许旺细胞的诱导激活和神经桥接体构建的实验研究

发布时间：2018-05-27 10:46

本文选题：许旺细胞 + 成纤维细胞　；参考：《第一军医大学》2005年硕士论文

【摘要】：外周神经损伤后自体神经移植是首选的神经重建方法,但存在着供体来源受限、供区感觉、运动功能障碍、手术时间延长的缺点,难以满足长段神经缺损的要求。因此,寻找理想的种子细胞和支架材料复合培养而构建出功能性的人工神经是解决神经缺损疾患的主要手段之一。如何在体外获得大量分裂增殖和分化成熟的许旺细胞(Schwann cells,SCs)并选择合适的支架材料对完成组织工程化神经桥接体的构建至关重要。本实验选择细胞培养级生物制剂IL-1β作为SC的诱导激活剂,摸索出诱导激活SC分裂增殖和分化成熟的最佳作用条件,获取理想化的种子细胞。与此同时,应用新型生物可降解支架材料—人发角蛋白(Human Hair Keratins,HHK)作为构建人工神经桥接体的支架材料来弥补目前医学上应用较多的合成的可降解聚合物—聚羟基乙酸(PGA)、聚乳酸(PLA)等所带来的降解快、易崩解、支架整体易塌陷、局部酸性产物浓度过高的不足。将构建后的组织工程化外周神经桥接体移植到动物外周神经缺损部位,摸索出激活后的SC与HHK复合培养构建外周神经桥接体的最佳实验条件。【目的】 1.检测IL-1β在体外培养的自体神经匀浆激活的巨噬细胞上的表达。 2.摸索细胞培养级生物制剂IL-1β对SC的诱导激活及其最佳作用浓度。 3.IL-1β诱导激活前后的SC与ECM凝胶修饰的HHK复合培养构建组织工程化外周神经桥接体的对比研究。【方法】 1.将1月龄SD幼鼠坐骨神经远端切断,预溃变2d后,制成神经匀浆液,回注自体腹腔3d后,抽取腹腔液体培养巨噬细胞即得自体神经匀浆激活的巨噬细胞的条件培养基。应用ELISA法检测该条件培养基中的IL-1β的含量。
[Abstract]:Autogenous nerve transplantation after peripheral nerve injury is the first choice for nerve reconstruction, but it is difficult to meet the requirements of long nerve defect due to the limitation of donor source, the defect of donor region sensation, motor dysfunction and the prolongation of operation time. Therefore, it is one of the main methods to find ideal seed cells and scaffold materials to construct functional artificial nerve. How to obtain a large number of Schwann cells (Schwann cells) and select suitable scaffolds in vitro is very important for the construction of tissue engineered nerve bridge. In this experiment, we selected IL-1 尾, a cell culture biological agent, as the inducer of SC, and found out the best conditions for inducing and activating the proliferation and differentiation of SC, and obtained the ideal seed cells. At the same time, A novel biodegradable scaffold, Human Hair Keratinshike, was used as the scaffold material for the construction of artificial nerve bridge to make up for the more synthetic degradable polymers, polylactic acid and polyglycolic acid, which have been widely used in medicine at present. The degradation brought by PLA, etc., is fast, It is easy to disintegrate, the whole scaffold collapses easily, and the concentration of local acidic products is too high. The constructed tissue engineered peripheral nerve bridging body was transplanted to the defect of peripheral nerve in animals, and the optimal experimental conditions for the construction of peripheral nerve bridging body were found out by co-culture of activated SC and HHK. [purpose] 1. The expression of IL-1 尾 in macrophages activated by autologous nerve homogenate was detected. 2. To explore the induction and activation of SC by IL-1 尾, a cell culture class biological agent, and its optimal concentration. A comparative study on the construction of tissue engineered peripheral nerve graft by co-culture of SC and ECM gel modified HHK before and after 3.IL-1 尾 -induced activation. [methods] 1. The distal sciatic nerve of 1 month old SD rats was cut off and predeformed for 2 days, then the nerve homogenate was prepared. After 3 days of retroperitoneal injection, macrophages were cultured in celiac fluid to obtain the conditioned medium for the activation of macrophages activated by autologous nerve homogenate. The content of IL-1 尾 in the conditioned medium was detected by ELISA method.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329

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