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Jodhbir S Mehta, Gary S L Peh, Muhammad Rizwan, Khadijah Adnan, Sascha Naso, Evelyn Yim; Evaluation of corneal endothelial cell monolayer formation on synthetic guttata micro-structures to study the feasibility of corneal endothelial cell regenerative therapy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5282. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
A common indication for corneal transplantation is a dysfunctional corneal endothelium due to Fuchs’ endothelial dystrophy (FED). A diagnostic clinical sign of FED is excrescences on Descemet membrane (DM), guttata (CG). Minimal invasive corneal endothelial cell regenerative medicine procedures such as endothelial cell injection therapy and Rho-kinase inhibitor pharmacotherapy have been proposed as alternative treatments for FED patients. However, published studies on cell injection therapy and pharmacotherapy have been performed in animal models with healthy DM, and hence, do not recapitulate the topographical micro-environment of an FED patient
In this study, we developed a synthetic guttata model by fabricating micro-structures based on clinical examination of reported guttata parameters. We then systematically investigated the formation of corneal endothelial monolayer on these synthetic guttata to evaluate and predict the success of corneal endothelial cell injection therapy and pharmacotherapy. Evalutaion on monlyaer was performed using Na/K ATPAse, ZO-1 staining and SEM. Synthetic guttata of different heights/diameter ranging from 5-20 microns were created. Experiments were performed with B4G12 Cells and then repeated with primary human corneal cells.
The results suggested that guttata dimensions, density and spacing could greatly affect the fate of corneal endothelial cells in terms of migratory behavior and monolayer reformation. Densely packed synthetic guttata mimicking late stage FED hindered the reformation of an intact monolayer, while synthetic guttata mimicking the early stage of FED with lower height and lower density showed significantly improved monolayer formation. These results indicate that depending on the severity of the onset of FED, the pre-existing guttata found on the DM could interfere with the injected cells and corneal endothelial monolayer formation.
The characterization of the types or the presence of guttata on corneal DM of corneal patients could also help to determine patient-specific therapy procedure.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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