Preservation of Preloaded DMEK Lenticules in Dextran and Non-Dextran-Based Organ Culture Medium

Abstract

Purpose. To determine the optimum preservation conditions for preloading DMEK lenticules using organ culture system. Methods. 8.5 mm DMEK lenticules were stripped and preserved with endothelium flap-in for 4 days at RT in an IOL cartridge that was blocked with rubber stoppers from each end. In C1, tissues were collected from tissue culture medium (TCM) and preserved in TCM. In C2, tissues were collected from transport medium (TCM + 6% dextran T500) (TM) and preserved in TM. In C3, tissues were collected from TCM and preserved in TM. Mortality, glucose uptake, histological staining, tight junctions and cell apoptosis were studied post-preservation. Results. Mortality in C1, C2, and C3 were 49.40%, 8.53%, and 27.74%, with 40.7%, 13%, and 41.8% uncovered areas. Glucose uptake (mg/mL) was 0.32, 0.43, and 0.56 in C1, C2, and C3. PAS staining showed presence of DM and endothelium in C2 but not in C1 and with fewer cells in C3. ZO-1 was expressed in all the conditions. Polymorphism was higher in C1 and C3. Mild apoptosis was observed in C3. Conclusions. Dextran may play an important role in preserving the endothelial cells before and after stripping for trifolded (endothelium-in) preloaded DMEK lenticules.

Comparison of preservation and transportation protocols for preloaded Descemet membrane endothelial keratoplasty

“…Our previous study has shown that excising a DMEK graft that has been stored in dextran-based medium is easier (100% success) compared with excision from organ culture media without dextran (76% success). 18 Standardised preparation methods may further reduce graft wastage. In our earlier studies we have shown that preloading a DMEK graft in an IOL cartridge does not lead to significant cell loss (<10%) when preserved in dextran-based medium at RT for 4 days.…”

Ultrastructural findings in graft failure after Descemet membrane endothelial keratoplasty (DMEK) and new triple procedure

“…[19] On the other hand, Yoeruek and Bayyoud reported only a moderate loss of endothelial cells in precut tissue and safe donor graft preparation with or without dextran. [20,21] Parekh et al [22] concluded that dextran should be used in precut tissues to prevent the loss of endothelial cells.…”

Extracellular Vesicles Derived From Human Corneal Endothelial Cells Inhibit Proliferation of Human Corneal Endothelial Cells

“…One separate positive sample was induced with apoptosis using TACS nuclease and all the samples were viewed and imaged using inverted light microscope (Nikon Eclipse TS100, Nikon, Surrey, UK). The apoptotic cells were manually counted, and an average was recorded from five random areas (36).…”