Cytotoxicity of Xenogeneic Pericardium Preserved by Epoxy Cross-Linking Agents

The aim of the study was to assess the cytotoxic effect of xenopericardial biomaterial treated with di- and pentaepoxides on the cell cultures in vitro. Materials and Methods. Samples of bovine and porcine pericardium were used in the work. Three different modes were employed for preservation: 1) 0.625% solution of glutaraldehyde (GA) and a two-fold change on days 2 and 7; 2) 5% solution of ethylene glycol diglycidyl ether (EGDE) changed on day 2; 3) 5% EGDE solution for 10 days, then 2% pentaepoxide solution also for 10 days. The cytotoxicity of the biomaterial was assessed by the extraction method. To determine the cytotoxicity of the biomaterial, EA.hy926 cells, multipotent mesenchymal stem cells (MMSCs), and fibroblasts were used. Cell viability was determined by the MTT test. The level of apoptosis and necrosis in the cell cultures was assessed by staining with acridine orange and ethidium bromide after cultivation with xenopericardial extracts employing different modes of preservation. Results. Extracts of bovine and porcine pericardium preserved with GA have been found to have the greatest toxic effect on the cell cultures showing 20-33% reduction of cell viability. Extracts from bovine and porcine pericardium preserved with di- and pentaepoxy compounds do not have a toxic effect on endothelial cells, MMSCs, and fibroblasts since cell viability reduction is by no more than 15%. The lowest level of apoptosis and necrosis is observed in the cell cultures under the influence of extracts from the pericardium, preserved with diepoxide and pentaepoxide compounds. Conclusion. According to the MTT test for cytotoxicity and determination of the level of apoptosis and necrosis in cell cultures, bovine and porcine pericardia treated with di- and pentaepoxides have been established to have no cytotoxic effect on the culture of endothelial EA.hy926 cells, MMSCs, fibroblasts in vitro, whereas GA, in comparison with di- and pentaepoxides, has a toxic impact on the cells.