Evaluation of the Physiological Corneal Intrastromal Riboflavin Concentration and the Corneal Elastic Modulus After Violet Light Irradiation

PURPOSE: KeraVio is a corneal crosslinking treatment modality that utilizes violet light (VL)-emitting glasses and topical epithelium-on riboflavin administration. We focus on the new KeraVio protocol without riboflavin. This study aims to quantify the physiological intrastromal concentrations of riboflavin in corneas without riboflavin decreases and evaluate the biomechanics of corneas after VL irradiation. METHODS: Twelve human donor corneas were included in this study and randomly categorized into four groups. The corneas underwent four imbibition techniques (physiological riboflavin without drops, epithelial [epi]-on with 0.05% flavin adenine dinucleotide [FAD], epi-off with FAD, and 0.1% riboflavin epi-off). Corneas in the FAD epi-on, FAD epi-off, and riboflavin epi-off groups were instilled with the respective solution every 2 minutes for 30 minutes. An ex vivo experiment was conducted with 24 porcine corneas arranged into three treatment groups and one control group. Corneas in the KeraVio with FAD epi-on group were treated with VL irradiation at 0.31 mW/cm(2) for 4.8 hours (5.4 J/cm(2)) and simultaneously received FAD drops every 30 minutes during the VL irradiation. Corneas in the group with KeraVio without FAD epi-on were only treated with VL irradiation (5.4 J/cm(2)). RESULTS: We identified the original physiological riboflavin of human corneal stroma at a concentration of 0.31 ± 0.03 µg/g, but its value was approximately 39-fold smaller than that in the 0.1% riboflavin epi-off group. The group with KeraVio without FAD and the standard corneal crosslinking group showed a significant increase in biomechanical stability compared with the controls, whereas the elastic modulus in the treated groups was equivalent. CONCLUSIONS: We preliminarily identified physiological riboflavin in human corneas without adding riboflavin drops. The VL exposure may strengthen the corneal biomechanics without requiring the use of additional riboflavin drops. TRANSLATIONAL RELEVANCE: We preliminarily identified physiological riboflavin in the human cornea without adding riboflavin drops. VL irradiation without riboflavin drops may increase the corneal stiffness using physiological riboflavin.