An Investigation of the Effects of Riboflavin Concentration on the Efficacy of Corneal Cross-Linking Using an Enzymatic Resistance Model in Porcine Corneas

PURPOSE: To investigate riboflavin concentration on enzymatic resistance following corneal cross-linking (CXL). METHODS: Ninety-six porcine eyes were divided into five groups in two treatment runs. Group 1 remained untreated. Group 2 received riboflavin 0.05%, group 3 riboflavin 0.1%, group 4 riboflavin 0.2%, and group 5 riboflavin 0.3%. Treated eyes underwent CXL with ultraviolet A at 9 mW/cm(2) for 10 minutes. Eight-millimeter discs from each cornea were submerged in pepsin digest solution. In the first run, disc diameters were measured daily. After 10 days, dry weights were recorded from five samples in each group. In the second run, dry weights were recorded in five samples in each group at 10 and 20 days. RESULTS: CXL-treated corneas took longer to digest than untreated (P < 0.001). Although eyes treated with higher riboflavin concentrations generally took longer to digest, there were no significant differences between groups (P = 0.3). Dry weights at 10 days demonstrated, with each increase in concentration, an increase in weight of residual undigested tissue (P < 0.001). In the second run, with each increase in riboflavin concentration there was an increase in weight of residual tissue (P < 0.001) at 10 days. At 20 days, the dry weight was lower with 0.05% riboflavin compared to 0.3% (P < 0.001) and 0.2% and 0.1% solutions (P < 0.05), with no other difference between groups. CONCLUSIONS: There is a consistent dose-response curve with higher concentrations of riboflavin achieving greater CXL efficacy, suggesting that manipulation of riboflavin dosage as well as the UVA protocol can be used to optimize CXL.