Decreased keratocyte death after laser-assisted subepithelial keratectomy and photorefractive keratectomy in rabbits

Purpose

To compare keratocyte loss in the corneal stroma after laser-assisted subepithelial keratectomy (LASEK) and photorefractive keratectomy (PRK) in rabbits.

Setting

Department of Ophthalmology, University of Essen, Essen, and the Institute of Anatomy, University of Bochum, Bochum, Germany.

Methods

Laser-assisted subepithelial keratectomy and PRK were performed in rabbits and studied 1, 3, 10, and 20 days after surgery. Excimer photoablation was done unilaterally with a 6.0 mm ablation zone and an 80 μm depth, equivalent to −6.0 diopters. Keratocyte death was analyzed using DNA fragmentation-detecting terminal deoxynucleotidyl transferase deoxy-UTR-nick end labeling (TUNEL) assay and transmission electron microscopy.

Results

Numerous TUNEL-positive keratocytes occurred 1 day after PRK; the number decreased significantly after 3 days. After LASEK, significantly fewer TUNEL-positive keratocytes were noted at the early time points (P<.001 at 1 day; P≤.05 at 3 days). At 10 days, the number of TUNEL-positive keratocytes decreased in both groups but remained significantly higher after PRK than after LASEK (P<.001). Twenty days after both procedures, no significant signs of keratocyte death were found in the corneal stroma. Transmission electron microscopy revealed few apoptotic keratocytes after LASEK. After PRK, apoptotic keratocytes, characterized by chromatin condensation, apoptotic bodies, and cell shrinkage, were scattered in the stroma. The ultrastructural findings confirmed the results obtained with the TUNEL assay.

Conclusions

Laser-assisted subepithelial keratectomy induced significantly less apoptotic keratocyte death than PRK and promoted wound healing in the acute phase after photoablation. This procedure may offer the possibility of treating higher myopia with a decreased risk for developing wound healing-related complications known to occur after PRK.