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Davine Sijnave, Tine Van Bergen, Evelien Vandewalle, Karolien Castermans, Nele Kindt, Lieve K M Moons, Ingeborg Stalmans; Inhibition of rho-associated kinase prevents excessive neovascularization after corneal trauma. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3234.
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The aim of this study was to investigate the effect of AMA0526, a selective and locally acting ROCK inhibitor on endothelial cells in vitro and corneal neovascularization (NV) in an in vivo corneal NV model.
In vitro, the effect of AMA0526 (0.1; 1 and 10 µM) on endothelial cell (HUVEC and HBMEC) proliferation (WST-1), FBS-stimulated migration (Boyden chamber) and viability (Caspase 3/7) was investigated. A mouse corneal micropocket (MCM) model was used to study the in vivo effect of the ROCK inhibitor on corneal neovascularization. Both eyes received a bFGF pellet and topical administration was applied once daily for 1 week post-implantation using AMA0526 (0.1%) in one eye and vehicle (PEG/H2O) in the contralateral eye. Treatment outcome was investigated by analysing vessel length, clock hours and NV area on digital photographs of the cornea taken at day 7 after pellet implantation. Histological outcome was evaluated by immunohistochemical staining for inflammation (CD45) and angiogenesis (CD31).
HBMEC and HUVEC proliferation was significantly (P<0.05) inhibited in a dose-dependent manner by ROCK inhibition. Moreover, AMA0526 (1 and 10 µM) also induced a dose-dependent reduction of FBS-stimulated endothelial cell migration, respectively by 29.12% and 40.17% (P<0.05 for both concentrations). The used doses did not affect the cell viability (P>0.05). In the MCM model, AMA0526 treatment significantly reduced NV area by 27.83% (P<0.05) and vessel length by 27.7% on day 7 (P<0.05), as compared to vehicle treated. These effects were associated with a decreased infiltration of inflammatory cells (P<0.005; 31.54% reduction, CD45) and a reduced blood vessel density (P<0.05; 40.14% reduction, CD31) in corneal sections taken at day 7.
The locally acting ROCK inhibitor, AMA0526 inhibits endothelial cell proliferation and migration in vitro and is efficacious in preventing corneal neovascularization after bFGF micropocket implantation. The results presented indicate that ROCK is an appealing target to treat and prevent corneal neovascularization and illustrate the potential therapeutic benefits of AMA0526.
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