiene Xilitolo e 1000
ppm di Fluoruro attraverso la cappetta sigillante in modo da
neutralizzare ogni possibile residuo di Ozono.
Figura 2 Manipolo, con cappetta applicata.
dott Cosimo Cigliola
Figura 1 Sistema di distribuzione dell’Ozono.
38

o L’ECM III (Lode Diagnostics BV, Groningen, Olanda)
La resistenza elettrica è stata presa nei punti centrale, mesiale, distale, occlusale e
linguale/palatale di ogni PRCL [Baysan et al., 2001a]. Il monitor registra il valore al termine del
tempo di essiccazione (valore finale) e l’area sotto la curva durante il periodo di essiccazione
stesso (valore integrato).
o Il DIAGNOdent
Il DIAGNOdent (KaVo, Germania) è stato utilizzato per
individuare e quantificare la severità delle PRCL.
Questo apparecchio misura la fluorescenza laser all’interno Figura 3 DIAGNOdent.
della struttura del dente. Quando la luce laser viene propagata dentro la lesione cariosa, le fibre
ottiche a due vie permettono all’unità di quantificare la luce laser riflessa. Una superficie di dente
sano presenta fluorescenza scarsa o nulla, visualizzando sul display letture in scala minime.
Tuttavia le lesioni cariose presentano alcuni livelli di fluorescenza proporzionali alla severità delle
lesioni, che risultano letture in scala sul display elevate. Il sistema DIAGNOdent registra i valori
istantanei e di picco. La lettura istantanea indica il valore in tempo reale che viene misurato dalla
punta della sonda, mentre il valore di picco si riferisce al livello più alto riportato sul dente.
L’apparecchio DIAGNOdent si accende premendo l’anello grigio sullo strumento da impugnare. La
calibrazione viene effettuata prima di ogni sessione secondo le istruzioni del produttore. Il valore
di picco è stato sottoposto ad analisi statistiche.
Materiale usato
- Sigillante per radici.
Il sigillante della radice (Seal & Protect, Dentsply, Germany) è autoadesivo e contiene una miscela
di resina dimetacrilata in acetone come solvente, con triclosan e fluoruro.
Scopo dello studio
Lo studio ha coinvolto 79 pazienti con 2 o 4 PRCL che sono stati suddivisi in modo casuale in
quattro gruppi.
Questi gruppi sono stati trattati nel modo seguente:
Gruppo 1. Gruppo con solo Ozono
1. All’inizio sono state effettuate le misurazioni ECM e DIAGNOdent.
2. Successivamente sono stati valutati i criteri clinici per individuare le PRCL.
3. Dopo averla pulita con uno spazzolino sterile e con acqua, la superficie del dente è stata isolata
e asciugata con un rotolo di lana e cotone sterile.
4. E’ stato emesso Ozono per una durata di 10 secondi nella cappetta, applicata serratamente ad
ogni PRCL.2
dott Cosimo Cigliola
39

5. L’aspirazione si è attivata in modo automatico per 10 s. Come parte del sistema di aspirazione,
il residuo di Ozono catturato è stato trasmesso dal sistema di distribuzione attraverso ioni di
Manganesio (II), che agivano come neutralizzatori di Ozono (consumatore).
6. Il liquido riducente ha riempito la cappetta per una durata di 5 s e la componente di aspirazione
del sistema ha rimosso ogni possibile traccia di Ozono mentre la cappetta era ancora applicata alla
PRCL.
7. Per riserva, è stato utilizzato in ogni occasione anche il sistema di aspirazione ad alta velocità
del riunito.
8. Dopo l’applicazione di Ozono la carie della radice non è stata rimossa.
9. Dopo un mese, le procedure applicate inizialmente sono state ripetute senza l’applicazione di
Ozono.
10. Dopo tre mesi, le stesse procedure sono state ripetute con l’applicazione di Ozono per una
durata di 10 s.
Gruppo 2. Gruppo senza Ozono e senza sigillante della radice (Gruppo di controllo)
1. All’inizio sono state effettuate le misurazioni ECM e DIAGNOdent.
2. Successivamente sono stati valutati i criteri clinici per individuare le PRCL, come per il gruppo
1.
3. Le PRCL hanno ricevuto solo il liquido, senza l’applicazione di Ozono per una durata di 5 s.
4. Le stesse procedure sono state ripetute dopo 1 e 3 mesi.
Gruppo 3. Gruppo con Ozono e sigillante
1. Al’inizio sono state effettuate le misurazioni di ECM e DIAGNOdent.
2. Successivamente sono stati valutati i criteri clinici per individuare le PRCL, come per il gruppo
1.
3. In questo gruppo è stato applicato un sigillante per radice (Seal & Protect, Dentsply, Germany)
sopra la PRCL senza rimuovere la dentina cariata e dopo l’applicazione di Ozono per 10 secondi
come per il gruppo 1.
4. Per precauzione, ad ogni trattamento è stato anche utilizzato il sistema di aspirazione ad alta
velocità del riunito.
5. Dopo un mese, le procedure applicate inizialmente sono state ripetute senza l’applicazione di
Ozono.
6. Sempre dopo un mese sono state eseguite anche delle valutazioni cliniche dei sigillanti secondo
i criteri USPHS modificati (Concilio dei Materiali dentali, Strumenti e attrezzature, 1994).
7. I sigillanti sono stati di nuovo applicati, ma solo in caso di sospetto di parziale o totale perdita
del sigillante.
8. Dopo tre mesi le stesse procedure sono state ripetute. In aggiunta, l’Ozono è stato applicato
per una durata di 10 s. Anche i sigillanti sono stati applicati, ma solo in caso di sospetto di parziale
o totale perdita del sigillante.
Group 4. Gruppo con solo sigillante per radici
dott Cosimo Cigliola
40

1. Inizialmente sono state eseguite misurazioni con resistenza elettrica e DIAGNOdent su ogni
lesione prima e dopo l’applicazione del sigillante alla radice.
2. Sono stati applicati i criteri clinici per individuare le PRCL.
3. In questo gruppo, a seguito dell’applicazione di liquido senza Ozono per una durata di 5 s, il
sigillante è stato messo solo sulla parte superiore delle PRCL, senza rimozione della dentina
cariata.
4. Anche in questo caso, dopo uno e tre mesi rispettivamente sono state eseguite misurazioni
della resistenza elettrica, valutazioni cliniche e i criteri USPHS modificati .
5. Sempre dopo uno o tre mesi è stato applicato il sigillante nel caso fossero andate parzialmente
o totalmente persi. Se i pazienti presentavano una qualunque forma di disagio, le PRCL di ogni
gruppo venivano immediatamente trattate con le procedure convenzionali di asportazione e
otturazione. I pazienti dei gruppi di cui sopra utilizzavano dentifricio standard, contenente 1,100
ppm di Fluoruro di Sodio (dentifricio sbiancante, Natural White, U.S.A) e spazzolini morbidi (Soft
bristle, Natural White, U.S.A). I dentifrici e gli spazzolini sono stati forniti durante il periodo di
studio, cioè dopo uno e tre mesi, o prima, per posta, se richiesto.
Analisi statistica
Le medie delle letture di resistenza e DIAGNOdent registrate all’inizio e rispettivamente dopo uno,
tre e sei mesi sono state utilizzate per l’analisi dei dati. Successivamente, le letture ECM sono
state trasformate con la funzione di logaritmo in base 10 in modo da normalizzare la varianza per
tutti i gruppi.
Sono state registrate per ogni variabile (cavità, dimensione, distanza dal margine gengivale e
indice di severità) media ed errore standard. Dove veniva eseguita l’analisi statistica, il gruppo con
Ozono è stato confrontato con il gruppo di controllo, mentre il gruppo con sigillante e Ozono è
stato confrontato con il gruppo con solo sigillante. Nei gruppi con Ozono e di controllo la variabile
principale risultante era invertita rispetto all’indice di severità, mentre per i gruppi con sigillante e
Ozono e con solo Ozono l’adattamento marginale del sigillante alla radice era la variabile
principale risultante.
- Indice di durezza e di severità
Sono state testate le differenze nel numero di lesioni che si induriscono il cui indice diventa meno
grave.
- Letture ECM e DIAGNOdent, distanza dal margine gengivale, cavità e dimensione.
Sono state calcolate le differenze di resistenza ECM (DR), misurazioni DIAGNOdent (DD), distanza
dal margine gengivale (DDGM), cavità (DC) e dimensione (DS) fra inizio dello studio, a uno, tre e
sei mesi, sottraendo i valori a uno, tre e sei mesi da quelli iniziali. Tutti i test statistici sono stati
eseguiti con il sistema statistico SPSS per Windows MS versione 6.1. e la soglia di significatività
statistica è stata fissata a 0.05.
dott Cosimo Cigliola
41

RISULTATI
Durezza All’inizio tutte le lesioni erano di consistenza coriacea. Le percentuali di durezza delle
PRCL nel gruppo dell’Ozono sono illustrate in Figura 3 (p < 0.001).
Figura 3 Percentuali di durezza delle PRCL nel gruppo di solo Ozono dopo uno, tre e sei mesi.
Cavità
40%
30%
20%
10%
0%
Lesioni inizialmente non incavate (

Valori ottenuti con l'ECM
I valori ECM medi sono mostrati in Tabella 1. All’inizio, i valori ECM erano simili per tutti i gruppi. I
valori ECM medi per il gruppo di controllo tendono a decrescere in tutti i controlli (p < 0.05). In
contrasto, I valori ECM medi delle lesioni dei gruppi con solo Ozono, con Ozono e sigillante e solo
con sigillante aumentano durante lo studio se confrontati con i valori iniziali. Si riscontrano
differenze statisticamente significative nei cambiamenti delle letture ECM per i gruppi con Ozono e
di controllo relativamente agli esami a uno, tre e sei mesi nei modelli di regressione (p < 0.001).
O3
Nessun
trattamento
O3 + sigillante Solo sigillante
Inizio 5.24 ± 0.04 5.20 ± 0.04 5.24 ± 0.31 5.25 ± 0.34
1 mese 5.78 ± 0.07 5.18 ± 0.03 6.30 ± 0.77 5.95 ± 0.93
3 mesi 5.63 ± 0.08 5.13 ± 0.03 5.75 ± 0.66 5.60 ± 0.71
6 mesi 5.62 ± 0.12 4.92 ± 0.21 5.56 ± 0.14 5.45 ± 0.16
Tabella 1 Medie (± errore standard) log10 dei valori ECM per tutti i gruppi.
dott Cosimo Cigliola
43

Valori ottenuti con il DIAGNOdent
I valori DIAGNOdent medi sono mostrati in Tabella 2. All’inizio, le letture con il DIAGNOdent erano
simili per tutti i gruppi. I valori DIAGNOdent medi per il gruppo di controllo tendono a decrescere
dopo uno, tre e sei mesi se confrontati con i valori iniziali (p

Il principale problema clinico degli approcci farmaceutici nel trattamento della carie radicolare è la
difficoltà nel sopprimere ed eliminare i microrganismi per un periodo piuttosto lungo di tempo.
Dopo il trattamento con farmaci selezionati, gli organismi possono proliferare e ricolonizzare le
PRCL. E’ interessante il fatto che il 38,1% delle lesioni in questo studio hanno avuto una
remineralizzazione dopo 6 mesi. Si possono fare congetture riguardo al fatto che la maggior parte
delle lesioni si demineralizzano. Questo è associato a svariati fattori, incluso il livello di riduzione
microbica e gli effetti ossidanti sulle PRCL. La drastica riduzione della flora microbica permette una
eradicazione della nicchia ecologica dei microrganismi acidogeni e acidurici. Questo spostamento
della flora microbica verso i normali commensali orali dovrebbe prevalentemente permettere alla
remineralizzazione di aver luogo durante il processo carioso. In aggiunta, un ossidante (ipoclorito
di sodio) precedentemente è stato dimostrato di migliorare il potenziale di remineralizzazione della
dentina demineralizzata. Inaba et al., [1995] hanno scoperto che l’utilizzo di un ossidante
(ipoclorito di sodio 10%) sulle lesioni demineralizzate della dentina radicolare migliora il loro
potenziale di remineralizzazione, perché l’ipoclorito di sodio è un agente proteolitico non specifico
ed è efficace nella rimozione di componenti organici delle lesioni. Successivamente Inaba et al.,
[1996] hanno dimostrato che quando dei campioni di dentina della radice vengono trattati con
questo ossidante per 2 min, aumenta la permeabilità degli ioni fluoruro (F - ) arrivando alla
conclusione che la rimozione di materiale organico dalle lesioni della dentina rappresenta una
tecnica accettabile per aumentare la remineralizzazione. Questo potrebbe essere un parziale
fattore degli ottimi risultati di remineralizzazione ottenuti in questo studio a seguito delle
applicazioni di Ozono. Potrebbe altresì essere indice del fatto che l’Ozono ha la capacità di
rimuovere le proteine nelle lesioni di carie e di agevolare la diffusione di ioni Calcio e di Fosfato
attraverso le lesioni, un fenomeno che in questo studio ha come risultato la remineralizzazione di
alcune PRCL a seguito dell’applicazione di Ozono. Dopo l’iniziale soppressione totale dei
microrganismi, la ricolonizzazione dei microrganismi potrebbe essere ritardata da una resistenza
della normale flora commensale orale all’intrusione di organismi all’interno delle lesioni. In
aggiunta, la nicchia ecologica di questi microrganismi acidogeni e acidurici verrebbe gravemente
disgregata, fatto che a sua volta potrebbe interferire con la ricolonizzazione e la ricrescita di
questa specifica microflora. Ciò potrebbe sfociare in una soppressione di lunga durata dei
microrganismi acidogenici e acidurici nelle PRCL. Anche Emilson [1981] ha riportato che dopo un
breve ed intensivo trattamento della bocca con clorexidina all’1%, l’S Mutans è stato soppresso in
vivo per un periodo di tempo significativamente lungo (14 settimane). E’ possibile che
l’ipermineralizzazione della superficie abbia minor probabilità di aver luogo al termine
dell’applicazione di Ozono. Dal momento che l’Ozono è un potente ossidante, indubbiamente
ossigenerà le biomolecole delle PRCL aprendo quindi i tubuli dentinali delle lesioni. L’Ozono
potrebbe anche favorire la diffusione all’interno della lesione di ioni Calcio e Fosfato. Inoltre, si
deve osservare, che i pazienti hanno utilizzato un dentifricio contenente una quantità standard di
fluoruro. Martens and Verbeeck [1998] hanno riportato che basse concentrazioni di fluoruro
hanno la capacità di remineralizzare lesioni cariose nella profondità. A questo riguardo, sono
necessari studi futuri per determinare il significato di questi studi. I materiali riempitivi falliscono
con frequenza allarmante e sono molto costosi in termini di dolore, disagio e fattori economici. In
dott Cosimo Cigliola
45

Inghilterra e nel Galles nel 2000 lo Stato ha speso 1.5 miliardi di sterline per l’odontoiatria. Ciò
non include gli studi privati, che attualmente si stima contribuisce al 50% sugli introiti dei dentisti.
Inoltre, ospedali e cliniche Odontoiatriche ricevono fondi molto sostanziosi per la cura dei pazienti.
Il costo totale di tutti i trattamenti Odontoiatrici in Inghilterra e nel Galles nel 2000 va
probabilmente oltre i 3 miliardi di sterline [General Dental Council, Annual Statistics, 2000].
La maggiorparte di queste spese sono da ascriversi a otturazioni, otturazioni della radice, protesi
dentarie, corone e ponti, ma almeno una parte dei costi associati a trattamenti odontoiatrici può
essere associata ad aspetti iatrogeni del trattamento della carie. Rapporti pubblicati indicano che
approssimativamente la metà dei restauri applicati nella pratica dentistica sono delle sostituzioni
di restauri precedenti e circa la metà di questi vengono sostituiti a causa di carie secondarie. Negli
USA si riscontra un trend simile, nonostante la prevalenza di carie sia diminuita. Il costo sanitario
relativo all’odontoiatria, negli USA è stimato per 52 bilioni di dollari l’anno, di cui la metà può
essere associato a trattamenti conservativi. E’ altresì riportato che vengono persi diversi giorni
lavorativi l’anno a causa di malattie orali. Nonostante progressi nella ricerca clinica e di
laboratorio, approssimativamente il 50% della popolazione americana di età superiore ai 65 anni
mostra evidenze di carie della radice. [Anusavice, 2000]. Chiaramente, esiste la possibilità di
risparmi considerevoli sia nei paesi avanzati, che nei paesi poveri e specialmente in quelli in via di
sviluppo. Questi potrebbero essere realizzati sfruttando il trattamento con Ozono e magari
combinando il trattamento con Ozono con l’uso di un dentifricio con elevato contenuto di fluoruro
(Lynch et al 2000, Baysan et al 2001a, Lynch and Baysan 2001). I risultati raccolti dallo
studio sull’Ozono sono molto promettenti. L’utilizzo di Ozono è sicuro ed efficace in termini di costi
e di tempo. Una corretta igiene orale e l’assistenza professionale per la cura della bocca diventano
difficili per le persone più anziane, perché condizioni somatiche e mentali diventano
compromettenti. Queste situazioni possono essere superate se si ricorre a strategie preventive di
intervento. A questo riguardo, può essere preso in considerazione l’utilizzo di Ozono, specie per
pazienti patologicamente compromessi, pazienti che necessitano di assistenza domiciliare, e
persone anziane costrette a rimanere a casa [Baysan et al., 2001b]. Per il trattamento con
Ozono non è necessaria alcuna iniezione e il sistema di distribuzione dell’Ozono è portatile. Per
questo, persone anziane che hanno accesso limitato ai servizi odontoiatrici possono avere
moltissimi beneficiare con questo trattamento.
In conclusione, questo nuovo regime di trattamento che utilizza l’Ozono è in grado di far
regredire clinicamente la PRCL persistenti e può essere considerato una rivoluzionaria
alternativa alla tecnica convenzionale di “trapanazione e otturazione”. Inoltre, il
sigillante radicolare ha una migliore adesione su PRCL persistenti trattate con Ozono.
dott Cosimo Cigliola
46

ENGLISH ABSTRACT
Sistematic review of the running literature on use of the
LASER, OZONE and new Technologies in Oral Hygiene
Treating open carious lesions in anxious children with ozone. A prospective controlled
clinical study.
Dahnhardt JE, Jaeggi T, Lussi A.
Department of Operative, Preventive and Pediatric Dentistry, School of Dental Medicine, University
of Bern, Switzerland. daehnhardt@zmk.unibe.ch
PURPOSE: To determine whether the treatment of dental caries with ozone was possible in
apprehensive children and to ascertain whether ozone reverses caries in open single-surface
lesions. Further, the influence of ozone on laser fluorescence was investigated.
METHODS: 82 lesions in 28 children with at least two open single-surface lesions were assessed.
The children were anxious and were judged by the referring dentist as non-treatable. For each
test lesion,
which was treated with ozone, a control lesion was left without ozone treatment. Hardness and
laser fluorescence values were assessed and the changes for hardness and laser fluorescence
values in the test lesion were compared with the values in the control lesion after 2, 4, 6, and 8
months.
RESULTS: 94 percent of the children were treatable and 93% lost their dental anxiety. The
hardness values improved significantly in the ozone-treated test lesions after 4, 6, and 8 months
(P< 0.05) compared with baseline while the control lesions had no significant change in hardness
at any recall interval. Comparing the differences between test and control teeth over time, the
laser fluorescence values improved, however the improvement was not statistically significant (P>
0.05). The use of ozone resulted in an average reduction of 13% of the laser fluorescence values
immediately after the ozone treatment.
PMID: 17073201 [PubMed - indexed for MEDLINE]
Am J Dent. 2005 Aug;18(4):223-8.
Effect of ozone on non-cavitated fissure lesions in permanente molar. A controlled
prospective clinical study.
dott Cosimo Cigliola
47

Huth KC, Paschos E, Brand K, Hickel R.
Department of Restorative Dentistry and Periodontology, Dental School, Ludwig-Maximilians-
University, Goethe Street 70, D-80336 Munich, Germany. khuth@dent.med.uni-muenchen.de
PURPOSE: To investigate, with a randomized controlled clinical study, the effect of ozone on non-
cavitated initial occlusal fissure caries compared with untreated contra-lateral control lesions (split
mouth) considering the patient's current caries risk.
METHODS: Forty-one patients with 57 pairs of lesions were enrolled in the study (mean age 7.7
+/- 2.2 years; upper jaw n=29, lower jaw n=28). Gaseous ozone (HealOzone) was applied once
for 40 seconds to the randomly assigned test molar of each pair without the use of remineralizing
solutions. Lesion progression or reversal was monitored by the laser fluorescence system
DIAGNOdent for up to 3 months and the deterioration or improvement compared between the
ozone-treated lesions and the untreated control lesions (in pairs). This was done for the whole
study population and a subgroup of patients with high current caries risk (lesion pairs n=26).
RESULTS: After 3 months, explorative data analysis revealed that the ozone-treated lesions
showed significantly more caries reversal or reduced caries progression than the untreated control
lesions within the group of patients at high current caries risk (Wilcoxon-Test, P= 0.035). There
was no statistical significance examining the whole study population. From the data it can be
concluded that ozone application significantly improved non-cavitated initial fissure caries in
patients at high caries risk over a 3-month period.
PMID: 16296426 [PubMed - indexed for MEDLINE]
Cochrane Database Syst Rev. 2004;(3):CD004153.
Ozone therapy for the treatment of dental caries.
Rickard GD, Richardson R, Johnson T, McColl D, Hooper L.
Old Sandstone Dental Practice, The Chine, Dorking, Surrey, UK, RH4 1QT.
BACKGROUND: Dental caries is a bacterially mediated disease characterised by demineralisation
of the tooth surface, which may lead to cavitation, discomfort, pain and eventual tooth loss.
Ozone is toxic to certain bacteria in vitro and it has been suggested that delivering ozone into a
carious lesion might reduce the number of cariogenic bacteria. This possibly could arrest the
progress of the lesion and may, in the presence of fluoride, perhaps allow remineralisation to
occur. This may in turn delay or prevent the need for traditional dental conservation by 'drilling
and filling'.
dott Cosimo Cigliola
48

OBJECTIVES: To assess whether ozone is effective in arresting or reversing the progression of
dental caries.
SEARCH STRATEGY: We searched the Cochrane Oral Health Group's Trials Register (to 7
November 2003); Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library
Issue 3, 2003); MEDLINE and PREMEDLINE (OVID, 1966 to November 2003); EMBASE (OVID,
1980 to November 2003); CINAHL (OVID, 1982 to November 2003); AMED (OVID, 1985 to
November 2003). Quintessence was handsearched through 2002 and KaVo were contacted as
manufacturers of the HealOzone apparatus for any additional published or unpublished trials.
SELECTION CRITERIA: Inclusion was assessed independently by at least two reviewers. Trials
were only included if they met the following criteria: randomisation in a controlled trial; single
surface in vivo carious lesion accessible to ozone application; clear allocation concealment; ozone
application to the lesions in the intervention group; no such application of ozone in the control
group; outcomes measured after at least 6 months.
DATA COLLECTION AND ANALYSIS: Reviewers independently extracted information in
duplicate. A paucity of comparable data did not allow meta-analytic pooling of the included
studies.
MAIN RESULTS: Three trials were included, with a combined total of 432 randomised lesions
(137 participants). Forty-two conference papers, abstracts and posters were excluded (from an
unknown number of studies). The risk of bias in all studies appeared high. The analyses of all
three studies were conducted at the level of the lesion, which is not independent of the person, for
this reason pooling of data was not appropriate or attempted. Individual studies showed
inconsistent effects of ozone on caries, across different measures of caries progression or
regression. Few secondary outcomes were reported, but one trial reported an absence of adverse
events.
REVIEWERS' CONCLUSIONS: Given the high risk of bias in the available studies and lack of
consistency between different outcome measures, there is no reliable evidence that application of
ozone gas to the surface of decayed teeth stops or reverses the decay process. There is a
fundamental need for more evidence of appropriate rigour and quality before the use of ozone can
be accepted into mainstream primary dental care or can be considered a viable alternative to
current methods for the management and treatment of dental caries.
PMID: 15266519 [PubMed - indexed for MEDLINE]
Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial.
Holmes J.
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49

UKSmiles Dental Practice, Wokingham, Berkshire, UK. julian@o3dc.co.uk
OBJECTIVE: To assess the effect of an ozone delivery system, combined with the daily use of a
remineralising patient kit, on the clinical severity of non-cavitated leathery primary root carious
lesions (PRCL's), in an older population group.
DESIGN: A total of 89 subjects, (age range 60-82, mean +/- SD, 70.8 +/- 6 years), each with
two leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for
treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were
recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft,
leathery or hard, scored with a validated root caries severity index.
RESULTS: There were no observed adverse events. After three months, in the ozone-treated
group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group,
four PRCL's (4%) had become worse (p

management of root caries. The best management strategy still remains to be developed. Initial
studies have indicated that an application of ozone for a period of either 10 or 20 seconds is
capable of clinically reversing leathery root carious lesions. It is suggested that, subject to
confirmation from extensive trials, this simple and non-invasive technique may benefit many
patients with root caries throughout the world since this approach to treat root caries can easily
be employed in primary care clinics and in the domiciliary treatment of home-bound elderly people
and immobile patients in hospices and hospitals.
PMID: 16393498 [PubMed - indexed for MEDLINE]
Stomatologiia (Mosk). 2005;84(6):20-2.
[Experience in medical ozone use for root canal treatment] [Article in Russian]
Bezrukova IV, Petrukhina NB, Voinov PA.
The results of clinical and laboratory assessment of effectiveness of PSR-diagnosis are presented.
The high efficacy of ozone therapy is revealed. The findings demonstrate the reduction of number
in the micro-organisms in root canal: Actinobacillus actinomycetemcomitans from 31.25% to
10.21%; Bacteriodes forsythus from 68.75% to 15.50%; Treponema denticola from 37.5% to
11.4%; Porphyromonas gingivalis from 56.25% to 45%; Prevotella intermedia from 16% to 0%.
PMID: 16353031 [PubMed - indexed for MEDLINE]
Dent Clin North Am. 2005 Oct;49(4):905-21, viii.
Nonsurgical treatment of incipient and hidden caries.
Thompson VP, Kaim JM.
Department of Biomaterials & Biomimetics, New York University College of Dentistry, 345 East
24th Street, 804S, New York, NY 10010, USA. van.thompson@nyu.edu
Traditionally, dentists have been trained in the surgical model for caries management whereby
detection is akin to diagnosis. This model unfortunately has been translated to patient
expectations. Nevertheless, a growing body of clinical evidence suggests that noncavitated
lesions, even those extending into dentin, can be managed by nonsurgical means with an
expectation for remineralization.
PMID: 16150323 [PubMed - indexed for MEDLINE]
Mediators Inflamm. 2004 Feb;13(1):3-11.
Ozone as Janus: this controversial gas can be either toxic or medically useful.
dott Cosimo Cigliola
51

Bocci V.
Department of Physiology, University of Siena, Italy. bocci@unsi.it
Ozone is an intrinsically toxic gas and its hazardous employment has led to a poor consideration of
ozone therapy. The aim of this review is to indicate that a wrong dogma and several
misconceptions thwart progress: in reality, properly performed ozone therapy, carried out by
expert physicians, can be very useful when orthodox medicine appears inadequate. The
unbelievable versatility of ozone therapy is due to the cascade of ozone-derived compounds able
to act on several targets leading to a multifactorial correction of a pathological state. During the
past decade, contrary to all expectations, it has been demonstrated that the judicious application
of ozone in chronic infectious diseases, vasculopathies, orthopedics and even dentistry has yielded
such striking results that it is deplorable that the medical establishment continues to ignore ozone
therapy.
PMID: 15203558 [PubMed - indexed for MEDLINE]
Toxicol Appl Pharmacol. 2006 Nov 1;216(3):493-504. Epub 2006 Jun 27.
Is it true that ozone is always toxic? The end of a dogma.
Bocci V.
Department of Physiology, University of Siena, via A. Moro 2, Siena 53100, Italy. bocci@unisi.it
There are a number of good experimental studies showing that exposure by inhalation to
prolonged tropospheric ozone damages the respiratory system and extrapulmonary organs. The
skin, if extensively exposed, may also contribute to the damage. The undoubtful strong reactivity
of ozone has contributed to establish the dogma that ozone is always toxic and its medical
application must be proscribed. Although it is less known, judiciously practiced ozonetherapy is
becoming very useful either on its own or applied in combination with orthodox medicine in a
broad range of pathologies. The opponents of ozonetherapy base their judgment on the ozone
chemistry, and physicians, without any knowledge of the problem, are often skeptical. During the
last 15 years, a clear understanding of the action of ozone in biology and medicine has been
gained, allowing today to argue if it is true that ozone is always toxic.
The fundamental points that are discussed in this paper are: the topography, anatomical and
biochemical characteristics of the organs daily exposed to ozone versus the potent antioxidant
capacity of blood exposed to a small and precisely calculated dose of ozone only for a few
minutes. It is becoming clear how the respiratory system undergoing a chronic oxidative stress
can release slowly, but steadily, a huge amount of toxic compounds able to enter the circulation
and cause serious damage. The aim of this paper is to objectively evaluate this controversial
issue.
PMID: 16890971 [PubMed - indexed for MEDLINE]
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52

J Dent. 2007 Mar;35(3):195-200. Epub 2006 Oct 6.
Assessment of the safety of two ozone delivery devices.
Millar BJ, Hodson N.
Primary Care Dentistry, King's College London Dental Institute at Guy's, King's College & St.
Thomas' Hospitals, Caldecot Road, London SE5 9RW, UK. brian.millar@kcl.ac.uk
OBJECTIVES: To evaluate the safety of an ozone gas device designed for use in dentistry.
METHODS: Two commercially available ozone applicators, Ozi-cure and HealOzone were used in a
clinical simulation using a phantom head while recordings of ozone levels were made in
pharyngeal and nasal regions of the patient and near the mouth of the operator. Clinical
simulations included ozone application for caries management and endodontic treatment.
Recordings were made five times with different levels of suction to assess the effect on ozone
levels.
RESULTS: The results with Ozi-cure on caries mode resulted in a peak ozone level in the pharynx
of 1.33+/-0.52 ppm when no suction was used. The use of suction nearby reduced the ozone level
to zero while suction on the opposite side of the mouth reduced the level to 0.22+/-0.04 ppm.
Used on endodontic mode the peak ozone level in the pharynx was 5.51+/-1.63 ppm when no
suction was used. The use of suction nearby reduced the ozone level to zero while suction on the
opposite side of the mouth reduced the level to 0.84+/-0.54 ppm. Recordings in the patient's
nasal region gave a peak of 0.22 ppm when using the Ozi-cure on endodontic mode with no
suction. At the operator's mouth the ozone level did not exceed 0.01 ppm although the
characteristic smell of ozone was detectable. All recordings with HealOzone were zero.
Concentrations of 15 ppm were recorded in a simulated tooth cavity with Ozi-cure and >20 ppm
with HealOzone.
CONCLUSIONS: The Ozi-cure device when used without adequate suction allows ozone to be
reach a concentration above permitted levels and therefore should not be used. The HealOzone
was safe to use.
PMID: 17030396 [PubMed - in process]
Res Rep Health Eff Inst. 1994 Nov;(65 Pt 7):3-26; discussion 27-34.
Consequences of prolonged inhalation of ozone on F344/N rats: collaborative studies.
Part VII: Effects on the nasal mucociliary apparatus.
Harkema JR, Morgan KT, Gross EA, Catalano PJ, Griffith WC.
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53

Inhalation Toxicology Research Institute, Lovelace Biomedical and Environmental Research
Institute, Albuquerque, NM.
Besides the centriacinar region of the lung, the nose is a principal target for ozone toxicity. Acute
exposures to concentrations of ozone in ambient air induce secretory cell metaplasia in the nasal
transitional epithelium of rats. This study examined the effects of chronic ozone exposure on the
structure and function of the nasal mucociliary apparatus of the rat. Male and female F344/N rats
were exposed to ozone concentrations of 0.0 (controls), 0.12, 0.5, or 1.0 parts per million (ppm),
six hours per day, five days per week, for 20 months. All rats were killed seven or eight days after
the end of the exposure. Immediately after death, mucous flow rates throughout the nasal
passages were determined using in vitro video motion analysis. Following assessment of
mucociliary function, the nasal tissues were processed for light microscopy and stained with Alcian
blue (pH 2.5)/periodic acid-Schiff to detect intraepithelial mucus. Image analysis was used to
quantitate the amount of mucus within the nasal transitional epithelium. In rats exposed to 0.5 or
1.0 ppm ozone, mucous flow rates were markedly slower over the lateral wall and turbinates of
the proximal third of the nasal airways than they were in rats exposed to 0.0 or 0.12 ppm ozone.
These intranasal regions in the rats exposed to 0.5 or 1.0 ppm ozone contained marked mucous
cell metaplasia and 25 to 300 times more mucus in nasal transitional epithelium than was found in
control rats. In addition, male and female rats exposed to 0.5 or 1.0 ppm ozone had marked
epithelial hyperplasia in nasal transitional epithelium, increases in eosinophilic globules in the
surface epithelium lining the distal nasal airways, and a mild to moderate inflammatory cell influx
in the nasal mucosa in the proximal and middle nasal passages. Male rats also had conspicuous
bony atrophy in maxilloturbinates and nasoturbinates. There were no significant decreases
between the mucous flow rates of rats exposed to 0.12 ppm ozone and those of control rats.
There were, however, mild increases in various flow rates in some areas of the nasal airways in
rats exposed to 0.12 ppm ozone compared with control rats. No significant morphologic
alterations were evident in the rats exposed to 0.0 or 0.12 ppm ozone. The results of this study
indicate that rats chronically exposed to 0.5 or 1.0 ppm ozone have significant alterations in the
function and structure of the nasal mucociliary apparatus. Though there was a mild increase in
mucous flow rates in a few nasal regions of some rats exposed to 0.12 ppm ozone, this functional
change was interpreted as a physiologic, rather than a pathologic, response to ozone at this
relatively low concentration.
PMID: 7888110 [PubMed - indexed for MEDLINE]
Am J Respir Cell Mol Biol. 1997 May;16(5):521-30.
Mucous cell metaplasia in rat nasal epithelium after a 20-month exposure to ozone: a
morphometric study of epithelial differentiation.
Harkema JR, Hotchkiss JA, Griffith WC.
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54

Department of Pathology, College of Veterinary Medicine, Michigan State University, East Lansing
48824, USA.
The present study was designed to examine the effects of long-term ozone exposure on nasal
epithelia and intraepithelial mucosubstances (IM) throughout the nasal airways of F344/N rats.
Animals were exposed to 0 (controls), 0.12, 0.5, or 1.0 ppm ozone, 6 h/day, 5 days/wk, for 20
mo. Rats were killed 1 wk after the end of the exposure, and nasal tissues were processed for
light and electron microscopy. Standard morphometric techniques were used to determine
epithelial cell densities and the amounts of IM in the surface epithelium lining the nasal airways.
No mucous cells or IM were present in the epithelia lining the nasal lateral meatus and maxillary
sinus of rats exposed to 0 or 0.12 ppm ozone. In contrast, rats exposed to 0.5 or 1.0 ppm ozone
had marked mucous cell metaplasia (MCM) with numerous mucous cells and conspicuous amounts
of IM in the surface epithelium lining these upper airways. Ozone-induced increases in total
epithelial cells (i.e., epithelial hyperplasia) were present only in rats exposed to 1.0 ppm. The
results of this study indicate that rats chronically exposed to 1.0 or 0.5 ppm, but not 0.12 ppm,
ozone can develop marked MCM with significant increases in IM in both proximal and distal nasal
airways. The epithelial changes observed throughout the nasal passages of ozone-exposed rats
may be adaptive responses in an attempt to protect the upper and lower respiratory tract from
further ozone-induced injury.
PMID: 9160834 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2006 Oct;114(5):435-40
Effect of ozone on oral cells compared with established antimicrobials.
Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K.
Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-University, Munich,
Germany. khuth@dent.med.uni-muenchen.de
Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its
antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous
ozone (4 x 10(6) microg m(-3)) and aqueous ozone (1.25-20 microg ml(-1)) exert any cytotoxic
effects on human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with
established antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl)
5.25%, 2.25%; hydrogen peroxide (H(2)O(2)) 3%], over a time of 1 min, and compared with the
antibiotic, metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and
apoptosis were evaluated. Ozone gas was found to have toxic effects on both cell types.
Essentially no cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic
to BHY cells, and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H(2)O(2) resulted
in markedly reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity
only to BHY cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of
the tested antiseptics.
dott Cosimo Cigliola
55

PMID: 17026511 [PubMed - indexed for MEDLINE]
Biofactors. 2006;27(1-4):5-18.
High resolution 1H NMR investigations of the oxidative consumption of salivary
biomolecules by ozone: relevance to the therapeutic applications of this agent in clinical
dentistry.
Grootveld M, Silwood CJ, Lynch E.
Department of Applied Science, London South Bank University, 103 Borough Road, London, UK.
grootvm@lsbu.ac.uk
High resolution proton (1H) nuclear magnetic resonance (NMR) spectroscopy was employed to
simultaneously evaluate the oxidising actions of ozone (O3) towards a wide range of salivary
biomolecules in view of its applications in dental practices, which may serve as a viable and
convenient means for the treatment of dental caries. Treatment of supernatants derived from
unstimulated human saliva specimens (n=12) with O3 (4.48 mmol) revealed that this reactive
oxygen species gave rise to the oxidative consumption of pyruvate (generating acetate and CO2
as products), lactate (to pyruvate and sequentially acetate and CO2), carbohydrates in general (a
process generating formate), methionine (giving rise to its corresponding sulphoxide), and urate
(to allantoin). Further, minor O3-induced modifications included the oxidation of trimethylamine
and 3-D-hydroxybutyrate, the fragmentation of salivary glycosaminoglycans to NMR-detectable
saccharide fragments, and the conversion of polyunsaturated fatty acids to their ozonides.
Moreover, evidence for the ability of O3 to induce the release of selected low-molecular-mass
salivary biomolecules from macromolecular binding-sites was also obtained. Since many of the
oxidation products detectable in O3-treated samples are identical to those arising from the attack
of *OH radical on biofluid components, it appears that at least some of the modifications observed
here are attributable to the latter oxidant (derived from O3*- generated from the single electron
reduction of O3).
PMID: 17012760 [PubMed - in process]
Braz Dent J. 2006;17(2):134-8
Antimicrobial potential of ozone in an ultrasonic cleaning system against
Staphylococcus aureus.
Estrela C, Estrela CR, Decurcio Dde A, Silva JA, Bammann LL.
Faculty of Dentistry, Federal University of Goias, Goiania, GO, Brazil. estrela3@terra.com.br
The aim of this study was to evaluate the antimicrobial potential of ozone applied to 3 different
solutions in an ultrasonic cleaning system against Staphylococcus aureus. A total of 120 mL of S.
aureus were mixed in 6 L of the experimental solutions (sterile distilled water, vinegar and sterile
dott Cosimo Cigliola
56

distilled water + Endozime AWpluz) used in a ultrasonic cleaning system (UCS). Ozone was
produced by an electric discharge through a current of oxygen and bubbling with flow rate at 7 g/
h ozone (1.2%) into the microbial suspensions. Ten mL of each experimental suspension were
collected and 5 fold dilutions were made in 9 mL of BHI and incubated at 37 degrees C for 48 h.
Bacterial growth was evaluated by turbidity of the culture medium. At the same time, 1 mL of
bacterial samples was collected and inoculated in BHIA plates. After incubation at 37 degrees C for
48 h, the number of colony forming units (cfu) per mL on BHIA surface was counted. In dilution
test in BHI tubes and in BHIA plates (cfu/mL), bacterial growth was not observed in any of the
experimental solutions when ozone was added. Under the tested conditions, it may be concluded
that the addition of ozone to a ultrasonic cleaning system containing different experimental
solutions resulted in antibacterial activity against S. aureus.
PMID: 16924341 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2006 Aug;114(4):349-53
Antibacterial effect of an ozone device and its comparison with two dentin-bonding
systems.
Polydorou O, Pelz K, Hahn P.
Department of Operative Dentistry and Periodontology, Dental School and Hospital, Albert-
Ludwigs-University Freiburg, Germany. olga.polydorou@uniklinik-freiburg.de
Microorganisms remaining beneath restorations can cause secondary caries and pulp damage.
Because of this, antimicrobial treatment could be useful. The aim of this study was to evaluate the
antibacterial effect of the HealOzone device on Streptococcus mutans and to compare it with the
already proven activity of two dentin-bonding systems. Thirty-five human molars were divided
into 5 groups. Cavities were then cut into the teeth (n = 28 cavities per group). After sterilization,
the teeth were left in broth cultures of 10(6) colony-forming units (CFU) ml(-1) of S. mutans at 36
degrees C for 48 h. The appropriate treatment followed (group A, control; group B, Clearfil SE
Bond; group C, Clearfil Protect Bond; group D, 40 s of treatment with ozone; and group E, 80 s of
treatment with ozone), and the cavities were then filled with composite resin. After 72 h, the
restorations were removed, dentin chips were collected with an excavator, and the total number
of microorganisms was determined. All treatments significantly reduced the number of S. mutans
present compared with the control group. The antimicrobial effect of both bonding systems and
treatment with 80 s of ozone was significantly higher than the 40 s ozone treatment. In
conclusion, HealOzone and the bonding systems show striking antimicrobial effects against S.
mutans.
PMID: 16911107 [PubMed - indexed for MEDLINE]
Quintessence Int. 2006 May;37(5):353-9
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57

The use of ozone in dentistry and maxillofacial surgery: a review.
Stubinger S, Sader R, Filippi A.
University Clinic for Reconstructive Surgery, Department of Cranio- and Maxillofacial Surgery,
University Hospital Basel, Switzerland. sstuebinger@uhbs.ch
Ozone has been successfully used in medicine because of its microbiologic properties for more
than 100 years. Its bactericide, virucide, and fungicide effects are based on its strong oxidation
effect with the formation of free radicals as well as its direct destruction of almost all
microorganisms. In addition, ozone has a therapeutic effect that facilitates wound healing and
improves the supply of blood. For medical purposes, ozone may be applied as a gas or dissolved
in water. Despite the advantages that the therapeutic use of ozone offers, reservations remain in
terms of its application in the oral and maxillofacial area. Particularly, the gaseous application of
ozone is critically evaluated because of its possible side effects on the respiratory system. The
objective of this article is to provide an overview of the current applications of ozone in dentistry
and oral surgery. Research was based on peer-reviewed sources found through a Medline/PubMed
search and other textbooks, reviews, and journals.
PMID: 16683682 [PubMed - indexed for MEDLINE]
Am J Dent. 2006 Feb;19(1):67-72
The impact of ozone treatment on enamel physical properties.
Celiberti P, Pazera P, Lussi A.
Department of Operative, Preventive and Pediatric Dentistry, School of Dental Medicine, University
of Bern, Freiburgstrasse 7, 3010 Bern, Switzerland. paulaceliberti@hotmail.com
PURPOSE: To assess the effects of the highly reactive molecule of ozone on sound enamel
physical properties and its effects on sealing ability.
METHODS: The effect of ozone on sealant tag length, microleakage and unfilled area proportion
were evaluated on intact and prepared sound molar fissures. Microhardness, contact angle and
acid resistance tests were performed on ground sound smooth surfaces. The samples were treated
with ozone for 40 seconds (HealOzone). Control samples were treated with air (modified
HealOzone) or left untreated.
RESULTS: No statistically significant difference was observed between the control and ozone
treated samples in all tests. Prepared fissures exhibited no unfilled areas and a statistically
significantly lower microleakage compared to intact fissures. Ozone was shown to dehydrate
enamel and consequently enhance its microhardness, which was reversible.
PMID: 16555661 [PubMed - indexed for MEDLINE]
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58

Ann Acad Med Stetin. 2005;51(2):39-42.
[Ozone therapy with the OzonyTron apparatus]
[Article in Polish]
Skomro P, Opalko K, Gadomska-Krasny J, Lietz-Kijak D, Perzanowska-Stefanska M.
Zaklad Propedeutyki i Fizykodiagnostyki Stomatologicznej Pomorskiej Akademii Medycznej al
Powstancow Wlkp 72, Szczecin.
PURPOSE: OzonyTron is one of the newest devices for the generation of ozone. The aim of this
study was to assess the benefits of ozone in some pathologies.
MATERIAL AND METHODS: Ozone therapy was used in 20 patients, including eight with lip
herpes, five after dental surgery and chiseling of third molars, six with seborrheal inflammation of
facial skin and one with mycosis of the toes.
RESULTS: Treatment with OzonyTron is noninvasive and painless. The oxidative properties of
ozone are exploited to combat microbial infection. Thus, the device is valuable primarily for its
antiseptic action. Following ozone therapy, a very good effect was observed in each case.
PMID: 16519095 [PubMed - indexed for MEDLINE]
Prim Dent Care. 2006 Jan;13(1):37-41.
The use of ozone in dentistry and medicine. Part 2. Ozone and root caries.
Baysan A, Lynch E.
Department of Fixed and Removable Prosthodontics, GKT Dental Institute, Guy's Tower Floor 25,
King's College London, Guy's Hospital, London SE1 9RT, UK. aylin.baysan@kcl.ac.uk
A previous paper, recently published in Primary Dental Care, gave an overview of the medical
uses of ozone and outlined some of its uses in dentistry. The current paper focuses on a
description of use of ozone in the management of root caries and considers recent studies in this
area. There has been relatively limited research into the non-invasive (pharmaceutical)
management of root caries. The best management strategy still remains to be developed. Initial
studies have indicated that an application of ozone for a period of either 10 or 20 seconds is
capable of clinically reversing leathery root carious lesions. It is suggested that, subject to
confirmation from extensive trials, this simple and non-invasive technique may benefit many
patients with root caries throughout the world since this approach to treat root caries can easily
be employed in primary care clinics and in the domiciliary treatment of home-bound elderly people
and immobile patients in hospices and hospitals.
PMID: 16393498 [PubMed - indexed for MEDLINE]
dott Cosimo Cigliola
59

Am J Dent. 2005 Aug;18(4):223-8.
Effect of ozone on non-cavitated fissure carious lesions in permanent molars. A
controlled prospective clinical study.
Huth KC, Paschos E, Brand K, Hickel R.
Department of Restorative Dentistry and Periodontology, Dental School, Ludwig-Maximilians-
University, Goethe Street 70, D-80336 Munich, Germany. khuth@dent.med.uni-muenchen.de
PURPOSE: To investigate, with a randomized controlled clinical study, the effect of ozone on non-
cavitated initial occlusal fissure caries compared with untreated contra-lateral control lesions (split
mouth) considering the patient's current caries risk.
METHODS: Forty-one patients with 57 pairs of lesions were enrolled in the study (mean age 7.7
+/- 2.2 years; upper jaw n=29, lower jaw n=28). Gaseous ozone (HealOzone) was applied once
for 40 seconds to the randomly assigned test molar of each pair without the use of remineralizing
solutions. Lesion progression or reversal was monitored by the laser fluorescence system
DIAGNOdent for up to 3 months and the deterioration or improvement compared between the
ozone-treated lesions and the untreated control lesions (in pairs). This was done for the whole
study population and a subgroup of patients with high current caries risk (lesion pairs n=26).
RESULTS: After 3 months, explorative data analysis revealed that the ozone-treated lesions
showed significantly more caries reversal or reduced caries progression than the untreated control
lesions within the group of patients at high current caries risk (Wilcoxon-Test, P= 0.035). There
was no statistical significance examining the whole study population. From the data it can be
concluded that ozone application significantly improved non-cavitated initial fissure caries in
patients at high caries risk over a 3-month period.
PMID: 16296426 [PubMed - indexed for MEDLINE]
Oral Microbiol Immunol. 2005 Aug;20(4):206-10
Microbicidal efficacy of ozonated water against Candida albicans adhering to acrylic
denture plates.
Arita M, Nagayoshi M, Fukuizumi T, Okinaga T, Masumi S, Morikawa M, Kakinoki Y, Nishihara T.
First Department of Prosthetic Dentistry, Kyushu Dental College, Kitakyushu, Japan.
BACKGROUND/AIMS: Ozone is known to act as a strong antimicrobial agent against bacteria,
fungi, and viruses. We examined the effect of ozonated water on Candida albicans on acrylic
denture plate.
dott Cosimo Cigliola
60

METHODS: The heat-cured acrylic resins were cultured with C. albicans. After treatment of
flowing ozonated water, the number of attached C. albicans was counted. In some experiments,
the test samples were treated with ozonated water in combination with ultrasonication.
RESULTS: After exposure to flowing ozonated water (2 or 4 mg/l) for 1 min, viable C. albicans
cells were nearly nonexistent. The combination of ozonated water and ultrasonication had a strong
effect on the viability of C. albicans adhering to the acrylic resin plates. There were no significant
differences in antimicrobial activity against C. albicans between plates immersed in ozonated
water with ultrasonication and those treated with commercially available denture cleaners. In
addition, electron microscopic analysis revealed that small amounts of C. albicans remained on the
plate after exposure to flowing ozonated water or immersion in ozonated water with
ultrasonication.
CONCLUSION: Our results suggest that application of ozonated water may be useful in reducing
the number of C. albicans on denture plates.
PMID: 15943763 [PubMed - indexed for MEDLINE]
Prim Dent Care. 2005 Apr;12(2):47-52
The use of ozone in dentistry and medicine.
Baysan A, Lynch E.
Department of Restorative Dentistry, GKT, King's College London, London SE1 9RT, UK.
aylin.baysan@kcl.ac.uk
There is growing interest in the use of ozone in oral healthcare and the National Institute for
Clinical Excellence (NICE) is at present reviewing the evidence for its effectiveness in the
management of occlusal and plain surface caries. These are only two of the clinical problems for
which ozone can, and has, been used; it has also been employed for a wide variety of other
purposes in both dentistry and medicine. This pale blue-coloured gas plays an important role as a
natural constituent in the higher layer of the Earth's atmosphere. There is growing evidence that it
can be employed as a useful therapeutic agent. This paper reviews its therapeutic uses to date
and suggests its possible future clinical applications. Consumer demands for this strong oxidant
may increase as the general public becomes increasingly aware of its therapeutic capacity and the
non-invasive manner in which it can be administered.
PMID: 15901432 [PubMed - indexed for MEDLINE]
Mediators Inflamm. 2004 Feb;13(1):3-11.
Ozone as Janus: this controversial gas can be either toxic or medically useful.
Bocci V.
dott Cosimo Cigliola
61

Department of Physiology, University of Siena, Italy. bocci@unsi.it
Ozone is an intrinsically toxic gas and its hazardous employment has led to a poor consideration of
ozone therapy. The aim of this review is to indicate that a wrong dogma and several
misconceptions thwart progress: in reality, properly performed ozone therapy, carried out by
expert physicians, can be very useful when orthodox medicine appears inadequate. The
unbelievable versatility of ozone therapy is due to the cascade of ozone-derived compounds able
to act on several targets leading to a multifactorial correction of a pathological state. During the
past decade, contrary to all expectations, it has been demonstrated that the judicious application
of ozone in chronic infectious diseases, vasculopathies, orthopedics and even dentistry has yielded
such striking results that it is deplorable that the medical establishment continues to ignore ozone
therapy.
PMID: 15203558 [PubMed - indexed for MEDLINE]
Int J Prosthodont. 1999 Mar-Apr;12(2):179-83.
Influence of ozone on oxidation of dental alloys.
Suzuki T, Oizumi M, Furuya J, Okamoto Y, Rosenstiel SF.
Department of Geriatric Dentistry, Tokyo Medical and Dental University Faculty of Dentistry,
Japan. t.suzuki.gero@dent.tmd.ac.jp
PURPOSE: The purpose of this study was to examine the influence of ozone on the surface of
removable partial denture (RPD) alloys to determine its usefulness as a cleaning method for RPDs,
since ozone has powerful sterilizing and deodorizing properties.
MATERIALS AND METHODS: Two types of ozone cleaning were used. The quantities of ozone
generated by both methods were the same (20 mg/h). In method A, ozone was generated for 10
minutes every 12 hours and in method B, ozone was generated over 24 hours a day. Test
specimens of 3 types of dental alloy (Co-Cr, Au-Ag-Pt, and Au-Cu-Ag-Pd) were subjected to
different cleaning methods for 7 days and measured in terms of reflectance, surface roughness,
and weight. Five different cleaning solutions (three commercial denture cleaners, acid-electrolyzed
water with a pH of 2.4, and pure water) were used for comparison with the ozone treatments.
RESULTS: No significant changes were detected after treatment of the Co-Cr and Au-Ag-Pt alloys
with ozone. Ozone caused a slight change in the Au-Cu-Ag-Pd alloy in terms of reflectance, but
the changes were significantly less than those caused by acid-electrolyzed water and one of the
commercial denture cleaners.
CONCLUSION: Ozone had little influence on the oxidation of dental alloys.
PMID: 10371921 [PubMed - indexed for MEDLINE]
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Stomatologiia (Mosk). 1998;77(6):7-8.
[Changes in the quantitative composition of the microbial flora of dental deposits during
the intensification of oral hygiene]
Lukinykh LM, Kosiuga SIu.
The efficacy of hygienic treatment of the oral cavity in combination with ozone therapy is
assessed. This combination mechanically removes soft dental deposit and decreases bacterial
contamination.
PMID: 10067405 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2007 Feb;115(1):77-80.
Efficacy of gasiform ozone and photodynamic therapy on a multispecies oral biofilm in
vitro.
Muller P, Guggenheim B, Schmidlin PR.
Clinic of Preventive Dentistry, Periodontology and Cariology, University if Zurich, Zurich,
Switzerland.
Ozone gas and photodynamic therapy (PDT) have been claimed to be antimicrobially effective.
This study assessed their antimicrobial potential in vitro. Mature six-species oral biofilms were
treated as follows (n = 9 per group):
(i) a 60-s application of gasiform vacuum-ozone or vacuum alone (on wet or air-dried biofilm
samples);
(ii) PDT (i.e. methylene blue in combination with or without a diode soft laser, and a soft laser
alone);
(iii) antimicrobial solutions: immersion of biofilms for 60 s in 0.2 and 2% chlorhexidine or in 0.5
and 5% hypochlorite solution. Treatment with chlorhexidine or hypochlorite served as a positive
control, whereas untreated samples served as negative controls. Colony-forming units on blood
agar were counted. Only the 5% hypochlorite solution was able to totally eliminate the
microorganisms in the biofilm. The observed reduction of viable counts by vacuum-ozone
application and PDT was less than one log(10) step. Under the conditions of the current study,
gasiform ozone and PDT had a minimal effect on the viability of microorganisms organized in a
cariogenic biofilm.
PMID: 17305720 [PubMed - in process]
Vopr Kurortol Fizioter Lech Fiz Kult. 2006 Nov-Dec;(6):34-7.
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[Ozone therapy in gastroduodenal pathology associated with Helicobacter pylori]
[Article in Russian]
Fedorov AA, Gromov AS, Sapronenok SV, Kurochkin VIu, Zhernakova ZM.
Three methods of ozone therapy (intravenous injection of ozonized physiologic saline, oral intake
of ozonized low-mineral water and combination of the two treatments) were compared by efficacy
in 215 patients with gastroduodenal pathology associated with Helicobacter pilori. All the
techniques of ozone therapy proved effective in relation to regress of clinical symptoms,
regeneration processes, eradication of Helicobacter pylori, but the combined regimen was the
best.
PMID: 17201222 [PubMed - indexed for MEDLINE]
Health Technol Assess. 2006 May;10(16):iii-iv, ix-80.
Systematic review of the effectiveness and cost-effectiveness of HealOzone for the
treatment of occlusal pit/fissure caries and root caries.
Brazzelli M, McKenzie L, Fielding S, Fraser C, Clarkson J, Kilonzo M, Waugh N.
Health Services Research Unit, Institute of Applied Health Sciences, University of Aberdeen, UK.
OBJECTIVES: To assess the effectiveness and cost-effectiveness of HealOzone (CurOzone USA
Inc., Ontario, Canada) for the management of pit and fissure caries, and root caries. The complete
HealOzone procedure involves the direct application of ozone gas to the caries lesion on the tooth
surface, the use of a remineralising solution immediately after application of ozone and the supply
of a “patient kit”, which consists of toothpaste, oral rinse and oral spray all containing fluoride.
DATA SOURCES: Electronic databases up to May 2004 (except Conference Papers Index, which
were searched up to May 2002).
REVIEW METHODS: A systematic review of the effectiveness of HealOzone for the management
of tooth decay was carried out. A systematic review of existing economic evaluations of ozone for
dental caries was also planned but no suitable studies were identified. The economic evaluation
included in the industry submission was critically appraised and summarised. A Markov model was
constructed to explore possible cost-effectiveness aspects of HealOzone in addition to current
management of dental caries.
RESULTS: Five full-text reports and five studies published as abstracts met the inclusion criteria.
The five full-text reports consisted of two randomised controlled trials (RCTs) assessing the use of
HealOzone for the management of primary root caries and two doctoral theses of three
unpublished randomised trials assessing the use of HealOzone for the management of occlusal
caries. Of the abstracts, four assessed the effects of HealOzone for the management of occlusal
dott Cosimo Cigliola
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caries and one the effects of HealOzone for the management of root caries. Overall, the quality of
the studies was modest, with many important methodological aspects not reported (e.g.
concealment of allocation, blinding procedures, compliance of patients with home treatment). In
particular, there were some concerns about the choice of statistical analyses. In most of the full-
text studies analyses were undertaken at lesion level, ignoring the clustering of lesions within
patients. The nature of the methodological concerns was sufficient to raise doubts about the
validity of the included studies' findings. A quantitative synthesis of results was deemed
inappropriate. On the whole, there is not enough evidence from published RCTs on which to judge
the effectiveness of ozone for the management of both occlusal and root caries. The perspective
adopted for the study was that of the NHS and Personal Social Services. The analysis, carried out
over a 5-year period, indicated that treatment using current management plus HealOzone cost
more than current management alone for non-cavitated pit and fissure caries (40.49 pounds
versus 24.78 pounds), but cost less for non-cavitated root caries ( 14.63 pounds versus 21.45
pounds). Given the limitations of the calculations these figures should be regarded as illustrative,
not definitive. It was not possible to measure health benefits in terms of quality-adjusted life-
years, due to uncertainties around the evidence of clinical effectiveness, and to the fact that the
adverse events avoided are transient (e.g. pain from injection of local anaesthetic, fear of the
drill). One-way sensitivity analysis was applied to the model. However, owing to the limitations of
the economic analysis, this should be regarded as merely speculative. For non-cavitated pit and
fissure caries, the HealOzone option was always more expensive than current management when
the probability of cure using the HealOzone option was 70% or lower. For non-cavitated root
caries the costs of the HealOzone comparator were lower than those of current management only
when cure rates from HealOzone were at least 80%. The costs of current management were
higher than those of the HealOzone option when the cure rate for current management was 40%
or lower. One-way sensitivity analysis was also performed using similar NHS Statement of Dental
Remuneration codes to those that are used in the industry submission. This did not alter the
results for non-cavitated pit fissure caries as the discounted net present value of current
management remained lower than that of the HealOzone comparator ( 22.65 pounds versus 33.39
pounds).
CONCLUSIONS: Any treatment that preserves teeth and avoids fillings is welcome. However, the
current evidence base for HealOzone is insufficient to conclude that it is a cost-effective addition
to the management and treatment of occlusal and root caries. To make a decision on whether
HealOzone is a cost-effective alternative to current preventive methods for the management of
dental caries, further research into its clinical effectiveness is required. Independent RCTs of the
effectiveness and cost-effectiveness of HealOzone for the management of occlusal caries and root
caries need to be properly conducted with adequate design, outcome measures and methods for
statistical analyses.
PMID: 16707073 [PubMed - indexed for MEDLINE]
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Systematic review of the effectiveness and cost-effectiveness of HealOzone ® for the
treatment of occlusal pit/fissure caries and root caries
M Brazzelli, 1* L McKenzie, 2 S Fielding, 3 C Fraser, 1 J Clarkson, 4 M Kilonzo 2 and N Waugh 3
1 Health Services Research Unit, Institute of Applied Health Sciences, University of Aberdeen, UK
2 Health Economics Research Unit, Institute of Applied Health Sciences, University of Aberdeen,
UK
3 Department of Public Health, Institute of Applied Health Sciences, University of Aberdeen, UK
4 Dental Health Services Research Unit, MacKenzie Building, Dundee, UK
* Corresponding author. Current affiliation: Department of Clinical Neurosciences, University of
Edinburgh, Western General Hospital, UK
Background
Dental caries is a chronic disease caused by the localised and progressive demineralisation of the
hard tissues of the coronal and root surfaces of the teeth. Caries location, development and
progression depend on a range of environmental, social and genetic factors, and vary greatly
among individuals.
Despite the decline in the prevalence of dental caries observed in the high-income countries
during the past few decades as a consequence of the increased availability of fluoride products
and improved oral hygiene, dental caries is still a common disease experienced by almost 80% of
children by the age of 18 years and by almost 90% of adults.
The current management of early non-cavitated occlusal and root caries, and cavitated root
caries, which are still accessible to cleaning, is based on non-operative preventive strategies that
include information on oral hygiene, dietary advice, use of topically applied fluorides and
application of sealants. For cavitated occlusal caries and cavitated root caries that are not easily
accessible to cleaning, restorative interventions are adopted (drilling and filling).
HealOzone ® (CurOzone USA Inc., Ontario, Canada) has recently been proposed as a novel method
for the treatment of dental caries. It is suggested that HealOzone may reverse, arrest or slow the
progression of dental caries. The complete HealOzone procedure involves the direct application of
ozone gas to the caries lesion on the tooth surface, the use of a remineralising solution
immediately after application of ozone and the supply of a ‘patient kit’, which consists of
toothpaste, oral rinse and oral spray all containing fluoride.
Objective
The review aims to assess the effectiveness and cost-effectiveness of HealOzone for the
management of pit and fissure caries, and root caries.
Methods
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Electronic searches were conducted to identify published and unpublished studies. The following
databases were searched: MEDLINE (1966 to May 2004), EMBASE (1980 to May 2004), MEDLINE
Extra (17 May 2004), Science Citation Index (1981 to May 2004), Biosis (1985 to May 2004),
AMED (1985 to May 2004), Cochrane Library (Issue 2, 2004) National Research Register (Issue 2,
2004), Current Controlled Trials (18 May 2004), Clinical Trials (18 May 2004), SCI Proceedings
(1991 to May 2004), Conference Papers Index (1982 to May 2002), ZETOC conferences (1993 to
May 2004) and IADR meeting abstracts (2002 to 2004). Two reviewers independently assessed
the methodological quality of included studies and extracted data. Criteria for assessment of study
quality included method and unit of randomisation, concealment of allocation, comparability of
groups at baseline, blinding procedures, number of withdrawals/dropouts and completeness of
assessment at follow-up.
A systematic review of the effectiveness of HealOzone for the management of tooth decay was
carried out. A systematic review of existing economic evaluations of ozone for dental caries was
also planned but no suitable studies were identified. The economic evaluation included in the
industry submission was critically appraised and summarised.
A Markov model was constructed to explore possible cost-effectiveness aspects of HealOzone in
addition to current management of dental caries.
Results
Number and quality of studies, and direction of evidence
Five full-text reports and five studies published as abstracts met the inclusion criteria. Of these,
only one was published in a refereed journal, but it lacked some study details. The remaining
studies were PhD theses, unpublished reports or conference proceedings. The five full-text reports
consisted of two randomised controlled trials (RCTs) assessing the use of HealOzone for the
management of primary root caries and two PhD theses of three unpublished randomised trials
assessing the use of HealOzone for the management of occlusal caries. Of the five studies
published as abstracts, four assessed the effects of HealOzone for the management of occlusal
caries and one the effects of HealOzone for the management of root caries.
Overall, the quality of the studies was modest, with many important methodological aspects not
reported (e.g. concealment of allocation, blinding procedures, compliance of patients with home
treatment). In particular, there were some concerns about the choice of statistical analyses. In
most of the full-text studies analyses were undertaken at lesion level, ignoring the clustering of
lesions within patients. The nature of the methodological concerns was sufficient to raise doubts
about the validity of the included studies’ findings. A quantitative synthesis of results was deemed
inappropriate.
Summary of benefits
Root caries
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Two studies (one published and one unpublished) assessing the use of HealOzone for the
management of primary non-cavitated root caries reported high success rates for ozone-treated
lesions and no significant changes in the control lesions, despite application of topical fluoride.
This is puzzling, since topical fluoride is known to be effective. Results of cavitated root lesions
were poorly defined and reported in one of these two studies. Cavitated lesions did not seem to
benefit from ozone application.
One unpublished study showed that fissure sealants preceded by the application of ozone for the
preventive treatment of non-cavitated root lesions were more likely to remain intact (61% versus
42%, p < 0.05).
Pit and fissure caries
One unpublished study did not show any significant benefits of HealOzone for the management of
non-cavitated pit and fissure lesions in the permanent dentition. Similarly, a small unpublished
pilot study did not show any significant differences between cavitated occlusal lesions treated with
or without ozone, apart from an improvement in the hardness and visual clinical indices. In
contrast, findings from conference proceedings (which provide little detail for the assessment of
their methodological quality and therefore are of little use in systematic reviews) reported very
high success rates (from 86.6% to 99% of reversal of caries).
Adding ozone to a fissure sealant did not seem to produce better sealant retention in occlusal
lesions extending 2–4 mm into dentine.
Data on the use of HealOzone for the treatment of occlusal lesion in the deciduous dentition were
available from only one unpublished study. An overall reduction in clinical severity scores was
reported for non-cavitated occlusal lesions in primary molars treated with ozone.
On the whole, there is not enough evidence from published RCTs on which to judge the
effectiveness of ozone for the management of both occlusal and root caries.
Costs
The perspective adopted for the study was that of the NHS and Personal Social Services. The
analysis, carried out over a 5-year period, indicated that treatment using current management
plus HealOzone cost more than current management alone for non-cavitated pit and fissure caries
(£40.49 versus £24.78), but cost less for non-cavitated root caries (£14.63 versus £21.45). Given
the limitations of the calculations these figures should be regarded as illustrative, not definitive.
Cost per quality-adjusted life-year
It was not possible to measure health benefits in terms of quality-adjusted life-years. This was
mainly due to uncertainties around the evidence of clinical effectiveness, and to the fact that the
adverse events avoided are transient (e.g. pain from injection of local anaesthetic, fear the drill).
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Sensitivity analyses
One-way sensitivity analysis was applied to the model. However, owing to the limitations of the
economic analysis, this should be regarded as merely speculative. For non-cavitated pit and
fissure caries, the HealOzone option was always more expensive than current management when
the probability of cure using the HealOzone option was 70% or lower. For non-cavitated root
caries the costs of the HealOzone comparator were lower than those of current management only
when cure rates from HealOzone were at least 80%. The costs of current management were
higher than those of the HealOzone option when the cure rate for current management was 40%
or lower.
One-way sensitivity analysis was also performed using similar NHS Statement of Dental
Remuneration codes to those that are used in the industry submission. This did not alter the
results for non-cavitated pit fissure caries as the discounted net present value of current
management remained lower than that of the HealOzone comparator (£22.65 versus £33.39).
Conclusions
Any treatment that preserves teeth and avoids fillings is welcome. However, the current evidence
base for HealOzone is insufficient to conclude that it is a cost-effective addition to the
management and treatment of occlusal and root caries.
Limitations of the calculations
The economic analysis was severely constrained by the uncertainty over clinical effectiveness, and
it could be argued that such analysis was inappropriate. It was done merely to illustrate the key
factors involved in economic modelling. The long-term effects of HealOzone are unknown and the
assumption that reversed caries remains inactive may not be reliable.
Recommendations for research
To make a decision on whether HealOzone is a cost-effective alternative to current preventive
methods for the management of dental caries, further research into its clinical effectiveness is
required. Independent RCTs of the effectiveness and cost-effectiveness of HealOzone for the
management of occlusal caries and root caries need to be properly conducted with adequate
design, outcome measures and methods for statistical analyses.
Publication
Brazzelli M, McKenzie L, Fielding S, Fraser C, Clarkson J, Kilonzo M, et al. Systematic review of the
effectiveness and cost-effectiveness of HealOzone ® for the treatment of occlusal pit/fissure caries
and root caries. Health Technol Assess 2006;10(16).
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NHS R&D HTA Programme
The research findings from the NHS R&D Health Technology Assessment (HTA) Programme
directly influence key decision-making bodies such as the National Institute for Health and Clinical
Excellence (NICE) and the National Screening Committee (NSC) who rely on HTA outputs to help
raise standards of care. HTA findings also help to improve the quality of the service in the NHS
indirectly in that they form a key component of the ‘National Knowledge Service’ that is being
developed to improve the evidence of clinical practice throughout the NHS.
The HTA Programme was set up in 1993. Its role is to ensure that high-quality research
information on the costs, effectiveness and broader impact of health technologies is produced in
the most efficient way for those who use, manage and provide care in the NHS. ‘Health
technologies’ are broadly defined to include all interventions used to promote health, prevent and
treat disease, and improve rehabilitation and long-term care, rather than settings of care.
The HTA Programme commissions research only on topics where it has identified key gaps in the
evidence needed by the NHS. Suggestions for topics are actively sought from people working in
the NHS, the public, service-users groups and professional bodies such as Royal Colleges and NHS
Trusts.
Research suggestions are carefully considered by panels of independent experts (including service
users) whose advice results in a ranked list of recommended research priorities. The HTA
Programme then commissions the research team best suited to undertake the work, in the
manner most appropriate to find the relevant answers. Some projects may take only months,
others need several years to answer the research questions adequately. They may involve
synthesising existing evidence or conducting a trial to produce new evidence where none currently
exists.
Additionally, through its Technology Assessment Report (TAR) call-off contract, the HTA
Programme is able to commission bespoke reports, principally for NICE, but also for other policy
customers, such as a National Clinical Director. TARs bring together evidence on key aspects of
the use of specific technologies and usually have to be completed within a short time period.
Criteria for inclusion in the HTA monograph series
Reports are published in the HTA monograph series if (1) they have resulted from work
commissioned for the HTA Programme, and (2) they are of a sufficiently high scientific quality as
assessed by the referees and editors.
Reviews in Health Technology Assessment are termed ‘systematic’ when the account of the
search, appraisal and synthesis methods (to minimise biases and random errors) would, in theory,
permit the replication of the review by others.
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The research reported in this monograph was commissioned and funded by the HTA Programme
on behalf of NICE as project number 03/50/01. The protocol was agreed in June 2004. The
assessment report began editorial review in June 2005 and was accepted for publication in
September 2005. The authors have been wholly responsible for all data collection, analysis and
interpretation, and for writing up their work. The HTA editors and publisher have tried to ensure
the accuracy of the authors’ report and would like to thank the referees for their constructive
comments on the draft document. However, they do not accept liability for damages or losses
arising from material published in this report.
The views expressed in this publication are those of the authors and not necessarily those of the
HTA Programme, NICE or the Department of Health.
Editor-in-Chief: Professor Tom Walley Series Editors: Dr Aileen Clarke, Dr Peter Davidson,
Dr Chris Hyde, Dr John Powell, Dr Rob Riemsma and Dr Ken Stein
Managing Editors: Sally Bailey and Sarah Llewellyn Lloyd
© 2006 Crown Copyright
Am J Dent. 2004 Feb;17(1):56-60.
Effect of ozone on the oral microbiota and clinical severity of primary root caries.
Baysan A, Lynch E.
The School of Dentistry, University of Birmingham, St Chad's Queensway, Birmingham B4 6NN,
UK. a.baysan@bham.ac.uk
PURPOSE: To assess the effect of ozone on the microbial flora and clinical severity of primary root
caries.
METHODS: 26 patients with 70 primary root carious lesions (PRCLs) were entered. Each PRCL
was classified in terms of color, cavitation, size, hardness, distance from the gingival margin and
severity. Overlying plaque was then removed and each lesion dried. A biopsy was taken from half
of each PRCL using a sterile excavator. Subsequently, the remaining lesions were exposed to
ozone gas for a period of either 10 seconds (n = 35) or 20 seconds (n = 35) and a further biopsy
was taken.
RESULTS: Using a paired Student t-test, a significant (P < 0.001) difference (mean +/- SE) in
total micro-organisms was observed in the ozone-treated samples after either a 10 seconds
(log10 4.35 +/- 0.49) or 20 seconds (log10 0.46 +/- 0.26) ozone application compared with the
control samples (log10 7.00 +/- 0.24) and (log10 6.00 +/- 0.21) respectively. Using Pearson's
correlation tests, there were significant correlations for the reduction in total micro-organisms
after 10 seconds of ozone application with cavitation, size, distance from gingival margin and
severity of PRCLs (P < 0.05). In conclusion, ozone application either for 10 or 20 seconds
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dramatically reduced most of the micro-organisms in PRCLs without any side effects recorded at
recall intervals between 3 and 5.5 months. Out of the 65 PRCLs reviewed, 33 lesions had become
hard, 27 lesions reversed to severity index 1 from severity index 2, and five lesions remained the
same following ozone application for a period of either 10 or 20 seconds.
PMID: 15241911 [PubMed - indexed for MEDLINE]
Oral Microbiol Immunol. 2004 Aug;19(4):240-6.
Efficacy of ozone on survival and permeability of oral microorganisms.
Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T.
Department of Operative Dentistry and Endodontics, Kyushu Dental College, Kitakyushu, Japan.
In the present study, we examined the effect of ozonated water on oral microorganisms and
dental plaque. Almost no microorganisms were detected after being treated with ozonated water
(4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial cells
were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Fluorescence microscopic analysis
revealed that S. mutans cells were killed instantaneously in ozonated water. Some breakage of
ozonated water-treated S. mutans was found by electron microscopy. When the experimental
dental plaque was exposed to ozonated water, the number of viable S. mutans remarkably
decreased. Ozonated water strongly inhibited the accumulation of experimental dental plaque in
vitro. After the dental plaque samples from human subjects were exposed to ozonated water in
vitro, almost no viable bacterial cells were detected. These results suggest that ozonated water
should be useful in reducing the infections caused by oral microorganisms in dental plaque.
PMID: 15209994 [PubMed - indexed for MEDLINE]
J Endod. 2004 Nov;30(11):778-81.
Antimicrobial effect of ozonated water on bacteria invading dentinal tubules.
Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M.
Department of Operative Dentistry and Endodontics, Kyushu Dental College, Kitakyushu, Japan.
Ozone is known to act as a strong antimicrobial agent against bacteria, fungi, and viruses. In the
present study, we examined the effect of ozonated water against Enterococcus faecalis and
Streptcoccus mutans infections in vitro in bovine dentin. After irrigation with ozonated water, the
viability of E. faecalis and S. mutans invading dentinal tubules significantly decreased. Notably,
when the specimen was irrigated with sonication, ozonated water had nearly the same
antimicrobial activity as 2.5% sodium hypochlorite (NaOCl). We also compared the cytotoxicity
against L-929 mouse fibroblasts between ozonated water and NaOCl. The metabolic activity of
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fibroblasts was high when the cells were treated with ozonated water, whereas that of fibroblasts
significantly decreased when the cells were treated with 2.5% NaOCl. These results suggest that
ozonated water application may be useful for endodontic therapy.
PMID: 15505509 [PubMed - indexed for MEDLINE]
J Med Dent Sci. 1998 Jun;45(2):135-9.
In vitro testing of a denture cleaning method using ozone.
Oizumi M, Suzuki T, Uchida M, Furuya J, Okamoto Y.
Department of Geriatric Dentistry, Faculty of Dentistry, Tokyo Medical and Dental University,
Japan. m.oizumi.gero@dent.tmd.ac.jp
The purpose of this study was to compare the microbicidal effect of gaseous ozone with that of
ozonated water in order to determine its usefulness as a method for disinfecting dentures.
Although a large number of research studies have been done on the bactericidal effect of ozone,
little is known about its microbicidal effects on oral microorganisms. Therefore, we tested the
effect of ozone on three standard strains of oral microorganisms: Streptococcus mutans (strain
IID 973), Staphylococcus aureus (strain 209-P), and Candida albicans (strain LAM 14322). When
the gaseous ozone injection method was used, the numbers of cells of all three strains decreased
to 1/10(5) at 1 min, and by 3 min they were below the detection limit. Thus, the microbicidal
effect of gaseous ozone was ascertained in a short time. In contrast, when ozonated water at 1
ppm and 3 ppm was used, C. albicans decreased to 1/10. A 700 mg/h ozone production level was
needed to prepare 1 ppm ozonated water, whereas 20 mg/h of ozone was required by the
gaseous ozone generator. These findings indicate that direct exposure to gaseous ozone seems to
be a more effective microbicide compared with ozonated water, and that gaseous ozone can be
clinically useful for disinfection of dentures.
PMID: 11186199 [PubMed - indexed for MEDLINE]
Arch Environ Health. 1989 Sep-Oct;44(5):311-6.
Oral and oronasal breathing during continuous exercise produce similar responses to
ozone inhalation.
Adams WC, Schelegle ES, Shaffrath JD.
Physical Education Department, University of California, Davis.
Breathing route has a profound effect on sulfur dioxide-induced pulmonary function response in
human subjects. There is comparatively little evidence of the effects of oral, nasal, and oronasal
breathing on ozone (O3)-induced responses in humans. In this study, six young adult males were
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exposed on five occasions to 0.40 parts per million (ppm) O3 while exercising continuously at one
of two workloads (minute ventilation, VE, of approximately 30 and 75 l/min). The VE exposure
time product was similar for all protocols. Four exposures were delivered randomly with a Hans-
Rudolph respiratory valve attached to a silicone facemask, with breathing route effected with and
without noseclip. A 2 x 2 analysis of variance revealed no statistically significant differences (p
less than .05) across conditions in pulmonary function, exercise ventilatory pattern, or subjective
symptoms responses. The fifth exposure, delivered via the same respiratory valve with
mouthpiece, but without facemask, revealed significantly greater forced expiratory volume in 1 s
(FEV1.0) impairment than that observed for the respiratory valve, facemask with noseclip
exposure (-20.4% and -15.9%, respectively). The latter suggests partial O3 reactivity to the
facemask and clean shaven facial surface of the subjects, although reduced oral scrubbing by
mouthpiece-induced bypassing of the oral vestibule might account, in part, for this difference.
Recent O3 uptake evidence from another laboratory, however, supports our conclusion that
breathing route during moderate and heavy continuous exercise does not affect acute physiologic
responses to 0.40 ppm O3.
PMID: 2817961 [PubMed - indexed for MEDLINE]
Eur J Oral Sci. 2006 Oct;114(5):435-40.
Effect of ozone on oral cells compared with established antimicrobials.
Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K.
Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-University, Munich,
Germany. khuth@dent.med.uni-muenchen.de
Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its
antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous
ozone (4 x 10(6) microg m(-3)) and aqueous ozone (1.25-20 microg ml(-1)) exert any cytotoxic
effects on human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with
established antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl)
5.25%, 2.25%; hydrogen peroxide H2O2) 3%], over a time of 1 min, and compared with the
antibiotic, metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and
apoptosis were evaluated. Ozone gas was found to have toxic effects on both cell types.
Essentially no cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic
to BHY cells, and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H2O2 resulted in
markedly reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity only
to BHY cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of the
tested antiseptics.
PMID: 17026511 [PubMed - indexed for MEDLINE]
Dtsch Zahnarztl Z. 1991 Jul;46(7):485-7.
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74

[Water disinfection of dental treatment units using ozone]
Filippi A, Tilkes F, Beck EG, Kirschner H.
Med. Zentrum fur Zahn-, Mund- und Kieferheilkunde der Justus-Liebig-Universitat Giessen.
The disinfecting effect of ozonized water has been investigated. Under the precondition that the
dental chair had been thoroughly sanitized, the system showed a good disinfecting effect. Finally,
for reasons of practical medical treatment, the ozone concentration in air and, for reasons of
hygiene in drinking water, the ozone concentrations in water were determined under various
conditions. In addition, the influence of continuous-flow water heaters is discussed. The problem
of continuous-flow water heaters regarding their effects on the colonisation of water by microbes
proves not to be significant. The exposure of patients to disinfectants is discussed.
PMID: 1817912 [PubMed - indexed for MEDLINE]
Evid Based Dent. 2005;6(2):34.
No reliable evidence that ozone gas stops or reverses tooth decay.
McComb D.
Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada.
DATA SOURCES: The Cochrane Oral Health Group's Trials Register, Cochrane Central Register of
Controlled Trials, Medline and Premedline, Embase, CINAHL (Cumulative Index to nursing and
Allied Health Literature)l and the Allied and Complementary Medicine Database were searched.
The journal Quintessence was searched by hand and KaVo Dental (Biberach, Germany)
manufacturers of HealOzone apparatus were contacted for any additional published or unpublished
trials.
STUDY SELECTION: Inclusion was assessed independently by at least two reviewers. Trials were
only included if they met the following criteria: randomisation in a controlled trial; single surface
in vivo carious lesion accessible to ozone application; clear allocation concealment; ozone
application to the lesions in the intervention group; no such application of ozone in the control
group; and outcomes measured after at least 6 months.
DATA EXTRACTION AND SYNTHESIS: Reviewers independently extracted information in
duplicate. A paucity of comparable data did not allow meta-analytic pooling of the included
studies.
RESULTS: Three trials were included, with a combined total of 432 randomised lesions (137
participants). Forty-two conference papers, abstracts and posters were excluded (from an
unknown number of studies). The risk of bias in all studies appeared high. The analyses of all
dott Cosimo Cigliola
75

three studies were conducted at the level of the lesion, which is not independent of the person.
For this reason, pooling of data was not appropriate or attempted. Individual studies showed
inconsistent effects of ozone on caries, across different measures of caries progression or
regression. Few secondary outcomes were reported, but one trial reported an absence of adverse
events.
CONCLUSIONS: Given the high risk of bias in the available studies and lack of consistency
between different outcome measures, there is no reliable evidence that application of ozone gas to
the surface of decayed teeth stops or reverses the decay process. There is a fundamental need for
more evidence of appropriate rigour and quality before the use of ozone can be accepted into
mainstream primary dental care or can be considered a viable alternative to current methods for
the management and treatment of dental caries.
PMID: 16208384 [PubMed]
Ozone therapy for the treatment of dental caries
GD Rickard, R Richardson, T Johnson, D McColl, L Hooper
Cochrane Database of Systematic Reviews 2007 Issue 1Copyright © 2007 The Cochrane
Collaboration. Published by John Wiley & Sons, Ltd.
DOI: 10.1002/14651858.CD004153.pub2 This version first published online: 19 July 2004 in
Issue 3, 2004
Date of Most Recent Substantive Amendment: 14 May 2004
This record should be cited as: Rickard GD, Richardson R, Johnson T, McColl D, Hooper L. Ozone
therapy for the treatment of dental caries. Cochrane Database of Systematic Reviews 2004, Issue
3. Art. No.: CD004153. DOI: 10.1002/14651858.CD004153.pub2.
Abstract
Background
Dental caries is a bacterially mediated disease characterised by demineralisation of the tooth
surface, which may lead to cavitation, discomfort, pain and eventual tooth loss. Ozone is toxic to
certain bacteria in vitro and it has been suggested that delivering ozone into a carious lesion
might reduce the number of cariogenic bacteria. This possibly could arrest the progress of the
lesion and may, in the presence of fluoride, perhaps allow remineralisation to occur. This may in
turn delay or prevent the need for traditional dental conservation by 'drilling and filling'.
Objectives
To assess whether ozone is effective in arresting or reversing the progression of dental caries.
dott Cosimo Cigliola
76

Search strategy
We searched the Cochrane Oral Health Group's Trials Register (to 7 November 2003); Cochrane
Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 3, 2003); MEDLINE
and PREMEDLINE (OVID, 1966 to November 2003); EMBASE (OVID, 1980 to November 2003);
CINAHL (OVID, 1982 to November 2003); AMED (OVID, 1985 to November 2003). Quintessence
was handsearched through 2002 and KaVo were contacted as manufacturers of the HealOzone
apparatus for any additional published or unpublished trials.
Selection criteria
Inclusion was assessed independently by at least two reviewers. Trials were only included if they
met the following criteria: randomisation in a controlled trial; single surface in vivo carious lesion
accessible to ozone application; clear allocation concealment; ozone application to the lesions in
the intervention group; no such application of ozone in the control group; outcomes measured
after at least 6 months.
Data collection and analysis
Reviewers independently extracted information in duplicate. A paucity of comparable data did not
allow meta-analytic pooling of the included studies.
Main results
Three trials were included, with a combined total of 432 randomised lesions (137 participants).
Forty-two conference papers, abstracts and posters were excluded (from an unknown number of
studies). The risk of bias in all studies appeared high. The analyses of all three studies were
conducted at the level of the lesion, which is not independent of the person, for this reason
pooling of data was not appropriate or attempted. Individual studies showed inconsistent effects
of ozone on caries, across different measures of caries progression or regression. Few secondary
outcomes were reported, but one trial reported an absence of adverse events.
Authors' conclusions
Given the high risk of bias in the available studies and lack of consistency between different
outcome measures, there is no reliable evidence that application of ozone gas to the surface of
decayed teeth stops or reverses the decay process. There is a fundamental need for more
evidence of appropriate rigour and quality before the use of ozone can be accepted into
mainstream primary dental care or can be considered a viable alternative to current methods for
the management and treatment of dental caries.
Plain language summary
There is no evidence that ozone therapy can reverse or stop tooth decay.
dott Cosimo Cigliola
77

Tooth decay can be painful, is expensive to treat and can lead to the loss of teeth. It has been
suggested that treatment of a decayed tooth with ozone will stop or reverse the decay process.
This review of trials found no sound evidence that ozone is capable of reversing or stopping the
progression of tooth decay. High quality research is needed to show whether or not it works.
Ozone should not be considered an alternative to current treatment methods in dental practices.
Ozone therapy for the treatment of dental caries
Rickard GD, Richardson R, Johnson T, McColl D, Hooper L
This is a Cochrane review abstract and plain language summary, prepared and maintained by The
Cochrane Collaboration. The full text of the review is available in The Cochrane Library (ISSN
1464-780X).
The Cochrane Database of Systematic Reviews 2007 Issue 1
Copyright © 2007 The Cochrane Collaboration. Published by John Wiley and Sons, Ltd.
Date of last Subtantial Update: May 14. 2004
Plain language summary
There is no evidence that ozone therapy can reverse or stop tooth decay.
Tooth decay can be painful, is expensive to treat and can lead to the loss of teeth. It has been
suggested that treatment of a decayed tooth with ozone will stop or reverse the decay process.
This review of trials found no sound evidence that ozone is capable of reversing or stopping the
progression of tooth decay. High quality research is needed to show whether or not it works.
Ozone should not be considered an alternative to current treatment methods in dental practices.
Abstract
Background
Dental caries is a bacterially mediated disease characterised by demineralisation of the tooth
surface, which may lead to cavitation, discomfort, pain and eventual tooth loss. Ozone is toxic to
certain bacteria in vitro and it has been suggested that delivering ozone into a carious lesion
might reduce the number of cariogenic bacteria. This possibly could arrest the progress of the
lesion and may, in the presence of fluoride, perhaps allow remineralisation to occur. This may in
turn delay or prevent the need for traditional dental conservation by 'drilling and filling'.
Objectives
To assess whether ozone is effective in arresting or reversing the progression of dental caries.
dott Cosimo Cigliola
78

Search strategy
We searched the Cochrane Oral Health Group's Trials Register (to 7 November 2003); Cochrane
Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 3, 2003); MEDLINE
and PREMEDLINE (OVID, 1966 to November 2003); EMBASE (OVID, 1980 to November 2003);
CINAHL (OVID, 1982 to November 2003); AMED (OVID, 1985 to November 2003). Quintessence
was handsearched through 2002 and KaVo were contacted as manufacturers of the HealOzone
apparatus for any additional published or unpublished trials.
Selection criteria
Inclusion was assessed independently by at least two reviewers. Trials were only included if they
met the following criteria: randomisation in a controlled trial; single surface in vivo carious lesion
accessible to ozone application; clear allocation concealment; ozone application to the lesions in
the intervention group; no such application of ozone in the control group; outcomes measured
after at least 6 months.
Data collection and analysis
Reviewers independently extracted information in duplicate. A paucity of comparable data did not
allow meta-analytic pooling of the included studies.
Main results
Three trials were included, with a combined total of 432 randomised lesions (137 participants).
Forty-two conference papers, abstracts and posters were excluded (from an unknown number of
studies). The risk of bias in all studies appeared high. The analyses of all three studies were
conducted at the level of the lesion, which is not independent of the person, for this reason
pooling of data was not appropriate or attempted. Individual studies showed inconsistent effects
of ozone on caries, across different measures of caries progression or regression. Few secondary
outcomes were reported, but one trial reported an absence of adverse events.
Authors' conclusions
Given the high risk of bias in the available studies and lack of consistency between different
outcome measures, there is no reliable evidence that application of ozone gas to the surface of
decayed teeth stops or reverses the decay process. There is a fundamental need for more
evidence of appropriate rigour and quality before the use of ozone can be accepted into
mainstream primary dental care or can be considered a viable alternative to current methods for
the management and treatment of dental caries.
Carcinogenesis. 2003 Mar;24(3):613-9.
dott Cosimo Cigliola
79

Combined oral benzo[a]pyrene and inhalatory ozone exposure have no effect on lung
tumor development in DNA repair-deficient Xpa mice.
Hoogervorst EM, de Vries A, Beems RB, van Oostrom CT, Wester PW, Vos JG, Bruins W,
Roodbergen M, Cassee FR, Vijg J, van Schooten FJ, van Steeg H.
National Institute of Public Health and the Environment, Laboratory of Toxicology, Center for
Environment and Health Research, Bilthoven, The Netherlands.
There is considerable concern about an enhanced risk of lung tumor development upon exposure
of humans to polycyclic aromatic hydrocarbons (PAHs), like benzo[a] pyrene (B[a]P), in
combination with induced lung cell proliferation by toxic agents like ozone. We studied this issue
in wild-type (WT) C57BL/6 mice, the cancer prone nucleotide excision repair-deficient Xeroderma
pigmentosum complementation group A mice (Xpa-/-) and the even more sensitive Xpa-/-/p53+/-
mice. The mice were treated with B[a]P through the diet at a dose of 75 p.p.m., in combination
with intermittent ozone exposures (0.8 p.p.m.). First, a dose-range finding study with WT and
Xpa-/- mice was conducted to determine the optimal ozone concentration giving high cell
proliferation and low toxic side effects. We show by BrdU incorporation that cell proliferation in the
lung was induced by ozone, with an optimal concentration of 0.8 p.p.m., which was subsequently
used in the (sub)chronic studies. In the subchronic study, in which lacZ mutant frequency and
BPDE-DNA adduct formation were measured, the mice were treated for 13 weeks with B[a]P and/
or ozone, whereas in the chronic study this treatment protocol was followed by a 6 month period
on control feed and filtered air. As expected, oral B[a]P exposure appeared to be highly
carcinogenic to Xpa-/- and Xpa-/-/p53+/- mice and to a lesser extent to WT mice. A high
incidence of forestomach tumors and some tumors of the esophagus were found. In the lung, a
clear genotoxic effect of B[a]P was found as shown by the presence of BPDE-DNA adducts.
However, these DNA adducts in combination with induction of cell proliferation did not result in
increased lacZ mutations, nor in lung tumor formation not even in the highly sensitive Xpa-/- and
Xpa-/-/p53+/- mice. The implication of these findings for tumor risk assessment will be discussed.
Publication Types:
Research Support, U.S. Gov't, P.H.S.
PMID: 12663525 [PubMed - indexed for MEDLINE]
European Journal of Oral Sciences
Volume 114 Issue 5 Page 435 - October 2006
To cite this article: Karin C. Huth, Franz M. Jakob, Bernd Saugel, Christian Cappello, Ekaterini
Paschos, Regina Hollweck, Reinhard Hickel, Korbinian Brand (2006) Effect of ozone on oral cells
dott Cosimo Cigliola
80

compared with established antimicrobials European Journal of Oral Sciences 114 (5), 435–440.
doi:10.1111/j.1600-0722.2006.00390.x
Prev Article Next Article
Effect of ozone on oral cells compared with established antimicrobials
Karin C. Huth 11
Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-
University, Munich, Germany, Franz M. Jakob 11
Department of Restorative Dentistry and
Periodontology, Ludwig-Maximilians-University, Munich, Germany, Bernd Saugel 22
Institute of
Clinical Chemistry and Pathobiochemistry, Christian Cappello 22
Institute of Clinical Chemistry and
Pathobiochemistry, Ekaterini Paschos 11
Department of Restorative Dentistry and Periodontology,
Ludwig-Maximilians-University, Munich, Germany, Regina Hollweck 33
Institute for Medical Statistics
and Epidemiology, Klinikum rechts der Isar, Technical University, Munich, Germany,
Reinhard Hickel 11
Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-
University, Munich, Germany, Korbinian Brand 22
Institute of Clinical Chemistry and
Pathobiochemistry
1 Department of Restorative Dentistry and Periodontology, Ludwig-Maximilians-University, Munich,
Germany; 2 Institute of Clinical Chemistry and Pathobiochemistry, and 3 Institute for Medical
Statistics and Epidemiology, Klinikum rechts der Isar, Technical University, Munich, Germany
Dr Karin C. Huth, Department of Restorative Dentistry & Periodontology, Dental School, Ludwig-
Maximilians-University, Goethe Street 70, 80336 Munich, Germany
Telefax: +49–89–51605344
E-mail: khuth@dent.med.uni-muenchen.de
Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K. Effect of
ozone on oral cells compared with established antimicrobials. Eur J Oral Sci 2006; 114: 435–440.
© 2006 The Authors. Journal compilation © 2006 Eur J Oral Sci
Abstract
Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its
antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous
ozone (4 × 10 6 µg m -3 ) and aqueous ozone (1.25–20 µg ml -1 ) exert any cytotoxic effects on
human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with established
antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl) 5.25%,
2.25%; hydrogen peroxide (H2O2) 3%], over a time of 1 min, and compared with the antibiotic,
metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and apoptosis
dott Cosimo Cigliola
81

were evaluated. Ozone gas was found to have toxic effects on both cell types. Essentially no
cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic to BHY cells,
and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H2O2 resulted in markedly
reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity only to BHY
cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of the tested
antiseptics.
dott Cosimo Cigliola
82

Efficacy of ozone on survival and permeability of oral microorganisms
M. Nagayoshi 1 , T. Fukuizumi 2 , C. Kitamura 1 , J. Yano 1 , M. Terashita 1 , T. Nishihara 2 ,
1 Department of Operative Dentistry and Endodontics, 2 Department of Oral Microbiology, Kyushu
Dental College, Kitakyushu, Japan
Tatsuji Nishihara, D.D.S., Ph.D., Department of Oral Microbiology, Kyushu Dental College, 2-6-1
Manazuru, Kokurakita-ku, Kitakyushu 803–8580, Japan Tel.: + 81 93 582 1131, ext. 5485; fax: +
81 93 581 4984; e-mail: tatsujin@kyu-dent.ac.jp
Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T. Efficacy of ozone on
survival and permeability of oral microorganisms. Oral Microbiol Immunol 2004: 19: 240–246.©
Blackwell Munksgaard, 2004.
Abstract
In the present study, we examined the effect of ozonated water on oral microorganisms and
dental plaque. Almost no microorganisms were detected after being treated with ozonated water
(4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial cells
were stained with LIVE/DEAD ® BacLight TM Bacterial Viability Kit. Fluorescence microscopic analysis
revealed that S. mutans cells were killed instantaneously in ozonated water. Some breakage of
ozonated water-treated S. mutans was found by electron microscopy. When the experimental
dental plaque was exposed to ozonated water, the number of viable S. mutans remarkably
decreased. Ozonated water strongly inhibited the accumulation of experimental dental plaque in
vitro. After the dental plaque samples from human subjects were exposed to ozonated water in
vitro, almost no viable bacterial cells were detected. These results suggest that ozonated water
should be useful in reducing the infections caused by oral microorganisms in dental plaque.
Efficacy of gasiform ozone and photodynamic therapy on a multispecies oral biofilm in
vitro.
Author: Müller P , Guggenheim B , Schmidlin PR
Source: Eur J Oral Sci, 115(1): 77-80 2007
Abstract: Ozone gas and photodynamic therapy (PDT) have been claimed to be antimicrobially
effective. This study assessed their antimicrobial potential in vitro. Mature six-species oral biofilms
were treated as follows (n = 9 per group):
1. a 60-s application of gasiform vacuum-ozone or vacuum alone (on wet or air-dried biofilm
samples);
2. PDT (i.e. methylene blue in combination with or without a diode soft laser, and a soft laser
alone); or
dott Cosimo Cigliola
83

3. antimicrobial solutions: immersion of biofilms for 60 s in 0.2 and 2% chlorhexidine or in 0.5
and 5% hypochlorite solution. Treatment with chlorhexidine or hypochlorite served as a
positive control, whereas untreated samples served as negative controls. Colony-forming units
on blood agar were counted. Only the 5% hypochlorite solution was able to totally eliminate
the microorganisms in the biofilm. The observed reduction of viable counts by vacuum-ozone
application and PDT was less than one log(10) step. Under the conditions of the current study,
gasiform ozone and PDT had a minimal effect on the viability of microorganisms organized in a
cariogenic biofilm.
The use of ozone in dentistry and medicine.
Author: Baysan A , Lynch E
Source: Prim Dent Care, 12(2): 47-52 2005
Abstract:
There is growing interest in the use of ozone in oral healthcare and the National Institute for
Clinical Excellence (NICE) is at present reviewing the evidence for its effectiveness in the
management of occlusal and plain surface caries. These are only two of the clinical problems for
which ozone can, and has, been used; it has also been employed for a wide variety of other
purposes in both dentistry and medicine. This pale blue-coloured gas plays an important role as a
natural constituent in the higher layer of the Earth's atmosphere. There is growing evidence that it
can be employed as a useful therapeutic agent. This paper reviews its therapeutic uses to date
and suggests its possible future clinical applications. Consumer demands for this strong oxidant
may increase as the general public becomes increasingly aware of its therapeutic capacity and the
non-invasive manner in which it can be administered.
Ozone therapy for the treatment of dental caries.
Author: Rickard GD , Richardson R , Johnson T , McColl D , Hooper L
Source: Cochrane Database Syst Rev, (3): CD004153 2004
Abstract:
BACKGROUND: Dental caries is a bacterially mediated disease characterised by demineralisation
of the tooth surface, which may lead to cavitation, discomfort, pain and eventual tooth loss.
Ozone is toxic to certain bacteria in vitro and it has been suggested that delivering ozone into a
carious lesion might reduce the number of cariogenic bacteria. This possibly could arrest the
progress of the lesion and may, in the presence of fluoride, perhaps allow remineralisation to
dott Cosimo Cigliola
84

occur. This may in turn delay or prevent the need for traditional dental conservation by 'drilling
and filling'.
OBJECTIVES: To assess whether ozone is effective in arresting or reversing the progression of
dental caries.
SEARCH STRATEGY: We searched the Cochrane Oral Health Group's Trials Register (to 7
November 2003); Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library
Issue 3, 2003); MEDLINE and PREMEDLINE (OVID, 1966 to November 2003); EMBASE (OVID,
1980 to November 2003); CINAHL (OVID, 1982 to November 2003); AMED (OVID, 1985 to
November 2003). Quintessence was handsearched through 2002 and KaVo were contacted as
manufacturers of the HealOzone apparatus for any additional published or unpublished trials.
SELECTION CRITERIA: Inclusion was assessed independently by at least two reviewers. Trials
were only included if they met the following criteria: randomisation in a controlled trial; single
surface in vivo carious lesion accessible to ozone application; clear allocation concealment; ozone
application to the lesions in the intervention group; no such application of ozone in the control
group; outcomes measured after at least 6 months.
DATA COLLECTION AND ANALYSIS: Reviewers independently extracted information in
duplicate. A paucity of comparable data did not allow meta-analytic pooling of the included
studies.
MAIN RESULTS: Three trials were included, with a combined total of 432 randomised lesions
(137 participants). Forty-two conference papers, abstracts and posters were excluded (from an
unknown number of studies). The risk of bias in all studies appeared high. The analyses of all
three studies were conducted at the level of the lesion, which is not independent of the person, for
this reason pooling of data was not appropriate or attempted. Individual studies showed
inconsistent effects of ozone on caries, across different measures of caries progression or
regression. Few secondary outcomes were reported, but one trial reported an absence of adverse
events.
REVIEWERS' CONCLUSIONS: Given the high risk of bias in the available studies and lack of
consistency between different outcome measures, there is no reliable evidence that application of
ozone gas to the surface of decayed teeth stops or reverses the decay process. There is a
fundamental need for more evidence of appropriate rigour and quality before the use of ozone can
be accepted into mainstream primary dental care or can be considered a viable alternative to
current methods for the management and treatment of dental caries.
Efficacy of ozone on survival and permeability of oral microorganisms.
Author: Nagayoshi M , Fukuizumi T , Kitamura C , Yano J , Terashita M , Nishihara T
Source: Oral Microbiol Immunol, 19(4): 240-6 2004
dott Cosimo Cigliola
85

Abstract: In the present study, we examined the effect of ozonated water on oral microorganisms
and dental plaque. Almost no microorganisms were detected after being treated with ozonated
water (4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial
cells were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Fluorescence microscopic
analysis revealed that S. mutans cells were killed instantaneously in ozonated water. Some
breakage of ozonated water-treated S. mutans was found by electron microscopy. When the
experimental dental plaque was exposed to ozonated water, the number of viable S. mutans
remarkably decreased. Ozonated water strongly inhibited the accumulation of experimental dental
plaque in vitro. After the dental plaque samples from human subjects were exposed to ozonated
water in vitro, almost no viable bacterial cells were detected. These results suggest that ozonated
water should be useful in reducing the infections caused by oral microorganisms in dental plaque
Ozone effect on dental plaque
14. Shargawi JM, Theaker ED, Drucker DB, MacFarlane T, Duxbury AJ.
Sensitivity of Candida albicans to negative air ion streams. J Appl Microbiol, 87:889-897;1999.
1999 O3 in Negative air ions (NAIs) at different emitter distances, exposure times, relative
humidities & under aerobic & oxygen-free conditions O3 levels increased with increasing exposure
times (P < 0. 01) but were significantly reduced as emitter distance increased (P < 0. 01). When
utilized in a nonventilated room, levels of O3 produced did not exceed recognized safety limits.
15. Grootveld M, Lynch E. Mills B, Smith C, Baysan A, Silwood C.
Therapeutic Oxidation Of Human Plaque Biomolecules by an Anti-Bacterial Ozone-Generating
Device. BSDR Abstract no. 292; 2001. 2001 12 10 seconds Analysis of plaque by high field 1H –
NMR spectroscopy to detect oxidation of biomolecules by O3 O3 treatment gave rise to the
oxidative decarboxylation of the electron donor pyruvate (generating acetate and CO2 as
products), and the oxidation of the volatile sulphur compound precursor methionine to its
corresponding sulphoxide. Moreover, evidence for the O3-mediated oxidation of 3-D-
hydroxybutyrate was also obtained.
16. Grootveld M., Baysan A, Silwood C, Lynch E.
Oxidation Of Human Plaque Biomolecules by an Anti-Bacterial Ozone-Generating Device. The First
Pan European Festival of Oral Sciences, Cardiff, UK. Abstract no. 595; 2002 2002 12 10 seconds
Analysis of plaque by high field 1H –NMR spectroscopy to detect oxidation of biomolecules by O3
O3 treatment gave rise to the oxidative decarboxylation of the electron donor pyruvate
(generating acetate and CO2 as products), and the oxidation of the volatile sulphur compound
precursor methionine to its corresponding sulphoxide. Moreover, evidence for the O3-mediated
oxidation of 3-D-hydroxybutyrate was also obtained.
dott Cosimo Cigliola
86

17. Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T.
Efficacy of Ozone on survival & permeability of oral microorganisms. Oral Microbiol Immunol,
19:240-246; 2004. 2004 ozonated water (4 mg/l) for 10 s Streptococcus mutans, And dental
plaque from human subjects 1. breakage S. mutans seen by electron microscopy . 2. Ozonated
water inhibited the accumulation of experimental dental plaque in vitro 3. Human dental plaque
exposed to ozonated water in vitro had no viable bacterial cells detected.
Assessment of Anxiety & Fear with Ozone Treatment v Traditional Dental Therapy
18. Al Shorman H, Abu-Naba'a L, Lynch E. Patient's Attitude to Treatment of Pit & Fissure Caries
with Ozone. Caries Res, 36:187; 2002. 2002 49 n/a n/a Pit & Fissure Caries 98% happy with
treatment, 94% happy with time, 94% would choose O3 even if higher fee, 94% would
recommend it for a friend or relative. 100% would choose O3 treatment again, & 100% not
anxious after O3 treatment same visit
19. Domingo H, Smith C, Freeman R, Lynch E.
Patients attitudes to managing caries with Ozone. J Dent Res, 81: A-183; 2002 2002 99 n/a n/a
Patients had drilling & fillings previously & now O3 treatment by the same dentist happy or
satisfied with 1. O3 treatment 99% 2. time 97% 3. if cost was more than conventional treatment
95% 4. Recommend to a friend/ Relative 97% 5.Receive treatment once more 100% 6. Patients
reported a reduction in anxiety associated with the O3 treatment (p

Combining Ozone Treatment with Other Preparation Systems
77. Clifford C.
Successful Use of Airbrasion in Conjunction with Ozone Treatment. J Dent Res, 82: B-2747; 2003.
2003 37 48 40 Seconds Approximal lesion requiring drilling and filling airabrasion and Ozone with
GI filling All lesions were successfully exposed and a seal established for the delivery of ozone.
Clinically acceptable seals have been achieved around all restorations. The airabrasion and Ozone
technique was significantly faster than conventional drilling and filling (P

Treatment of Primary/Deciduous Teeth
73. Abu-Salem OT.
Management of Occlusal Caries in Primary Teeth Using Ozone. Mphil thesis, Queens University
Belfast, 2004.
74. Abu-Salem OT, Marashdeh MM, Lynch E .
Immediate Effect of Ozone on Occlusal Caries of Primary Teeth. The First Pan European Festival of
Oral Sciences, Cardiff, UK. Abstract no. 432; 2002 and J Dent Res, 82: C-535; 2003 2002 12
invitro 10 seconds Occlusal Carious Lesions DIAGNOdent and standard scale ECM readings
correlated significantly with the clinical classification (rs=0.675, p = 0.016 and rs= -0.697,
p=0.012 respectively). Ozone application reduced the DIAGNOdent readings after 10s application
(t = 2.891, p =0.015), and after 20s (t = 5.313, p < 0.001).
75. Abu-Salem OT, Marashdeh MM, Lynch E Ozone Efficacy in Treatment of Occlusal Caries in
Primary Teeth. J Dent Res, 82: B-136; 2003 2003 16 42 10 Seconds Occlusal Carious Lesions At
six months ECM readings were improved significantly (p

treated restorations were symptom less throughout the 9 months. All 34 Fuji 7 restorations,
removed after 3 months recall, showed hard ‘caries' to exploration suggesting remineralisation
was successful. All lesions were successfully exposed and a seal established for the delivery of
ozone at baseline. The airbrasion and Ozone technique was significantly faster (lesions were
exposed, ozonated and sealed in under 7 minutes) than conventional drilling and filling (P

Reduction in treatment time with combined air abrasion & Ozone compared to traditional ‘Drill &
Fill’ & cost comparison. IADR abstract 2004. 2004 64 128 40 Seconds Primary Root Caries AA &
Ozone was faster than Drill & Fill. AA was more profitable than D&F
Ozone Treatment of Endodontic Pathologies
84. Schwan L, Bamfaste M.
[Experiences with the use of chlorine gas and ozone in the treatment of root gangrene and dental
granuloma]. Dtsch Zahnarztl Z, 6:301-308; 1951. 1951
85. Brunel A, Vannier R, Archinet F.
[Sterilization of minute endodontic material by the combination of ethylene oxide and ozone.
Experimental evaluation of its effectiveness]. Acta Stomatol Belg 62:355-359;1965. 1965
86. Sandhaus S.
[Ozone therapy in odontostomatology, especially in treatments of infected root canals]. Rev Belge
Med Dent, 20:633-646;1965. 1965.
87. Haimovici A, Lacatusu S, Irjicianu A, Joan E.
[Ozone in endodontic therapy]. Stomatologia (Bucur ),; 17:303-307; 1970. 1970
88. Chang H, Fulton C, Lynch E.
Antimicrobial Efficacy of Ozone on Enterococcus faecalis. J Dent Res, 82: B-220; 2003. 2003 n/a
n/a 60, 30, 20, 10, 0 seconds 108 solution E Faecalis 60= 0 cfu's, 30= 0 cfu's, 20= 0 cfu's, 10= 0
cfu's at 106 & 105 concentrations. Higher concentrations were markedly reduced
89. Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M.
Antimicrobial effect of ozonated water on bacteria invading dentinal tubules. J Endodontics,
778-781; 2004. Part 2 2004 Enterococcus faecalis & Streptcoccus mutans infections in vitro in
bovine dentin. When the specimen was irrigated with sonication, ozonated water had nearly the
same antimicrobial activity as 2.5% sodium hypochlorite (NaOCl). After irrigation with ozonated
water, the viability of E. faecalis and S. mutans invading dentinal tubules significantly decreased
Use of ozone in treatment associated with Dental and maxillofacial surgery
90. Minguez F, Gomez-Lus ML, Andre J, Cabronero MJ, Prieto J.
[Antimicrobial activity of ozonized water in determined experimental conditions]. Rev Sanid Hig
Publica (Madr ), 64:415-423; 1990. 1990 Disinfection of Buccal flora and hand flors Antimicrobial
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activity in ozonized water on bacterial suspensions and contaminated materials was meaningful
and depended fundamentally on concentration and time of exposure. On buccal flora, one rinse
alone had no effect, but various successive rinses led to substantial reductions in the number of
colonies of bacteria. Ozone had a similar effect, although more pronounced, on the flora of the
hands. Ozonized water placed in an open dish kept up antimicrobial activity for the first 20
minutes, but after 30 minutes this activity decreased substantially.
91. Kiniapina ID, Durnovo EA
[The efficacy of using ozone in the combined treatment of disseminated odontogenic phlegmons of
the maxillofacial area]. Stomatologiia (Mosk), Spec No:60-6; 1996. 1996
92. Lazutikov OV, Lunev BV.
[The use of ozonized solutions in the combined treatment of odontogenic putrefactive-necrotic
phlegmons of the maxillofacial area and neck]. Stomatologiia (Mosk), Spec No:64-65;1996. 1996
93. Malanchuk VA, Gorshevikova EV, Kopchak AV.
[Antimicrobial action of ozone in the treatment of mandibular fracture]. Klin Khir, 3:43-46;2000.
2000.
94. Korotkikh NG, Lazutikov OV, Dmitriev VV.
[The effect of ozone on the microbiological characteristics of the oral fluid in patients with
mandibular fractures]. Stomatologiia (Mosk), 79:20-21;2000. 2000
95. Korzhachkina NB, Radzievskii SA, Olesova VN.
[Preventive use of ozone, short waves, and laser therapy alone and in combination in early
postoperative period after dental implantation]. Vopr Kurortol Fizioter Lech Fiz Kult,6:17-19;
2002. 2002
Ozone treatment for Gingivitis and Periodontitis
96. Brauner A.
[Clinical studies of therapeutic results from ozonized water for gingivitis and periodontitis].
Zahnarztl Prax , 42:48-50;1991. 1991
OHManagement Software for Patient Management
97. Scholz V.
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OHManagement Software for quality management in an Ozone treatment practice. IADR Absrtact
no. 715; 2004. 2004 n/a n/a n/a 10 clinics operating with new OHManagement Software,
compared to no OHM Patient recall attendence (53% v 44%) & compliance (84% v 75%) were
better with the new OHM
Ozone Treatment of Dentinal sensitivities
98. Ciriello G.
[Ozone and dentinal sensitivity.]. Riv Ital Stomatol, 10:159-164;1955. 1955
Ozone treatment of Pulpal pathologies
99. Dechaume M.
[The use of ozone in the local treatment of caries, pulpitis and periapical osteitis.]. Suom
Hammaslaak Toim, 48:61-66; 1952. 1952
1H NMR Studies on Tooth Whitening
100. Holmes J, Grootveld M, Smith C, Claxson AWD, Lynch E.
Bleaching of Components Responsible for Extrinsic Tooth Discoloration by Ozone. AADR Abstract
no. 615; 2003. 2003 n/a n/a 5,10,15, 20 seconds tooth stain removal (Melanoidins) Stains were
generated via the reaction of L-lysine (1.25-250.0 x 10-3 mol. dm-3) with an equivalent
concentration of alpha-D-glucose in phosphate buffer (pH 7.00) at 80oC for a period of 240 hr.
Substantial bleaching of melanoidins following treatment with O3 [e.g., at an O3 delivery level of
4.48 mmol, the decrease in absorbance at 366 nm was 90 ± 4% (mean ± s.e.) for the 1.25 x
10-3 mol.dm-3 reaction mixture, and 28 ± 3% for that initially containing 250.0 x 10-3 mol.dm-3
reactants]. The extent of the bleaching process observed increased with increasing levels of O3
treatment The mechanism of this process may involve the ozonation of (>C=C

In vitro testing of a denture cleaning method using Ozone. J Med Dent Sci, 45:135-139; 1998.
1998 700 mg/h gas for 1 & 3 minutes ozonated water at 1 ppm & 3 ppm Streptococcus mutans
(IID 973), Staphylococcus aureus (209-P), & Candida albicans (LAM 14322). direct exposure to
gaseous O3 seems to be a more effective microbicide compared with ozonated water, & that
gaseous O3 can be clinically useful for disinfection of dentures.
103. Krozer A, Hall J, Ericsson I.
Chemical treatment of machined titanium surfaces. An in vitro study. Clin Oral Implants Res,
10:204-11; 1999. 1999 O3 made by commercial mercury lamp in ambient air Implant material
rinsed with a disinfectant solution with amino-alcohol 1. Amino alcohols forms a stable & dense
film in vitro. Which most likely prevents re-integration to occur at the implant-tissue interface in
vivo 2 Rrinsing in water, saline solution, & 5% H2O2 did not remove the amino-alcohol from the
surface. O3 complete removal of the adsorbed amino-alcohol adherent film.
104. Suzuki T, Oizumi M, Furuya J, Okamoto Y, Rosenstiel SF.
Influence of Ozone on oxidation of dental alloys. Int J Prosthodont, 12:179-83; 1999. 1999 O3
gas 20 mg/h dental alloys (Co-Cr, Au-Ag-Pt, & Au-Cu-Ag-Pd). reflectance, surface roughness, &
weight were measured O3 caused a slight change in the Au-Cu-Ag-Pd alloy in terms of measured
reflectance, but the changes were significantly less than those caused by acid-electrolyzed water
& one of the commercial denture cleaners.
105. Zhao H, Zheng D, Hong L.
The disinfection efficiency comparison of different treatments on dental impression & gypsum
casts. Hua Xi Kou Qiang Yi Xue Za Zhi, 18:332-335;2000. 2000 Gypsum casts O3 treatment is an
effective method in disinfecting the gypsum casts.
106. Hussey D, Armstrong C,Lynch E.
Bond strengths of composite to enamel /dentine treated with ozone. The First Pan European
Festival of Oral Sciences, Cardiff, UK. Abstract no. 697; 2002. 2002 n/a 40 teeth enamel and
dentine samples 10 Seconds 2.7min diameter tubes of composite resin (Esthet-X, Dentsply,
&bonding agent (Prime and Bond NT, Dentsply) The mean loads (Newtons) to debond the
specimens were as follows: enamel without ozone 116.4 (sd 50. 1), enamel with ozone 128.6 (sd
49.4), dentine without ozone 54.7 (sd 23.6), dentine with ozone 51.6 (sd 15.6). Wilcoxon signed
rank test revealed no statistically significant difference between the groups with and without the
ozone treatment (Enamel z= -1.05, p=0.29, Dentine z-- -0.15, p=0.88).
107. Baysan A, Lynch E.
Management of Primary Root Caries using Ozone The First Pan European Festival of Oral Sciences,
Cardiff, UK. Abstract no 195; 2002. 2002 79 220 10 seconds Fissure sealant applied over Root
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caries 61% of intact sealants in the O3 & sealant group & 42% of intact sealants in the sealant
only group (p < 0. 05). After 3 6 & 9 months, O3 & sealant group also had greater improvements
in the ECM & DIAGNOdent values when compared to the sealant only group (p < 0. 05).
108. Matsumura K, Ikumi K, Nakajima N, Peng C, Hyon SH, & Tsutsumi S,
A Trial of Regeneration of Periodontal Ligament around Dental Implants J Dent Res, 81: A-101;
2002. 2002 titanium implant PDL cell taken from dog’s a. titanium implant was oxidized by
30%H2O2 b. poly(ethylene-co-vinyl alcohol)(EVA) was coated onto the implant & O3 c. Third, the
collagen sponge was immobilized around the implant by freeze-drying method. d. PDL cells were
seeded onto the implant and then implanted into dog's alveolar bone. After 3 months of
implantation, the implants and their surrounding tissues were removed. The specimens were
stained with Masson Trichrome stain. From the results, the tissue around implant was observed to
be similar to the one around natural tooth.
109. Murakami H, Mizuguchi M, Hattori M, Ito Y, Kawai T, Hasegawa J.
Effect of denture cleaner using ozone against methicillin-resistant Staphylococcus aureus and E.
coli T1 phage. Dent Mater J, 21:53-60; 2002. 2002 10 ppm Methicillin-resistant Staphylococcus
aureus (MRSA) and T1 phag virus bacteria was 3.1 x 10(3) CFU/mL at the beginning of the
experiment, fell to 1.0 x 10(0) CFU/mL 10 min later, and was 1.0 x 10(0) CFU/mL or less
afterwards. In contrast, when the ozone supply was cut off (air bubble only), the number of
bacteria was 3.4 x 10(3) CFU/mL at the beginning of the experiment, and had fallen to 3.0 x
10(3) CFU/mL 60 min later (no statistically significant difference). In the virucidal activity test, the
number of phages was 1.2 x 10(6) PFU/mL before ozone treatment, fell to about 1/10 of that
number 10 min later, and was 6.1 x 10(0) PFU/mL 40 min later.
110. Campbell D, Hussey D, Cunningham L, Lynch E.
Effect of Ozone on Surface Hardness of Restorative Materials. J Dent Res,82:B-262; 2003. 2003
n/a 18 x 3 10 Seconds Restorative Materials Statistical analysis using a 2-way ANOVA did not
reveal any difference in surface hardness following the treatment with O3 (p >0.15)
111. Baysan A, Lynch E.
12-month Assessment of Ozone on Root Caries J Dent Res, 82; B-311; 2003. 2003 79 220 10
seconds Fissure sealant applied over Root caries Modified USPHS criteria revealed that there were
61% of intact sealants in the ozone and sealant group and 26.1% of intact sealants in the sealant
only group (p < 0.05). After 1, 3, 6, 9 and 12 months, the ozone and sealant group also had
greater improvements in the ECM and DIAGNOdent values when compared to the sealant only
group (p < 0.05). Conclusions: Leathery root caries can be treated non-operatively with ozone
112. Hiller Ka, Federlin M, Mackow A, Redlich M, And Schmalz G.
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Influence of ozone treatment on marginal adaptation of fissure sealing Continental NOF Divisions
of the IADR Abstract no. 62; 2004. 2004 120 extracted human molars Art + Group I: O3 40s, seal
Tetric Flow, Helioseal Clear or Fuji VII Group II: restorations were placed immediately after
preparation. Before TC, the frequency of silver-staining was 26-50% (group I) and 23-50% (group
II), TF revealing the lowest (26%) and FU the highest frequency (50%) (group I). After TC, the
frequency of silver-staining was 8-46% (group I) and 10-50% (group II), TF revealing the lowest
(8%/10%) and FU the highest frequency (46%/50%) in groups I and II. Before and after TC,
microleakage was limited to enamel with TF and FU, whereas with HC silver-staining included
enamel and dentin. The Error Rates Method revealed a significant influence of the sealing material
upon the frequency of silver-staining but no difference was determined with respect to ozone
treatment. thermocycling 60s/cycle), stored in saline at 37°C for one week.
113. Czarnecka B, Deegowska-Nosowicz P, Prylinski M, Limanowska-Shaw H.
Bond strength of glass-ionomer's to dentine after Heal Ozone treatment Continental NOF Divisions
of the IADR Abstract no. 63; 2004. 2004 60 extracted bovine teeth n/a group Ozone then
conditioned with conditioner group B were treated with Ozone and Reductant then conditioned.
Group C (controls) were conditioned. samples of glass ionomer Fuji Fast (GC International-X) and
20 similar samples of Ketac Molar Aplicap (3M-ESPE, Seefeld, Germany-Y) were bonded to the
prepared surfaces; Group A gave the lowest values of SBS in both cases though this was
statistically significant (p

Ozone & Sealant Treatment of Root Caries after 12-Months. J Dent Res, IADR Abstract 2005. 2005
62 124 40 seconds flowable root sealant Primary Occlusal Fissure Carious Lesions and 60/61
arrested in treatment group, 24/61in control at one month. retention of sealants on these
arrested lesions is very promising 2/55 Tt; 8/32 in control lost retention completely or partially 1
& 12 month.
117. Abu-Naba'a L, Al Shorman H, Lynch E.
Fissure sealant retention over Ozone-treated occlusal pit & fissure caries: 12-months results. J
Dent Res, IADR Abstract 2005. 2005 53 132 10 seconds Primary Occlusal Fissure Carious Lesions
& fissure sealant O3 treatment followed by immediate sealant placement was not detrimental to
retention rate compaired to the control group. 0% secondery caries, 0% complete loss of fissure
sealant retention after one year.
118. Abu-Naba'a L, Al Shorman H, Hayajneh R, Lynch E.
Ozone effects on denture acrylic surface. J Dent Res, IADR Abstract 2005. 2005 40 samples 1 & 2
hours Heat cured acrylic with a polished & glazed surface O3 didn’t produce any change in the
roughness of the surface of treated samples.
Treatment of Dental Unit Water Lines (DUWL)
119. Filippi A, Tilkes F, Beck EG, Kirschner H.
[Water disinfection of dental treatment units using ozone]. Dtsch Zahnarztl Z, 46:485-487; 1991.
1991 Dental unit Under the precondition that the dental chair had been thoroughly sanitized, the
system showed a good disinfecting effect. Finally, for reasons of practical medical treatment, the
ozone concentration in air and, for reasons of hygiene in drinking water, the ozone concentrations
in water were determined under various conditions. In addition, the influence of continuous-flow
water heaters is discussed. The problem of continuous-flow water heaters regarding their effects
on the colonisation of water by microbes proves not to be significant. The exposure of patients to
disinfectants is discussed
120. Al Shorman, Coulter W, Lynch E Claxson AWD, Silwood CJL,Grootveld M
Use of Ozone to Treat Dental Unit Water Lines. BSDR Abstract no. 219; 2001. 2001 n/a n/a 10
seconds DUWL samples the biomolecules in the DUW were acetate, proprionate, formate, the
amino acid glycine, aromatic compounds & occasionally ethanol. O3 treatment of the DUW gave
rise to oxidation of ethanol & an increase in formate levels presumably due to oxidation of
carbohydrate.
121. Walker JT, Bradshaw DJ, Fulford MR, Martin MV, Marsh PD.
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Control of planktonic and biofilm contamination in a laboratory dental unit water system J Dent
Res, 81:A-445; 2002. 2002 200mg/hour Combizyme (1.25%) Tegodor (1%), Spor-Klenz (Neat),
Dialox (Neat), Tegodor (1%), Bleach (0.5%), Ozone (200mg/hour), Chlorhexidine (0.2%),
Ultrakleen (powder) and Betadene (10%) were tested overnight (16h). Flushing did not reduce
the viability or the extent of the biofilm on DUWS tubing. Only Combizyme did not completely
reduce the total viable counts of planktonic or biofilms cells. However, the efficiency of biofilm
removal was: Dialox > Bleach > Ultrakleen > Betadine > Spor-Klenz > Bio2000.
122. Cardon B, Eleazer P, Miller R, Staat R.
Low concentration Ozone treatment insufficient to control DUWL biofilm. AADR Abstract no. 714;
2002. 2002 n/a n/a O3 conc 0. 05ppm DUWL samples The O3 concentrations at the handpiece
during recirculation was 0.01 to 0.06 ppm;. Microbiological data indicated that water samples
taken 30 or more minutes after the O3 treatment cycles all exceeded 10,000 CFU. After 10 weeks
of O3treatment, microscopic analysis showed biofilm formation on the interior of all tubing
sections.
123. Al Shorman, Abu-Naba'a, Coulter W, Lynch E.
Ozone efficacy in the treatment of Dental Unit Water Lines. J Dent Res, 81: B 299. 2002. 2002 2
dental units n/a 1 & 3 minutes / unit/ days 1,2,3,4,& 7 DUWL samples after flushing for 2 minutes
Reduction of bacteria 1000 fold on day 7 for the 1 min application. Sterile water on day 2 for the 3
min application, sterile water followed up till 5 weeks.
124. Al Shorman, Coulter W, Abu-Naba'a, Mohan G, Boyle C, Lynch E.
Effect of Ozone on biofilms in Dental Unit Water Lines. AADR Abstract 2002. 2002 4 dental units
n/a 1,2,3 minutes & 15 min 1,2,3 minutes application 2 min flushing, 15 min application & 15 min
flushing 10 fold increase for 1 min, 10,000 fold decrease of bacterial counts for 2 & 3 min
application times. Biofilm layer seen to be reduced at 15 min application time & 10 min flushing,
but completely removed at 15 minutes application & flushing time 7 days for units 1,2 & 3
125. Al Shorman, Abu-Naba'a, Coulter W, Lynch E.
Ozone, An Effective Treatment For Dental Unit Water Lines. J Dent Res 81:A-112; 2002. 2002 1
dental units n/a 3 minutes, at 2100ppm O3, 615 ml/min DUWL flushed for 2 minutes O3
treatment showed reduction from 5. 2*103 CFU/ml to 300 CFU/ml after first application, then 0
CFU/ml at the second & subsequent (1000 fold reduction)
126. Al Shorman, Abu-Naba'a, Coulter W, Lynch E.
Ozone, An Effective Treatment For Dental Unit Water Lines. J Dent Res 81:A-112, 2002. 2002 1
dental units n/a 3 minutes, at 2100ppm O3, 615 ml/min DUWL flushed for 2 minutes O3
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treatment showed reduction from 5. 2*103 CFU/ml to 300 CFU/ml after first application, then 0
CFU/ml at the second & subsequent (1000 fold reduction)
127. Smith C, Al Shorman H, Grootveld M, Silwood C, Lynch E, Mills B, Silwood C.
Rapid Detection of Microbial-Derived Components in Dental Unit Water Lines by NMR Analysis. J
Dent Res, 81:A-112, 2002. 12 dental units Multicomponent 1H NMR investigations of DUWLs
Results acquired revealed that many biomolecules were detectable in the samples examined,
including a wide range of microbial fermentation products (MFPs). For example, the organic acid
anions acetate, formate, lactate, propionate and succinate, and occasionally ethanol were present
in the samples examined. Further components detectable included the amino acids alanine and
glycine, and also a series of aromatic compounds. Treatment of DUWLs with the powerful
microbicidal agent ozone gave rise to a substantial reduction in many of the MFPs detectable in
samples collected 18 hr. after treatment (p < 0.01).
128. Walker JT, Bradshaw DJ, Fulford MR, Marsh PD.
Microbiological evaluation of a range of disinfectant products to control mixed-species biofilm
contamination in a laboratory model of a dental unit water system. Appl Environ Microbiol,
69:3327- 32; 2003. A model in-vitro O3 was applied for 10 min In-vitro model Low concentration
of O3 did not completely reduce the biofilm total bacterial count at this concentration nor reduce
the percentage biofilm coverage 129. Smith C, Al Shorman H, Abu-Naba'a L, Grootveld M, Silwood
C and Lynch E
Detection of Microbial-Derived Components in Dental Unit Water Lines using NMR. The First Pan
European Festival of Oral Sciences, Cardiff, UK. Abstract no. 477; 2002 & J Dent Res, 82: C-542;
2003. 16 dental units 10 seconds Multicomponent 1H NMR investigations of DUWLs signals
presented in control samples by microbial-derived organic acid anionsused for chemotaxonomic
‘markers’ of, notably acetate, formate , lactate, propionate and succinate. Others are the amino
acid glycine, a number of aromatic compounds and occasionally ethanol. O3 caused reduction in
many of the microbial fermentation products detectable in samples collected 18 hr. after
treatment (p < 0.01).
130. Al Shorman, Abu-Naba'a, Coulter W, Lynch E.
Primary Colonization of DUWL by P. aeruginosa & its Eradication by Ozone. J Dent Res, 82: B-284;
2003. 5 minutes ozone then 10 minutes flushing Dental Unit Water Lines treated either with
continuous H2O2 (Oxygenal, KaVo, Germany) or ozone Oxygenal treatment continuously
produced water with TVC of less than 100 CFU/mL. TVC of water from the control unit was 2.3 x
104 and 3.4 x 104 CFU/mL after 1 and 2 weeks of installation. The primary coloniser was
identified (API 20 NE kit) as pure P.A. After the first O3 treatment the TVC was reduced to 60
CFU/mL and rose to 3.9 x 104 CFU/mL after a week with few Pseudomonas colonies. After two
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weeks, TVC was 2.8 x 103CFU/mL CFU/mL with no detected P.A. and became 0 CFU/mL after the
treatment. Repeated sampling of the unit for 9 weeks showed no re-growth of P.A.
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Studies on Ozonated Oils
131. Lynch E, Grootveld M, Holmes J, Silwood CJ, Claxson AWD, Prinz J, Toms H.
Analysis of Ozone-treated Grapeseed, Olive, & Sunflower Seed Oils. AADR Abstract no. 182; 2003.
Ozonated Oils Comparison Treatment of each vegetable oil with O3 gave rise to the consumption
of polyunsaturated fatty acids present (i.e. significant reductions in their mono- and bis-allylic-
CH2 group resonances located at 2.06 and 2.76 ppm respectively, and also that of their vinylic
protons at 5.38 ppm), consistent with their ozonation. Indeed, signals present in the 5.10-5.25
ppm regions of the ozonated GO and SO spectra are assignable to the ring protons of ozonides.
Further O3-induced modifications to the oils included the production of aldehydes, i.e. -CH2CHO
aldehydic group triplet resonances at 9.65 (ozonated GO and SO) and 9.74 ppm (all ozonated
oils), terminal products arising from the decomposition of ozonides.
Other References
132. Baysan A, Lynch E, Grootveld M.
The use of Ozone for the management of primary root carious lesions. Tissue Preservation &
Caries Treatment. . Quintessence Book 2001, Chapter 3, 49-67. 2001 50 100 10 Seconds Primary
Root Caries
Ozone safety
1. Baysan A, Lynch E.
Safety of an Ozone delivery system during caries treatment in-vivo. J Dent Res, 80: 1159; 2001.
2001 40 80 10 & 20 Seconds Root Caries Both application times produced O3 aroungd the
application cup below the FDA & EU permissible O3 levels in air
2. Ebensberger U, Pohl Y, Filippi A.
PCNA-expression of cementoblasts & fibroblasts on the root surface after extraoral rinsing for
decontamination. Dent Traumatol,18:262-266;2002. 2002 O3 in water Extracted teeth,
cementobalsts & fibroblasts Marked by Proliferating Cell Nuclear Antigen (PCNA): Irrigation with
ozonized water showed higher labelling indices in comparison with saline, but this could not be
statistically substantiated (P = 0. 24). Ozonized water, not being isotonic, had no negative effect
on periodontal cells remaining on the tooth surface after irrigation for 2 min.
3. Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M.
Antimicrobial effect of ozonated water on bacteria invading dentinal tubules. Journal of
Endodontics, 778-781;2004. Part 1 2004 O3 in water L-929 mouse fibroblasts The metabolic
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activity of fibroblasts was high when the cells were treated with ozonated water, whereas that of
fibroblasts significantly decreased when the cells were treated with 2. 5% NaOCl
1H NMR Studies on Caries Biomolecules
4. Smith C, Lynch E, Baysan A, Silwood CJ, Mills B, Grootveld M.
Oxidative consumption of root caries biomolecules by a novel anti bacterial Ozone delivery
system, J Dent Res, 80:1178;2001. 2001 Analysis of root caries by high field 1H –NMR
spectroscopy to detect oxidation of biomolecules by O3. O3 caused oxidative decarboxylation of
pyruvate & oxidative attack of carbohydrates to generate formate. O3 also oxidised PRCL lactate,
urate, glycosaminoglycans & methionine to yield acetate & CO2, allantoin, low-molecular-mass
saccharide fragments & methionine sulphoxide, respectively.
5. Lynch E, Silwood CJL, Smith C, Grootveld M.
Oxidising actions of an Anti-Bacterial Ozone-Generating Device towards Root Caries Biomolecules.
J Dent Res, 81:A-138;2002. 2002 20 20 5 Seconds Analysis of 12 soft root caries by high field 1H
–NMR spectroscopy to detect oxidation of biomolecules by O3. Results obtained revealed that in
addition to giving rise to the oxidative decarboxylation of pyruvate (generating acetate and CO2
as products), and the attack of carbohydrates to produce formate, O3 also oxidised PRCL lactate,
urate, glycosaminoglycans and methionine to yield acetate and CO2 (via pyruvate), allantoin, low-
molecular-mass saccharide fragments and methionine sulphoxide, respectively.
6. Lynch E, Silwood C, Smith C, Grootveld M.
Oxidising actions of Ozone towards Root Caries Biomolecules. The First Pan European Festival of
Oral Sciences, Cardiff, UK. Abstract no 197; 2002. 2002 18 10 seconds Analysis of Soft root caries
by high field 1H –NMR spectroscopy to detect oxidation of biomolecules by O3. Bacterial- or yeast-
derived organic acid anions (formate, acetate, propionate, n-butyrate, lactate, and pyruvate),
amino acids (predominantly alanine and glycine) and carbohydrates such as glucose were present.
O3 gave rise to the oxidative decarboxylation of pyruvate (generating acetate and CO2),
carbohydrates to produce formate, also oxidised lactate, urate and glycosaminoglycans to yield
acetate and CO2, allantoin and low-molecular-mass saccharide fragments respectively. VSC
precursor methionine was oxidised to sulphoxide. O3-mediated oxidation of 3-D-hydroxybutyrate .
7. Lynch E, Silwood CJ, Abu-Naba'A L, Al Shorman H, Baysan A, Holmes J, Grootveld M.
Oxidative Consumption of Root Caries Biomolecules using Ozone. Journal Caries Res,
38:364;2004. 2004 30 30 30 Seconds Analysis of root caries by high field 1H -NMR spectroscopy
to detect oxidation of PRCL biomolecules by O3. O3 caused oxidative decarboxylation of pyruvate
& oxidative attack of carbohydrates to generate formate. O3 also oxidised PRCL lactate, urate,
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glycosaminoglycans & methionine to yield acetate & CO2, allantoin, low-molecular-mass
saccharide fragments & methionine sulphoxide, respectively.
1H NMR Studies on Saliva Biomolecules
8. Lynch E, A Baysan A, Silwood C, Grootveld M.
Therapeutic oxidising activity of a novel anti-bacterial Ozone-generating device on saliva. J Dent
Res, 77: 1187; 1998. 1998 20 30 seconds Analysis of saliva by high field 1H -NMR spectroscopy
to detect oxidation of biomolecules by O3 O3 produced oxidative de- carboxylation of pyruvate, &
oxidation e sulphur compound methionine to its corresponding sulphoxide, oxidative consumption
of polyunsaturated fatty acids O3. Moreover, -mediated oxidation of 3-D-hydroxybutyrate was
also obtained.
9. Lynch E, Smith E, Baysan A, Silwood CJL, Mills B, Grootveld M.
Salivary Oxidising Activity of a Novel Anti-bacterial Ozone-generating Device. J Dent Res,
80:13;2001. 2001 20 10 seconds Analysis of saliva by high field 1H –NMR spectroscopy to detect
oxidation of biomolecules by O3 Oxidation e sulphur compound methionine to its corresponding
sulphoxide, oxidative consumption of polyunsaturated fatty acids O3.
10. Claxson AWD, Smith C, Turner MD, Silwood CJL, Lynch E, Grootveld M.
Oxidative Modification of Salivary Biomolecules with Therapeutic Levels of Ozone. J Dent Res,
81:A-502; 2002. 2002 20 20 10 seconds Analysis of saliva by high field 1H –NMR spectroscopy to
detect oxidation of biomolecules by O3. In addition to the complete oxidative consumption of
salivary pyruvate (mean ? s.e. salivary level prior to treatment 1.75 ? 0.62 mM) and partial
oxidation of methionine, results acquired revealed (1) marked elevations in the concentration of
formate (a product derived from the oxidation of carbohydrates), (2) substantial reductions in
lactate level (up to 92%) via its oxidation to pyruvate, which in turn is oxidised to acetate and
CO2, (3) the oxidation of 3-D-hydroxybutyrate to acetoacetate, which liberates acetone on
decomposition, (4) the generation of allantoin from salivary urate (up to levels of 40??M), (5) the
production of low-molecular-mass saccharide fragments from hyaluronate, and (6) oxidation of
malodorous trimethylamine (initial salivary concentration 78 ? 23 ?M) to its corresponding, non-
malodorous N-oxide.
11. Turner M, Silwood CJL, Grootveld M, Lynch E.
Oxidative Consumption of Biomolecules using Ozone. The First Pan European Festival of Oral
Sciences, Cardiff, UK. Abstract no. 594; 2002. 2002 10 seconds 5.00 ml of aqueous solutions:
sodium pyruvate, D-glucose, L-cysteine and L-methionine (5.00 mM) were prepared in 40.0 mM
phosphate buffer (pH 7.00)(rigorously deoxygenated with argon gas) As expected, treatment of
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aqueous solutions of L-methionine with ozone confirmed oxidation to its corresponding sulphoxide.
H NMR analysis demonstrated that exposure of aqueous solutions of L-cysteine to ozone
generated its corresponding disulphide, cystine, as a major product. H NMR analysis also showed
that ozonolysis of D-glucose generated formate as a major reaction product, i.e., a concentration
of 1.29 mM was produced from the 5.00 mM glucose substrate, an observation consistent with
previous studies conducted on the interactions of ROS (e.g., radiolytically-generated .OH radical)
with carbohydrates in general. Treatment of pyruvate with this oxidant produced acetate and CO2
via an oxidative decarboxylation process immediate.
12. Turner M, Grootveld M, Silwood C, Lynch E.
Oxidative Consumption of Biomolecules by Therapeutic Levels of Ozone J Dent Res,
81:A-272;2002. 2002 10 seconds Aqueous solutions containing sodium pyruvate, ?-D-glucose, L-
cysteine and L-methionine (5.00 mM) were prepared in 40.0 mM phosphate buffer (pH 7.00)
which was rigorously deoxygenated with argon gas prior to use. Attack of O3 on ?-D-glucose gave
rise to formate as a major product, i.e. 1.21 ? 0.11 mM (mean ? s.e.) generated, and treatment of
pyruvate with this oxidant produced acetate and CO2 via an oxidative decarboxylation process (93
? 4 % yield under our experimental conditions). Moreover, the amino acid volatile sulphur
compound (VSC) precursors cysteine and methionine were converted to their corresponding
primary oxidation products cystine (100% yield) and methionine sulphoxide (98 ? 2 % yield)
respectively.
13. Silwood C, Smith C, Turner M, Grootveld M, Lynch E.
Oxidative Modification of Salivary Biomolecules with Ozone. The First Pan European Festival of
Oral Sciences, Cardiff, UK. Abstract no. 593; 2002. 2002 16 0.60 ml saliva 20 seconds Analysis of
saliva by high field 1H –NMR spectroscopy to detect oxidation of biomolecules by O3. After
complete oxidative consumption of salivary pyruvate and partial oxidation of methionine, results
acquired revealed (1) marked elevations in the concentration of formate (a product derived from
the oxidation of carbohydrates), (2) substantial reductions in lactate level via its oxidation to
pyruvate, which in turn is oxidised to acetate and CO2, (3) the oxidation of 3-D-hydroxybutyrate
to acetoacetate, which liberates acetone on decomposition, (4) the generation of allantoin from
salivary urate, (5) the production of low-molecular-mass saccharide fragments from hyaluronate,
and (6) oxidation of malodorous trimethylamine to its corresponding, non-malodorous N-oxide.
Efficacy of ozone on survival and permeability of oral microorganisms. Oral Microbiol
Immunol 2004: 19: 240–246.
Key words: antimicrobial activity; dental plaque biofilm; disinfectant; oral microorganisms;
ozone
Abstract
In the present study, we examined the effect of ozonated water on oral microorganisms and
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dental plaque. Almost no microorganisms were detected after being treated with ozonated water
(4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial cells
were stained with LIVE/DEAD BacLight TM Bacterial Viability Kit. Fluorescence microscopic analysis
revealed that S. mutans cells were killed instantaneouslyin ozonated water. Some breakage of
ozonated water-treated S. mutans was found byelectron microscopy. When the experimental
dental plaque was exposed to ozonated water, the number of viable S. mutans
remarkablydecreased. Ozonated water stronglyinhibited the accumulation of experimental dental
plaque in vitro. After the dental plaque samples from human subjects were exposed to ozonated
water in vitro, almost no viable bacterial cells were detected. These results suggest that ozonated
water should be useful in reducing the infections caused byoral microorganisms in dental plaque.
Efficacy_of_ozone.pdf (460 KB)
Ozone Treatment of Primary Occlusal Pit and Fissure Caries (POPFC): 12-Months Clinical
Severity Changes
by L Abu-Naba'a, H Al Shorman and E Lynch
School of Dentistry, Queen's University, Belfast, Northern Ireland, UK
Conclusion: Clinical changes were significant from baseline for both groups. Detailed clinical
scoring should be developed to refine this validated diagnostic tool to monitor remineralisation. As
Ozone-treated lesions stabilised using a clinical severity score system from 6-months, the control
lesions steadily deteriorated. (*, p

improved or remained stable in Trt than in Ctrl at all recalls. 6 lesions in Ctrl changed score from 1
to 3 compared to only 1 in Trt. Different patterns of change suggested dissimilar mechanisms
within the groups. As Ctrl steadily increased, most probably by further lesion progression, the Trt
stabilisation may have reflected what was clinically noticed by lesions getting darker (score 1
changes to 2 but score 2 remains stable with more staining). More surface destruction (score 3)
was noticed in Ctrl.
Conclusion
Clinical changes were significant from baseline for both groups. Detailed clinical scoring should be
developed to refine this validated diagnostic tool to monitor remineralisation. As Ozone-treated
lesions stabilised using a clinical severity score system from 6-months, the control lesions steadily
deteriorated. (*, p

Objectives
To assess and monitor the progression of carious lesions using both DIAGNOdent® (KaVo,
Germany) and Electrical Caries Monitor (ECM, LODE BV, Netherlands) standard scale readings
after Ozone treatment in primary teeth.
Methods
17 patients with 50 non-cavitated occlusal carious lesions were recruited. All teeth were cleaned
by an air-abrasive system called Prophyflex 2® (KaVo, Germany) before DIAGNOdent, ECM
standard scale readings and clinical classifications (Ekstrand, 1998) were recorded. After
randomisation, half of the lesions were treated with Ozone using a silicone-sealed cup via a hand
piece (HealOzone unit, CurOzone USA) for 10 seconds, whereas, the other half was reserved as a
control group. The DIAGNOdent, ECM standard scale readings and clinical classifications readings
were repeated after 3 months.
Results
ECM readings were increased significantly (p

criteria for sealant quality. The changes in readings were tested from baseline by t-test of (loge
recall value/base value), and paired t-test between various groups and 2-way analysis for block
and treatment effects. The effect of co-variables was tested.
Results
No adverse effect was recorded. Mean change in ECM was 0.337 and -0.065 for the treatment and
control groups respectively. Difference between groups was better in treatment group from the
first month recall (p60, mean 70.8yrs, SD 7). A total of 68 patients with 3
lesions each (204 PRCL’s in total) were recruited. The lesions were randomly assigned into three
groups; treatment with Ozone, or air abrasion and filling or traditional drilling and filling (D&F).
Results: 64 patients completed this study. There were no observed adverse events. Air abrasion
decreased the mean time for cavity preparation by 4 times when compared to traditional rotary
drills (67 seconds (SD 12) compared to 4.5 minutes (SD 1.75 minutes)(p < 0.05)). The mean
time required for ozone treatment was 40 seconds (SD 5 seconds) for each lesion, whilst for
traditional D&F, the average time needed was 27 minutes (1620 seconds) (SD 8 minutes) (p <
0.01). Ozone treatment was faster by a factor of 40. The cost of the treatment was assessed. The
average cost to treat a lesion with ozone was 3.6 times less compared to the average cost for
each traditional filling.
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Conclusions: The new technologies of air abrasion and Ozone offer economic savings for dental
practices and patients. Compared with conventional drilling and filling of root caries, Ozone
treatment is 40 times faster and 3.6 times cheaper.
Keywords: Gerontology, Health services research, Cariology, Therapeutics, Quality of life.
First Author
Presenting
Helene Domingo Health Services Research Queen's University of Belfast Dental School
Clinical reversal of root caries using ozone
By E Lynch, N Johnson and J Johnson
Objectives: The objective was to assess the safety and efficacy of a novel ozone delivery system
(1) for the management of soft primary root caries as well as non cavitated leathery primary root
caries in the least severe category.
Methods: A total of 260 subjects, each with 2 primary root carious lesions (PRCLs) were
recruited. 60 Subjects had each 2 soft PRCL's and 200 subjects had each 2 leathery non cavitated
PRCL's which were in the least severe leathery category (J Dent Res 1993,73: 623-629). The
lesions were randomly assigned into two groups; treated with ozone, or no ozone. The Ozone
treatment was applied by a different operator than the one recording the clinical criteria used to
define the severity of the lesions. When no Ozone was applied the cup and suction system were
placed on the control lesion for the same time interval as on the test lesions. At baseline, 3 and 6
months, the lesions were clinically assessed as soft, leathery or hard, scored with the validated
root caries severity index and received treatment as at baseline.
Results: After 6 months, there were no observed adverse events, 48 of the ozone treated soft
PRCL’s had reversed from severity index 4 to 3, whilst the control soft lesions did not significantly
change (p < 0.01), 189 of ozone treated leathery PRCL’s had reversed from severity index 1 to 0
(i.e., hard and arrested), whilst the control leathery lesions did not significantly change (p <
0.01).
Conclusions: Leathery non cavitated primary root caries can be arrested non-operatively with
ozone. This novel treatment regime using ozone is an effective alternative to conventional
management methods including "drilling and filling". 1 HealOzone unit, CurOzone USA and KaVo
Keywords: Gerontology, Cariology, Therapeutics, Clinical trials, Remineralization
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First Author
Presenting
Edward Lynch, BDentSc, FDS, Ph Second Author Newton Johnson, BDS, FDS Third Author Judith
Johnson, MSC
In-Vivo Reversal of Occlusal Caries with Ozone after Six Months
By G Megighian and L Bertolini Private practice, Verona, Italy The use of ozone using HealOzone
technology allows the treatment of a vast range of dental caries at a very early stage and to
induce lesion remineralisation from saliva. Such methodology has the potential to be the
alternative to the conventional drilling and filling of dental caries.
Objectives: The aim of this randomised, double blind clinical trial was to longitudinally monitor
pit and fissure carious lesions (PFCLs) using light-induced fluorescence readings using a clinical
classification of PFCLs as well as the DIAGNOdent (KaVo Germany), and to quantify the effects of
Ozone application on these readings after six months.
Methods: A total of 300 PFCLs were entered in 80 patients. After cleaning each lesion using air
abrasion with 27.5 micron particles, a clinical severity index and DIAGNOdent (KaVo, Germany)
standard scale readings were recorded. 220 lesions were treated at baseline with Ozone (using
the HealOzone unit from CurOzone USA and KaVo Germany) for 20, 30 or 40 seconds according to
their clinical severity, after one other lesion (receiving no Ozone) was randomised as control in
each Patient. The clinical severity indices and DIAGNOdent readings were repeated after six
months to evaluate changes on both the test and control lesions.
Results: Ozone treated lesions clinically reversed (P

Ability of Therapeutic Levels of Ozone to Oxidatively Consume Biomolecules
C Smith, M Grootveld, C Silwood, L Abu-Naba'a, H Al Shorman and E Lynch Queen's ngdom
Objectives: The powerful microbiocidal activity of ozone (O3)is well known and previous
investigations have demonstrated the potential employment of this reactive oxygen specis
(ROS)as a therapeutic agent in the treatment of dental caries. This ROS is extremely reactive
towards selected carious dentine biomolecules, and such reactions are likely to be of relevance to
its microbiocidal activity. Therefore, in this study we have employed high resolution proton (1H)
nuclear magnetic resonance (NMR) spectroscopy to determine the nature and extent of the
oxidation of biomolecules present in carious dentine, plaque and saliva.
Methods: Aqueous solutions containing sodium pyruvate, a-D-glucose, L-cysteine and L-
methionine (4.00 mM) were prepared in 40.0 mM phosphate buffer (pH 7.00) which was
rigorously deoxygenated with argon gas prior to use. 5.00 ml aliquots of these solutions were
treated with O3 [1] for a period of 20 s (equivalent to a delivery of 8.96 mmol. of this oxidant).
These experiments were conducted in triplicate. Matching de-oxygenated solutions of these
biomolecules untreated with O3 served as controls.
Results: Attack of O3 on a-D-glucose gave rise to formate as a major product, i.e. 1.69 ± 0.1
mM (mean ± s.e.) generated, and treatment of pyruvate with this oxidant produced acetate and
CO2 via an oxidative decarboxylation process (98 ± 2% yield under our experimental conditions).
Moreover, the amino acid volatile sulphur compound (VSC) precursors cysteine and methionine
were converted to their corresponding primary oxidation products cystine (100% yield) and
methionine sulphoxide (99 ± 1% yield) respectively.
Conclusions: Multicomponent analysis of appropriate chemical model systems provides valuable
molecular information regarding the reactivity of O3 towards oral environment biomolecules. For
example, oxidation of cysteine and methionine provides evidence for the ability of O3 to combat
oral malodour since these biomolecules are precursors to volatile sulphur compounds (VSCs). [1]
HealOzone Unit, CurOzone, U.S.A. and KaVo
Topic: Cariology Research
Keywords: Cariology, Chemistry, Therapeutics
Oxidative Consumption of Root Caries Biomolecules by Ozone
Objectives: Restorative treatment of primary root carious lesions (PRCLs) represents a major
challenge. This investigation compared a unique method of non-restorative management of PRCLs
using ozone gas (O3). Hence, a multicomponent evaluation of the oxidative consumption of PRCL
biomolecules by O3 has been conducted using high-field proton (1H) nuclear magnetic resonance
(NMR) spectroscopy. O3 was generated by the commercially-available HealOzone Unit [1].
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Methods: 24 soft PRCLs requiring restoration from teeth were used since these are the severest
type of lesions found in humans. After plaque removal and drying, a sample of PRCL was taken
using a sterile excavator from half of the most active part of lesion. After this collection process,
O3 was applied to the lesion for 20 s (equivalent to a delivery of 8.96 mmol. of this oxidant) and
another sample was taken from the other half of the most active part of the lesion. Each sample
was then accurately weighed on a microbalance and perchloric acid extracts derived therefrom
were subjected to 1H-NMR analysis at an operating frequency of 600 MHz.
Results: Results obtained revealed that O3 gave rise to the oxidative decarboxylation of pyruvate
(generating acetate and CO2 as products), and its oxidative attack of carbohydrates to generated
formate. O3 also oxidised PRCL lactate, urate, glycosaminoglycans and methionine to yield
acetate and CO2 (via pyruvate), allantoin, low-molecular-mass saccharide fragments and
methionine sulphoxide, respectively.
Conclusions: Multicomponent analysis of root caries by high field 1H-NMR spectroscopy provides
useful information regarding the oxidation of PRCL biomolecules by O3. For example, pyruvate is
an extremely strong acid and its consumption by O3 may offer protection against tooth
demineralisation. Moreover, oxidation of the volatile sulphur compound precursor methionine to
its corresponding sulphoxide may serve as a useful means of blocking the development of oral
malodour. [1] CurOzone U.S.A. and KaVo Germany
Topic: Geriatric Oral Research
Keywords: Gerontology, Cariology, Chemistry, Diagnosis, Therapeutics
First Author
Presenting
Martin Grootveld, BSc, PHD,Health Services Research,Queen's University of Belfast Dental School
Ozone Treatment of Root Caries after 21-Months
By J C Holmes Queen's University of Belfast, United Kingdom
Objectives: The objective was to assess the efficacy of an ozone delivery system1 combined with
the daily use of a remineralising toothpaste, mouthrinse and spray(KaVo Gmbh) for the
management of non cavitated leathery primary root carious lesions (PRCL’s) in an ageing patient
group (>60, mean 70.8yrs, SD+ 6).
Methods: A total of 89 patients with 2 leathery lesions each (178 PRCL’s in total) in the middle
severity lesion group 2 (J Dent Res 1993,73: 623-629) were recruited. The lesions were randomly
assigned into two groups; treated with ozone, or no ozone and received a professionally applied
remineralising solution1. The Ozone treatment was applied by a different operator than the one
recording the clinical criteria used to define the severity of the lesions. When no Ozone was
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applied the cup and suction system were placed on the control lesion for the same time interval as
on the test lesions. Remineralising pastes, mouthrinses and sprays were also dispensed to each
patient with instructions. At baseline, 3, 6, 12, 18 and 21 months, the lesions were clinically
assessed as soft, leathery or hard, scored with the validated root caries severity index and
received treatment as at baseline.
Results: After 21 months, 81 patients completed the study. There were no observed adverse
events. At 21 months, 100% of ozone treated PRCL’s had reversed, whilst only 8% of the control
lesions reversed (p < 0.001). At 21 months, in the control group,12% of the PRCL’s had
progressed from severity index 2 to 3 (p < 0.001); i.e. they had become more severe.
Conclusions: Leathery non cavitated primary root caries can be treated non-operatively with
ozone. This novel treatment regime using ozone is an effective alternative to conventional "drilling
and filling".
Restoration of ART and Ozone treated primary root carious lesions
By Julian Holmes Queen's University of Belfast, United Kingdom
Objectives: This study assessed placement time required and durability of restorations placed on
ART and Ozone treated primary root carious lesions (PRCL) in the most severe category (J Dent
Res 1993;73: 623-629) compared to conventional treatment. A previous study showed that 100%
caries reversal with ozone treated PRCL’s can be obtained (Gerodontology, 2003: 20, 106-114) in
the middle severity lesion group 2.
Methods: 60 subjects, each with 2 soft PRCL were selected. After randomisation, each subject
had one PRCL made caries free using conventional local analgesia (LA), drilling and filling using
Optibond Solo Plus and Point 4 (KerrHawe) composite resin. 11 pulp exposures were detected
during cavity preparation and RCT was also performed on these teeth. The remaining 60 PRCL in
the 60 subjects received only the ART technique and Ozone treatment for 20 seconds (HealOzone,
KaVo) without LA, followed by filling using Optibond Solo Plus and Point 4.. Up to 1 mm of
softened carious dentine was left overlying the pulpal floor prior to Ozone treatment. The mean
(SD) time required for ART, Ozone treatment and placement of a restoration was 10 (2) minutes,
whilst the conventional technique, including LA, required 23 (4) minutes (P

Restoration of Ozone treated carious lesions
By L Steier Dental Clinic, Mayen, Germany Some Dentists are currently treating stained carious
lesions with Ozone without placing any restoration whilst others are bonding composite resin on
top of the Ozone treated stained lesion.
Objectives: The aim of this study was to assess the placement time required and durability of
restorations placed on Ozone treated carious lesions as well as to assess the aestetics of these
restorations from a Patients perspective.
Methods: 100 lesions in 50 subjects (2 per subject) were treated with Ozone from the HealOzone
system (KaVo, Germany) for 40 seconds. Immediately after Ozone treatment, 50 randomly
chosen lesions (1 per subject) were etched for 15 seconds with 37% Phosphoric Acid Etching Gel
and sealed with Nano-Bond Adhesive (Pentron Clinical Technologies,Wallingford, CT USA 06492),
cured using Ultralume 5 (Ultradent Products, South Jordan, Utah 84095) for 20 seconds, and
Simile Nano-hybrid Composite (Pentron) was placed and cured for 20 seconds. The remaining 50
Ozone treated lesions served as controls.
Results: The mean (SD) time required to Ozone treat and place a restoration was 3 (1) minutes.
After 6 weeks, 100% of all restorations scored alpha in every recorded USPHS criteria. The test
lesions were scored aesthetically as excellent in appearance in 100% of cases whilst the control
lesions were scored aesthetically as excellent in only 10% of cases (P

Alena Knezevic
University of Zagreb
School of Dental Medicine
Department of Endodontics and Restorative Dentistry
Gunduliceva 5, 10000 Zagreb, Croatia
Tel: 00385 (0)1 48 02 113
Fax: 00385 (0)1 48 02 159
e-mail: ma505ak@yahoo.com
Received: October 19, 2006
Accepted: January 10, 2007
Available online: March 1, 2007
Acta Stomatol Croat. 2007;41(1):31-8
Original scientific article
Abstract
Minimally invasive dentistry holds an important place in the treatment of caries lesion.
Introduction of laser fluorescence as a diagnostic and ozone as a therapeutic means finds more
frequent usage in both preventive and curative dentistry. DIAGNOdent device (KaVo, Germany) is
based on laser fluorescence which enables early diagnosis as well as monitoring the development
of caries lesion while use of ozone with HealOzone device (KaVo, Germany) provides painless
treatment. Even though there are number of indications for use of ozone in dentistry, most
important is therapy of caries lesions. Purpose of this investigation was to observe the effect of
ozone applied via HealOzone device on the total of 70 primary fissure lesions previously detected
with DIAGNOdent device and to determine whether there is a decrease in lesion demineralization
values between first visit, first and second recall after one and two month period. Results showed
statistically significant difference in demineralization values recorded on DIAGNOdent scale
between first visit and every repeated recall (p

The most often used conventional caries diagnostic methods such as probing and radiography are
based primarily on subjective evaluations, which may lead to large diagnostic variations among
different examiners. These methods have qualitative, limited value for monitoring lesion
development and evaluating the effectiveness of preventive procedures. The ideal diagnostic
method should be safe for both patients and users, enabling lesion detection at an early stage.
Also it should be objective and quantitative, non invasive and cost effective and the
measurements observed with this method should be reproduciable (2).
One of the most popular methods for caries detection today is the laser fluorescence based device
DIAGNOdent (Kavo, Biberach, Germany). DIAGNOdent contains a laser diode (655 nm,
modulated, 1 mW peak power) as the excitation light source and a photo diode combined with a
long pass filter (transmission >680 nm) as detector. Fluorescence spectroscopic investigations
revealed considerable contrast between sound and carious tooth tissues when excited by red light,
655 nm (3), fluorescence was found to be more intense in carious tissue than sound tooth tissue.
Due to the good reproducibility, the device should be useful for longitudinal monitoring of the
carious process and thus for assessing the outcome of preventive interventions (4).
Technology development has not only resulted in caries detection methods improvement, but also
in non-invasive, painless caries treatment methods. One of the most popular is caries treatment
with ozone. Ozone has been used for many years in medicine, and within recent years in dentistry
in the management of occlusal and plain surface caries. These are only two of the clinical
problems for which ozone can, and has, been used in dentistry (5).
Ozone is a very powerful antimicrobial agent, and commercially is available as HealOzone
(CuroOzone, USA and KaVo, Germany). Recently, Baysan et al. reported that ozone application
either for 10 or 20 seconds was effective to kill the great majority of microorganisms in carious
lesions (>99% microbial killing after 10 seconds ozone application) (6,7). Prskalo et al. also
confirmed in their study the antimicrobial effect of HealOzone on carious lessions (8).
The HealOzone has been proven to reverse primary root caries in longitudinal clinical trials (9,10).
Bysan and Lynch (9) found that ozone application either for 10 or 20 seconds dramatically
reduced most of the micro-organisms in primary root caries lesions without any side effects.
Holmes (10) concluded that non-cavitated primary root caries can be arrested non-operatively
with ozone and remineralising products. This treatment regime is an effective alternative to
conventional „drilling and filling“.
Baysan and Lynch (11) also reported that this simple and non-invasive technique may benefit
many patients with root caries throughout the world since this approach to treat root caries can
easily be employed in primary care clinics and in the domiciliary treatment of home-bound elderly
people and immobile patients in hospices and hospitals (12).
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The aim of this pilot study was to detect initial carious lesions with laser fluorescence device and
to assess the effect of an ozone delivery system, combined with the daily use of a remineralizing
patient kit, on the initial fissure caries in premolars and molars.
Materials and methods
Study population
Ethical approval was obtained by the Ethics Committe of School of Dental Medicine, University of
Zagreb, Croatia. Each subject had given their informed consent for both dental examinations and
ozone treatment to be undertaken at the Department of Endodontics and Restorative Dentistry,
School of Dental Medicine, Zagreb. A total of 70 lesions in 39 subjects, males and females, aging
from 22 to 60 years, have been selected.
Equipment used
The DIAGNOdent was used to detect and quantify the severity of fissure carious lesions. The
instant reading indicated the real time value that the probe tip was measuring, whilst the peak
value refered to the highest level scanned on the lesion. The peak value was used in statistical
analyses.
HealOzone – ozone delivery system. The HealOzone is a device that takes in air and produces
ozone gas which is then delivered via a hose into a disposable sterile cup at a concentration of
2,100 ppm±10%. The ozone gas is refreshed in this disposable cup at a rate of 615 cc/minute
changing the volume of gas inside the cup over 300 times every second. The cup forms a seal
around the lesion being treated so that ozone cannot leak into the oral cavity.
Protocol treatment
After inspection (Figure 1), tooth cleaning with PROPHYflex and recording of demineralisation
values in fissures of premolars and molars using the DIAGNOdent (Figure 2), treatment with
HealOzone was used as shown in Table 1 (13,14). After treatment with ozone, 5 drops of a
remineralizing solution containing xylitol, fluoride, calcium, phosphate and zinc (HealOzone
remineralizing solution) was applied to the lesion. To each subject instructions were given for
using the HealOzone remineralizing toothpaste twice a day, and the HealOzone mineral mouth
wash once a day. Subjects were recalled at one and two months after the first visit. At each recall
the same protocol was repeated.
Descriptive statistics, t-test and Pearson Correlations were used for data analyses.
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Figure 1 Suspected caries lesion on premolar
Figure 2 Use of DIAGNOdent for determining demineralization value
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118

Table 1 Treatment duration with ozone according to DIAGNOdent values (6)
Results
The results are shown in tables 2, 3, 4. As it can be seen, there was a significant difference
between the first visit and the first recall, the first visit and the second recall, and between the
first and the second recall: t-test revealed a signifficant difference between all three pairs
(combinations) at the level of p

Table 4 Correlation between tested parameters
Discussion
Early diagnosis of primary fissure caries with the laser fluorescence device DIAGNOdent is of great
importance because of the low sensitivity of visual, probing and bitewing examination which leads
to a significant number of teeth with undetected dentinal caries. High reproducibility of
DIAGNOdent has previously been reported under both in vitro and in vivo conditions (15,16).
Thus, this method is suitable for longitudinal monitoring of carious lesions and also enables
preventive interventions. Longitudinal monitoring of remineralization of demineralized lesions is of
great important in the case of ozone therapy.
The goal of treatment of carious lesions with ozone is to reduce the causative microbiota and
contributing risk factors to halt the caries decay process and stimulate remineralization. Research
has shown that ozone breaks up acid products of cariogenic bacteria, which may be important in
the aetiology of the developing carious lesion (14). Research by Baysan and others (5) has shown
that an application of ozone for a period of 10 seconds was capable of reducing the Streptococcus
Mutans and Streptococcus Sobrinus in vitro. Further research by OT Abu-Salem (17) have shown
that occlusal caries in deciduous teeth can be effectively controlled with ozone. The main problem
of non-invasive pharmaceutical approaches in clinical practice for management of caries reversal
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and remineralization of the lesion is the difficulty in suppressing or eliminating micro-organisms
for long periods of time required for remineralization.
Ozone enables the shifting of microbial flora from acidogenic and aciduric micro-organisms to
normal oral commensals which will allow the remineralization to occur within the carious process
(6). Ozon has the ability to remove proteins in carious lesions, and to enable calcium and
phosphate ions to diffuse through the lesions, which results in remineralization of the lesion after
ozone application.
Ozone is extensively used in general practice followed by sealing of the carious lesions.
Fortunately this ozone treatment has been proven not to be detrimental to the bond. Ozone did
not influence the studied enamel physical properties and neither enhanced nor harmed the sealing
ability (18). Thus, adhesive restoration placement should be possible immediately after ozone
application for cavity disinfection (19).
In this study, there were highly significant decreases in DIAGNOdent values between the first
baseline visit and the first/second recall. After the initial elimination of microorganisms,
decolonisation of microorganisms may be reinforced by lack of available organic substrate
essential for the metabolism of cariogenic bacteria (20). Within the limitations of this study it can
be concluded that the use of ozone in primary fissure caries not only preserves the tooth structure
by avoiding the use of invasive techniques for the removal of carious tissue, but also promotes the
caries reversal and tooth remineralization.
Acknowledgments
This study was supported by Ministry of Science, Education and Sports, Grant No. 0065007,
Zagreb, Croatia.
References
1. Featherstone JD. The caries balance: the basis for caries management by risk assessment. Oral
Health Prev Dent. 2004;2 Suppl 1:259-64.:. ABSTRACT :.
2. Heinrich-Weltzien R, Weerheijm KL, Kuhnisch J, Oehme T, Stosser L. Clinical evaluation of
visual, radiographic, and laser fluorescence methods for detection of occlusal caries. ASDC J Dent
Child. 2002;69(2):127-32, 123.:. ABSTRACT :.
3. Hibst R, Paulus R, Lussi A. Detection of occlusal caries by laser fluorescence. Basic and clinical
investigations. Med Laser Appl. 2001;16:205–13.
4. Astvaldsdottir A, Holbrook WP, Tranaeus S. Consistency of DIAGNOdent instruments for clinical
assessment of fissure caries. Acta Odontol Scand. 2004;62(4):193-8.:. ABSTRACT :.
5. Baysan A, Lynch E. The use of ozone in dentistry and medicine. Prim Dent Care. 2005;12(2):
47-52.:. ABSTRACT :.
dott Cosimo Cigliola
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6. Baysan A, Whiley RA, Lynch E. Antimicrobial effect of a novel ozone- generating device on
micro-organisms associated with primary root carious lesions in vitro. Caries Res. 2000;34(6):
498-501.:. ABSTRACT :.
7. Baysan A, Lynch E. Management of root caries using ozone in vivo. J Dent Res. 2001;
80:37.
8. Prskalo K, Perinić M, Šutalo J, Kalenić S, Tarle Z, Knežević A. Antimicrobial effect of HealOzone
in class I caries lesions. J Dent Res. 2005; 63.
9. Baysan A, Lynch E. Effect of ozone on the oral microbiota and clinical severity of primary root
caries. Am J Dent. 2004;17(1):56-60.:. ABSTRACT :.
10. Holmes J. Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial. Gerodontology. 2003;20(2):106-14.:. ABSTRACT :.
11. Baysan A, Lynch E. The use of ozone in dentistry and medicine. Part 2. Ozone and root caries.
Prim Dent Care. 2006;13(1):37-41.:. ABSTRACT :.
12. Huth KC, Paschos E, Brand K, Hickel R. Effect of ozone on non-cavitated fissure carious lesions
in permanent molars. A controlled prospective clinical study. Am J Dent. 2005;18(4):223-8.:.
ABSTRACT :.
13. Holmes J, Lynch E. News From the HealOzone Users Congress & The HealOzone User Group
[document on the Internet]. Yorkshire: The First HealOzone User Group Congress Meetings; 2003
[cited 2006 April 3]. Available from: http://www.the-o-zone.cc/docs/TxO3A01.pdf
14. Lynch E. Ozone: The revolution in dentistry. Surrey: Quintessence Publishing, 2004.
15. Knezevic A, Tarle Z, Negovetic Mandic V Brzovic V, Panduric V, Prskalo K. Use of DIAGNOdent
and HealOzone for detecting and treatment of fissure initial caries lesions. Ital J Oper Dent. 2006;
Abstr. R33., 259.
16. Lussi A, Hibst R, Paulus R. DIAGNOdent: an optical method for caries detection. J Dent Res.
2004;83:C80-3.:. ABSTRACT :.
17. Abu Salem OT, Marashed MM, Lynch E. Immediate effect of ozone on occlusal caries of
primary teeth. J Dent Res. 2002;82:C-535.
18. Celiberti P, Pazera P, Lussi A. The impact of ozone treatment on enamel physical properties.
Am J Dent. 2006;19(1):67-72.:. ABSTRACT :.
19. Schmidlin PR, Zimmermann J, Bindl A. Effect of ozone on enamel and dentin bond strength. J
Adhes Dent. 2005;7(1):29-32.:. ABSTRACT :.
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20. Inaba D, Ruben J, Takagi O, Arends J. Effect of sodium hypochlorite treatment on
remineralization of human root dentine in vitro. Caries Res. 1996;30(3):218-24.:. ABSTRACT :.
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Gerodontology
Volume 20 Issue 2 Page 106 - December 2003.
Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial
• J Holmes 11
UKSmiles Dental Practice, Wokingham, Berkshire, UK
•
1 UKSmiles Dental Practice, Wokingham, Berkshire, UK
Address for correspondence: Dr Julian Holmes, BDS UKSmiles Dental Practice, 3 Old Row Court,
Rose Street,Wokingham, Berkshire, RG40 1XZ, United Kingdom Tel: +44 1189795559 e-mail:
julian@o3dc.co.uk
Abstract
Objective: To assess the effect of an ozone delivery system, combined with the daily use of a
remineralising patient kit, on the clinical severity of non-cavitated leathery primary root carious
lesions (PRCL's), in an older population group.
Design: Atotal of 89 subjects, (age range 60–82, mean ± SD, 70.8 ± 6 years), each with two
leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for
treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were
recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft,
leathery or hard, scored with a validated root caries severity index.
Results: There were no observed adverse events. After three months, in the ozone-treated
group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group,
four PRCL's (4%) had become worse (p

This article is cited by:
This article is cited by the following articles in Blackwell Synergy and CrossRef
• J Holmes. (2006) Missing the point. British Dental Journal 200:6, 305
• Dorothy McComb. (2005) No reliable evidence that ozone gas stops or reverses tooth decay.
Evidence-Based Dentistry 6:2, 34
• D Richards. (2005) Ozone revolution? Or emperor's new clothes?. British Dental
Journal 199:8, 485
Gerodontology
Volume 20 Issue 2 Page 106 - December 2003
To cite this article:
J Holmes (2003) Clinical reversal of root caries using ozone, double-blind, randomised, controlled
18-month rial Gerodontology 20 (2), 106–114. doi:10.1111/j.1741-2358.2003.00106.x
Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial
• J Holmes 11
UKSmiles Dental Practice, Wokingham, Berkshire, UK
•
1 UKSmiles Dental Practice, Wokingham, Berkshire, UK
Address for correspondence: Dr Julian Holmes, BDS UKSmiles Dental Practice, 3 Old Row Court,
Rose Street,
Wokingham, Berkshire, RG40 1XZ, United Kingdom Tel: +44 1189795559 e-mail:
julian@o3dc.co.uk
Abstract
Objective: To assess the effect of an ozone delivery system, combined with the daily use of a
remineralising patient kit, on the clinical severity of non-cavitated leathery primary root carious
lesions (PRCL's), in an older population group.
Design: Atotal of 89 subjects, (age range 60–82, mean ± SD, 70.8 ± 6 years), each with two
leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for
treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were
recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft,
leathery or hard, scored with a validated root caries severity index.
Results: There were no observed adverse events. After three months, in the ozone-treated
group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group,
four PRCL's (4%) had become worse (p

and 18 months, 87 Subjects attended. In the ozone group at 12 months, two PRCL's remained
leathery, compared to 85 (98%) that had hardened, whilst in the control group 21 (24%) of the
PRCL's had progressed from leathery to soft, i.e. became worse, 65 PRCL's (75%) were still
leathery, and one remained hard (p

Design: Atotal of 89 subjects, (age range 60–82, mean ± SD, 70.8 ± 6 years), each with two
leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for
treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were
recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft,
leathery or hard, scored with a validated root caries severity index.
Results: There were no observed adverse events. After three months, in the ozone-treated
group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group,
four PRCL's (4%) had become worse (p

Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial
• J Holmes 11
UKSmiles Dental Practice, Wokingham, Berkshire, UK
•
1 UKSmiles Dental Practice, Wokingham, Berkshire, UK
Address for correspondence: Dr Julian Holmes, BDS UKSmiles Dental Practice, 3 Old Row Court,
Rose Street, Wokingham, Berkshire, RG40 1XZ, United Kingdom Tel: +44 1189795559 e-mail:
julian@o3dc.co.uk
Abstract
Objective: To assess the effect of an ozone delivery system, combined with the daily use of a
remineralising patient kit, on the clinical severity of non-cavitated leathery primary root carious
lesions (PRCL's), in an older population group.
Design: Atotal of 89 subjects, (age range 60–82, mean ± SD, 70.8 ± 6 years), each with two
leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for
treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were
recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft,
leathery or hard, scored with a validated root caries severity index.
Results: There were no observed adverse events. After three months, in the ozone-treated
group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group,
four PRCL's (4%) had become worse (p

• D Richards. (2005) Ozone revolution? Or emperor's new clothes?. British Dental
Journal 199:8, 485
CrossRef
Miracle Ozone Dentistry. A New Anti-Decay Dental Practice
By Dr Julian Holmes 1-31-7
Let me paint you a picture. Every Monday morning, dental practices over the world power up for a
week's worth of drilling and filling, tooth removal, and reconstruction work. Queues of fearful
patients, tearful children being comforted by anxious parents, people in considerable pain, line up
outside the doors to their dental surgeries. Once inside, these patients are subjected to the
traditional smells of oil of cloves, disinfectants, and the noises of the high pitch whine of the
dental drill; the odd scream or two filters through the hushed, usually silent waiting room into the
street outside. It is a sad fact of life that every dentist is trained that if there is an area of decay
in your tooth, the only way to treat this is to drill the decay out or amputate it, and then place a
filling that will have to be replaced at some stage.
For a small minority of patients, where their dental practices have chosen to invest in a new
technology, the opposite happens almost every day. The queue is one of bright, cheerful adults
and children; there are few smells to associate this practice with the traditional one down the
road. The noise of the drill is seldom heard, and happy smiling faces emerge from the treatment
room.
Since 1998 researchers, lead by Professor Edward Lynch from Queen's Dental Hospital and Belfast
University, Ireland, have opened a radically and revolutionary way forward. The dental profession
no longer has to destroy tooth tissue to eliminate bacteria. A simple 60 second (average
treatment time) treatment with a device that delivers a burst of ozone will destroy all the bacteria
that caused the infection and the decay. It destroys all the organic effluents that are produced by
these bacteria; this is shown in Fig 1 opposite. The top data shows the spread of organic bio-
molecules produced by an active carious lesion. The lower data shows the effect of 10 seconds of
ozone; the profile now consists of a single main spike of acetate acid, and the other oxidation by-
product is carbon dioxide. This H MNR research proved that ozone worked as a pharmaceutical
approach to caries in-vitro. By effectively sterilising the lesion, minerals from the patients own
saliva will re-enter the areas of mineral loss to harden them. Once hardened, it is more resistant
to future bacterial attack and mineral loss.
There is in all our mouths a natural balance. Your tooth surface losses minerals into your saliva at
certain times, usually just after you start to eat. These acidic conditions favour mineral loss. The
normal acid/alkaline balance exists in the oral cavity. When bacteria attach themselves to a tooth
surface, they set up a complex community of some 450 different bacterial types over a period of
time. If patients skimp on using a tooth brush, or forget to use dental floss on a regular basis,
these communities of bacteria evolve into one which produces large volumes of acids. These acids
attack the tooth surface, dissolving out the minerals, leaving a hole or cavity. As this cavity now
dott Cosimo Cigliola
129

gives the bacterial colonies a degree of protection, tooth brushing cannot remove the bacteria and
so the process of decay accelerates.
Yet decay is only an infection process that leads to the softening of the tooth, and the formation of
a cavity, so could this infection be treated with, say, antibiotics? The bacteria that cause tooth
decay are often found deep within the structure of the tooth, so their removal has to date only
been by amputating the infected part of the tooth. Antibiotics and other pharmaceutical agents
cannot penetrate deep enough through bacterial pellicle and tooth structure to eliminate acid
niche environment. This teaching and technique is based on sound engineering principles that
originate from the Victorians! Despite modern advances, there is no simple test that can be
applied to a cavity to tell the dentist if they have removed all the infection! or enough tooth
material! And if areas of infection are left behind, there is a good chance that the filling placed will
fail at some time in the future.
In an attempt to prevent further infection and to restore the tooth to its original shape and
function, a filling is then placed. Studies over the years have shown that fillings do not last very
long, any where from 6 months to several years. But once a tooth has had part of it amputated,
there is no going back. For each time the filling needs to be removed and replaced, there is a little
less of the original tooth left and a larger filling. A point is reached where there is no option but to
opt for expensive reconstruction work with advanced dental care or have the tooth removed.
The dental professions goal is to help and educate the patient, how to avoid them entering into
this cycle of tissue amputation and periodic filling replacement. Oral care education and modern
toothpastes have helped reduce the number of cavities, but in poorer communities, those with
disabilities and in long term institutions, as well as our aging population, decay is still prevalent.
And diet advice is often lacking, so although patients may think they are doing well at home, the
process of decay continues! The days of 'Wait & Watch' are over the profession is not entirely
sure of what it is watching unless clinicians are using advanced clinical diagnostic tools.
Dental Ozone is a completely new way to look at decay. In the early stages of mineral loss, no use
of the drill is required. The treatment is simple, inexpensive (certainly less than the cost of a
filling), and requires no injection of anaesthetics. This does of course depend on the use of
modern diagnostic equipment, such as the DIAGNOdent (KaVo GmbH). This laser is more accurate
than x-rays, and far superior to the traditional mirror and probe that dentists often use. The
mirror and probe are tools that can find holes, not diagnose areas of first stage decay. And
research has shown that x-rays are very poor to visualise decay in a tooth surface, until it is 2-3
mm inside the inner layer of the tooth.
If the area of decay is deeper, and more extensive, Ozone still has a role to play. The dental drill
may need to be used to remove the cover of enamel over the decay, but this can also be carried
out with air abrasion. No local anaesthetics are required, and Ozone is used to sterilise the area of
dott Cosimo Cigliola
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decay, without the need to amputate a large volume of tooth structure. In this way, damage to
the tooth is limited, and the inherent strength is preserved. Even in really deep areas of decay,
Ozone can be used to preserve tooth tissue.
When Ozone is combined with traditional care, then there are a number of advantages that
patients and the dental profession can make use of. Ozone can be used to sterilise a cavity before
a filling is placed, so there will be virtually no sensitivity after the local anaesthetic wears away. It
can be used to eliminate sensitivity after new crowns or veneers are placed. And where wear
facets have produced sensitive areas at the necks of teeth, in most cases a simple 40 second
application with ozone can eliminate this sensitivity. There are many more applications for Ozone
in a general dental or medical practice, and the studies for some of these can be found on
www.the-o-zone.cc. This www site not only allows you to find a dental practice which has and
uses Ozone, but also allows you to view the research papers that have been published from
centres around the world.
There is not a single facet of a medical, dental or veterinary practice where ozone cannot be used
in some form for the benefit of patients from the general to specialist practice and health care
centre all can take advantage of this new technology.
So, perhaps a Monday morning at a dental practice that has invested in modern technology is no
longer the stressful, painful and anxious visit that it used to be. The waiting room in this practice
are full of smiles, people chatting to the dental team members as they wait with happy
anticipation at being called through for their turn in the treatment room. Mums and dads with
children have no fear or anxiety, as they are reassured that modern technology has opened a door
for them, that most of the older population had never believed possible. For both the patient and
the dental practice, it is a technology that has a winning solution for both; the treatment is fast, it
is predictable, it is painless, and also reduces the long-term cost of the treated tooth. For the
dental practice, the treatment times are reduced, it is profitable, and the treatment less stressful.
For the patient, modern technology has allowed them to have a 21st century treatment, rather
than one that is old fashioned, and out-dated.
In a lead article published in 2003 I wrote;
'In the vast majority of dental practices - not just in the United Kingdom and Europe, but
throughout the world - the primary method to reverse the effects of decay remains 'drill and fill'.
The entry of a patient into the cycle of drill and fill is irreversible. Once a hole is drilled into a
tooth, the patient always will have it; and no matter how good a clinician each dentist perceives
themselves to be, any restorative material will fail at some time.'
The dental profession has not kept up with their medical counterparts in finding a holistic
treatment method for what is a prevalent infection caries. The Victorian principles of amputation
to establish a sound foundation for restorative care no longer are valid in the light of new
published research and clinical experience.
dott Cosimo Cigliola
131

Ozone at last offers the dental profession this new approach instead of the out-dated and Victorian
amputational model.
Dr Julian Holmes, 2007.
The Ozi-cure is a self-contained device to generate ozone gas from air for the use in Dental,
Medical or Veterinary Practice for surface application.
The Ozi-cure Dental Unit manufactured by O3 and distributed in Germany by American Dental
GmbH. The Ozi-cure achieved CE and Medical Directive Certificates in November 2006.
The treatment times used in dental ozone treatment are short from 10 to 60 seconds (Baysan and
Lynch, 2001). The area to be treated and the application of ozone is carefully controlled by
localised 'bagging' and high-volume suction.
The use of ozone in dental practice takes dental care into the 21st Century. It sets the standard of
a modern pharmaceutical method to treat dental and medical patients;
Studies from Europe (Abu-Salem et al, 2003; Baysan and Lynch 2001; Holmes, 2003; Holmes and
Lynch, 2003) have shown conclusively that the use of ozone in dental care is effective as a non-
destructive method to manage decay and its destructive effects. The use of ozone has been shown
to be the ideal way to manage anxiety of patients young and old - and their carers (Dahnhardt et
al, 2003; Domingo et al, 2004).
The effects of ozone reduce tooth destruction in routine preparation (Clifford, 2004; Holmes,
2004; Holmes and Lynch, 2004) and ozone reduces the time and the cost of dental care (Domingo
and Holmes, 2004; Johnson et al, 2003) and raises the practice income. In Endodontics, ozone is
effective against Enterococcus faecalis (Chang et al, 2003).
Professor Velio Bocci from Milan University (1994) has emphasised that the potential toxicity of O3
should not preclude its employment for medical, dental & veterinary purposes. This statement has
been echoed by thousands of health professionals who use ozone in clinical practices around the
world, and millions of patients that have been treated.
The correct operation of the Ozi-cure meets all current Health and Safety Regulations in all
countries. The Ozi-cure is completely safe when used according to the Ozi-cure Operating
Instructions.
Ozone is perceived to be a dangerous gas; let's put this into a scientific context. In 1978 an FDA
Report showed that 1.5 million people were hospitalised by pharmaceutical reactions, and there
were 140,000 deaths from prescription drug usage.
In stark contrast, a 1980 German Medical Society Report for Ozone Therapy cited 5.6 million
ozone treatments carried out for that year. Of the 5.6 million ozone treatments, there were just
40 reported cases of side effects (0.000007%), and 4 deaths from inappropriate administration of
ozone gas. Ozone remains the safest and effective pharmaceutical treatment.
In World War I, ozone was used to treat wounds, burns and infections. The modern development
of ozone's application to Medicine began in the 1950s in Europe, Australia, Israel, Cuba, Brazil and
Columbia. Today, over 9000 doctors, dentists & vets worldwide now routinely use ozone in their
clinical management.
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Research in Cuba, Europe, the USA and South Africa concerning the anti-microbial efficacy of
ozone has continued over the last twenty years and has conclusively shown the ability of both
gaseous and dissolved ozone to eradicate a wide range of bacteria, bacterial spores and viruses
(Baysan and Lynch, 2001; Ishizaki, 1986; Katzeneleson, 1974; Vaughan, 1987; Whistler and
Sheldon, 1989).
A clinical guide for the use of ozone in dental and medical practice is included with every Ozi-cure
device sold.
References;
Abu-Salem OT, Marashdeh MM, Lynch E: Ozone Efficacy in Treatment of Occlusal Caries in
Primary Teeth. IADR Abstract 2003
Baysan A and Lynch E: Management of root caries using ozone in-vivo. Journal of Dental Research
2001; 80:37
Bocci V: Autohaemotherapy after treatment of blood with ozone, a reappraisal. Int Med Res 1994;
22: 131-144.
Chang H, Fulton C, Lynch E: Antimicrobial Efficacy of Ozone on Enterococcus faecalis. IADR
Abstract 2003.
Clifford C: Reversal of Caries Using Airbrasion and Ozone- Nine Month Results. IADR Abstract
2004
Dahnhardt JE, Jaeggi T, Scheidegger N, Kellerhoff N, Francescut P, Lussi A: Treating Caries in
Anxious Children with Ozone: Parents' Attitudes after the First Session. IADR Abstract 2003
Domingo H, Abu-Naba'a L, Al Shorman H, Holmes J, Marshdeh MM, Abu-Salem AT, Freeman R,
Lynch E: Reducing Barriers to Care in Patients Managed with Ozone. IADR Abstract 2004.
Domingo H and Holmes J: Reduction in treatment time with combined air abrasion and ozone
compared to traditional 'Drill & Fill'. IADR abstract 2004.
Holmes J: Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-
month trial. Gerodontol 2003: 20 (2): 106-14.
Holmes J: Restoration of ART and Ozone treated primary root carious lesions. IADR Abstract 2004.
Holmes J and Lynch E: Arresting Occlusal Fissure Caries Using Ozone. IADR Abstract 2003.
Holmes J and Lynch E: Reversal of Occlusal Caries using Air Abrasion, Ozone, and Sealing. IADR
Abstract 2004
Ishizaki K: Inactivation of bacillus spores by gaseous ozone. J Applied Bacteriol 1986; 60: 67-72.
Johnson N, Johnson J, Lynch E: Cost Benefit Assessment of a Novel Ozone Delivery System vs.
Conventional Treatment. IADR Abstract 2003.
Katzeneleson E: Inactivation of viruses and bacteria by ozone. In chemistry of water supply,
treatment and disinfection. Ann Arbor Science Publishers Inc., Ann Arbor, Mich, 1974.
: J Adhes Dent. 2005 Spring;7(1):29-32
Effect of ozone on enamel and dentin bond strength.
• Schmidlin PR,
Zimmermann J, Bindl A.
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Clinic for Preventive Dentistry, Periodontology and Cariology, Center for Dental Medicine,
University of Zurich, Switzerland. patrick.schmidlin@zzmk.unizh.ch
PURPOSE: To evaluate the influence of direct high-dose gaseous ozone application (2100 ppm)
on dentin and enamel shear bond strength.
MATERIALS AND METHODS: Ten bovine enamel and dentin samples per group were pretreated
as follows: (I) ozone application (Healozone, KaVo) for 60 s alone or (II) with subsequent
application of a fluoride- and xylitol-containing antioxidant (liquid reductant), (III) light-activated
bleaching with 35% hydrogen peroxide for 5 min serving as negative control (Hi-Lite, Shofu), and
(IV) untreated enamel and dentin (positive control). Specimens were bonded with a functional 3-
step adhesive system (Syntac Classic, Ivoclar Vivadent) and restored with a composite (Tetric
Ceram, Ivoclar Vivadent) according to the Ultradent method. After storage in water at 37 degrees
C for 24 h, shear bond strength was measured using a Zwick universal testing machine. Data
were analyzed using ANOVA and Scheffe's post hoc analysis. Results: In concordance with the
existing literature, bleaching resulted in significantly decreased bond strength (p < 0.05) on
enamel specimens. No decrease in shear bond strength was detected for ozone-pretreated
specimens compared to untreated controls.
CONCLUSION: Despite a possible retention of surface and subsurface oxide-related substances
during high-dose ozone application, shear bond strength was not impaired. Thus, adhesive
restoration placement should be possible immediately after ozone application for cavity
disinfection.
PMID: 15892361 [PubMed - indexed for MEDLINE]
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International Association for Dental Research IADR 81st General
Session 2nd Meeting of the Pan European Federation
Göteborg, Sweden , June 25-28, 2003
Abstracts
2755 Comparison of Conventional Treatment vs. Ozone for Occlusal Caries with Ozone
Therapy
N. JOHNSON1, J. JOHNSON1, H. DOMINGO2, and E. LYNCH2,
1 Private practice,Wales, United Kingdom, 2 Queen's University Belfast, United Kingdom
Traditional restorative treatment for a carious lesion involves several invasive steps. The
treatment of a primary carious lesion by means of ozone does not require any invasive treatment,
simply the isolation of the tooth and the application of the ozone via the novel ozone delivery
system1.
Objectives: This study aimed to assess the differences in time expenditure in providing
treatment with ozone compared with traditional ‘drill and fill’ methods by measuring the actual
time taken to deliver both treatment modalities.
Method: 40 adult patients judged to have an early primary fissure carious lesion were randomly
divided into two equal groups. The first group had traditional ‘drill and fill’ treatment; the second
group was treated by the application of ozone. Both groups were treated by the same dentist and
dental nurse. Each patient’s treatment was timed, in minutes, from the patient sitting in the
dental chair until the completion of the procedure.
Results: The conventionally treated (i.e. ‘drill and fill’) patients averaged 35 minutes per patient
with a SD of 10 minutes whilst the group treated by therapeutic ozone averaged 8 minutes with a
SD of 2 (P

2408 12-month Assessment of Ozone on Root Caries
A. BAYSAN, Birmingham University, Dental School, United Kingdom, and E. LYNCH,
Queen's University, Belfast, United Kingdom
Objectives: The objective was to assess the safety and efficacy of a novel ozone delivery
system1 with or without a root sealant2 for the management of root caries.
Methods: A total of 79 patients with 220 PRCLs were recruited. Group 1 had only ozone
treatment, Group 2 had neither ozone nor root sealant, whilst Group 3 had both ozone and
sealant and Group 4 had only sealant. At baseline, and 1, 3, 6, 9 and 12 months, the Electrical
Caries Monitor3(ECM)and DIAGNOdent4 were employed. Lesions were clinically assessed. Modified
USPHS criteria were also performed.
Results: After 12 months, 74 patients completed. There were no observed adverse events. At 12
months, 47% of PRCLs reversed from severity index 1 to 0 (i.e., hard) in the ozone only group,
whilst none of the lesions became hard in the control group (p < 0.001) and 52% of lesions
reversed from severity index 2 to 1 in the ozone group compared to only 11.6% in the control
group (p < 0.001). Modified USPHS criteria revealed that there were 61% of intact sealants in the
ozone and sealant group and 26.1% of intact sealants in the sealant only group (p < 0.05). After
1, 3, 6, 9 and 12 months, the ECM and DIAGNOdent readings showed improvements in the ozone
only group when compared to the control group (p < 0.001). The ozone and sealant group also
had greater improvements in the ECM and DIAGNOdent values when compared to the sealant only
group (p < 0.05).
Conclusions: Leathery root caries can be treated non-operatively with ozone. This novel
treatment regime using ozone is an effective alternative to conventional "drilling and filling". The
sealant was retained better on ozone treated lesions. 1HealOzone unit, CurOzone USA,2Seal and
Protect, Dentsply, Germany,3LODE Diagnostics, Netherlands, 4Kavo,Germany
Seq #247 - Oral Function, Caries and Health Education (Morita Junior Investigator Award)
9:00 AM-11:00 AM, Saturday, 28 June 2003 Svenska Massan F5
Back to the Geriatric Oral Research Program Back to the 81st General Session of the International
Association for Dental Research (June 25-28, 2003)
2008 Effect of Ozone on Surface Hardness of Restorative Materials
D. CAMPBELL1, D.L. HUSSEY2, L. CUNNINGHAM2, and E. LYNCH2,
1 Royal Group of Hospitals, Belfast, United Kingdom, 2 Queen's University, Belfast, UK
Objective: There has been much recent interest in the use of ozone as an interceptive treatment
in dentistry. The residual effect of such treatment on the surface hardness of restorative materials
has not been reported. The aim of this study was to investigate the effect of ozone treatment on
the surface hardness of a range of commonly used restorative materials.
Method: Six prefabricated hardness testing disks containing 3 wells were each filled with
commonly used restorative materials: hybrid composite (Spectrum), glass ionomer (Chemflex),
compomer (Dyract AP), amalgam (Disperalloy), flowable composite (Revolution) and resin
dott Cosimo Cigliola
136

modified glass ionomer (Fuji II LC). The materials were packed against a glass slab to produce a
flat surface. 24 hours later the specimens (18 of each) were subjected to Micro Vickers Hardness
Testing (Mitutoyo) using a load of 1Kg for 10 seconds. Ozone was then applied for 10 seconds
(Healozone, Curozone, USA and Kavo, Germany) and the surface hardness test was repeated.
Results: The Vickers numbers obtained for the samples was as follows:
Material Mean surface hardness ± s.d. Mean surface hardness after
dott Cosimo Cigliola
Ozone treatment ± s.d.
Composito ibrido 46.82± 2.82 46.33 ± 3.07
Vetro-ionomerico 38.40 ± 10.85 37.02 ± 4.75
Compomero 27.32 ± 2.77 27.64 ± 2.93
Amalgama 127.13 ± 15.65 124.30 ± 14.02
Composito fluido 14.59 ± 2.11 12.61 ± 1.41
Resina modificata
G.I.
32.74 ± 3.18 30.06 ± 2.14
137

Statistical analysis using a 2-way ANOVA did not reveal any difference in surface hardness
following the treatment with Ozone (p >0.15)
Conclusion: It is concluded that the application of Ozone for 10 seconds has no detrimental effect
on the surface hardness of the materials tested and should therefore not affect the clinical
performance of such materials in-vivo.
Seq #209 - Polymer Materials: Properties, Clinical Variables
3:45 PM-5:00 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
2034 Treating Caries in Anxious Children with Ozone: Parents' Attitudes after the First
Session
J.E. DÄHNHARDT, T. JAEGGI, N. SCHEIDEGGER, N. KELLERHOFF, P.
FRANCESCUT, and A. LUSSI,
University of Bern, Switzerland Treating anxious children remains a challenge in pediatric
dentistry. Many times the only options are the treatment with sedatives or in general anesthesia
which includes high cost,
certain risks and a child that will stay anxious.
Objective: The goal was to evaluate parents’ attitudes towards the treatment of caries with
ozone (Heal Ozone Unit, CurOzone, U.S.A.) in anxious children.
Methods: After the first treatment of 20 children from age 2-10 years (which included brushing
the teeth, hand excavation (Neos Vanadium Excavator No. 591/3, Hawe Neos Dental,
Switzerland), DIAGNOdent measurement (KaVo, Germany) and ozone application), the father
and/or mother was questioned about their attitudes toward the ozone treatment.
Results: All parents answered the questionnaire but most children were accompanied by their
mothers only (90%). Seventy-five per cent of the children were afraid of going to the dentist
according to their parents prior to the ozone treatment but this group lost some of their fear and
all children were happy to come back to the next appointment. Forty per cent of the parents were
afraid that the ozone treatment could be hazardous for their child but all of them were happy that
they started the ozone treatment. After the first session 75% of the parents would recommend
the ozone treatment to family or friends and would agree to have an ozone treatment again.
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138

Eighty per cent of the parents were willing to pay more for the ozone treatment than for
conventional „drilling and filling“.
Conclusions: After the first session, the treatment with ozone seems to be well accepted by
children and by parents and decreases anxiety in children.
Seq #211 - Fear, Anxiety, and Behavior
3:45 PM-5:00 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
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(June 25-28, 2003)
0989 Ozone Efficacy in Treatment of Occlusal Caries in Primary Teeth
O. ABU-SALEM, M. MARASHDEH, and E. LYNCH,
Queen's University, Belfast, United Kingdom
Ozone is now being investigated as a pharmaceutical treatment for occlusal caries in permanent
and primary teeth.
Aims : To assess and monitor the progression, stabilization or reversal of occlusal carious lesions
using DIAGNOdent, Electrical Caries Monitor (ECM) standard scale readings, clinical classification
(Ekstrand, 1998), Stained enamel (mm), Colour index, Texture index, Frosted enamel and
perceived treatment need index after Ozone treatment in primary teeth.
Methods : In vivo study where 16 patients with 42 non-cavitated occlusal carious lesions were
reviewed at the 6- month stage. At baseline, three months and six months All teeth were cleaned
by an air-abrasive system called Prophyflex 2® (KaVo, Germany). Then, DIAGNOdent ® (D)
(KaVo, Germany), ECM (LODE BV, Netherlands) standard scale readings and clinical classification
(Ekstrand, 1998) were taken. Half of the lesions were treated with Ozone at each of these three
visits, using a silicone-sealed cup via a hand piece (HealOzone unit, CurOzone USA and KaVo,
Germany) for 10 seconds, whereas, the other half of the lesions was reserved as a control group.
Results : At six months ECM readings were improved significantly (p

1671 Antimicrobial Efficacy of Ozone on Enterococcus faecalis
H.H. CHANG, C. FULTON, and E. LYNCH,
Queen's University of Belfast, United Kingdom
Aim: To investigate the bactericidal efficacy of Ozone on different concentrations of Enteroccocus
faecalis
Methods: Enterococcus faecalis was cultured overnight on blood agar plates. A suspension of the
micro-organisms was made up to an approximate concentration 108 using McFarland Standards in
phosphate buffered saline (PBS). Serial dilutions to 107, 106, 105 were made. From these 50 µl of
each concentrate was pipetted into each well of a microtitre plate. Each well of concentrate was
treated in turn with Ozone (HealOzone Unit, CurOzone, USA and KAVO, Germany) at different
exposure times of 10s, 20s, 30s and 60s. Samples were taken from the wells, spiral plated,
incubated and enumeration of the colony forming units(cfu) performed.
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140

Results:
Exp Time ? 60s 30s 20s 10s Trattamento
Concentrazione?
10 8 cfu 2 x 103cfu 4.0 x
105cfu
2.0 x
107cfu
10 7 cfu 0 cfu 0 cfu 3.4 x
105cfu
5.5 x
107cfu
4.4 x
105cfu
in no-PBS
6.0 x 108cfu
5.6 x 107cfu
10 6 cfu 0 cfu 0 cfu 0 cfu 0 cfu 6.5 x 106cfu
10 5 cfu 0 cfu 0 cfu 0 cfu 0 cfu 5.2 x 105cfu
Exp Time=time exposed to Ozone PBS = phosphate buffered saline
Conc.= approximate concentration of the bacterial suspension
Conclusions: Ozone kills all Enterococcous faecalis when its concentration in suspension is 106
and lower, even at exposure times of only 10 seconds.
Seq #174 - Oral Microbiology & Immunology I
11:00 AM-12:15 PM, Friday, 27 June 2003 Svenska Massan Exhibition Hall B
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(June 25-28, 2003)
2753 Efficacy of Ozone to Reverse Occlusal Caries
R. MORRISON, Harbour Dental Practice, Donaghadee, United Kingdom, and E. LYNCH, Queen's
University Belfast, United Kingdom
Ozone has been shown to clinically reverse primary root carious lesions and early occlusal pit and
fissure caries.
Objectives: This study assessed the effect of a novel ozone delivery system1 on primary pit and
fissure carious lesions over a 13-week period in a general dental practice.
Methods : subjects were randomly allocated to one of 2 groups. 145 patients were entered with
240 test carious lesions whilst another 60 subjects were entered with 60 control lesions, which did
not receive any treatment. Each carious lesion had been deemed to require conventional drilling
and filling. Ozone was applied to each test lesion for 40 seconds. After 13 weeks, patients were
recalled and the lesions were clinically re-assessed for severity.
Results: 89 test subjects wit h 141 test lesions have to date attended the recall visit. There were
no observed adverse events. 123 of the ozone treated primary pit and fissure carious lesions had
dott Cosimo Cigliola
141

clinically reversed based on the clinical measurement of lesion severity whilst the other 18 lesions
remained stable and none became worse (P

(June 25-28, 2003)
2069 Patients' Attitudes toward and Satisfaction with Managing Caries with Ozone as a
Routine Treatment in Dental Private Practice
G.D. MEGIGHIAN, and M.V. DAL VERA, Private Practice, Verona, Italy
Ozone with the HealOzone (KaVo, Germany) is currently being used as an alternative treatment of
dental decay to local anaesthesia, drilling and filling.
Objectives: To assess the patients' attitudes to the treatment with ozone in a private practice
environment.
Methods : Patients (n=250) who had a carious lesion treated with conventional minimally
invasive drilling and/or air abrasion and filling within the previous 6 months with a similar lesion
requiring treatment by the same dentist took part in the study. Attitudes, dental anxiety and
satisfaction, one month after treatment, were assessed on a 5-point Likert scale questionnaire.
100% of patients participated and 94% completed the questionnaire within three months after
treatment. A similar questionnaire was administered to patients (n=45) who had undergone
conventional minimally invasive treatment within the last 6 months but who did not require ozone
treatment.
Results: All patients were happy or satisfied with the Ozone treatment they received and were
happy or satisfied with the amount of time the Ozone treatment required. 85% considered the
check up appointments to monitor the progression of clinical reversal of lesions a minor draw
back. 55% were satisfied to choose this treatment even if the Ozone treatment costs more than
regular treatment, 100% would recommend this treatment to a friend or close relative and 100%
would like to receive this treatment again. 80% of patients reported a reduction in anxiety (p

2754 Effective Treatment of Occlusal Fissure Caries Using Ozone
N. JOHNSON1, J. JOHNSON1, K. JOHNSON1, and E. LYNCH2,
1 Private practice, Wales, United Kingdom, 2 Queen's University Belfast, United Kingdom
Ozone has been shown to clinically reverse primary root carious lesions and early occlusal fissure
caries.
Objectives: This study assessed the effect of a novel ozone delivery system (HealOzone unit,
CurOzone USA) on primary occlusal fissure carious lesions over a 1- month period.
Methods: 105 patients attending a general dental practice were entered with 300 primary
occlusal fissure carious lesions. Each carious lesion had been deemed to require drilling and filling
and had a DIAGNOdent (KaVo, Germany) reading between 20 and 60 at baseline. After
randomisation, lesions were assigned to either receiving no treatment or ozone treatment with
each subject having at least one control lesion. Ozone was applied to each test lesion for 20
seconds. After 1 month, patients were recalled and clinically re-assessed for lesion severity. The
DIAGNOdent was again employed to objectively quantify the primary occlusal fissure carious
lesions.
Results: After 1 month, patients were recalled for revaluation. There were no observed adverse
events. Based on the clinical measurement of lesion severity, the ozone treated lesions showed
significant signs of reversal (P

airbrasion delivered at 80 psi using 27-micron aluminium oxide as the cutting agent (Abradent
DV1). Once access to the caries was established, ozone was applied for 40 seconds using a
HealOzone unit (CurOzone USA and KaVo Germany). In the treatment of interproximal caries,
techniques have been explored to ensure an effective seal. Glass ionomer restorative (Fuji 9 (GC)
or Diamond Carve (Kemdent)) restored the cavities.
Results: All lesions were successfully exposed and a seal established for the delivery of ozone.
Clinically acceptable seals have been achieved around all restorations. The airabrasion and Ozone
technique was significantly faster than conventional drilling and filling (P

Conclusion: The results demonstrate the effectiveness of Ozone to reduce the QLF readings at
two months after application in more than 90% of the lesions. Control showed increased QLF
readings . Any effect will be monitored over time.
Seq #277 - Miscellaneous Caries Clinical Studies
11:00 AM-12:15 PM, Saturday, 28 June 2003 Svenska Massan Exhibition Hall B
Back to the Cariology Research Program
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(June 25-28, 2003)
0983 6-month Clinical Indices Changes after Ozone Treatment of Pit and Fissure Caries
(PFC)
L. ABU-NABA'A, H. AL SHORMAN, and E. LYNCH,
Queen's University Belfast, United Kingdom
Ozone treatment of PFC is investigated in a controlled clinical trial.
Objectives: to quantify changes in clinical indices after Ozone treatment.
Methods: 34 non-cavitated PFC lesions (8 patients) were enrolled. After cleaning (Prophyflex 2®;
KaVo, Germany), the following were recorded: clinical classification (Ekstrand, 1998), the
DIAGNOdent® (D) (KaVo, Germany) readings, Hardness index (hard, leathery, soft), Colour index
(normal, yellow, light brown, grey, dark brown, black), Frosted enamel (mm), Stained enamel
(mm), Perceived treatment need index (requiring no intervention, requiring a pharmaceutical
approach but not drilling and filling (D&F) and not preventive resin restoration (PRR), possibly
requiring D&F or PRR, and definitely requiring D&F), visual index (Sound, arrested, active). 17
lesions were treated with Ozone (HealOzone unit; CurOzone & KaVo, USA & Germany) for 40
seconds and the other 17 were reserved as controls. After 1, 3 & 6- months (m), the procedure
was repeated. The change in the clinical indices values was tested from baseline by t-test, and
between groups by univariate analysis of the mean change over the 6m.
Results: The lesions in the treatment group showed a significant reduction in hardness, frosted
enamel and perceived treatment need indices at 1,3 and 6m and visual index at 6m (p0.05). No significant
changes were recorded for the control group for the hardness index at any recall visit.
Conclusion: Treating PFC lesions with Ozone significantly produced clinical changes that can be
detected in a regular dental setting and consistent at recall visits.
Seq #109 - Clinical and Restorative Studies
3:45 PM-5:00 PM, Thursday, 26 June 2003 Svenska Massan Exhibition Hall B
Back to the Cariology Research Program
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(June 25-28, 2003)
dott Cosimo Cigliola
146

2750 Treatment of Primary Occlusal Pit and Fissure Caries with Ozone: Six-month
Results
M.A. CRONSHAW, Private Practice, Isle of Wight, United Kingdom
Ozone has been shown to be effective in the arrest and reversal of primary pit and fissure as well
as root carious lesions.
Objectives: This study aimed at assessment of the effect of an ozone application on the clinical
status as well as the DIAGNOdent readings of occlusal pit and fissure caries over a six- month
period.
Methods : 18 subjects with 49 carious lesions deemed to require drilling and filling were entered.
31 teeth were treated with 30 seconds Ozone (HealOzone, CurOzone and KaVo, USA and
Germany) per test lesion. Each of the subjects had one control lesion which received no
treatment. All lesions were reassessed after a six- month period. Determination of the severity of
the lesions was made by measurement with an index of clinical severity and calibrated
DIAGNOdent (KaVo, Germany) readings.
Results : Of the 31 teeth treated, 25 showed improvements in DIAGNOdent measurements whilst
6 remained the same (P

lesions remained stable or progressed (P

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L.A.S.E.R.
LASER è l'acronimo inglese di Light Amplification by the Stimulated Emission of Radiation, ovvero
Amplificazione di Luce tramite Emissione Stimolata di Radiazioni.
La radiazione laser proviene dunque interamente dal processo di emissione stimolata:
M * + hν → M + 2hν
In condizioni di equilibrio del sistema questo è un processo sfavorito rispetto all'assorbimento.
Infatti se ci rifacciamo all'analisi di Einstein, le probabilità che avvengano l'emissione stimolata e
l'assorbimento sono date da:
Pes = BN2ρ(ν12)
Pass = BN1ρ(ν12)
dove B è il coefficiente di Einstein;
N1 è la popolazione dello stato a energia E1;
N2 è la popolazione dello stato a energia E2; (E2 > E1);
ρ(ν12) è la densità del campo di radiazione alla frequenza ν12 = (E2 - E1)/h;
Dal confronto tra Pes e Pass risulta evidente che è la popolazione dei livelli a determinare il processo
dominante. In condizioni di equilibrio N1 è sempre maggiore di N2 (perché le popolazioni dei due
livelli sono descritte dalla distribuzione di Boltzmann:
N2 =N1e -(E2-E1)/KT
da notare l'esponente negativo) e quindi per ottenere prevalenza dell'emissione stimolata è
necessario mantenere il sistema lontano dall'equilibrio, attuando l'inversione di popolazione.
Caratteristiche della radiazione laser
Direzionalità
Al contrario delle sorgenti tradizionali il laser permette di emettere la radiazione in un'unica
direzione. Più precisamente l'angolo solido sotteso da un fascio laser è estremamente piccolo.
Questa caratteristica viene sfruttata in diversi ambiti, per esempio permette di trattare le superfici
in maniera estremamente accurata (litografia, CD, etc.). In spettroscopia si ha la possibilità di
aumentare notevolemente il cammino ottico e quindi la sensibilità usando una sorgente laser che
attraversa il campione con una traiettoria a zig-zag grazie a un sistema di specchi.
Monocromaticità
L'allargamento della banda di emissione è dato dalla larghezza naturale e dall'effetto Doppler (che
può essere eliminato o comunque contenuto parecchio). In spettroscopia si sfrutta questa
caratteristica per ottenere spettri ad alta risoluzione. Gli spettri Raman non potrebbero essere
ottenuti senza questa caratteristica dei laser.
Brillanza
Nei laser la quantità di energia emessa per unità di angolo solido è incomparabilmente più elevata
rispetto alle sorgenti tradizionali. In particolare è elevato il numero di fotoni per unità di
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152

frequenza. Questa caratteristica è diretta conseguenza delle due precedentemente citate. Grazie a
questa caratteristica si ha la possibilità di osservare fenomeni particolari, come per esempio
l'assorbimento a molti fotoni. L'elevata intensità ha trovato anche diverse applicazioni
tecnologiche, per esempio nel taglio dei metalli.
Coerenza
Mentre nell'emissione spontanea ogni fotone viene emesso in maniera casuale rispetto agli altri,
nell'emissione stimolata ogni fotone ha la stessa fase del fotone che ha indotto l'emissione. La
fase viene mantenuta nel tempo e nello spazio. Questa caratteristica ha permesso lo sviluppo
della tecnica CARS.
Impulsi ultra-brevi
Con diverse tecniche è possibile costruire laser che emettano pacchetti d'onde estremamente
stretti nel dominio del tempo, attualmente si è giunti allo sviluppo di impulsi dell'ordine del
femtosecondo. Questi laser hanno trovato impieghi in diversi ambiti di ricerca, hanno per esempio
permesso la nascita di una nuova disciplina, che è stata chiamata femtochimica.
Il laser in medicina
Successivamente alla sua invenzione nel 1960, il laser è stato usato diffusamente per scopi
medici. La risposta terapeutica dipende in maniera complessa dalla scelta della lunghezza d'onda,
della durata di irradiazione e della potenza del laser. Combinazioni diverse di questi parametri
sono impiegate per trasformare l'energia luminosa in energia meccanica, termica o chimica.
Generalmente, gli effetti meccanici sono prodotti dall'applicazione di brevi impulsi (dell'ordine dei
nanosecondi) e alte energie.
In questo modo onde di stress meccanico possono essere prodotte con sufficiente forza per
disintegrare calcoli urinari. Gli effetti termici si ottengono abbassando la potenza del laser. Brevi
impulsi laser vengono usati per ablare sottili strati di tessuto in chirurgia rifrattiva, utilizzando luce
laser che penetra solo alcuni micrometri nel tessuto. La lunghezza d'onda della luce laser può
essere scelta in modo tale che la luce sia assorbita selettivamente dal bersaglio. La coagulazione
selettiva delle vene varicose in chirurgia estetica può essere compiuta usando luce laser assorbita
selettivamente dall'emoglobina. L'impulso è scelto allora sufficientemente breve così da non
arrecare danno al tessuto normale circostante, ma anche lungo a sufficienza da permettere la
coagulazione sull'intero diametro del vaso.
Oftalmologia
Un altro importante uso medico del laser è la cura di difetti alla visione, in particolare la correzione
della miopia, ottenuta modificando con varie tecniche (PRK, LASIK) il profilo della cornea.
L'asportazione di un nanometro di superficie cornea corregge un difetto di movimento del muscolo
oculare con l'introduzione di un difetto organico nell'occhio.
Dal 2005 è utilizzato anche per la correzione di un difetto visivo più raro, l'astigmatismo.
Sostanzialmente, l'errore non è nell'occhio, ma nel modo in cui viene mosso. Poiché è difficile
correggere il movimento e rieducare la muscolatura, si deforma lievemente (ma in modo
permanente) la cornea per riequilibrare il difetto e acquisire una visione perfetta (10/10). Il
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153

macchinario è derivato da quelli utilizzati nell'industria per l'asportazione di truciolo (invece del
truciolo si asporta una parte di cornea), con la precisione che caratterizza una macchina a
controllo numerico.
L'intervento non è così banale, poiché il macchinario non opera sulla superficie corneale dell'occhio
che è necessario sollevare e richiudere dopo avere operato su uno strato di cornea sottostante.
Tali macchine elaborano delle curve statistiche per sezionare correttamente una superficie sottile.
L'interpolazione è buona di solito ed ha successo (un buon taglio) anche nel singolo caso.
Le statistiche però non dicono nulla sul singolo caso, e non si può assicurare un 100% dei
successi. L'insuccesso in genere non dipende da un'imprecisione del macchinario, quanto dal fatto
che la cornea del paziente ha una forma particolare (un difetto organico) sulla quale macchinari di
quel produttore o concorrenti hanno operato in pochi casi e il macchinario non è in grado di
correggere. L'esperienza accumulata e le curve interpolate sono inserite di serie su tutti i
macchinari anche nuovi. Non sempre i produttori condividono l'apprendimento delle macchine di
produzione. Ciò rallenta l'estendibilità del laser a tutti i tipi di cornee. L'intelligenza del
macchinario dipende totalmente dall'esperienza acquisita in interventi precedenti; la sua
precisione è nell'applicare nella cornea esattamente le misure di taglio calcolate, che però non
sempre sono adatte con precisione alla cornea operata. Tuttavia, prima dell'intervento si può
capire se la cornea è operabile col macchinario in dotazione. Accade che la macchina è tarata per
ampiezze della pupilla (che sono dell'ordine del micron) leggermente inferiori alla massima
estensione da misurare che è quella raggiunta dalla pupilla di notte.
Alla "pupilla grossa" è attribuito talvolta un errore di decentramento della macchina durante
l'operazione.
Non è idonea per un intervento del genere una cornea "pulsante", in altre parole predisposta di
suo a degli "spostamenti" che si sarebbero tradotti in continue fluttuazioni e cambiamenti di
gradazione.
L'intervento ha successo in un buon 90% dei casi; rarissimi i casi di cecità; diversamente, si
mantiene un livello di vista simile a quello precedente l'intervento con l'onere di portare tutta la
vita occhiali o lenti a contatto.
Il macchinario consente il day-hospital e richiede (per l'elevato costo da ammortizzare) interventi
con una certa frequenza (da tre a trenta minuti per intervento). Non c'è un macchinario così
flessibile da poter operare tutti i tipi di cornee, ma una varietà di macchinari in commercio per
ogni tipo di cornea. Spesso però il macchinario ultramoderno in dotazione è uno solo.
Arrivati alla quarta generazione di macchinari attuale, gli "effetti collaterali", assenti o non troppo
evidenziati nel "consenso informato", che lamenta un non trascurabile 7% degli operati, sono:
secchezza degli occhi (assenza di lacrime), abbassamento della visione notturna, fotofobia,
astigmatismo irregolare che fa esplodere le luci in tante macchie indefinite, sdoppiamento delle
immagini, aloni, forte bruciore nei locali chiusi, gli occhi perennemente arrossati. Le complicanze
possono essere gravi, impedire la guida notturna (insufficienza e fastidio dell'illuminazione
stradale), il lavoro in ambienti a forte luminosità (ogni luce diviene un'esplosione di luci). Le
complicazioni tendono a manifestarsi fin dai primi giorni dopo l'intervento e ad aggravarsi con gli
anni.
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Se l'occhio è molto scavato e il lembo da rimuovere per l'incisione laser è cicatrizzato
definitivamente, oppure se il laser ha commesso errori nel sollevare e riporre la superficie
corneale per asportare microon dello strato sottostante, l'occhio non è rioperabile e il danno è
permanente.
Talora si ricorre al trapianto di cornea, dall'esito incerto e meno promettente rispetto alle attese
per un intervento al laser. Nel caso poi di errori del laser restano pieghe sulla cornea che diviene
di forma irregolare per cui non esistono occhiali e lenti a contatto in commercio in grado di
correggerlo.
Forti dolori post-operatori sono dovuti talvolta ad allergia al cortisone contenuto nei colliri.
Nel caso in cui venga firmata una liberatoria prima dell'intervento, non vi sono responsabilità né
estremi di reato. Il "consenso informato" in vari casi viene firmato pochi minuti prima
dell'intervento, sotto anestesia locale degli occhi. Non è viceversa semplice trovare uno specialista
che certifichi con la firma su una cartella clinica il danno eventualmente subito da un collega, che
sarebbe equivalente ad una denuncia penale. Per ottenere un ricarcimento in tribunale e una
radiazione dall'albo è appunto necessaria una perizia tecnica.
Terapia fotodinamica contro alcuni tumori
Il laser viene utilizzato come tecnica non invasiva per la completa rimozione di tumori allo stadio
iniziale.
Nelle cellule viene iniettato un farmaco innocuo che riconosce e si "incolla" alle sole cellule malate,
sensibile alla luce. Al passaggio del laser, il farmaco attiva una reazione che ha per protagonista
l'ossigeno, ossida e distrugge le sole cellule malate. Il fatto eccezionale è che il farmaco agisce
selettivamente e le cellule sane non vengono minimamente intaccate, come purtroppo avviene
durante un asportazione chirurgica.
Per tumori più estesi, serve a circoscrivere la metastasi, ma non guarisce la malattia.
Altri utilizzi
Dei laser vengono utilizzati per realizzare effetti speciali
durante uno spettacolo
Il laser viene utilizzato nella tecnica in una gran varietà di
apparecchiature:
Nelle telecomunicazioni e nelle reti di computer viene
utilizzato per trasferire enormi quantità di dati attraverso le
fibre ottiche. Viene utilizzato come elemento di lettura nei
player di CD e DVD e per la scrittura nei masterizzatori. In ambito industriale il laser viene
utilizzato per tagliare o saldare lamiere in metallo anche di elevati spessori. In metrologia grazie ai
laser si possono effettuare delle misure di estrema precisione nel campo che va dai micron alle
decine di metri. In campo edile vengono utilizzate sempre più spesso livelle laser. Si realizzano
puntatori per armi, o più pacificamente, per conferenze. Enormi laser permetteranno forse in un
prossimo futuro di ottenere reattori nucleari a fusione efficienti.
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Nel settore militare i laser vengono utilizzati come componente dei sistemi di puntamento ma il
loro uso come arma offensiva o difensiva non è diffuso. I laser hanno ricevuto spesso ingenti fondi
ma ottenendo risultati piùttosto modesti. I comandi militari hannno richiesto sistemi laser di
elevata potenze (100 KiloWatt almeno) e maneggevoli, cioè apparecchiature trasportabili su mezzi
cingolati o su gomma. I ricercatori sono stati in grado di realizzare laser di notevole potenza
(anche diversi MegaWatt) e laser portatili ma non sono stati in grado di realizzare sistemi che
riunissero entrambi le caratteristiche.
Nel febbraio 2007 utilizzando un laser SSHCL (Solid State Heat Capacity Laser) ricercatori
statunitensi hanno dichiarato di aver raggiungo potenze di 67 KiloWatt con un dispositivo
trasportabile.
I ricercatori ritengono di poter raggiungere potenze di 100 KiloWatt entro il 2008. [1]
Il Laser viene utilizzato anche per manipolare la materia a livello atomico. Il laser può essere
utilizzato per saldare, dividere o forare elementi a livelli atomici, inoltre viene spesso utilizzato per
raffreddare i composti a temperature prossime allo zero assoluto (-273,16 ° C). Il raffreddamento si
ottiene illuminando la materia con i fotoni, sotto opportune condizioni gli atomi assorbono il fotone
e ne emettono uno a energia superiore perdendo di conseguenza energia. Si sta studiando la
possibilità di utilizzare queste tecniche per raffreddare i semiconduttori. [2]
Tipi di laser
- A rubino - A gas - A giunzione - A coloranti organici
Riferimenti
1. Un laser da guerre spaziali contro missili e carri armati
2. Semiconduttori raffreddati con la luce. Le Scienze. URL consultato il 09-05-2007.
Bibliografia
• Orazio Svelto. Principi dei laser (Tamburini, 1970).
• (EN) Anthony E. Siegman. Lasers (University Science Books, 1986).
• Mario Bertolotti. Storia del laser (Bollati Boringhieri, 1999). ISBN 8833911985
Voci correlate
• Maser
• Laser a eccimeri
• Laser atomico
Altri progetti
• Commons contiene file multimediali su Laser
Collegamenti esterni
• (EN) Simulatore interattivo per il laser al rubino
• (EN) Simulatore interattivo per il laser all'elio neon
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• (EN) Simulatore interattivo per il diodo laser
• Per una fantasiosa storia del laser
Storia del Laser
1917: Il fisico tedesco Albert Einstein descrive il principio teorico del laser: è possibile stimolare la
materia in modo da farle emettere luce con particolari caratteristiche.
1951: I fisici americani Charles Townes e Joseph Weber e i russi Alexander Prokchorov e Nicolai
Basov , separatamente, studiano il maser, progenitore del laser, in cui al posto della luce sono
usate le microonde.
1957: I fisici americani Charles Townes e Arthur Schawlow iniziano a lavorare sui principi del
laser.
1960: Il fisico americano Theodor e Maiman costruisce il primo laser mettendo in pratica il
principio descritto da Einstein.
1961: Gli oftalmologi americani Charles Campbell e Charles Koester applicano per la prima volta il
laser sull'uomo rimuovendo un tumore retinico.
1964: Nicolai Basov , Alexander Prokhorov , Charles Townes ricevono il premio Nobel in Fisica per
lo studio del maser e delle applicazioni del laser.
Anni 60-70: I. Kaplan impiega i laser per la prima volta in chirurgia plastica, Leon Goldman in
dermatologia, Billie Aronoff e R.eza Jako in Otorinolaringoiatria, Alfons Hofstetter in urologia,
Peter Kiefhaber , R. Verschueren e R. Dwyer in gastroenterologia ed endoscopia, J. Bellina in
ginecologia ed ostetricia, G. Abela in cardiochirurgia, P.W. Ascher in neurochirurgia, M. W. Berns
per lo studio delle cellule in vitro, G . Lynn-Powell in odontoiatria, D. Apfelberg per la cura degli
angiomi e delle lesioni vascolari.
Seconda metà degli anni '60
II secondo impiego fu quello del laser Nd-Yag come supporto alla chirurgia endoscopica.
Successivamente fu il laser ad Argon a trovare largo impiego nel l'oculistica per la correzione del
distacco della retina, grazie alla sua capacità di essere assorbito dall'emoglobina.
1966: E. Mester si accorge che il laser rosso a bassa energia accelera la ricrescita del pelo nei
ratti depilati e successivamente lo impiega per stimolare la cicatrizzazione delle ulcere da
decubito.
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1967: T. Dougherty utilizza sperimentalmente i laser per attivare alcune sostanze fotosensibili che
identificano le cellule tumorali, si legano ad esse e le distruggono selettivamente: si parla di
diagnosi e terapia fotodinamica.
Anni '70
È con l'introduzione del laser Co2 che il laser entra prepotentemen te nello studio del medico. È del
1972 la prima unità laser ad anidride carbonica per uso chirurgico realmente funzionante
realizzata da Isaac Kaplan, chirurgo plastico e dermatologo israeliano.
1972: J. Walker , P.R. Chekurov , T. Oshiro e M. Trelles utilizzano gli effetti
antidolorifici/antinfiammatori dei laser nella cura delle malattie osteo-reumatiche e nella
traumatologia sportiva. J. Walker nella nevralgia da arto fantasma.
1975: Primo impiego italiano dei laser in Chirurgia plastica, da parte del Prof. Riccardo Pariente ,
a Roma.
Anni 75-80: Rosario Brancato utilizza i primi laser per oftalmologia in Italia, a Trieste., e
Pasquale Spinelli li utilizza in endoscopia, a Milano, Gaetano Bandieramonte (Milano) e Marcello
D'Ovidio (Roma) in Urologia, Fausto Chiesa (Milano) e G. Motta (Napoli) in Otorinolaringoiatria,
Anton Giulio Sesti e Leonardo Longo per la guarigione delle ferite e delle ulcere cutanee e per la
cura delle malattie osteo-muscolo-tendinee (reumatismi fibromiositici , traumatologia sportiva), D.
Dioguardi , G. Micali e A. Musajo-Somma per la cura degli angiomi, V. Fasano (Torino) in
neurochirurgia, F. Buttafarro (Torino) e P. Santoianni (Napoli) in dermatologia, E. Panzoni e C.
Clauser in odontoiatria e chirurgia maxillo-facciale , H. Emanuelli in chirurgia generale, S.
Cavaliere in broncologia , L. Roccia in agopuntura.
1976: primo caso di ulcera da decubito trattato in Italia con laser non chirurgici, a Firenze, presso
l' Ospedale INRCA di Poggiosecco , dall'equipe del Prof. A.G. Sesti.
1977-1983: Il CNR Italiano promuove un progetto di ricerca sulle Applicazioni biomediche dei
laser , diretto dal Prof. Alberto Sona (Milano).
1979: nasce la prima Societa' Italiana di Laser Chirurgia e Medicina, a cui ne seguono altre
trentadue.
1980: R. Rox- Anderson e John Parrish elaborano la teoria della fotolisi selettiva, su cui si basa
l'80% degli impieghi dei laser in medicina: in poche parole i laser riconoscono i tessuti dal colore
di essi e si assorbono selettivamente solo nei tessuti di colore complementare.
1981: Nicolaas Bloembergen , Arthur Schawlow , Kai Siegbahn ricevono il premio Nobel in Fisica
per lo sviluppo della spettroscopia laser.
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1983: T Oshiro e K. Atsumi descrivono gli impieghi dei laser non chirurgici ed i possibili
meccanismi d'azione. N. Passerella studia l'effetto dei laser sui mitocondri cellulari. In varie parti
del mondo si inizia a trattare l'iperglicemia dei diabetici insulino-dipendenti con i laser
ipoglicemizzanti.
1984: GD Baxter descrive gli impieghi dei laser in fisioterapia.
1984: L' FDA traccia le linee guida per gli impieghi dei laser per le lesioni vascolari e
dermatologiche, e le aggiorna tutti gli anni.
1985: P.W. Ascher e D. Choy impiegano i laser per la decompressione delle ernie del disco
vertebrale senza ricorrere alla chirurgia.
1986: primo caso di Induratio Penis Plastica Calcifica trattato nel mondo con un laser fotoacustico
, da A. Durval , P. Peruzzi e L. Longo , a Firenze.
1987: Viene proposta la Laserscleroterapia combinata per il trattamento delle telangiectasie
(capillari) degli arti inferiori, metodica tutt' ora insuperata.
Anni '90
Nell'ultimo decennio le fonti laser si sono moltiplicate, ottenendo risultati assolutamente unici in
molti ambiti della medicina: un esempio tra tutti il laser ad eccimeri per la correzione della miopia.
1990: primi casi di Laser-resurfacing del viso (R. Gregory ed altri) e si inizia a parlare di saldatura
laser dei tessuti (Laser Tissue Welding , Dix Poppas ).
I laser per il ringiovanimento
1993: CO2. Nasce il primo laser per il ringiovanimento del viso. Agisce vaporizzando i tessuti, cioè
creando un'ustione che, distruggendo le vecchie cellule, permette la "risalita" di una pelle nuova
durante il processo di guarigione. Il laser CO2 non solo elimina gli strati più superficiali ma è anche
in grado di determinare una importante contrazione delle fibre collagene ed elastiche presenti nel
derma e di rendere perciò la pelle molto più tonica ("effetto lifting"). Il periodo post-operatorio è
abbastanza impegnativo e l'arrossamento che ne consegue permane a lungo (circa 3 mesi).
1995: primi casi di laser hair removal.
1997:primi casi di ringiovanimento cutaneo con laser e luce pulsata.
1997: nasce Laser Florence, “Una finestra sul mondo della Laser Medicina”, dove vengono
presentate, discusse e selezionate le principali novita' mondiali del settore. Tale conferenza ha
cadenza annuale.
1998: Laser Erbium. Molto più 'dolce' del CO2, funziona causando delle microesplosioni
(fotoablazione) delle cellule superficiali della pelle, senza surriscaldare gli strati più profondi del
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derma. Ciò consente un periodo postoperatorio più rapido (10/15 giorni di lontananza dalla vita
sociale, mentre solo dal 5° giorno ci si può truccare o radere). Dunque un arrossamento più
limitato nel tempo, ma anche un risultato inferiore.
2000: Laser non ablativi. Con l'arrivo del nuovo secolo arrivano i laser non ablativi. Alla loro
base un raggio freddo, emesso ad una potenza molto bassa, associato all'emissione di un getto di
gas refrigerato per evitare ogni surriscaldamento, in grado di sollecitare il collagene e rigenerare i
tessuti sottostanti l'epidermide, senza "toccare" la superficie della pelle. Un trattamento soft, con
scarsissimi effetti collaterali e la possibilità di tornare subito al lavoro o alle proprie occupazioni
abituali. I risultati sono meno evidenti, incostanti e non prevedibili, non certo paragonabili a quelli
di un CO2 e nemmeno di un laser Erbium.
2003: La 'Luce Pulsata'. Nata per l'uso dermatologico vascolare negli anni '90 solo con i primi
anni del nuovo secolo ottiene quella precisione tecnica che permette la sua diffusione. Si tratta di
una sorgente a luce pulsata che emette un flash con un ampio spettro luminoso. La luce si
trasforma in calore che innalza la temperatura e provoca una leggera lesione controllata dello
strato superiore del derma senza causare alcun danno all'epidermide. Il processo di guarigione
determina la neoformazione di collagene migliorando l'aspetto generale della pelle. Clinicamente
l'effetto della neoformazione di collagene si evidenzia alcuni mesi dopo il ciclo di trattamenti (circa
8-10 distanziati di 10 giorni l'uno dall'altro). Il risultato migliore si ottiene sulle microrughe e sullo
stato generale (texture, luminosità e turgore) della cute. Il rischio di bruciature è più elevato con i
fototipi più scuri (V-VI).
2006: Fraxel. Dopo un anno di studi e sperimentazione negli USA, arriva in Italia Fraxel: un
laser a fibre ottiche di tipo 'non ablativo', dunque una metodica con minime conseguenze post-
trattamento, in grado però di offrire i risultati di un laser CO2, in modo più sicuro e prevedibile, su
una serie di inestetismi sino ad oggi affrontabili solo in parte e con metodiche assai più invasive.
Bibliografia:
Istituto Laser Medicina
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