组织工程血管基质的制备与改性

The preparation and modification of acellular vascular matrix

目的:完全脱除猪胸主动脉细胞,对脱细胞血管基质进行改性,增强基质的力学强度,制备组织工程血管支架材料。方法:取家猪的新鲜去除外膜胸主动脉20根,随机分成4组,分别采用胰蛋白酶、TritonX-100及十二烷基硫酸钠(SDS)作为脱细胞试剂对猪胸主动脉进行脱细胞处理,并对脱细胞后的血管基质进行改性,采用HE染色、弹力纤维染色、力学性能测试,以评价脱细胞效果以及材料的力学性能。结果:采用1%TritonX-100单独作用84h既能完全脱除细胞,同时又可完整保留血管基质的三维结构,对胶原纤维、弹力纤维的损伤小,是一种较理想的脱细胞方法。对脱细胞后的基质进行冷冻干燥及真空热交联处理,能有效提高材料的机械强度,冷冻干燥24h后真空120℃下热交联处理12h所获得的材料的机械强度最好,断裂强度可达到1.70MPa。结论:以脱细胞血管基质经冷冻干燥和真空热交联处理后,可以作为血管组织工程的支架材料。

Objective： In order to prepare the scaffolds for tissue engineering blood vessel, we fully extracted the porcine aorta cells, modificated the acellular matrix in vessles, and intensified the mechanical strength of the matrix. Methods： 20 aorta of flesh pig were selected and randomly separated into 4 groups. 0.1% trypsin, 1% Triton X-100, and 0.5% sodium dodecyl sulfate （SDS）were separately applied on them to extract the cells of porcine thoracic aorta, the vessels matrix after extracted were also modified. Using HE specimens staining, elastic protein fiber dyeing, and the biomechanical tests to assess the extracting results and mechanical characterization of the materials. Results： after 1% Triton X-100 individually applied on the cells for 84 hours, the cells can be fully extracted, and at the same time the three-dimensional structure of fiber of matrix can also be well kept. It is an relatively ideal cell-extracted method with slightest injures on coliagenous fibre and the elastic fibers. Modification of the acellular matrix by freeze-drying for 24 hours and then thermal cross-linking under vacuum at 120 ℃ for 12 hours, The acellular matrix under tensile strength test could reach the maximum of 1.70 MPa. Cclusion： These experimental results demonstrate that porcine aortic acellular matrix are good vascular tissue engineering scaffolds by freeze-drying and thermal cross-linking methods to change the mechanical characterization of the materials.