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H. Takahashi, N. Koizumi, N. Okumura, M. Ueno, Y. Sakamoto, J. Hamuro, S. Kinoshita; The Efficacy of A ROCK Inhibitor Containing Corneal Storage Medium for Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2829.
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The appropriate storage of donated human cornea is crucial for the surgical outcome following keratoplasty. We have previously demonstrated that Rho kinase (ROCK) inhibitor Y-27632 suppresses apoptosis in primate corneal endothelial cells (CECs) in vitro (Okumura, et al. IOVS 2009). The purpose of this current study was to demonstrate the usefulness of the Y-27632 as a supplement for corneal storage medium.
Corneas obtained from Japanese white rabbits were stored in Optisol-GS® (Bausch & Lomb, Inc., Rochester, NY) corneal storage media supplemented by 10 µM of Y-27632 at 4°C for 14 days. Optisol-GS® without Y-27632 was used as the control. Annexin V-positive apoptotic cells and PI-positive dead cells in CECs were evaluated by immunohistochemistry. Hoechst 33258 was used for counterstaining.
The mean percentage of Annexin V-positive cells and PI-positive cells in the Y-27632 group (2.0±0.7% and 12.0±7.9%, respectively) were significantly suppressed compared with those in the control group (4.8±1.5% and 29.1±24.2%, respectively) (p<0.01, p<0.05) after 14 days.
The results of this study demonstrate that Y-27632-supplemented corneal storage medium suppresses the apoptosis and cell death of CECs during corneal graft storage. We theorize that our results will also be applicable for human donor corneas in the clinical setting.
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