钛表面粗糙度和官能团对内皮细胞细胞外基质原位构建的影响

发布时间：2018-03-05 23:12

本文选题：细胞外基质　切入点：碱活化　出处：《西南交通大学》2014年硕士论文　论文类型：学位论文

【摘要】：钛是一种具有良好生物相容性、机械性能及抗腐蚀性能的生物材料,广泛应用于研究心血管疾病领域。目前,生物材料表面改性成为加速内皮化,预防血管再狭窄的一大热题。研究表明生物材料的表面形貌、官能团结构对内皮细胞的粘附及增殖有促进作用,并且能够诱导细胞分泌更多的细胞外基质,从而改善材料表面生物相容性。内皮细胞自分泌的细胞外基质含有对细胞粘附、生长、增殖有促进作用的多种成份,因此将钛碱活化、等离子体聚合烯丙胺、表面沉积多巴胺与细胞自分泌的细胞外基质相结合应用于钛表面改性,充分改善材料表面的生物相容性。本实验分别在钛箔(Ti)以及碱活化钛箔(Ti-OH)、等离子体聚合烯丙胺薄膜(Ti-PPAam)、聚多巴胺薄膜(Ti-DA)改性的钛箔表面,由内皮细胞(EC)原位沉积细胞外基质(ECM)。接触角检测结果表明,沉积ECM使得Ti-OH和Ti-PPAam的接触角均有所提高,Ti-DA的接触角则减小19°。扫描电镜结果显示,除了光滑钛箔外,其它钛箔界面均形成一层均匀的交联状的ECM结构。傅里叶变换红外光谱(FTIR)检测结果表明,Ti-OH表面富含大量羟基,Ti-PPAam及Ti-DA表面含大量-NH2,各组样品经ECM改性后,都在1650cm-1处有-C=O官能团吸收峰,表明细胞外基质的成功构建。千万分之一天平精确称量并计算出各组钛箔表面ECM改性前后的质量差,发现多孔亲水性的Ti-OH表面ECM含量最多,达到0.075mmg,与其他两组相比具有显著性差异(P0.05)。体外EC粘附及增殖结果表明：Ti-OH表面、Ti-PPAam表面对EC有一定的抑制作用,经EC-ECM改性后,细胞相容性有一定的提高,而ECM改性的Ti-DA表面相比其他两组,细胞相容性更好(P0.05)。体外平滑肌细胞(SMC)的粘附及增殖结果表明：经EC-ECM改性后,各组样品的SMC粘附数量及活性均有显著性提高。内皮细胞NO释放结果表明：各组样品经EC-ECM改性后均促进EC的一氧化氮(NO)释放,且聚多巴胺薄膜与EC-ECM共同促进EC释放NO,有利于保持EC的功能和表型。体外血小板粘附结果表明：碱活化表面改性前后均明显抑制血小板粘附和聚集(P0.05),等离子体聚合烯丙胺表面血小板粘附和激活较严重,而聚多巴胺薄膜在抑制血小板粘附和激活的同时具有相对良好的细胞相容性,并且经细胞外基质改性后,进一步提高了血液相容性和细胞相容性。本研究结果表明,在以聚多巴胺为基底的钛表面原位构建EC-ECM,是改善材料表面生物相容性的有效方法。
[Abstract]:Titanium is a kind of biomaterial with good biocompatibility, mechanical properties and corrosion resistance, which is widely used in the field of cardiovascular disease. Studies have shown that the surface morphology and functional group structure of biomaterials can promote the adhesion and proliferation of endothelial cells, and induce the cells to secrete more extracellular matrix. In order to improve the biocompatibility on the surface of the material, the extracellular matrix secreted by endothelial cells contains many components which can promote cell adhesion, growth and proliferation, so the titanium base was activated and plasma polymerized allylamine. Dopamine deposited on the surface was combined with extracellular matrix (ECM) to improve the biocompatibility of titanium. In this experiment, the extracellular matrix (ECM) was deposited in situ from endothelial cells on the surface of titanium foil modified by titanium foil (Ti), alkaline activated titanium foil (Ti-OHN), plasma polymerized allylamine film (Ti-PPAamn) and polydopamine film (Ti-DA). The results of contact angle measurement showed that, The contact angle of Ti-OH and Ti-PPAam was increased by deposition of ECM, and the contact angle of Ti-DA decreased by 19 掳. The SEM results showed that, except for smooth titanium foil, The results of Fourier transform infrared spectroscopy (FTIR) show that the surface of Ti-OH is rich in a large amount of hydroxyl group Ti-PPAam and the surface of Ti-DA contains a large amount of -NH _ 2. The samples were modified by ECM. At 1650 cm ~ (-1), the absorption peak of C _ (10) O functional group was found, which indicated that the extracellular matrix was successfully constructed. The accurate weighing of 1/10000000 scales and the calculation of the mass difference before and after modification of ECM on each group of titanium foil surfaces showed that the surface ECM content of porous hydrophilic Ti-OH was the highest. Compared with the other two groups, there was a significant difference between the two groups (P 0.05). The results of EC adhesion and proliferation in vitro showed that Ti-PPAam surface of W Ti-OH surface could inhibit EC to some extent, and the cytocompatibility of Ti-DA modified by EC-ECM was improved to a certain extent, while that of Ti-DA modified with ECM was higher than that of the other two groups. The results of adhesion and proliferation of smooth muscle cells (SMC) in vitro were as follows: modified by EC-ECM, The number and activity of SMC adhesion were significantly increased in each group. The results of no release from endothelial cells showed that the release of nitric oxide (no) from EC was promoted by EC-ECM modification in each group. Poly (dopamine) film and EC-ECM can promote the release of no from EC, which is beneficial to maintain the function and phenotype of EC. The results of platelet adhesion in vitro showed that platelet adhesion and platelet aggregation (P0.05N) were significantly inhibited before and after surface modification by alkaline activation, and plasma aggregation was observed. The adhesion and activation of platelet on the surface of allylamine were serious. The membrane of polydopamine has good cytocompatibility while inhibiting platelet adhesion and activation, and after modified by extracellular matrix, it can further improve the blood compatibility and cytocompatibility. The results show that in situ construction of EC-ECM on the surface of titanium based on polydopamine is an effective method to improve the biocompatibility of the surface.
【学位授予单位】：西南交通大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R318.08

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