【作者】 刘欣欣；

【导师】 段潜； 李艳辉；

【作者基本信息】 长春理工大学 ， 无机化学， 2011， 硕士

【摘要】 本文为载基因的壳聚糖胶原凝胶基因治疗提供了前期准备工作。选用酶消化法,以牛腱为原料提取胶原,保证提取的胶原具有三股螺旋结构。并对提取的胶原进行的相应的表征工作。测定了胶原提取液的粘度和固体含量。研究了胃蛋白酶消化法中酶与材料的最佳配比为E：B=1：10。经红外、紫外光谱分析确定提取物为胶原。同时用提取的胶原制备胶原凝胶,测定胶原凝胶中固体含量。在胶原凝胶中培养细胞,用光学显微镜进行观察细胞生长状态,证明胶原凝胶可以作为细胞生长的立体三维生长支架。另外,对天然的壳聚糖进行了一系列生物学表征工作,凝胶阻滞电泳实验证明壳聚糖具有一定的DNA结合能力。粒径电位分析表明壳聚糖与DNA形成了粒径在100-300nm,表面电位稳定的纳米复合物颗粒。MTT实验测试了壳聚糖的没有细胞毒性。证明了壳聚糖可以作为基因载体。更多还原

【Abstract】 This paper provides prerequisite preparation for gene therapy of Collagen gel containing chitosan gene.The method of enzyme digestion was used to extract collagen from bovine tendon so that triple helix of collagen molecules can be maintained. And the extracted collagen was characterized. The extracted collagen viscosity and solid content were measured. In pepsin digest method, the best ratio for enzymes and material is that E to B equals 1 to 10. The extracted protein was confirmed to be collagen by FTIR and UV. At the same time, collagen gel was prepared by the extracted collagen, and collagen gel solid content was measured. The cell was seeded in collagen gel. The cell growth state was observed by optical microscope and the results show that the collagen gel can be used as 3D cell scaffold.In addition, a series of biology characterizations of natural chitosan were made. The results of gel retardation experiment indicated the chitosan had strong affinity to DNA. CS-DNA could form composite nano-particles with diameters from 100 to 300 nm and the surface potential is stable. The cytotoxicity was measured by MTT experiment that the results showed chitosan had no cytotoxicity and proved that chitosan can be used as gene carrier.更多还原