Photochemical Crosslinking as an Adjunct for Repairing Corneal Perforations in Rabbits

Objectives To assess the efficacy and tolerability of Rose Bengal‐induced photochemical crosslinking (RB‐PCL) as an adjunct to suturing for corneal perforation repair in rabbits. Methods A 4.5 mm corneal wound was created in New Zealand White Rabbits (right eyes), followed by suturing and RB‐PCL treatment using 532 nm green light (0.6 W/cm²) and 0.01% RB photosensitizer. A control group received sutures alone. Healing was evaluated at Weeks 1, 2, and 4 using slit‐lamp biomicroscopy, anterior segment optical coherence tomography (AS‐OCT), RT‐PCR for α‐SMA gene expression, and immunofluorescence for myofibroblast activation. Histopathological changes were assessed via Hematoxylin and Eosin (H&E) staining, and corneal endothelial cell counts were taken to assess cytotoxicity. Results RB‐PCL accelerated suture removal by Day 5, while sutures in the control group required longer retention. At Weeks 1 and 2, RB‐PCL demonstrated lower inflammation and reduced corneal opacity. By Weeks 2 and 4, α‐SMA expression was significantly reduced in the RB‐PCL group, indicating reduced myofibroblast activation and fibrosis. H&E staining showed complete healing in the RB‐PCL group by Week 2, whereas the control group had incomplete repair. By Week 4, the RB‐PCL group had better fiber arrangement, while the control group showed persistent fibrosis. Immunofluorescence confirmed reduced α‐SMA expression in the RB‐PCL group. No retinal damage was observed, and endothelial cell counts were similar between groups, indicating RB‐PCL's tolerability. Conclusions RB‐PCL improved corneal wound healing, reduced fibrosis and opacity, and allowed earlier suture removal without phototoxicity. These results suggest RB‐PCL is a promising adjunct for corneal perforation repair, enhancing both efficacy and tolerability.