Preparation and endothelialization of decellularised vascular scaffold for tissue-engineered blood vessel

The proliferation of cells on the decellularised tissues fixed by chemical crosslinking agent is retarded for cytotoxicity of crosslinked tissues. To overcome this disadvantage, we prepared the decellularised vascular scaffold through fixing the porcine thoracic arteries with 40 mL/L ethylene glycol diglycidyl ether (EGDE), and reduced the cytotoxicity of this scaffold by treating it with lysine and coating it with type I collagen, finally endothelialized it in vitro. The EGDE-fixed porcine thoracic arteries were examined morphologically. The fixation index determination and the biomechanics test were also performed. Human umbilical vein endothelial cells (HUVECs) were seeded on the type I collagen-coated surface of different modified vascular tissues (fixed with glutaraldehyde or EGDE or EGDE + lysine), and the growths of HUVECs on the specimens were demonstrated by means of MTT test. Finally, HUVECs were seeded on the luminal surface of the modified porcine vascular scaffolds which were respectively treated in the same manner described above, and then cultured for 7 days. On the seventh day, the HUVECs on the specimens were examined by means of light microscopy, scanning electron microscopy and transmission electron microscopy (TEM). The antigenicity of the vascular tissues can be diminished by EGDE through getting rid of cell in the vascular tissues or reducing the level of free amino groups in the vascular tissues. In this study, it was also found that the EGDE-fixed porcine vascular tissues appeared similar to the native porcine vascular tissues in color and mechanical properties. After treated by 2% lysine and coated with type I collagen, the EGDE-fixed porcine vascular tissues were characterized by low cytotoxicity and good cytocompatible. The HUVECs can proliferate well on the modified vascular tissues, and easily make it endothelialized. The results showed that the modified porcine vascular scaffolds should be a promising material for fabricating scaffold of tissue-engineered blood vessel.