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Published on: July 10, 2019
Effect of Rho-associated kinase inhibitor on human corneal endothelial cell apoptosis
Asaf Achiron1, Anna Feldman, Lily Karmona
1From the Department of Ophthalmology, Edith Wolfson Medical Center (Achiron, Karmona, Pe'er, Avizemer, Bartov, Burgansky), Holon, Sackler Faculty of Medicine, Tel Aviv University (Achiron, Feldman, Karmona, Pe'er, Avizemer, Bartov, Burgansky, Rosner, Vishnevskia-Dai), Tel Aviv, The Multiple Sclerosis Center, Sheba Medical Center (Feldman), and The Goldschleger Eye Institute, Sheba Medical Center (Rosner, Vishnevskia-Dai), Tel Hashomer, Israel.
Purpose:
To evaluate whether exposure to Rho-associated protein kinase (ROCK) inhibitor will promote human-cultured corneal endothelial cells (CECs) survival in a commercial storage medium.
Setting:
Edith Wolfson Medical Center, Holon, and Sheba Medical Center, Tel Hashomer, Israel.
Design:
Experimental study.
Methods:
Fragments of human donor corneolimbal rings were stored in commercial storage media for 1 week, half with the addition of 10 μM ROCK inhibitor (Y-27632). Evaluation of CECs for early and late apoptosis\necrosis rates was performed using anti-human CD166 antibody and flow cytometric double staining analysis of propidium iodide and Annexin V.
Results:
CECs of 6 corneolimbal rings demonstrated a reduced early apoptosis rate (4.35% ± 1.07% vs 12.18% ± 5.5%, P = .026) and a reduced late apoptosis\necrosis rate (5.5% ± 2.39% vs 9.43% ± 2.61%, P = .004) compared with control. Subsequently, the rate of apoptotic CECs expressing ROCK was significantly lower in cells exposed to ROCK inhibitor compared with cells that were not (19.01% ± 4.17 vs 30.42% ± 4.27, P < .001).
Conclusions:
ROCK inhibitor reduced endothelial cell loss in vitro and might be used to limit or slow CEC loss in donor corneal tissue during eye banking. This might be a promising new method for promoting future graft survival.
