家蚕再生丝素蛋白管状支架的制备及性能研究
发布时间：2018-11-22 20:37
【摘要】：小口径人造血管近年来一直是研究的重要课题,如何解决小口径人造血管力学性能不足及凝血问题是关注的焦点。本文采用冷冻、风干方法,选择具有良好生物相容性的家蚕再生丝素蛋白(以下简称丝素蛋白)与作为交联剂的聚乙二醇双环氧丙烷醚(PEG-DE)复合作为内膜和外膜、以蚕丝编织物作为中膜、模拟天然血管组织结构构建直径小于6mm的丝素蛋白管状支架。详细探索了交联比例(SF:PEG-DE,w/w)、SF浓度和冷冻温度对管状支架制备的影响,研究了管状支架的形态结构,评价了PEG-DE作为交联剂的丝素蛋白复合材料的细胞毒性。进一步研究了管状支架对SD大鼠成纤维细胞(L929)和人脐静脉内皮细胞(HUVECs)的生长支持作用。 以SF:PEG-DE=1:0.2～1:1.5制备的丝素蛋白管状支架,随着交联比例中PEG-DE比重的增加,管状支架成型更佳,管状支架丝素蛋白结合率增加,当SF:PEG-DE=1:1时管状支架成型最好,丝素蛋白管状支架中未交联的丝素蛋白溶失最少,丝素蛋白分子充分发生了交联反应,交联程度达到最佳,支架内表面微孔分布均匀。随着丝素蛋白浓度由3%增加到6%,管状支架成型更佳,丝素蛋白结合率上升,壁厚增加,孔径变小。丝素蛋白浓度为3%时,管状支架内表面不能形成良好内膜。随着冷冻温度(-20℃、-40℃、-80℃)的下降,管状支架孔径增大,孔壁因疏松使壁厚略有增加,对管状支架中丝素蛋白的结合率没有影响。 制备的丝素蛋白管状支架松弛外径约4.4～4.7mm,松弛内径为3.0～3.2mm,壁厚1mm,弯折直径5.8～6.7mm,具有较好的弯曲性能。丝素蛋白管状支架的轴向拉伸断裂强度90MPa,轴向拉伸断裂伸长率84%,径向拉伸断裂强度1000MPa,断裂伸长率160%,同时具有良好的径向压缩回复率。结果表明,本文制备的管状支架具有非常优越的力学性能。另外,丝素蛋白管状支架在37℃中的热水溶失率1%。 MTT和细胞DNA总含量分析结果显示,以PEG-DE作为交联剂制备的丝素蛋白复合材料没有明显细胞毒性,L929细胞在共混膜上呈长梭形,铺展紧密,细胞增殖活性强。采用荧光倒置显微镜、SEM、激光共聚焦显微镜观察及MTT法测试结果显示,丝素蛋白管状支架能很好的支持L929细胞的粘附和增殖。进一步研究了HUVECs在管状支架内表面的生长,同样显示了良好的粘附和增殖。
[Abstract]:In recent years, small diameter artificial blood vessel has been an important research topic, how to solve the problem of mechanical properties and coagulation of small diameter artificial blood vessel is the focus. In this paper, the regenerated silkworm silk fibroin with good biocompatibility and polyethylene glycol dioxide propane ether (PEG-DE) as crosslinking agent were selected as inner membrane and outer membrane by freezing and air drying methods. Silk fibroin tubular scaffolds with diameter less than 6mm were constructed by simulating the tissue structure of natural blood vessels. The effects of crosslinking ratio (SF:PEG-DE,w/w), SF concentration and freezing temperature) on the preparation of tubular scaffolds were investigated in detail. The cytotoxicity of fibroin composite with PEG-DE as crosslinking agent was evaluated. The effects of tubular scaffolds on the growth of SD rat fibroblasts (L929) and human umbilical vein endothelial cells (HUVECs) were further studied. With the increase of the proportion of PEG-DE in the crosslinking ratio of fibroin tubular scaffolds prepared by SF:PEG-DE=1:0.2~1:1.5, the tubular scaffolds were better formed, and the binding ratio of fibroin proteins in the tubular scaffolds increased. When SF:PEG-DE=1:1 was used, the tubular scaffold had the best shape, and the uncrosslinked fibroin in the fibroin tubular scaffold had the least dissolved loss, and the fibroin molecule had the best cross-linking reaction, the cross-linking degree was the best, and the distribution of micropores on the inner surface of the scaffold was uniform. With the increase of fibroin concentration from 3% to 6%, the tubular scaffold is better, the binding ratio of silk fibroin increases, the wall thickness increases, and the pore size becomes smaller. When fibroin concentration was 3, the inner surface of the tubular scaffold could not form a good intima. With the decrease of freezing temperature (-20 鈩，
本文编号：2350522
[Abstract]:In recent years, small diameter artificial blood vessel has been an important research topic, how to solve the problem of mechanical properties and coagulation of small diameter artificial blood vessel is the focus. In this paper, the regenerated silkworm silk fibroin with good biocompatibility and polyethylene glycol dioxide propane ether (PEG-DE) as crosslinking agent were selected as inner membrane and outer membrane by freezing and air drying methods. Silk fibroin tubular scaffolds with diameter less than 6mm were constructed by simulating the tissue structure of natural blood vessels. The effects of crosslinking ratio (SF:PEG-DE,w/w), SF concentration and freezing temperature) on the preparation of tubular scaffolds were investigated in detail. The cytotoxicity of fibroin composite with PEG-DE as crosslinking agent was evaluated. The effects of tubular scaffolds on the growth of SD rat fibroblasts (L929) and human umbilical vein endothelial cells (HUVECs) were further studied. With the increase of the proportion of PEG-DE in the crosslinking ratio of fibroin tubular scaffolds prepared by SF:PEG-DE=1:0.2~1:1.5, the tubular scaffolds were better formed, and the binding ratio of fibroin proteins in the tubular scaffolds increased. When SF:PEG-DE=1:1 was used, the tubular scaffold had the best shape, and the uncrosslinked fibroin in the fibroin tubular scaffold had the least dissolved loss, and the fibroin molecule had the best cross-linking reaction, the cross-linking degree was the best, and the distribution of micropores on the inner surface of the scaffold was uniform. With the increase of fibroin concentration from 3% to 6%, the tubular scaffold is better, the binding ratio of silk fibroin increases, the wall thickness increases, and the pore size becomes smaller. When fibroin concentration was 3, the inner surface of the tubular scaffold could not form a good intima. With the decrease of freezing temperature (-20 鈩，
本文编号：2350522
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