Rho-associated kinase inhibitor decreases human corneal endothelial cell apoptosis

Session Details

Session Title: Moderated Poster Session: Cornea

Venue: Poster Village: Pod 1

First Author: : A.Achiron ISRAEL

Co Author(s): :    A. Feldman   L. Peer   L. Karmona   H. Avizemer   Z. Burgansky   V. Vishnevskia-Dai        

Abstract Details

Purpose:

Corneal endothelial cells (CEC) are essential for corneal function and clear vision. Several recent studies have demonstrated that selective Rho-associated kinase inhibitor (ROCKi) decreased apoptosis and promoted migration of both primate- and human-cultured CEC. Our aim was to evaluate whether exposure to ROCKi will promote human CEC survival in a commercial storage medium or following exposure to phacoemulsification

Setting:

The Edith Wolfson Medical Center, Holon, Israel

Methods:

Human-donor corneo-limbal rings were divided into fragments which were stored in commercial storage media with or without addition of 10µM ROCKi (Y-27632 dihydrochloride, AdooQ Bioscience,CA) for one week (natural decay study) and subject to phacoemulsification (stress exposure study) . Samples were dissociated into single cells by 0.25% trypsin digestion. CEC were targeted using the Anti-Human CD166 antibody. CEC survival was evaluated for early and late apoptosis rate with flow cytometric analysis of Annexin-V and propidium iodide (PI) double staining.

Results:

1.Natural decay study: Following 1 week of incubation with ROCKi, CEC demonstrated 64% reduced early apoptosis rate control (4.35%±1.07 vs. 12.18%±5.5, p=.026, mean difference=7.8%, and 41% reduced late apoptosis rate (5.5%±2.39 vs. 9.43%±2.61, p=.004, mean difference=3.9%), compared to control. 2.Stress exposure study: Following phacoemulsification, CEC exposed to ROCKi demonstrated a 37.06% reduction in early apoptosis rate (29.36%±4.33 vs. 46.65%±1.51,p=0.006) and 45.27% reduction in late apoptosis rate (17.6%±16.81 vs. 32.16%±26.30,p=0.007), compared to control. Subsequently, ROCK levels in apoptotic CEC were significantly lower in cell incubated with ROCKi than the control medium.

Conclusions:

This is the first report to study the effect of ROCKi on CEC loss in human tissue stored in storage media and following exposure to phacoemulsification. ROCKi reduced endothelial loss and thus may be used to limit or slow down CEC loss in donor corneal tissue during eye banking. ROCKi may also be used prior to cataract surgery, especially in high risk patients. ROCKi use may be a promising new method for promoting future graft survival and for preventing pseudophakic bullous keratopathy.

Financial Disclosure:

None