Abstract
Purpose :
Proliferative vitreoretinopathy (PVR) is an intractable disease with poor visual prognosis due to contraction of the proliferative membrane. Ripasudil that has antifibrotic properties has potential applications in intraocular disease. The purpose of this study was to evaluate the effects of ripasudil in an in vitro PVR model.
Methods :
The retinal pigment epithelium (RPE) sheet was harvested from swine eyes using 2% Dispase. RPE sheets were cultured and passaged three times. Ripasudil was dissolved in 10% fetal bovine serum (FBS), and concentrations of 100 µM, 50 µM and 10 µM were prepared. Control medium contained only 10% FBS. The effect of ripasudil on RPE sheet growth was determined by expansion of cultured RPE sheet in the presence or absence of ripasudil. Wound scratch assay was used to examine the migration of confluent RPE cells cultured in 6-well dishes. A scratch was made using a 200-µL pipette tip. RPE cell proliferation was examined using Cell Counting kit-8. In vitrocontractile membrane formation assay was used to determine the effect of ripasudil in preventing contraction of type I collagen gel in the presence of cultured RPE cells. 2×10000/well RPE cells were seeded on collagen gel, and ripasudil was added to the medium after 48 h. After further incubation for 24 h, the collagen gel was peeled gently by 26G needle, and contraction was measured 4 hours later using Image J.
Results :
Ripasudil had no inhibitory effects in the migration assay and RPE sheet growth assay. However, inhibition by ripasudil was observed in the proliferation assay, although there was no significant statistical difference compared to control. In vitro contractile membrane formation showed that ripasudil at concentrations of 100 µM and 50 µM significantly inhibited contraction of collagen gel with cultured RPE cells (p < 0.05).
Conclusions :
Ripasudil did not inhibit RPE cell migration or proliferation, but significantly inhibited proliferative membrane contraction, which is the most important pathogenetic factor in PVR.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.