Endothelial Safety and Efficacy of Ex Vivo Collagen Cross-linking of Human Corneal Transplants

Abstract

To assess endothelial safety and efficacy of exvivo corneal collagen cross-linking (CXL) in human corneal transplants stored in 2 different culture media. Fellow-eye controlled laboratory study of exvivo human donor corneas. Three sets of paired human donor corneas, 5 pairs each, were stored in organ culture medium before deswelling either at 31 C or at room temperature. One eye of each pair was cross-linked by 0.1% riboflavin in hydroxylpropyl methylcellulose (HPMC) instillation for 10minutes followed by 10minutes of ultraviolet-A (9 mW/cm2) irradiation while contralateral eyes served as controls. In Set 1, endothelial cell densities were determined. In Set 2, paired samples were assigned to the 2 deswelling media and CXL efficacy was assessed comparing to untreated controls using collagenase-A-assisted enzymatic digestion. In Set 3, biomechanical testing was performed in the eye pairs (treated vs control) by stress/strain measurements. There was no difference in endothelial cell counts between CXL samples and controls (P= .21). No statistically significant difference in digestion dynamics was found between tissues stored in the 2 different culture media. Complete enzymatic digestion was slowed down by 3 hours in the cross-linked samples (P= .036). Stress needed for a 12% strain was increased by 34% in the treatment group compared to control (P= .04). Exvivo CXL of human donor tissue is an effective and safe procedure with no difference regarding efficacy between 2 commercially available deswelling media. Biochemical and biomechanical resistance were significantly increased after CXL. Patients requiring keratoplasty owing to corneal melting might benefit from the strengthening effect of preoperative CXL of donor tissue.