發(fā)布時(shí)間：2018-05-27 19:50

  本文選題：碳納米管 + 絲素蛋白　； 參考：《浙江大學(xué)》2016年碩士論文

【摘要】：目的以市售的納米羥基磷灰石/聚酰胺66(納艾康R)骨植入材料為基礎(chǔ),制備生物相容性、成骨誘導(dǎo)性良好的載藥碳納米管復(fù)合絲素蛋白修飾的納米羥基磷灰石/聚酰胺66骨組織工程支架,促進(jìn)骨髓間充質(zhì)干細(xì)胞的成骨分化。方法1.考察不同類型的多壁碳納米管(MWCNT):未經(jīng)修飾的多壁碳納米管(p-MWCNT)、羧基化多壁碳納米管(MWCNT-COOH)、羥基化多壁碳納米管(MWCNT-OH)與骨髓間充質(zhì)干細(xì)胞(bone mesenchymal stem cells, BMSCs)的細(xì)胞相容性。從細(xì)胞表面標(biāo)記、細(xì)胞骨架、細(xì)胞粘著斑蛋白、細(xì)胞增殖、細(xì)胞體外遷移和體內(nèi)分布等方面進(jìn)行研究,篩選與BMSCs相容性較好的碳納米管類型；2.配制羧基化碳納米管溶液和絲素蛋白溶液,得到不同含量MWCNT-COOH和絲素蛋白(Silk Fibroin, SF)溶液混合液；采用冷凍干燥法結(jié)合化學(xué)交聯(lián)法,對(duì)納米羥基磷灰石/聚酰胺66 (nHA/PA66)多孔支架進(jìn)行CNT/SF表面修飾,制備碳納米管復(fù)合絲素蛋白修飾的納米羥基磷灰石/聚酰胺66(CNT/SF-nHA/PA66)多孔骨組織工程支架；3.對(duì)支架的表面性質(zhì)、力學(xué)性能、孔隙率、降解性能進(jìn)行考察,并研究了BMSCs在支架上的細(xì)胞粘附與細(xì)胞增殖行為；4.為改善支架對(duì)BMSCs的成骨誘導(dǎo)性,使用MWCNT-COOH攜載地塞米松(Dexamethasone, DEX),制備了載藥的CNT/SF-nHA/PA66多孔支架,以考察載藥支架誘導(dǎo)BMSCs成骨分化的性能,同時(shí)檢測(cè)藥物在不同釋放介質(zhì)中的體外釋放行為。結(jié)果1.篩選獲得與BMSCs相容性較好的碳納米管類型-MWCNT-COOH; 2.使用冷凍干燥的方法成功地在羥基磷灰石/聚酰胺66表面修飾上了碳納米管-絲素蛋白；構(gòu)建得到的CNT/SF-nHA/PA66多孔支架具有較為理想的孔隙結(jié)構(gòu)和貫通性；3.當(dāng)CNT/SF混合液中CNT的含量為0.2 mg/ml時(shí),修飾的復(fù)合支架更利于BMSCs的生長(zhǎng)。生物樣本的SEM表明,BMSCs能在修飾后的支架表面粘附、鋪展和生長(zhǎng)；4.制備的攜載地塞米松的CNT/SF-nHA/PA66具有緩釋功能,能促進(jìn)BMSCs的成骨分化。結(jié)論采用冷凍干燥-化學(xué)交聯(lián)方法制備的載地塞米松碳納米管復(fù)合絲素蛋白修飾的納米羥基磷灰石/聚酰胺66骨組織工程支架,能滿足骨組織工程的要求,并能夠促進(jìn)骨髓間充質(zhì)干細(xì)胞的成骨分化,有望為載藥骨組織工程提供理論與實(shí)驗(yàn)依據(jù)。
[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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R318.08

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本文編號(hào)：1943528