Safety of a novel dual-chamber vial for corneal preservation

Our group developed a novel dual-chamber corneal storage vial (DCV) comprised of two compartments that become isolated by the corneal button and prevent epithelial and endothelial interaction. We hypothesize that the DCV results in similar levels of corneal swelling and corneal endothelial cell (EC) death compared to the standard vial (SCV) and is safe for corneal storage. To assess corneal thickness and EC density, human cornea pairs (N = 18) were recovered by the Florida Lions Eye Bank, with one cornea from each pair stored in the SCV and the other in the DCV. All cornea pairs were preserved in Optisol-GS media and stored at 2-8 °C for 2 weeks. Corneal thickness and EC density were evaluated on days 1, 7, and 14. Annexin V levels were measured in the corneal tissue (N = 12) to assess for cytotoxicity and apoptosis on day 14. Histological evaluation was performed on 3 additional cornea pairs on day 14. Three additional cornea pairs were collected to assess markers of endothelial cell death (casp3 and BID; N = 3) and endothelial cell function (ATP1A1, AQP1, and SLC4A4) were measured using RT-qPCR on 7 days (N = 3 for ATP1A1, AQP1 and N = 2 for SLC4A4). Preliminary results revealed no significant differences in central corneal thickness at all time points between corneal button stored in the SCV compared to the DCV (p = 0.648, p = 0.295, p = 0.180). There was no significant difference in EC density between chambers at all time points (p = 0.451, p = 0.573, p = 0.666). After 14 days of storage in the DCV and SCV, histopathology slides demonstrated no difference in stromal swelling or endothelial cell loss upon examination under light microscopy. No differences were found in markers of endothelial cell death and function. Given that the DCV demonstrates no difference in central corneal thickness, endothelial cell loss, or stromal swelling on histopathological examination during a two-week period, the DCV is a safe and viable alternative for corneal button storage. The DCV provides an alternative platform for human corneal storage for transplantation and allows the use of customized media in each compartment. The DCV can serve as a platform for future ex-vivo research, including the exploration and development of customized preservation solutions, such as the selective use of anti-fungal additives and other targeted interventions.