Endothelial Wound Repair of the Organ-Cultured Porcine Corneas

ABSTRACTPurpose: To assess whether injured porcine endotheliumof small and large corneoscleral disc differ in its reparative/regenerative capacity under various conditions of organ culturestorage.Material and methods: 166 paired porcine corneas were trephinedto obtain tissues with diameter 12.0 mm and 17.5 mm (with area neighboring endothelial periphery). In tested discs, central endotheliumwas mechanically wounded. Density of live endothelial cells (LECD), percentage of dead cells (%DC), coefficient of variation and cell hexagonality were assessed in central and paracentral endotheliumfollowing 5- or 9-day incubation in medium with 2% or 10% fetal bovine serum. The parameters were assessed also in fresh and intact cultured discs. Dead endothelial cells (EC) were visualized by trypan blue, cell borders by AlizarinRed S dye. Endothelial imprints were immunoassayed for the proliferation marker Ki-67 and the nucleolar marker fibrillarin.Results: In fresh corneas, the LECD/mm2 (mean ± standard deviation...