染料介导光氧化固定牛带瓣颈静脉的生物特性评价

Biostability and Calcification Resistance of Valved Bovine Jugular Veins Stabilized by Dye-Mediated Photooxidation

为了解染料介导光氧化固定牛颈静脉的生物稳定性,抗钙化性和免疫原性,新鲜的牛颈带瓣静脉经染料介导光氧化法固定,分别进行化学消化和胃蛋白酶消化,上清液进行梯度聚丙烯酰胺电泳来了解其生物稳定性;将氧化固定的材料埋植于SD大鼠皮下,3周后取出,测钙含量、进行钙染色以及组织学检查了解其抗钙化性和免疫原性;采用戊二醛固定以及新鲜的牛带瓣颈静脉作为对照。染料介导光氧化固定的牛带瓣颈静脉经化学和酶消化之后的上清液电泳之后出现极淡的蛋白条带,明显淡于新鲜的未处理的材料,戊二醛固定的材料几乎不出现蛋白条带。皮下埋植3周后,钙含量测定,染料介导光氧化组显著低于戊二醛组,统计学上差异有显著性意义;钙染色,戊二醛组可见大量的钙沉积,新鲜组也可见钙沉积,但比戊二醛组少,染料介导光氧化组只见少量钙沉积;组织学检查提示光氧化组细胞浸润明显少于其他两组。体外动物实验提示,染料介导光氧化固定的牛带瓣颈静脉具有生物稳定性,低的钙化性和低免疫原性。

This study was conducted to make an in vitro evaluation of the biological characteristic of valved bovine jugular veins stabilized by dye-medlated photooxidation. The fresh valved bovine jugular veins were stabilized by dye-mediated photooxidation. The glutaraldehyde-treated veins and fresh veins served as controls. The samples were subjected to protein extraction assay and pepsin digestion assay, and were analyzed by polyacrylamide gel electrophoresis. The rat subcutaneous model for assessment of tissue calcification and immunologic properties was used. Calcium levels were determined by elemental analysis on dried samples and von Kossa staining. Both photooxidized and glutaraldehyde-treated tissues were resistant to protein extraction and pepsin digestion, compared to fresh tissue. Calcification levels were much lower for photooxidized tissues when compared with those for both glutaraldehyde-treated and fresh tissues. The perigraft infiltrate was lower in the photooxidized tissues than in the fresh and glutaraldehyde-treated tissues. Valved bovine jugular veins stabilized by dye-mediated photooxidation were biostable and resistant to calcification, and the immunologic property was well.