title: 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.
authors.affiliation: International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy.
authors.forename: Mohit
authors.identifier: 0000-0002-5186-068X
authors.initials: M
authors.lastname: Parekh
authors.affiliation: International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy.
authors.forename: Alessandro
authors.identifier: 0000-0003-4582-1948
authors.initials: A
authors.lastname: Ruzza
authors.affiliation: International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy.
authors.forename: Stefano
authors.identifier: 0000-0003-3983-3328
authors.initials: S
authors.lastname: Ferrari
authors.affiliation: International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy.
authors.forename: Diego
authors.identifier: 0000-0002-6038-3977
authors.initials: D
authors.lastname: Ponzin