Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - Corneamay be responsible, in part, for the delayed epithelial wound closurein diabetic rat corneas.Commercial Relationships: HAIJING SUN, None; Feng Wang,None; Ilham Bettahi, None; Fushin X. Yu, NoneSupport: R01 EY017960Program Number: 2566 Poster Board Number: D0366Presentation Time: 2:45 PM - 4:30 PMClinical Factors Can Predict the Outcome of AutologousCultivated Limbal Epithelial TransplantationAnupam Bagdi 1 , Sayan Basu 1 , Hasnat Ali 2 , Virender S. Sangwan 1 .1 Cornea, L V Prasad Eye Institute, Hyderabad, India; 2 Biostatisticsand Clinical Epidemiology, L V Prasad Eye Institute, Hyderabad,India.Purpose: It is difficult to prognosticate the outcome of stem cellbased therapy for ocular surface disease because the risk factorspredisposing to failure of surgery are not clearly known.To addressthis issue,this study aimed to identify the clinical Indicatorsassociated with failure of autologous cultivated limbal epithelialtransplantation for the treatment of Limbal Stem Cell Deficiency(LSCD).Methods: This retrospective study included 526 eyes of patientssuffering from unilateral LSCD following ocular surface burns whounderwent autologous ex-vivo cultivated limbal epithelialtransplantation between 2001 and 2011. This procedure involvedobtaining a one-clock hour wide limbal biopsy from the healthyfellow eye and cultivating the limbal epithelial cells on a deepithelisedhuman amniotic membrane graft for 10-14 days. Aconfluent epithelial sheet along with the amniotic membrane graftwas then transplanted onto the affected eye after clearing away thepathologic tissue covering the cornea. Post-operatively success oftransplantation was defined as a completely epithelized, avascularand clinically stable corneal surface. A multivariate analysis wasperformed using multiple logistic regression to analyze the strengthof association between the pre-operative, intra-operative and postoperativeclinical factors and recurrence of LSCD.Results: Of the 526 eyes, the ocular surface was successfullyrestored in 292 (55.5%) eyes at a mean follow-up of 1.4 years.Among all the pre-, intra- and post-operative clinical factors thatwere assessed: presence of symblepharon (OR=1.2, P< 0.001),previous ocular surgeries (OR= 1.3, P= 0.02), pediatric age group(OR=1.4, P= 0.04) and simultaneous keratoplasty (OR=3.2, P= 0.02)were found to be associated with greater chances of failure ofautologous cultivated limbal epithelial transplantation.Conclusions: Patients with extensive symblepharon, patientspreviously having undergone multiple ocular reconstructiveprocedures, children and patients requiring keratoplasty along withautologous limbal transplantation are at higher risk of recurrence ofLSCD and need to be counseled accordingly.Commercial Relationships: Anupam Bagdi, None; Sayan Basu,None; Hasnat Ali, None; Virender S. Sangwan, NoneSupport: Partnership grant from Champalimaud Foundation, Lisbon,Portugal; Hyderabad Eye Research Foundation, Hyderabad, India;Department of Biotechlonology, New Delhi, India(BT/01/COE/06/02/10)Program Number: 2567 Poster Board Number: D0367Presentation Time: 2:45 PM - 4:30 PMUtilizing contact lenses as carriers for human corneal limbalepithelial and induced pluripotent stem cellsNir Erdinest, Abraham Solomon. Ophthalmology, Hadassah HebrewUniv Med Ctr, Jerusalem, Israel.Purpose: To delineate the best technique for culturing and carryinghuman corneal limbal epithelial cells and induced pluripotent stem(iPS) cells on contact lenses (CLs) for the purpose of celltransplantation for limbal stem cell deficiency (LSCD).Methods: Limbal explant sections from corneoscleral rims,remaining from corneal transplantation, or induced pluripotent stem(iPS) cells were placed on the inner aspect of 5 types of CLs coatedwith or without 0.1% gelatin. The CLs used included lotrafilcon A(Air Optix Night & Day® Aqua), vifilcon A (Focus Monthly®),lotrafilcon A (Focus Night & Day® Aqua), etafilcon A (1-DayAcuvue®) and nelfilcon A (Focus Dailies®). Plastic culture platesand the human amniotic membrane were used, respectively, asgrowth platform controls for the CLs. Comparison between thegrowth patterns on the different CLs and the gelatin coating wasmade with regard to epithelial proliferation rate and epithelial cellmorphology utilizing light and electron microscopy.Results: All the growth platforms yield monolayers culture cells withuniform epithelial cell morphology. There were marked differencesin growth patterns between the different CLs. The explants placed onsilicone hydrogel CL lotrafilcon A (CIBA Vision Corporation,Duluth, GA, USA) showed the fastest proliferative and migratoryrates, with CLs covered with gelatin. Explants applied on lotrafilconA were tightly attached to it, and growth was detected first on day 4.Lotrafilcon A (Air Optix Night & Day® Aqua) reached 91.2±8.5%confluence by day 16±2, and Lotrafilcon A (Focus Night & Day®)reached 93.7±6.2% confluence by day 14±3. iPS cells which wereseeded on CLs were attached to lotrafilcon A by day 3. Cellsexamined under light microscopy were morphologically similar tocorneal epithelium grown on 6 wells culture plate and amnioticmembrane. Electron microscopy revealed cell-to-cell links andmultiple cell layers. Microvilli and cell projections were also evidentand most probably served as anchorage points on the CLs surface.Conclusions: Silicone hydrogel CL lotrafilcon A was advantageousover the other types of CLs for culturing and carrying corneal limbalepithelial and iPS cells. These CLs may be used as possible carriersfor cultured human ocular surface epithelial cell sheets as a part of acell therapy strategy in LSCD.Commercial Relationships: Nir Erdinest, None; AbrahamSolomon, NoneProgram Number: 2568 Poster Board Number: D0368Presentation Time: 2:45 PM - 4:30 PMLumican Is Required for Epithelium Migration during Healingof Corneal Epithelium DebridementJianhua Zhang, Vivien J. Coulson-Thomas, Yong Yuan, OsamuYamanaka, Hongshan Liu, Winston W. Kao. Ophthalmology,University of Cincinnati, Cincinnati, OH.Purpose: Purpose: Lumican, a member of small leucine richproteoglycan (SLRP), is a matrikine besides serving as anextracellular matrix ECM component and regulator of collagenfibrillogenesis. Previous studies have shown that lumican promoteshealing of corneal epithelial debridement via enhanced epithelial cellproliferation. The current study attempts to determine whetherlumican plays an additional role in promoting corneal wound healingby enhancing epithelial cell migration.Methods: Methods: Adult lumican null (Lum-/-), hemizygous(Lum+/-) and wild type (Lum+/+) mice were anesthetized andsubjected to circumvent corneal epithelium debridement (2 mm indiameter) with Agerbrush®. Corneas were isolated at differentintervals (0 through 6 h) and subjected to whole mount IF in order tocharacterize the epithelial cell migration by analyzing the expressionand localization of cytoskeletal and adherens components, e.g.,paxillin, etc. The role of lumican on stress fiber and filopodia©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.