生物性组织工程血管支架的制备及其内皮化研究

研究经乙二醇缩水甘油醚（EX-810）处理的猪动脉的性能特点,为构建组织工程化血管提供理想支架材料,并在体外实现其内皮化.用EX-810处理猪动脉7 min～72 h.所获材料进行形态学观察并测定交联指数和力学性能.在体外培养、扩增人脐静脉内皮细胞,随后种植培养于四组不同改性的材料表面并用MTT法检测细胞活性;最后用加压铺坪法将其种植到经赖氨酸处理并复合I型胶原的EX-810交联血管支架材料内表面,体外静态培养7天,取样行形态学检测及第Ⅷ因子相关抗原检测.结果显示EX-810既能有效去除和降低组织中产生抗原性的细胞成分和自由氨基,又保持了血管总体组织构架完整;经它处理的血管组织柔韧性好、接近天然血管组织,在各项力学性能方面也与天然血管组织相似;本材料经用赖氨酸处理并复合I型胶原后细胞毒性小,与细胞亲和性高,人脐静脉内皮细胞在其内表面能较好的生长增殖,容易内皮化.

Preparation of Biological Tissues Scaffold for Tissue-engineered Blood Vessel and Its Endothelialization

The porcine thoracic arteries were treated with 40 ml/L ethylene glycol diglycidyl ether(EX-810),and then some relevant examination and test were performed.Human umbilical vein endothelial cells(HUVECs) were seeded on the surface of different modified vascular tissues,and the growths of HUVECs on specimens were demonstrated by means of MTT test.Finally,HUVECs were seeded the internal surface of the modified EX-810-fixed porcine vascular tissues which were treated by 2% lysine,coated with collagen,and cultured for 7 days.The HUVECs on specimens were examined morphologically.The antigenicity of vascular tissues can be diminished by EX-810 through getting rid of cell in the vascular tissues or reducing the level of free amino groups in the vascular tissues.The structural integrity of vascular tissues can be preserved after treating with EX-810.The EX-810-fixed porcine vascular tissues also had the color and mechanical properties similar to the native vascular tissues.After treated by 2% lysine and coated with collagen,the EX-810-fixed porcine vascular tissues were characterized by low cytotoxicity and good cytocompatible.The collagen-coated EX-810-fixed porcine thoracic arteries treated with lysine showed low cytotoxicity,low antigenicity and favorable characteristic similar to nature vessel.