A refractive and histopathologic study of excimer laser keratectomy in primates.

Using a 193-nm excimer laser, we produced wide-area, refractive keratectomies on 18 cynomolgus monkey corneas and followed them up for up to 18 months. All corneas developed some subepithelial haze by one month. Electron microscopy disclosed epithelial thickening, absence of Bowman's layer, and subepithelial activated fibroblasts surrounded by disorganized collagen. By six months, the haze faded to a variable degree, the epithelium regained normal thickness, and the collagen was more organized. Persistent corneal haze at 12 months in some corneas correlated with electronlucent spaces in the subepithelial zone. Corneas were 90 microns thinner centrally two weeks after myopic ablation, but returned to preoperative thickness by six months. Myopic flattening and hyperopic steepening of 6 diopters were targeted, and over 7 diopters of each were achieved initially. Regression of induced curvature stabilized over several months. At 18 months, 4.4 diopters of myopic flattening and 5.2 diopters of hyperopic steepening remained.

Binding of Pseudomonas aeruginosa to neutral glycosphingolipids of rabbit corneal epithelium.

35S-labeled Pseudomonas aeruginosa isolates were shown to bind to neutral glycosphingolipids (NGSLs) of rabbit corneal epithelia in culture by a thin-layer chromatogram overlay procedure. The lipids of the corneal epithelial cells grown in culture were extracted and partitioned into a chloroform-rich lower phase containing NGSLs and an aqueous upper phase containing gangliosides. By using a dot-blot assay, at least six times more radiolabeled P. aeruginosa isolates were shown to bind to the lipids in the lower phase compared with those in the upper phase. Thin-layer chromatography of the lower-phase lipids followed by staining with an orcinol spray revealed at least 10 NGSL components and several fast-migrating, nonglycosylated neutral lipid components (including cholesterol). 35S-labeled P. aeruginosa was shown to bind to NGSL components 1, 2, 5, 6, and 9. P. aeruginosa-reactive NGSL components 6 and 9 migrated with chromatographic mobilities similar to those of the standards ceramide trihexoside (CT) and ceramide monohexoside, respectively. Components 1 and 2 migrated slightly ahead of asialo GM1, and component 5 migrated faster than globoside but slower than CT. Among the various standards tested, P. aeruginosa bound to asialo GM1 and, to a lesser extent, to ceramide dihexoside and CT but not to GM1, GD1A, GM3, or ceramide monohexoside. It remains to be determined whether any of the five P. aeruginosa-reactive NGSL components of corneal epithelium identified in this study plays a role in the development of corneal infection. However, we have previously shown that component 9, one of the five P. aeruginosa-reactive NGSL components identified in this study, is present in significantly greater amounts in migrating epithelia than it is in nonmigrating epithelia (N. Panjwani, G. Michalopoulos, J. Song, G. Yogeeswaran, and J. Baum, Invest. Ophthalmol. Vis. Sci., in press). This may prove to be of biological significance because it is generally believed that traumatized (migrating) epithelia are more susceptible to infection than normal (nonmigrating) epithelia are.

Experimental corneal studies with the excimer laser.

The excimer laser is potentially capable of achieving wide area central corneal reprofiling because of its extreme precision and limited penetration into adjacent tissues. A beam modifying system designed for this application is described. Initial clinical studies in monkeys and in ten human patients with blind eyes were performed. Long-term clinical data and interim histologic analyses are available from these studies. The results indicate that following ablation with an ultraviolet laser in both humans and primates, the ablated tissue shows a normal healing reaction resulting in a mild to moderate stromal interface haze. The effects of this healing on best corrected vision must be elucidated through additional research. Some loss of refractive effect was seen early in the healing process with apparent stabilization.

Human excimer laser lamellar keratectomy. A clinical study.

The first ten blind human eyes in the United States to receive excimer laser (ArFl 193 nm) lamellar keratectomy (reprofiling) are presented. Seven of these patients were followed 6 to 12 months after ablation. All eyes are grossly clear in the region of ablation. Results of slit-lamp examination of all flattened ablated areas show mild superficial haze at the epithelial/stromal interface. This haze might not interfere significantly with vision in patients 7 to 10. Serial pachymetry and keratometry measurements, refraction, and digital keratoscopy show a progressive filling in of the excavated area by approximately two thirds but a loss of initial diopteric correction of only one third. Histopathologic analysis was obtained for four eyes. Transmission electron microscopy of three eyes enucleated 3 to 12 days after ablation shows 40-microns ablation depths through Bowman's layer and superficial stroma with minimal adjacent tissue damage and no inflammatory cells. The epithelium is increased in thickness by 50%, and firmly attached to the underlying stroma. A 4-month postablation specimen shows keratocyte activation with increased protein synthesis (presumed collagen and ground substance).

Human excimer laser corneal surgery: preliminary report.

The first human trial utilizing the argon fluoride excimer laser at 193 nm to produce a superficial keratectomy in ten human eyes has been described with the histopathological evaluation of four eyes and the longer gross appearance of six eyes at intervals extending to 10 months post-excimer laser treatment. The process of laser superficial keratectomy has proved to be one of the promising areas of surgical intervention for reconstructive or refractive keratoplasty in the future. Intensive investigations need to be undertaken on the corneal wound healing process following laser ablation as well as the nature, and long-term stability of the corneal excisions or induced refractive corrections. It is essential that the optimal laser parameters be established for the various refractive corrections and other corneal surgical techniques, and that pathophysiologic and histopathologic changes that have been induced by the excimer laser-corneal tissue interaction in animals and humans be critically and extensively analyzed.