Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaResults: The optical density of the cells is measured via an MTTassay and compared to controls (cells in PBS alone) to obtain the cellviability after treatment with UVA-RF combination. Dose responsecurves are plotted against exposure time for low and high irradiancesand the time required to get 50% cell viability (=EC50) is estimatedusing a non-linear regression model and compared for high and lowirradiance treatments. Results show cell viability decreasing as afunction of increasing energy dose applied.Conclusions: This custom chamber allows for the comparison ofcytotoxicity levels of UVA-RF combination on human cornealendothelial cells while controlling the gaseous composition andtemperature of the chamber close to in vivo conditions.Commercial Relationships: Radha Pertaub, Avedro Inc (E); MarcD. Friedman, Avedro Inc (E); David Muller, Avedro Inc (E)Program Number: 1654 Poster Board Number: D0289Presentation Time: 8:30 AM - 10:15 AMA Nonsynthetic, Biological Carrier for Cultivated HumanCorneal Endothelial Cells (HCECs) for potential therapeuticpurposesJesintha Navaratnam, Eli Gulliksen, Kristine Ustgaard-Andersen,Jon K. Slettedal, Liv Drolsum, Bjorn Nicolaissen, AboulghassemShahdadfar. Center for Eye Research, Oslo University Hospital,Oslo, Norway.Purpose: The aim of our ongoing study is to establish a carrier forHCECs for therapeutic purposes. In the present study we investigatethe feasibility of using nonsynthetic, biological carrier for cultivatedHCECs.Methods: Descemet’s membrane with the attached endothelial cellswas carefully dissected from human corneas in small strips. One partharvested as non-cultured cells and the other cultivated in cornealendothelial cell growth medium for 6 weeks at 37 degree Celsiuswith 5% CO2 in a humidified atmosphere and the medium waschanged every 2-3 days. The cultivated HCECs were seeded onacellular, nonsynthetic carrier and cultivated for further 3 weeks incorneal endothelial cell expansion medium that was changed every 2-3 days. Cultivated HCECs on nonsynthetic carrier and non-culturedHCEC were comparatively analyzed by qRT-PCR, electronmicroscopy (EM) and immunohistochemistry.Results: In our study the cultivated HCECs seeded on nonsyntheticcarrier formed a stable monolayer. Our results show that thecultivated HCECs seeded on nonsynthetic carrier and the nonculturedHCECs are functional and express stem cell markers whenanalyzed by qRT-PCR. The expression levels of markers associatedwith neural crest, stem cells and corneal endothelial function (SNAI1,SNAI2, SOX9, NES, ZO-1, CX43, VADC2 and VADC3) are higherin cultivated HCECs seeded on nonsynthetic carrier compared tonon-cultured HCECs. The structure of cultivated HCECs seeded onnonsynthetic carrier on transmission EM is very similar compared toex vivo HCECs.Conclusions: The preliminary results show that nonsynthetic carrierused in this study can potentially be used in therapeutic purposes inthe future.Commercial Relationships: Jesintha Navaratnam, None; EliGulliksen, None; Kristine Ustgaard-Andersen, None; Jon K.Slettedal, None; Liv Drolsum, None; Bjorn Nicolaissen, None;Aboulghassem Shahdadfar, NoneProgram Number: 1655 Poster Board Number: D0290Presentation Time: 8:30 AM - 10:15 AMDifferences in corneal endothelial abnormalities in the centraland intermediate zones in Fuchs’ corneal dystrophyHisataka Fujimoto, Takeshi Soma, Yoshinori Oie, Shizuka Koh,Motokazu Tsujikawa, Naoyuki Maeda, Kohji Nishida. OsakaUniversity, Suita, Osaka, Japan.Purpose: Fuchs’ corneal dystrophy is a progressive cornealendothelial dystrophy that causes irreversible endothelial change andbullous keratopathy. To investigate the regional differences in theabnormality, we examined the corneal endothelium at multiple sites,including the periphery.Methods: 17 eyes of 9 patients (6 women and 3 men; 30-80 yearsold) with Fuchs’ corneal dystrophy were studied at Osaka UniversityHospital. We used a newly developed non-contact specularmicroscope (NIDEK CEM-530) to examine the intermediate zone 27degrees in the periphery from the corneal center in addition to thecenter and the peripheral 5 degrees from the center. We measured thesize of the degenerative area with guttata and the cellular density inthe residual intact area.Results: The abnormal areas in the central zone (center and 5 degreesfrom the center) represented 65.4 ± 33.7% of the total area, and thosein the 27 degrees of the peripheral zone represented 27.1 ± 34.2% ofthe total area. The results differed significantly (P < 0.001). Theabnormal areas in the 27 degrees of the peripheral zone of 16 of 17eyes were significantly (P< 0.05) larger than those in the center. Inthe 6 points 27 degrees of the peripheral zones, the abnormal areasinferiorly were significantly (P < 0.001) larger than superiorly, by theusage of z score analysisConclusions: In Fuchs’ corneal dystrophy, the corneal endothelialcells degenerate more rapidly in the central zone than theintermediate zone. This finding might offer new information tofacilitate an understanding of the disease mechanisms.Commercial Relationships: Hisataka Fujimoto, None; TakeshiSoma, HOYA corporation (R), Santen Pharmaceutical Co., Ltd (F),Otsuka Pharmaceutical Co., Ltd (F); Yoshinori Oie, Santen (F),HOYA (F); Shizuka Koh, Santen, Inc. (R), Johnson & Johnson (R),Topcon (R), Otsuka Pharmaceutical Co. (R); Motokazu Tsujikawa,Shionogi & Co. (C), Daiichi Sankyo Co. (F), Daiichi Sankyo Co. (R),Santen Co. (R), AMO Co. (R); Naoyuki Maeda, Topcon (F), Santen(R), Otsuka (R), Oculus (R), HOYA (R); Kohji Nishida, Alcon (C),Alcon (F), HOYA (F), Senju (F), Pfizer (F), Santen (F), OsakaUniversity (P)Program Number: 1656 Poster Board Number: D0291Presentation Time: 8:30 AM - 10:15 AMDevelopment and Characterization of Decellularized HumanCorneal Stroma as a Scaffold for Tissue EngineeringRadhika Tandon 1 , Sujata Mohanty 2 , Himi Singh 1 , Deepika Gupta 3 ,Seema Sen 4 , Seema Kashyap 4 , Amit K. Dinda 5 , Manjeet Jassal 3 ,Ashwini K. Agrawal 3 . 1 Department of Ophthalmology, Dr. RajendraPrasad Centre for Ophthalmic Sciences, All India Institute of MedicalSciences, New Delhi, India; 2 Stem Cell Facility, All India Institute ofMedical Sciences, New Delhi, India; 3 SMITA Research Labs,Department of Textile, Indian Institute of Technology, New Delhi,India; 4 Department of Ocular Pathology,Dr. Rajendra Prasad Centre©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.