磁性形状记忆聚氨酯复合栓塞材料的生物学评价

发布时间：2018-06-01 13:43

  本文选题：血管平滑肌细胞 + Fe3O4/PCU复合材料　； 参考：《武汉理工大学》2014年硕士论文

【摘要】：近年来，经导管栓塞动静脉畸形、动静脉瘘、动脉瘤等血管疾病成为介入治疗术中最重要的技术，但是，单纯的机械性栓塞易导致管腔再通。因此，如果引入一种可促进分泌细胞外基质的活性物质，使机械性栓塞与生物栓塞共同作用，便形成永久性栓塞。本文就课题组研究的Fe3O4/PCU复合微球进行了生物相容性评价，探讨其作为血管栓塞材料的可行性。 为了获得可靠来源的目的细胞，首先探讨了血管平滑肌细胞的分离与纯化的方法。以组织块贴壁法从大鼠胸腹主动脉提取了血管平滑肌细胞，用自然纯化法进行纯化处理，并采用平滑肌细胞特有的肌动蛋白作为抗体以免疫组织化学方法鉴定所提细胞的特性。结果表明采用改良的主动脉分离法提取的血管平滑肌细胞数量多、纯度高，是一种简单可行、经济实用、操作性强的方法。 将培养至第三代的血管平滑肌细胞分别与PCU材料和不同比例的Fe3O4/PCU材料浸提液作用，采用倒置荧光显微镜下观察细胞迁移情况，MTT法评价材料的细胞毒性；将细胞分别接种在不同的材料表面共培养，扫描电镜观察细胞在材料表面的黏附状态，检测细胞与材料作用后细胞胰岛素样生长因子-1蛋白含量，考察材料对细胞增殖的影响。结果表明，细胞毒性等级为Ⅰ级，Fe3O4/PCU复合材料可促进细胞迁移与黏附，细胞生长良好，在细胞增殖方面与实验的对照组无显著性差异，初步判定Fe3O4/PCU具有良好的细胞相容性。 将富血小板血浆（Platelet Rich Plasma, PRP）与材料共同培养，扫描电镜观察到Fe3O4/PCU复合膜材料表面较PCU膜材料表面血小板的粘附数量较少，，聚集程度与扩展程度低，且无形态变化；通过血红蛋白吸光度值检测内源性凝血因子被激活的程度，Fe3O4/PCU复合材料的抗凝血性能优于PCU材料；按照ISO10993-4：2002标准，评价材料的体外溶血性能，证明Fe3O4/PCU复合材料无明显的溶血反应，溶血率为0.26%，符合医疗器械国家标准中关于栓塞材料的应用的要求，具有良好的血液相容性。
[Abstract]:In recent years transcatheter embolization of arteriovenous malformations arteriovenous fistula aneurysm and other vascular diseases has become the most important technique in interventional therapy. However simple mechanical embolization can easily lead to recanalization. Therefore, if the introduction of an active substance that can promote the secretion of extracellular matrix, mechanical embolization and biological embolization will form permanent embolization. In this paper, the biocompatibility of Fe3O4/PCU composite microspheres studied by our research group was evaluated and the feasibility of using them as vascular embolization materials was discussed. In order to obtain a reliable source of target cells, the methods of isolation and purification of vascular smooth muscle cells (VSMC) were studied. Vascular smooth muscle cells (VSMCs) were extracted from rat thoracic and abdominal aorta by tissue mass adherence method and purified by natural purification. The specific actin of smooth muscle cells was used as antibody to identify the characteristics of the cells by immunohistochemical method. The results showed that the improved aortic separation method was a simple, economical and practical method for the extraction of vascular smooth muscle cells (VSMCs) with large number and high purity. The third passage of vascular smooth muscle cells were treated with PCU material and different proportion of Fe3O4/PCU material respectively. The cell migration was observed under inverted fluorescence microscope and the cytotoxicity of the material was evaluated by MTT method. Cells were co-cultured on different materials. The adhesion of cells on the surface of materials was observed by scanning electron microscope (SEM). The content of insulin-like growth factor-1 (IGF-1) protein was detected after the interaction between cells and materials, and the effect of materials on cell proliferation was investigated. The results showed that the cytotoxic grade 鈪

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