Analysis of aqueous humor total antioxidant capacity and its correlation with corneal endothelial health

Corneal endothelial decompensation is a serious condition that frequently requires treatment via corneal transplantation which contributes to a global shortage in donor corneas. Therefore, the purpose of this study was to analyze the influence of aqueous humor total antioxidant capacity (TAC) on corneal endothelial health. There is an urgent need for discovering protective factors to combat corneal endothelial cell (CEC) loss. For methods, we developed a cupric ion-based TAC (CuTAC) assay to analyze TAC level in a small volume of aqueous humor, that is, 10 μL per test, and examined the influences of ascorbic acid (AA) and antioxidant proteins on aqueous humor TAC. To broaden the investigation, we conducted a case-control study with patients classified into two groups, an insufficient endothelial cell density (ECD < 2100 cells/mm²) group, and a control group. These groups were formed based on baseline ECD values and were used to evaluate the influence of aqueous humor TAC and AA on overall corneal endothelial health. A CuTAC assay was used to accurately measure aqueous humor TAC without the need for sample dilution. After analyzing a total of 164 human aqueous humor samples, we found that AA was the major contributor to aqueous humor TAC (73.2%). In addition, TAC and AA levels in the IECD and control groups were both found to be significantly different (1.168 vs. 1.592 mM, p = 0.009 and 0.856 vs. 1.178 mM, p = 0.016). TAC and AA were considered independent protective factors against IECD with adjusted odds ratios of 0.02 (p = 0.017) and 0.023 (p = 0.033), respectively. In conclusion, aqueous humor TAC and AA contribute to the maintenance of sufficient corneal ECD, and our CuTAC assay can be a useful tool for analyzing TAC using only a small aqueous humor sample volume.