Cross‐linking of scleral collagen in the rabbit using riboflavin and UVA

PURPOSE: Scleral biomechanical weakness and thinning is known to be one of the main factors in the pathogenesis of progressive myopia. We tried to strengthen rabbit sclera by cross-linking scleral collagen using ultraviolet A (UVA) and the photosensitizer riboflavin. METHODS: Circumscribed 10 x10 mm sectors of the posterior--equatorial sclera of six chinchilla rabbit eyes were treated in vivo using a UVA double diode with 4.2 mW/cm(2) UVA at 370 nm and applying 0.1% riboflavin-5-phosphate drops as photosensitizer for 30 min. 1 day postoperatively biomechanical stress--strain measurements of three treated scleral strips were performed using a microcomputer-controlled biomaterial testing device and compared to non-treated contralateral control sclera. In addition, three treated eyes were examined histologically by light microscopy, TUNEL staining and electron microscopy to evaluate side-effects. RESULTS: Following the cross-linking treatment, the ultimate stress was 11.87+/-1.8 MPa versus 3.63+/-0.40 in the controls (increase of 227.9%, p=0.014), Young's modulus 27.67+/-4.16 MPa versus 4.9+/-.15 MPa in the controls (increase of 464.7%, p=0.021) and ultimate strain 92.2+/-7.43% versus 165.63+/-19.09% in the controls (decrease of 54.52%, p=0.012). Histologically, serious side-effects were found in the entire posterior globe with almost complete loss of the photoreceptors, the outer nuclear layer and the retinal pigment epithelium (RPE). CONCLUSIONS: Our new method of scleral collagen cross-linking proved very effective in increasing the scleral mechanical strength; the new treatment may represent an option for strengthening scleral tissue in progressive myopia. However, serious side-effects were observed in the outer retina. In future studies these side-effects could be avoided by reducing the irradiation dose below the cytotoxic level of the retina. Before its clinical application, the new method should be tested in a myopia animal model.