EEBA Program (PDF/3MB) - EEBA - Annual Meeting

ROCK INHIBITOR ENHANCES ADHESION AND WOUND HEALING ON HUMAN CORNEALENDOTHELIAL CELLS EX VIVO AND IN VITROM. Nicolas¹, A. Pipparelli¹, G. Thuret², P. Gain², F. Majo¹¹Jules-Gonin Eye Hospital, Switzerland; ²Faculty of Medicine, University of Saint Etienne, FrancePurpose: Maintenance of corneal transparency is crucial for vision. When endothelial cell density fallsbelow a critical threshold, the barrier and “pump” functions of the endothelium are compromised andthis results in the formation of a corneal oedema and loss of visual acuity. The conventional treatmentfor such severe disorder is transplantation of cornea. Unfortunately, there is a worldwide shortage ofdonor corneas. Recently it was reported that the ROCK inhibitor Y-27632 promotes adhesion, inhibitsapoptosis, increases the number of proliferating monkey corneal endothelial cells in vitro and enhancecorneal endothelial wound healing both in vitro and in vivo in animal models. Here, we proposed toassess the potential of this compound to increase the number of corneal graft available for the clinic.Methods: Using organ-culture human cornea (N=34), the effect of ROCK inhibitor was evaluatedeither in vitro or ex vivo. Toxicity, ECD, cell proliferation, apoptosis, cell morphometry, adhesionand wound healing process were evaluated by standard cell counting method, EdU labelling, Ki67,Caspase3, Zo-1 and Actin immunostaining.Results: In our study, we demonstrated for the first time in human endothelial cells ex vivo and invitro, that ROCK inhibitor did not induce any toxicity effect and did not alter cell viability. Compared toanimal model, ROCK inhibitor treatment did not induce human endothelial cell proliferation. However,ROCK inhibitor significantly enhances corneal endothelial cell adhesion and wound healing.Conclusions: The present study shows that Y-27632, a selective ROCK inhibitor, has no effect onhuman corneal endothelial cells proliferative capacities, but alters cellular behaviours. It induces achange in cell shape, increases cell adhesion and enhances wound healing ex vivo and in vitro. Eventhe results were promising in animal models; this inhibitor is not able to induce human endothelial cellproliferation of organ-culture human cornea.54