Endothelial reaction to perforating and non-perforating excimer laser excisions in rabbits

With an ArF excimer laser (193 nm, 750 mJ/cm2, 20 Hz) and a special slit-mask system, perforating and non-perforating linear keratectomies were performed in 55 rabbit corneas with a follow-up from 1 hour to 6 months. Varying the pulse number according to ablation rate (0.8 micron/pulse) and corneal thickness, four linear radial excisions (3 mm length, 70 microns width) of increasing depth (70%, 80%, 90%, 100% perforation) were produced. The corneas were processed for light microscopy, scanning and transmission electron microscopy, and vital staining of the endothelium. Except for mild cell contact alterations and discrete single cell damage in the 90% deep excisions, no endothelial damage could be detected after non-perforating keratectomies. Minute (less than 20 microns) and small (20 to 100 microns maximal diameter) perforations induced cell enlargement, formation of pseudopodia, rosette-like figures, multi-nucleated giant cells, and ultimately uniform reformation of the cell pattern (1 hour to 7 days postoperatively). Larger excimer laser defects of Descemet's membrane (greater than 100 microns) were overgrown by dedifferentiated endothelial cells producing a new PAS-positive basement membrane. Vital staining revealed the complete and stable reorganization of the endothelium over these lesions within 6 months. Our observations are similar to those reported on the endothelial repair process following other surgical manipulations (knife incisions, direct Nd:YAG-laser trauma) and support the applicability of excimer lasers for corneal trephination in patients.