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ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - Corneamelting. Early identification of corneal/scleral thinning and meltingin Boston type I KPro patients allows for early intervention and maylead to improved outcomes.Commercial Relationships: David Sola-Del Valle, None;Bernardo M. Cavalcanti, None; Andrea Cruzat, None; Claes H.Dohlman, None; Pedram Hamrah, NoneSupport: Keratoprosthesis Manufacturing Fund, NIH K08-EY020575, Research to Prevent Blindness Career DevelopmentAward, Falk Medical Research TrustProgram Number: 3467 Poster Board Number: D0094Presentation Time: 11:00 AM - 12:45 PMScanning Laser Ophthalmoscope Microperimetry through theBoston KeratoprosthesisRony R. Sayegh, Claes H. Dohlman, Mary Lou Jackson.Ophthalmology, Massachusetts Eye & Ear Infirmary, Boston, MA.Purpose: Scanning laser ophthalmoscope (SLO) microperimetry isuseful in accurately mapping macular function. We explore thefeasibility and potential use of this method in eyes implanted with aBoston keratoprosthesis.Methods: Retrospective review of eyes with a type I Bostonkeratoprosthesis tested with OPKO SLO microperimetry. Bestcorrectedvisual acuity, contrast sensitivity and complete slit-lampand posterior segment examination findings were noted. Visual fieldand ocular coherence tomography results were collected whenavailable.Results: Five eyes were included in this study. Mean age was 66years (range: 54 to 82). Mean LogMAR best-corrected visual acuitywas 0.76 (range: 0.30 to 1.20). Contrast sensitivity was decreased inall eyes at a mean of 0.90 log units (range: 0.45 to 1.5). SLOmicroperimetry could be performed on all 5 eyes. Accurate functionalmapping of the macula and preferred retinal locus was obtained in allcases and was not affected by media opacities or decreased contrastsensitivity. The results correlated well with the findings on Goldmannperimetry and OCT in cases in which these were available.Conclusions: SLO microperimetry through the Bostonkeratoprosthesis is possible. It is a good adjunct to standard perimetrywith involvement of central fixation for accurate macular mapping. Itmay also allow better assessment of macular pathology unveiled byclearing of the visual axis with a keratoprosthesis.Commercial Relationships: Rony R. Sayegh, None; Claes H.Dohlman, None; Mary Lou Jackson, Nonehistologic appearance (H&E staining) and presence of inflammation(immunolocalization of CD45 positive cells) in rabbit corneal tissues50 days after an intralamellar implant of the modified Ti. Surfacecharacterization of the modified Ti included assessment by scanningelectron microscopy (SEM), X-ray diffraction crystallography(XRD), Fourier transform infrared spectroscopy (FTIR) andchemical/color stability was determined after exposure to 70%alcohol for three months.Results: Blue and brown coloration of Ti was achieved byanodization at 33 and 13.4 volts, respectively with a current supply of3 Amps, (Figure 1). The in vitro biocompatibility assays showed nosignificant differences in cell proliferation, cytotoxicity or migrationbetween HCLE cells co-cultured with surface modified or nonmodifiedTi (p> 0.745 for all group comparisons. Analysis of cornealtissues that had harbored the Ti implants showed normal cellappearance, and lack of CD45 or TUNEL positive cells. SEMshowed the presence of a nano porous surface and similarcrystallographic prints were observed with XRD and FTIR for thenon-coated and the modified Ti, (Figure 2). Three months exposureto alcohol had no effect on the color or color homogeneity of theoxidized surface.Conclusions: Ti backplate coloring was achieved by altering thevoltage of the electrochemical anodization and controlling thethickness of the Ti oxidation. In vitro and in vivo results suggest thatthe modified Ti is equally biocompatible and as safe as the standardnon-coated Ti. The color modification of the BKpro may improve thecosmesis and acceptance of the KPro by patients.*Supported by The Boston Keratoprosthesis Fund at theMassachusetts Eye and Ear Infirmary.Program Number: 3468 Poster Board Number: D0095Presentation Time: 11:00 AM - 12:45 PMAssessment of Titanium Modification for Coloring the Backplateof Boston KeratoprosthesisEleftherios I. Paschalis 1, 2 , James Chodosh 1 , Sandra J. Spurr-Michaud 2 , Andrea Cruzat 1 , Allyson Tauber 1 , Irmgard Behlau 1 , IleneK. Gipson 2 , Claes H. Dohlman 1 . 1 Ophthalmology, Massachusetts Eyeand Ear Infirmary - Harvard Medical School, Boston, MA;2 Ophthalmology, Schepens Eye Research Institute - Harvard MedicalSchool, Boston, MA.Purpose: Recent use of a titanium (Ti) backplate has improved thedesign and biocompatibility of the Boston Keratoprosthesis (BKPro).Titanium’s shiny metallic appearance, however, makes itcosmetically less favorable. The purpose of this study was to developand test a coloring, surface modification of Ti.Methods: Ti coloring was achieved electrochemically by controlledanodization of the Ti surface, which creates an oxide film on the Ti.Biocompatibility was assessed in vitro by co-culture of the modifiedTi with human corneal limbal epithelial cells (HCLE) followed byassays of cell proliferation, cytotoxicity and migration, and in vivo byCommercial Relationships: Eleftherios I. Paschalis, None; JamesChodosh, Alcon (C), Allergan (C), 3-V Biosciences (C), Novabay(C); Sandra J. Spurr-Michaud, None; Andrea Cruzat, None;Allyson Tauber, None; Irmgard Behlau, None; Ilene K. Gipson,None; Claes H. Dohlman, NoneProgram Number: 3469 Poster Board Number: D0096Presentation Time: 11:00 AM - 12:45 PM©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.
ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaFemto-second-laser and eximer-laser assisted preparation ofbiosynthetic corneal collagen donor implants and the recipientbedRaphael T. Neuhann 1 , Kerstin M. Wand 1 , Karin Kobuch 1 , MichaelBaumann 4 , May Griffith 2 , Mohammad M. Islam 2 , Johannes Junger 4 ,Roland Ritter 3 , Chris Lohmann 1 . 1 Ophthalmology, Klinkum Rechtsder Isar, Munich, Germany; 2 Regenerative Medicine, LinkopingUniversity, Linkoping, Sweden; 3 Technolas Perfect Vision, Munich,Germany; 4 MLase AG, Germering, Germany.Purpose: The goal was to achieve a setup that allows precise dockingof the laser as well as easy handling of the recipient and donorcorneas for an ALK (anterior lamellar keratoplasty) usingbiosynthetic corneal collagen implants. Before starting in vivo trailsin the rabbit eye, the trial setup was tested ex vivo on porcine andrabbit eyes. Furthermore the diameter, depth and surface of therecipient bed as well as the diameter and thickness of the biosyntheticcornea were examined.Methods: We used two different laser platforms on eyes of twodifferent species (porcine eyes and rabbit eyes). One was the VictusFemto-laser-System (Technolas Perfect Vision, Germany) and aPrototype Excimer Laser (M-Lase, Germany). On both lasers thebiosynthetic donor tissue was cut first (Diameter 600µm). The donortissue (thickness 350µm) was placed on a porcine eye to facilitateplacement under the laser aperture. The recipient porcine and rabbiteyes were treated with the same parameters (Diameter 600µm, depth350µm).In case of the eximer laser two specially designed aluminum maskshad to be used to achieve the desired diameter and shape of the cuts.The diameter, depth(stromal bed), thickness (donor tissue) andsurface structure were evaluated histologically (paraffin embedded,HE staining) and by OCT Imaging as well as electronmikroskopy.Results: Both laser systems allowed precise cutting of thebiosynthetic corneal implants as well as the donor tissue. Theexcimer laser treatment provided a smoother surface structureregarding the stromal bed, but the treatment duration wassignificantly longer than the treatment duration of the femto-secondlaser laser.Conclusions: Both laser platforms offer precise cutting options of thedonor and recipient tissue. Although both treatment setups in the exvivo animal trials showed promising results, the in vivo setup(presented separately) might be a greater challenge regardingpositioning, docking and cutting parameters.Commercial Relationships: Raphael T. Neuhann, None; KerstinM. Wand, None; Karin Kobuch, None; Michael Baumann, None;May Griffith, Univ. of Ottawa - OHRI (P); Mohammad M. Islam,None; Johannes Junger, MLase AG (E); Roland Ritter, TechnolasPerfect Vision (E); Chris Lohmann, NoneSupport: Euro Nanomed I-CARE, Transnational collaborativeprojectProgram Number: 3470 Poster Board Number: D0097Presentation Time: 11:00 AM - 12:45 PMImaging of Surface Defects and Biofilm Formation of ExtrudedKeratoprostheses Using Confocal MicroscopyHeather A. Durkee 1, 4 , Darlene Miller 2 , Victor L. Perez 2 , YohSawatari 3 , Aleksandra V. Rachitskaya 2 , Audina M. Berrocal 2 ,Eduardo C. Alfonso 2 , Jean-Marie A. Parel 1, 2 . 1 OphthalmicBiophysics Center, Bascom Palmer Eye Institute, University ofMiami Miller School of Medicine, Miami, FL; 2 Department ofOphthalmology, Bascom Palmer Eye Institute, University of MiamiMiller School of Medicine, Miami, FL; 3 Department of MaxillofacialSurgery, University of Miami Miller School of Medicine, Miami, FL;4 Department of Biomedical Engineering, College of Engineering,University of Miami, Coral Gables, FL.Purpose: The goal of the study was to examine surface structure andbiofilm deposits on keratoprostheses (KPro) that extruded frompatients secondary to Streptococcal Endophthalmitis. Confocalmicroscopy studies allow for fresh cultures in contrast to biofilmdetection using SEM which was presented previously (De La Cruz etal, IOVS Meeting abstracts 2011 52:345).Methods: Three keratoprostheses (2 Boston Type I and 1 ModifiedOsteo Odonto Keratoprothesis (MOOKP)) that extruded due toStreptococcal Endophthalmitis were stained with BacLightLive/Dead fluorescence assay and imaged with a Leica 5PS confocalmicroscope with 63x objective lens. Surface features of thekeratoprostheses were visualized using bright field illumination. Thebiofilm and bacteria distribution across the entire KPro wereevaluated using volumetric image sets that were captured fromdifferent regions of the keratoprostheses.Results: Biofilm deposits were present on all three of the KPros (seeFigure). Bacteria were found in higher quantities in areas of the opticwith surface scratches. Surface features corresponding to machiningmarks were found on the 2 Boston KPros and surface scratches onboth of the optical surfaces of all KPros.Conclusions: Surface irregularities on the KPros created duringfabrication allowed for more bacterial and biofilm growth. Improvingsurface quality of the implants might reduce the areas where bacteriaand biofilm can attach to the keratoprothesis.Figure 1: Image A (left) bright field of scratches on cylindrical faceof MOOKP (right) live dead stain of same section, biofilm depositsappear in dome-like shapes; Image B (left) bright field of deepscratch in cylindrical face of MOOKP, (right) bacteria deposit increvices of scratch; Image C (left) bright field of center anterior faceof KPro#1, machining marks can been identified as concentric rings(right) organic matter deposited on anterior face of optic; Image Dunidentified mark on posterior face of optic (inside anterior chamber)Commercial Relationships: Heather A. Durkee, None; DarleneMiller, None; Victor L. Perez, Alcon (C), Bausch & Lomb (C),Genentech (C), Cleveland Clinic Foundation (P), Alcon (F), Alcon(R); Yoh Sawatari, University of MIami (P); Aleksandra V.Rachitskaya, None; Audina M. Berrocal, thrombogenics (C),genentech (C); Eduardo C. Alfonso, Bio Tissue (C); Jean-Marie A.Parel, CROMA (F), InnFocus (F), Abeamed (F), University ofMiami (P)Support: USAMRMC Department of Defense W81XWH-09-1-0674; Florida Lions Eye Bank; NIH Center Grant P30EY14801; anunrestricted grant from Research to Prevent Blindness; Henri andFlore Lesieur Foundation (JMP)Program Number: 3471 Poster Board Number: D0098Presentation Time: 11:00 AM - 12:45 PM©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.
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