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Michael Nicolas, Aurélien Pipparelli, Yvan Arsenijevic, Gilles Thuret, Philippe Gain, Francois Majo; ROCK inhibitor enhances adhesion and wound healing on human corneal endothelial cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1688. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Recently it was reported that the ROCK inhibitor Y-27632 promotes adhesion, inhibits apoptosis, increases the number of proliferating monkey corneal endothelial cells in vitro and enhance corneal endothelial wound healing both in vitro and in vivo. Here, we proposed to evaluate the effects of ROCK inhibitor on HCEC either in vitro or ex vivo, firstly to assess the potential of this compound to increase the number of corneal graft available for the clinic and secondly to validate the previous results obtained in animal models, a step required before potential clinical application.
Using organ culture human cornea (N=34), the effect of ROCK inhibitor was evaluated either in vitro or ex vivo. Toxicity, endothelial cell density, cell proliferation, apoptosis, cell morphometry, adhesion and wound healing process were evaluated by live/dead assay standard cell counting method, EdU labelling, Ki67, Caspase3, Zo-1 and Actin immunostaining.
In our study, we demonstrated for the first time in human endothelial cells ex vivo and in vitro, that ROCK inhibitor did not induce any toxicity effect and did not modulate metabolism activity. Compared to animal model, ROCK inhibitor treatment did not induce human endothelial cell proliferation. However, ROCK inhibitor significantly enhances corneal endothelial cell adhesion and wound healing.
These results strongly suggest that ROCK inhibitor is a promising and safe compound to improve the treatment of corneal endothelial dysfunction in human. ROCK inhibitor could be a potential therapeutic strategy in order to improve adhesion of transplanted human cultured endothelial cells. Furthermore, ROCK inhibitor treatment can increase in human the closure of endothelial cell defect.
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