本文選題：血管平滑肌細(xì)胞 + Fe3O4/PCU復(fù)合材料��； 參考：《武漢理工大學(xué)》2014年碩士論文

【摘要】：近年來(lái)，經(jīng)導(dǎo)管栓塞動(dòng)靜脈畸形、動(dòng)靜脈瘺、動(dòng)脈瘤等血管疾病成為介入治療術(shù)中最重要的技術(shù)，但是，單純的機(jī)械性栓塞易導(dǎo)致管腔再通。因此，如果引入一種可促進(jìn)分泌細(xì)胞外基質(zhì)的活性物質(zhì)，使機(jī)械性栓塞與生物栓塞共同作用，便形成永久性栓塞。本文就課題組研究的Fe3O4/PCU復(fù)合微球進(jìn)行了生物相容性評(píng)價(jià)，探討其作為血管栓塞材料的可行性。 為了獲得可靠來(lái)源的目的細(xì)胞，首先探討了血管平滑肌細(xì)胞的分離與純化的方法。以組織塊貼壁法從大鼠胸腹主動(dòng)脈提取了血管平滑肌細(xì)胞，用自然純化法進(jìn)行純化處理，并采用平滑肌細(xì)胞特有的肌動(dòng)蛋白作為抗體以免疫組織化學(xué)方法鑒定所提細(xì)胞的特性。結(jié)果表明采用改良的主動(dòng)脈分離法提取的血管平滑肌細(xì)胞數(shù)量多、純度高，是一種簡(jiǎn)單可行、經(jīng)濟(jì)實(shí)用、操作性強(qiáng)的方法。 將培養(yǎng)至第三代的血管平滑肌細(xì)胞分別與PCU材料和不同比例的Fe3O4/PCU材料浸提液作用，采用倒置熒光顯微鏡下觀察細(xì)胞遷移情況，MTT法評(píng)價(jià)材料的細(xì)胞毒性；將細(xì)胞分別接種在不同的材料表面共培養(yǎng)，掃描電鏡觀察細(xì)胞在材料表面的黏附狀態(tài)，檢測(cè)細(xì)胞與材料作用后細(xì)胞胰島素樣生長(zhǎng)因子-1蛋白含量，考察材料對(duì)細(xì)胞增殖的影響。結(jié)果表明，細(xì)胞毒性等級(jí)為Ⅰ級(jí)，F(xiàn)e3O4/PCU復(fù)合材料可促進(jìn)細(xì)胞遷移與黏附，細(xì)胞生長(zhǎng)良好，在細(xì)胞增殖方面與實(shí)驗(yàn)的對(duì)照組無(wú)顯著性差異，初步判定Fe3O4/PCU具有良好的細(xì)胞相容性。 將富血小板血漿（Platelet Rich Plasma, PRP）與材料共同培養(yǎng)，掃描電鏡觀察到Fe3O4/PCU復(fù)合膜材料表面較PCU膜材料表面血小板的粘附數(shù)量較少，，聚集程度與擴(kuò)展程度低，且無(wú)形態(tài)變化；通過(guò)血紅蛋白吸光度值檢測(cè)內(nèi)源性凝血因子被激活的程度，F(xiàn)e3O4/PCU復(fù)合材料的抗凝血性能優(yōu)于PCU材料；按照ISO10993-4：2002標(biāo)準(zhǔn)，評(píng)價(jià)材料的體外溶血性能，證明Fe3O4/PCU復(fù)合材料無(wú)明顯的溶血反應(yīng)，溶血率為0.26%，符合醫(yī)療器械國(guó)家標(biāo)準(zhǔn)中關(guān)于栓塞材料的應(yīng)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

本文編號(hào)：1964380