Preloaded descemet membrane endothelial keratoplasty grafts with endothelium outward: a validation study of the DMEK rapid device

Abstract

AbstractPurposeTo validate a device (DMEK Rapid, Geuder AG, Germany) for preloading and transporting DMEK grafts with endothelium facing outwards.MethodsStripped DMEK tissues were divided into 2 groups: internal transportation simulations (study A) and cross‐country transport study (Study B). Study A was designed to investigate the influence of 2 types of media, tissue culture medium (TCM; n = 12) or transport medium (TM; n = 12), on DMEK tissues preloaded in the DMEK Rapid device. The tissues were placed on the laboratory rocker for 4 days at room temperature. For Study B, DMEK tissues (n = 9), preloaded with TM were shipped from Italy to the UK and analysed within 72 hr. For both studies, endothelial cell loss (ECL – trypan blue staining), morphology (alizarin red staining), viability (live/dead staining) and expression of biomarker (ZO‐1 immunostaining) were performed.ResultsPost‐stripping ECL was 7.5 ± 66.2% in TCM group, compared to 9.1 ± 64.9 from the TM group (p = 0.54). Post‐simulation ECL was 14.3 ± 63.8% from the TCM group and 11.5 ± 64.6% from the TM group (p = 0.08). Alizarin red (n = 4 in each group) showed the low polymorphism in both media groups. Weka segmentation of the live/dead analysis showed 78.83 ± 62.78% viability in TCM group and 80.83 ± 63.48% viable cells in TM group (p = 0.60). ZO‐1 immunostaining (n = 4 in each group) showed the presence of tight junctions in both the groups. In study B (n = 9), post‐stripping ECL was 10.1 ± 64.5% and 3.3 ± 66.6% after transporting the tissues from Italy to the UK (p = 0.05). Alizarin red (n = 3) showed proper tissue morphology and viability of 85.16 ± 65.33% was detected (n = 3). ZO‐1 (n = 3) was expressed in all tested grafts.ConclusionsPreloading DMEK tissue with endothelium outwards and transported within 72 hr using the DMEK Rapid device is safe and can be pursued for clinical application.