EEBA Program (PDF/3MB) - EEBA - Annual Meeting
EEBA Program (PDF/3MB) - EEBA - Annual Meeting EEBA Program (PDF/3MB) - EEBA - Annual Meeting
VALIDATION OF AN AUTOMATED TEST SYSTEM FOR STERILITY CONTROLS OF AMNIOTICMEMBRANE FOR CLINICAL APPLICATIONSH. Thomasen, K. P. Steuhl, D. MellerCornea Bank Essen, Department of Ophthalmology, University Hospital Essen, GermanyPurpose: Cryopreserved amniotic membrane (AM) is widely used in ophthalmology because of itsbeneficial clinical capabilities. For clinical use a careful testing for microbial contamination is essentialto ensure a safe application. Up to now there has been no validated automated test system accordingto EU regulations to perform this task. In our study we evaluated the use of the BacT/Alert® test systemfor the screening of microbial growth in AM.Methods: Sections of fresh AM which were cleaned of placental blood and tissue residues,cryopreserved AM (each about 5 cm2) were injected separately in BacT/Alert culture media testbottles. BacT/Alert i-AST® bottles were used to evaluate the growth of fungi and aerobic bacteriaand BacT/Alert i-NST® bottles to evaluate anaerobic bacteria. Several bacterial and fungal test strainsaccording to European Pharmacopaea monograph 2.6.27 were applied to test the performance ofthe system (Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans,Aspergillus brasiliensis, Propioniobacterium acnes and clostridium sporogenes). About 10 to100 colony forming units of each germ were applied on the samples prior to injection in thecorresponding test bottles. For each strain about four bottles with AM samples were inoculated, twoadditional battles only contained the germ to verify its growth while bottles with samples lackinggerms served as negative controls. Bottles were incubated at 37°C for a period of seven days. In case ofmicrobial growth within a test bottle the germ was subcultivated and identified.Results: Growth of the test strains was detected in all inoculated samples within the seven daysincubation period. Neither non-processed nor cryopreserved AM inhibited the growth of the germs,despite the standardised application of antibiotics in cryopreserved AM.Conclusions: Since all applied test strains were detected, we conclude that the BacT/Alert® incombination with the i-NST and i-AST bottles system is suitable for testing of the microbial safety ofamniotic membrane in clinical practice according to EU regulations.74
INFLUENCE OF STORAGE CONDITIONS OF PLACENTAL TISSUE ON STERILITY ANDHISTOLOGIC PROPERTIES OF AMNIOTIC MEMBRANED. Meller, H. Thomasen, K. P.SteuhlCornea Bank Essen, Department of Ophthalmology, University Hospital Essen, GermanyPurpose: Cryopreserved amniotic membrane (AM) is a common tool for the treatment of severalophthalmologic applications because it provides beneficial clinical effects. The preparation of AMneeds to follow stringent standard operating procedures to ensure, quality and safety of the tissuefor clinical application. Part of the SOPs is to define how long and under which conditions donorplacentas can be stored without compromising clinical efficacy of the prepared tissue. Our aim was toevaluate the impact of storage time and temperature on clinical important features of AM.Methods: After informed consent placentas were obtained and stored for different periods of timeat various temperature conditions. In total there were six groups , each n=3, specified by the storagetimes and conditions: 1h at 8°C, 20°C and 40°C, 6h at 8°C, 1h at 40°C followed by 5h at 20°C and 1h at20°C followed by 5h at 8°C. Afterwards AM was prepared from the placentas and examined in regardof tissue sterility, histologic appearance and the basement membrane composition. Sterility controlswere performed at the department of microbiology. The AM was stained with hematoxilin/eosin andthe presence of the basement membrane components Laminin, fibronectin, collagen IV and collagenVII was analyzed by means of immunohistochemistry.Results: Only tissue from the group of samples which was stored for 1h at 40°C showedmicrobiological contamination with the bacterium Ralstonia pickettii whiles all other samples werenegative for microbial growth. The histologic appearance of the tissue did not differ within or betweenthe particular groups. Neither the general staining, nor the immunhistologic staining displayed arecognizable difference.Conclusions: The detected contamination is most likely an artifact which resulted from bacterialcontamination of the water bath used to hold the temperature of 40°C. There was no link between theexamined storage conditions of the placenta prior to preparation and microbial growth on the tissue.The examined storage conditions also did not seem to have a significant effect on the histologicalstructure of AM. Therefore we conclude, that storing the placenta up to 6h at these temperaturesbefore preparation is tolerable for tissue quality of the AM.XXV ANNUAL MEETING OF THE EUROPEAN EYE BANK ASSOCIATION Zagreb, Croatia 18/19 January 2013 75
- Page 23 and 24: Accommodation & TravelingHow to get
- Page 25 and 26: Table of contentsINVITED LECTURE: T
- Page 27 and 28: INVITED LECTURE: THOMAS REINHARD, G
- Page 29 and 30: EUROCET 128: SUPPORTING THE IMPLEME
- Page 31 and 32: MENTAL ATTITUDES CONCERNING CORNEA
- Page 33 and 34: AN EVALUATION OF TOTAL BLOOD AND PL
- Page 35 and 36: USE OF FIVE-YEAR CORNEAL GRAFT SURV
- Page 37 and 38: PRECUTTING OF DONOR CORNEAS FOR POS
- Page 39 and 40: STUDY OF STROMAL FEMTOSECOND LASER
- Page 41 and 42: ULTRA- THIN DESCEMET STRIPPING AUTO
- Page 43 and 44: POSTERIOR LAMELLAR KERATOPLASTY- PO
- Page 45 and 46: ENDOTHELIAL CELL DENSITY AFTER DESC
- Page 47 and 48: INVITED LECTURE: DONALD TAN, SINGAP
- Page 49 and 50: INTEGRITY OF HUMAN CORNEAL EPITHELI
- Page 51 and 52: A NOVEL OBJECTIVE METHOD TO EVALUAT
- Page 53 and 54: EVALUATION OF DONOR CORNEAS DURING
- Page 55 and 56: DELIVERY OF MOLECULES INTO CORNEAL
- Page 57 and 58: VISIONGRAFT® STERILE CORNEA - NEW
- Page 59 and 60: EMERGENCY PREPAREDNESSP. DahlThe Ey
- Page 61 and 62: MANAGEMENT IN NON-TRAUMATIC CORNEAL
- Page 63 and 64: CATARACT AND FUCH’S DYSTROPHY: DS
- Page 65 and 66: RECURRENCE OF ANTERIOR CORNEAL DYST
- Page 67 and 68: USE OF ANTI-VEGF (BEVACIZUMAB) AFTE
- Page 69 and 70: INVITED LECTURE: KEVIN CORCORAN, US
- Page 71 and 72: IDENTIFICATION OF LABEL-RETAINING E
- Page 73: EPITHELIAL AND PROGENITOR CELL MARK
- Page 77 and 78: PostersRESULTS OF MORPHOLOGICAL EXA
- Page 79 and 80: KERATOPLASTY “A CHAUD“D. Saric,
- Page 81 and 82: CELL-BY-CELL ALIGNMENT OF REPEATED
- Page 83 and 84: RIGID GAS-PERMEABLE CONTACT LENS CO
- Page 85 and 86: DEEP ANTERIOR LAMELLAR KERATOPLASTY
- Page 87 and 88: CORECTION OF POST-KERATOPLASTY ASTI
- Page 90: yChoose the highestquality for your
VALIDATION OF AN AUTOMATED TEST SYSTEM FOR STERILITY CONTROLS OF AMNIOTICMEMBRANE FOR CLINICAL APPLICATIONSH. Thomasen, K. P. Steuhl, D. MellerCornea Bank Essen, Department of Ophthalmology, University Hospital Essen, GermanyPurpose: Cryopreserved amniotic membrane (AM) is widely used in ophthalmology because of itsbeneficial clinical capabilities. For clinical use a careful testing for microbial contamination is essentialto ensure a safe application. Up to now there has been no validated automated test system accordingto EU regulations to perform this task. In our study we evaluated the use of the BacT/Alert® test systemfor the screening of microbial growth in AM.Methods: Sections of fresh AM which were cleaned of placental blood and tissue residues,cryopreserved AM (each about 5 cm2) were injected separately in BacT/Alert culture media testbottles. BacT/Alert i-AST® bottles were used to evaluate the growth of fungi and aerobic bacteriaand BacT/Alert i-NST® bottles to evaluate anaerobic bacteria. Several bacterial and fungal test strainsaccording to European Pharmacopaea monograph 2.6.27 were applied to test the performance ofthe system (Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans,Aspergillus brasiliensis, Propioniobacterium acnes and clostridium sporogenes). About 10 to100 colony forming units of each germ were applied on the samples prior to injection in thecorresponding test bottles. For each strain about four bottles with AM samples were inoculated, twoadditional battles only contained the germ to verify its growth while bottles with samples lackinggerms served as negative controls. Bottles were incubated at 37°C for a period of seven days. In case ofmicrobial growth within a test bottle the germ was subcultivated and identified.Results: Growth of the test strains was detected in all inoculated samples within the seven daysincubation period. Neither non-processed nor cryopreserved AM inhibited the growth of the germs,despite the standardised application of antibiotics in cryopreserved AM.Conclusions: Since all applied test strains were detected, we conclude that the BacT/Alert® incombination with the i-NST and i-AST bottles system is suitable for testing of the microbial safety ofamniotic membrane in clinical practice according to EU regulations.74
Change language