Endothelial viability after DSAEK cutting of corneas stored in Tissue‐C organoculture medium and deswelled in Carry‐C medium (Alchemia)

Abstract

AbstractPurposeIn France, the adoption by an eyebank of a new technique for graft preparation and/or a new CE‐marked storage medium requires the submission of a “process” file to the competent health authority. This file must include pre‐clinical data on corneas stored in this medium or prepared with the new technique.AimTo analyze the endothelial alterations induced by the pre‐cutting with the microkeratome of corneas stored in Tissue‐C organoculture medium and deswelled in Carry‐C medium.MethodsTen pairs of corneas (donation for scientific purposes) were retrieved by in situ corneo‐scleral cutting and immersed in 100mL of Tissue‐C (Alchemia, Italy) and stored at 31°C in a dry incubator for 30 days (D). At the beginning (D2 to D5) and end (D30) of storage, the usual quality control was performed: central endothelial cell density (ECD) after incubation with 0.9% NaCl, transparency, thickness (OCT) and microbiological control. At D30, the corneas were transferred to a deswelling Dextran‐containing medium (Carry‐C, Alchemia) for 24 H. After randomization, one cornea of each pair was pre‐cut with the MORIA linear microkeratome (DSAEK, single pass, objective #150 µm) and put back in Carry‐C for 72H. The endothelial viability (viable ECD) of the all corneas was then measured by triple Hoechst‐Ethidium‐Calcein‐AM staining as we previously described (IOVS2011, Cornea2014).ResultsThe initial ECD was 2401 ± 815 for the DSAEK group and 2298 ± 805 cells/mm2 for the control group (p = 0.386) and at D30, respectively, from 1945 ± 587 versus 1868 ± 577 cells/mm2 (p = 0.421). One cornea was perforated. Graft thickness was 187 ± 112 um. For the other 9 pairs, the viable ECD (by definition always lower than bank ECD) was 1061 ± 448 for the DSAEKs and 1149 ± 409 cells/mm2 for the controls (p = 0.066). The pre‐cut grafts had fewer endothelial folds.ConclusionsPre‐cutting by bank of corneas stored in Tissue‐C/Carry‐C does not induce significant additional cell loss compared to control.