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  本文選題：許旺細胞 + 成纖維細胞��； 參考：《第一軍醫(yī)大學(xué)》2005年碩士論文

【摘要】：外周神經(jīng)損傷后自體神經(jīng)移植是首選的神經(jīng)重建方法,但存在著供體來源受限、供區(qū)感覺、運動功能障礙、手術(shù)時間延長的缺點,難以滿足長段神經(jīng)缺損的要求。因此,尋找理想的種子細胞和支架材料復(fù)合培養(yǎng)而構(gòu)建出功能性的人工神經(jīng)是解決神經(jīng)缺損疾患的主要手段之一。如何在體外獲得大量分裂增殖和分化成熟的許旺細胞(Schwann cells,SCs)并選擇合適的支架材料對完成組織工程化神經(jīng)橋接體的構(gòu)建至關(guān)重要。本實驗選擇細胞培養(yǎng)級生物制劑IL-1β作為SC的誘導(dǎo)激活劑,摸索出誘導(dǎo)激活SC分裂增殖和分化成熟的最佳作用條件,獲取理想化的種子細胞。與此同時,應(yīng)用新型生物可降解支架材料—人發(fā)角蛋白(Human Hair Keratins,HHK)作為構(gòu)建人工神經(jīng)橋接體的支架材料來彌補目前醫(yī)學(xué)上應(yīng)用較多的合成的可降解聚合物—聚羥基乙酸(PGA)、聚乳酸(PLA)等所帶來的降解快、易崩解、支架整體易塌陷、局部酸性產(chǎn)物濃度過高的不足。將構(gòu)建后的組織工程化外周神經(jīng)橋接體移植到動物外周神經(jīng)缺損部位,摸索出激活后的SC與HHK復(fù)合培養(yǎng)構(gòu)建外周神經(jīng)橋接體的最佳實驗條件。 【目的】 1.檢測IL-1β在體外培養(yǎng)的自體神經(jīng)勻漿激活的巨噬細胞上的表達。 2.摸索細胞培養(yǎng)級生物制劑IL-1β對SC的誘導(dǎo)激活及其最佳作用濃度。 3.IL-1β誘導(dǎo)激活前后的SC與ECM凝膠修飾的HHK復(fù)合培養(yǎng)構(gòu)建組織工程化外周神經(jīng)橋接體的對比研究。 【方法】 1.將1月齡SD幼鼠坐骨神經(jīng)遠端切斷,預(yù)潰變2d后,制成神經(jīng)勻漿液,回注自體腹腔3d后,抽取腹腔液體培養(yǎng)巨噬細胞即得自體神經(jīng)勻漿激活的巨噬細胞的條件培養(yǎng)基。應(yīng)用ELISA法檢測該條件培養(yǎng)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.
【學(xué)位授予單位】：第一軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R329

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