A COMPARISON OF ENDOTHELIAL REGENERATION WITH TWO DIFFERENT ROCK INHIBITORS: AN EX-VIVO STUDY

Abstract

Purpose : To evaluate the efficacy of endothelial wound closure of two ROCKi from different classes in a “peel” and “scratch” wound model.

Methods : Research grade cornea pairs with a donor age ranging from 61 to 65 years old were procured from Lions Eye Bank. Peel and scratch wounds of 3 mm were created in corneas from the same donor pairs. The corneas were randomly assigned for treatment with either 10 µM of Y27632 or 1 µM of AR13324 and maintained in a stabilization culture media system for 21 days. Endothelial wound area and healing rates (WHR) were assessed with 0.2% trypan blue after wounding and weekly thereafter. Data were analyzed by ImageJ color threshold method and recovered areas were compared. All samples were also analyzed by alizarin red staining and scanning electron.

Results : In both groups, ROCKi treatment reduced the mean wounded area to <20% for scratched (p<0.001) and <60% for peeled (p<0.05). The mean WHR was 1.72 times higher for AR13324 and 2.15 times higher for Y27632 for scratched wounds when compared to peeled wounds. There was no significant difference between groups regarding WHR or final healed area. Alizarin Red staining and SEM showed cells indicative of enlarged hCEC over both scratched and peeled areas.

Conclusions : The use of either Y27632 or AR13324 in our ex vivo model induced significant endothelial wound recovery of both peeled and scratched wounds. However, this was more evident for scratched wounds. Our results showed that both AR13324 and Y27632 promoted endothelial wound closure. These ROCKi classes hold clinical potential in endothelial regenerative treatments.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.