Abstract
Purpose :
To assess the effect of Rho-Kinase Inhibitor (Ripasudil) to inhibit corneal neovascularization and lymphangionesis on murine corneal transplantation.
Methods :
Donor corneas (C57BL6) were transplanted onto BALB/c recipient mice. Placebo instillation (P group), 0.04% Ripasudil (R1 group), 0.4% Ripasudil (R2 group), 2.0% Ripasudil (R3 group) were instilled three times a day post transplantation immediately. Recipient’s corneas were harvested at day 14 post-transplantation (n=5 mice/group). The expression of VEGF-A, VEGF-C, VEGFR-1, VEGFR-2, VEGFR-3 and IFNγ were analyzed by quantitative real-time PCR. Immunohistochemistry was used to measure the area of CD31high (blood vessels) and LYVE-1high (lymphatic vessels). The graft infiltration of CD45+leukocytes were analyzed using flow cytometry.
Results :
The expression of VEGF-A, VEGF-C, VEGFR-1, VEGFR-2, VEGFR-3 and IFNγ were significantly decreased in R1, R2 and R3 group compared to P1 group. The area of angiogenesis in the graft was significantly reduced in R2 and R3 compared to P1 (P: 6.3±1.8%, R1: 6.6±2.4%, R2: 2.6±0.9%, R3: 1.6±1.1%, p=0.002). The area of lymphangionesis in the graft was significantly reduced in R2 and R3 group compared to P group (P: 6.3±1.8%, R1: 6.6±2.4%, R2: 2.6±0.9%, R3: 1.6±1.1%, p＜0.001). The frequencies of CD45+ leukocyte were significantly decreased in R2 and R3 group compared to P group (P: 16.4±0.46%, R1: 15.8±0.48%, R2: 6.7±2.6%, R3: 7.0±2.9%, p＜0.001).
Conclusions :
Ripasudil instillation could inhibit the immune rejection by suppressing neovascularization and inflammation in corneal transplantation.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.