三维多孔炭支架材料的制备及其对成骨细胞相关功能基因影响的研究
发布时间：2018-09-15 19:57
【摘要】：骨缺损的修复与替代一直是医学上的难题之一。常规治疗骨缺损的方法如自体骨移植、异体骨移植等都存在一定的弊端，而组织工程技术构建的组织工程骨是解决这些弊端的一个新方案。在组织工程中，体外构建组织工程骨时需要大量高活性的种子细胞，而细胞在体外平面培养时存在增殖速率低、易退化等问题，因此研制出一种生物相容性良好的三维细胞培养支架以满足细胞高活性高密度的增殖可以解决组织工程的这个难题。本文研究了具有三维网络连通孔结构的多孔炭支架的制备与性能，并初步讨论了该多孔炭支架对MG-63细胞的生物相容性以及生长、分化相关基因转录的影响。 本研究分两个部分，第一部分是研究如何制备出机械性能合适、开孔率高的三维多孔炭支架。本文选用聚氨酯泡沫为模板，，酚醛树脂为先驱体，采用模板法制备多孔炭支架。首先尝试将酚醛树脂逐滴滴加到模板上，然后经过120℃固化、900℃碳化制备出多孔炭支架材料。结果表明该方法制备出的炭支架开孔率较高，表观密度较大，但其机械性能无法满足细胞培养的需求。通过研究聚氨酯泡沫的预处理方法，酚醛树脂的浸渍、固化方法和碳化工艺对炭支架的机械性能和开孔率的影响，本文建立了一套制备具有较好机械性能和较高开孔率的多孔炭支架的方法：聚氨酯泡沫模板首先经过30min无水乙醇浸泡处理，接着需要重复酚醛树脂浸渍聚氨酯泡沫模板、固化这两个步骤4-5次，最后将固化的材料在900℃下碳化。通过上述方法可以得到开孔率在80%以上、机械性能合适的炭支架材料。 第二部分是研究用本文所述方法制备的多孔炭支架的生物相容性及其对MG-63细胞生长、分化的影响。首先将MG-63细胞接种在多孔炭支架上，通过环境扫描电镜观察细胞生长、增殖情况。结果表明MG-63细胞增殖明显；经过6d培养后MG-63细胞基本长满材料表面，说明炭支架材料具有优越的生物相容性。本文采用Alamar Blue法定量地测定MG-63细胞在该多孔炭支架的增殖状况和在培养板平面的增殖状况。检测结果表明：MG-63细胞在多孔炭支架材料上的增殖速度明显高于培养板上的增殖速度，说明多孔炭支架材料相比平面培养更利于MG-63细胞的生长和增殖。为了探索多孔炭支架材料促进MG-63细胞的生长和增殖的分子生物学机理，本文采用了RT-PCR方法来检测与生长、分化相关基因的转录水平。通过与二维平面培养的MG-63细胞对比，在多孔炭支架上培养的MG-63细胞内ColI、BGN、ON等基因的转录水平有明显的不同。
[Abstract]:Bone defect repair and replacement has been one of the medical problems. The conventional methods of treating bone defects, such as autogenous bone transplantation and allograft bone transplantation, have some drawbacks, but the tissue engineering bone constructed by tissue engineering technology is a new scheme to solve these problems. In tissue engineering, a large number of highly active seed cells are needed to construct tissue engineering bone in vitro, but the proliferation rate is low and the cells are easy to degenerate in plane culture in vitro. Therefore, the development of a biocompatible three-dimensional cell culture scaffold to meet the high activity and high density of cell proliferation can solve the problem of tissue engineering. In this paper, the preparation and properties of porous carbon scaffold with three-dimensional network connected pore structure were studied, and the effects of the scaffold on the biocompatibility, growth and differentiation related gene transcription of MG-63 cells were preliminarily discussed. This study is divided into two parts. The first part is to study how to prepare three-dimensional porous carbon scaffolds with suitable mechanical properties and high porosity. In this paper, polyurethane foam was used as template, phenolic resin as precursor, and porous carbon scaffold was prepared by template method. Firstly, the phenolic resin was added to the template one by one, and then the porous carbon scaffold was prepared by curing at 120 鈩
本文编号：2244354
[Abstract]:Bone defect repair and replacement has been one of the medical problems. The conventional methods of treating bone defects, such as autogenous bone transplantation and allograft bone transplantation, have some drawbacks, but the tissue engineering bone constructed by tissue engineering technology is a new scheme to solve these problems. In tissue engineering, a large number of highly active seed cells are needed to construct tissue engineering bone in vitro, but the proliferation rate is low and the cells are easy to degenerate in plane culture in vitro. Therefore, the development of a biocompatible three-dimensional cell culture scaffold to meet the high activity and high density of cell proliferation can solve the problem of tissue engineering. In this paper, the preparation and properties of porous carbon scaffold with three-dimensional network connected pore structure were studied, and the effects of the scaffold on the biocompatibility, growth and differentiation related gene transcription of MG-63 cells were preliminarily discussed. This study is divided into two parts. The first part is to study how to prepare three-dimensional porous carbon scaffolds with suitable mechanical properties and high porosity. In this paper, polyurethane foam was used as template, phenolic resin as precursor, and porous carbon scaffold was prepared by template method. Firstly, the phenolic resin was added to the template one by one, and then the porous carbon scaffold was prepared by curing at 120 鈩
本文编号：2244354
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