Abstract
Purpose :
To compare the effect of three commonly used anti-inflammatory eye drops: cyclosporin (0.05%), lifitegrast (5%), and tacrolimus (0.1%) on porcine corneal endothelial cell viability.
Methods :
We harvested porcine eyes and separated their corneascleral rims. Corneal endothelial cells were scraped from Descemet membrane after 0.5% trypsin treatment for 15 min. Cells derived from pig eyes were first plated into one well of a 12 well plate, proliferated to confluency in a low growth factor medium, passaged to a well of 6-well plate, and then passaged to a 96 well plate for drug treatment. Wells were separated into the following three treatment groups: cyclosporine 0.05%; lifitegrast 5%; tacrolimus 0.1%, as well as two control groups: a balanced salt solution (BSS) group and no treatment group. The cells were exposed to medium containing 20% cyclosporin (original 0.05% diluted to 0.01%), 20% lifitegrast (original 5% diluted to 1%), and 10% tacrolimus (original 0.1% diluted to 0.01%) and washed out after 1h, 2h, 4h, and 24h (N = 6 wells for each condition derived from 3 pig eyes). The survival rates of corneal endothelial cells were assessed by ATP quantification 48 h after the beginning of eye drop application.
Results :
Measurements of porcine endothelial cell viability after application of cyclosporine, lifitegrast, and tacrolimus revealed a statistically significant decrease down from 100±10% survival in the untreated group to 75±6%, 42±21%, and 0±0% (mean ± SD, N = 6), respectively. Compared to non-treated control group, decreases in viability after application of cyclosporine became statistically significant at 24h [75±6% (mean ± SD, N = 6)]. After application of lifitegrast, decreases became statistically significant at 1h and 24 hours [67±22% at 1h, 42±21% at 24h (mean ± SD, N = 6)]. After application of tacrolimus, decreases became statistically significant at 1h, 2h, 4h and 24h [58±21% at 1 h, 61±10% at 2h, 16±13% at 4h, 0±0% at 24h (mean ± SD, N = 6)].
Conclusions :
Porcine endothelial cell viability decreased progressively across application of cyclosporin, lifitegrast, and tacrolimus containing anti-inflammatory eye drops. These findings may serve as a resource for appropriate selection of anti-inflammatory eyedrops in the clinical setting and provide further insight into the study of inflammatory signaling and cell death pathways.
This is a 2021 ARVO Annual Meeting abstract.