载药碳纳米管复合骨组织工程支架的制备及其与骨髓间充质干细胞的相容性评价

发布时间：2018-05-27 19:50

本文选题：碳纳米管 + 丝素蛋白　；参考：《浙江大学》2016年硕士论文

【摘要】：目的以市售的纳米羟基磷灰石/聚酰胺66(纳艾康R)骨植入材料为基础,制备生物相容性、成骨诱导性良好的载药碳纳米管复合丝素蛋白修饰的纳米羟基磷灰石/聚酰胺66骨组织工程支架,促进骨髓间充质干细胞的成骨分化。方法1.考察不同类型的多壁碳纳米管(MWCNT):未经修饰的多壁碳纳米管(p-MWCNT)、羧基化多壁碳纳米管(MWCNT-COOH)、羟基化多壁碳纳米管(MWCNT-OH)与骨髓间充质干细胞(bone mesenchymal stem cells, BMSCs)的细胞相容性。从细胞表面标记、细胞骨架、细胞粘着斑蛋白、细胞增殖、细胞体外迁移和体内分布等方面进行研究,筛选与BMSCs相容性较好的碳纳米管类型；2.配制羧基化碳纳米管溶液和丝素蛋白溶液,得到不同含量MWCNT-COOH和丝素蛋白(Silk Fibroin, SF)溶液混合液；采用冷冻干燥法结合化学交联法,对纳米羟基磷灰石/聚酰胺66 (nHA/PA66)多孔支架进行CNT/SF表面修饰,制备碳纳米管复合丝素蛋白修饰的纳米羟基磷灰石/聚酰胺66(CNT/SF-nHA/PA66)多孔骨组织工程支架；3.对支架的表面性质、力学性能、孔隙率、降解性能进行考察,并研究了BMSCs在支架上的细胞粘附与细胞增殖行为；4.为改善支架对BMSCs的成骨诱导性,使用MWCNT-COOH携载地塞米松(Dexamethasone, DEX),制备了载药的CNT/SF-nHA/PA66多孔支架,以考察载药支架诱导BMSCs成骨分化的性能,同时检测药物在不同释放介质中的体外释放行为。结果1.筛选获得与BMSCs相容性较好的碳纳米管类型-MWCNT-COOH; 2.使用冷冻干燥的方法成功地在羟基磷灰石/聚酰胺66表面修饰上了碳纳米管-丝素蛋白；构建得到的CNT/SF-nHA/PA66多孔支架具有较为理想的孔隙结构和贯通性；3.当CNT/SF混合液中CNT的含量为0.2 mg/ml时,修饰的复合支架更利于BMSCs的生长。生物样本的SEM表明,BMSCs能在修饰后的支架表面粘附、铺展和生长；4.制备的携载地塞米松的CNT/SF-nHA/PA66具有缓释功能,能促进BMSCs的成骨分化。结论采用冷冻干燥-化学交联方法制备的载地塞米松碳纳米管复合丝素蛋白修饰的纳米羟基磷灰石/聚酰胺66骨组织工程支架,能满足骨组织工程的要求,并能够促进骨髓间充质干细胞的成骨分化,有望为载药骨组织工程提供理论与实验依据。
[Abstract]:Objective to prepare biocompatibility of nano-hydroxyapatite / polyamide 66 (NAA) bone implant material. The bone tissue engineering scaffold of nano-hydroxyapatite / polyamide 66 modified by drug-loaded carbon nanotubes and fibroin modified bone tissue engineering scaffolds can promote the osteogenic differentiation of bone marrow mesenchymal stem cells. Method 1. The cytocompatibility of different types of multiwalled carbon nanotubes (MWCNT) with bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells) was investigated, such as unmodified multiwalled carbon nanotubes (MWCNT), carboxylated multiwalled carbon nanotubes (MWCNT-COOHH), hydroxylated multiwalled carbon nanotubes (MWCNT-OH) and bone marrow mesenchymal stem cells (mesenchymal stem cells, BMSCs). The cell surface labeling, cytoskeleton, cell adhesion protein, cell proliferation, cell migration in vitro and distribution in vivo were studied, and the type of carbon nanotubes with good compatibility with BMSCs was selected. Carboxylated carbon nanotube solution and silk fibroin solution were prepared to obtain the mixed solution of MWCNT-COOH and silk fibroin (SF) solution, and the freeze-drying method combined with chemical crosslinking method was used. Nano-hydroxyapatite / polyamide 66 / PA66 porous scaffold was modified with CNT/SF to prepare nano-hydroxyapatite / polyamide 66 CNT / SF-NAA / PA66 porous bone tissue engineering scaffold. The surface properties, mechanical properties, porosity and degradation properties of the scaffolds were investigated, and the cell adhesion and cell proliferation behavior of BMSCs on the scaffolds were studied. In order to improve the osteoblastic induction of BMSCs by stents, the porous CNT/SF-nHA/PA66 scaffolds loaded with Dexamethasone (DEXN) were prepared with MWCNT-COOH to investigate the osteogenic differentiation of BMSCs in vitro and the release behavior of BMSCs in different release media. Result 1. The type of carbon nanotubes (MWCNT-COOH) with good compatibility with BMSCs was obtained. 2. Carbon nanotube fibroin was successfully modified on the surface of hydroxyapatite / polyamide 66 by freeze-drying method, and the CNT/SF-nHA/PA66 porous scaffold had ideal pore structure and transfixibility. When the content of CNT in CNT/SF mixture is 0. 2 mg/ml, the modified composite scaffold is more favorable to the growth of BMSCs. SEM of biological samples showed that BMSCs could adhere, spread and grow on the surface of modified scaffolds. The prepared CNT/SF-nHA/PA66 with dexamethasone has the function of slow release and can promote the osteogenic differentiation of BMSCs. Conclusion the nano-hydroxyapatite / polyamide 66 bone tissue engineering scaffold with dexamethasone carbon nanotube composite fibroin modified by freeze-drying and chemical crosslinking can meet the requirements of bone tissue engineering. It can promote osteogenic differentiation of bone marrow mesenchymal stem cells and provide theoretical and experimental basis for bone tissue engineering.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R318.08

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