Biomechanical evidence of the distribution of cross-links in corneastreated with riboflavin and ultraviolet A light

Abstract

PURPOSE: To examine to which depth of the cornea the stiffening effect is biomechanically detectable. SETTING: Department of Ophthalmology, University of Dresden, Dresden, Germany. METHODS: Of 40 enucleated porcine eyes, 20 eyes were treated with the photosensitizer riboflavin (0.1%) and ultraviolet A (UVA) light (370 nm, 3 mW/cm2, 30 minutes); the other 20 eyes served as control. From each eye, 2 flaps of 200 microm thickness were cut with a microkeratome, and strips of 5 mm width and 7 mm length were prepared. Stress-strain behavior was measured with a material tester to characterize the stiffening effect. Five pairs of human donor eyes were tested in the same way. RESULTS: In porcine corneas, the stiffening effect was stronger in the anterior-treated flaps than in the posterior-treated flaps and the control flaps (P = .001). A 5% strain was achieved at a stress of 261.7 +/- 133.2 x 10(3) N/m2 in the anterior-treated flaps and 104.1 +/- 52.7 x 10(3) N/m2 in the anterior control flaps. The posterior-treated flaps (105.0 +/- 55.8 x 10(3) N/m2) and the posterior control flaps (103.7 +/- 61.8 x 10(3) N/m2) showed no difference (P = .95). A similar stiffening effect was observed in human eyes, but contrary to findings in porcine corneas, in human corneas the anterior control flaps were stiffer than the posterior control flaps (P = .027). CONCLUSIONS: Treatment of the cornea with riboflavin and UVA significantly stiffened the cornea only in the anterior 200 microm. This depth-dependent stiffening effect may be explained by the absorption behavior for UVA in the riboflavin-treated cornea. Sixty-five percent to 70% of UVA irradiation was absorbed within the anterior 200 microm and only 20% in the next 200 microm. Therefore, deeper structures and even the endothelium are not affected.