Rho Kinase Inhibitor AMA0526 improves surgical outcome in a rabbit model of glaucoma filtration surgery

Abstract

AbstractPurpose To elucidate the effect of the ROCK inhibitor AMA0526 on the wound healing process and surgical outcome of glaucoma filtration surgery.Methods The in vitro effect of ROCK inhibitor AMA0526 on human brain microvascular endothelial cells (HBMEC) and human Tenon fibroblasts (HTF) was determined using a proliferation assay. Secondly the in vivo effect of topical AMA0526 0.3% TID was investigated in a rabbit model of glaucoma filtration surgery (n=5/time point). Treatment outcome was studied by clinical investigation of the bleb area as well as immunohistological analyses for inflammation (CD45), angiogenesis (CD31) and collagen deposition at day 8, 14 and 30 after surgery. Contralateral eyes were used as control and were treated with vehicle.Results A dose‐dependent reduction of HBMEC and HTF proliferation was measured after incubation with AMA0526. Incubation with the highest concentration of AMA0526 reduced proliferation of HBMEC and HTF to 22 and 35%. In the surgery model, AMA0526 significantly improved bleb area and survival compared to vehicle treated eyes. Immunohistological analyses showed significant reduction of inflammation, angiogenesis and collagen deposition after treatment with the ROCK inhibitor. Compared to vehicle, inflammation was decreased by 33% at 8 days and angiogenesis by 52% on day 8 and by 29% at 14 days. Collagen deposition was significantly reduced by 11 and 42% on day 14 and 30.Conclusion This study shows that AMA0526 is able to inhibit proliferation of microvascular endothelial cells and Tenon fibroblasts in vitro, and to improve glaucoma surgery outcome in rabbits. In addition to improved bleb area, AMA0526 led to decreased inflammation, angiogenesis and fibrosis.