發(fā)布時間：2018-09-15 19:57

【摘要】：骨缺損的修復與替代一直是醫(yī)學上的難題之一。常規(guī)治療骨缺損的方法如自體骨移植、異體骨移植等都存在一定的弊端，而組織工程技術(shù)構(gòu)建的組織工程骨是解決這些弊端的一個新方案。在組織工程中，體外構(gòu)建組織工程骨時需要大量高活性的種子細胞，而細胞在體外平面培養(yǎng)時存在增殖速率低、易退化等問題，因此研制出一種生物相容性良好的三維細胞培養(yǎng)支架以滿足細胞高活性高密度的增殖可以解決組織工程的這個難題。本文研究了具有三維網(wǎng)絡連通孔結(jié)構(gòu)的多孔炭支架的制備與性能，并初步討論了該多孔炭支架對MG-63細胞的生物相容性以及生長、分化相關(guān)基因轉(zhuǎn)錄的影響。 本研究分兩個部分，第一部分是研究如何制備出機械性能合適、開孔率高的三維多孔炭支架。本文選用聚氨酯泡沫為模板，，酚醛樹脂為先驅(qū)體，采用模板法制備多孔炭支架。首先嘗試將酚醛樹脂逐滴滴加到模板上，然后經(jīng)過120℃固化、900℃碳化制備出多孔炭支架材料。結(jié)果表明該方法制備出的炭支架開孔率較高，表觀密度較大，但其機械性能無法滿足細胞培養(yǎng)的需求。通過研究聚氨酯泡沫的預處理方法，酚醛樹脂的浸漬、固化方法和碳化工藝對炭支架的機械性能和開孔率的影響，本文建立了一套制備具有較好機械性能和較高開孔率的多孔炭支架的方法：聚氨酯泡沫模板首先經(jīng)過30min無水乙醇浸泡處理，接著需要重復酚醛樹脂浸漬聚氨酯泡沫模板、固化這兩個步驟4-5次，最后將固化的材料在900℃下碳化。通過上述方法可以得到開孔率在80%以上、機械性能合適的炭支架材料。 第二部分是研究用本文所述方法制備的多孔炭支架的生物相容性及其對MG-63細胞生長、分化的影響。首先將MG-63細胞接種在多孔炭支架上，通過環(huán)境掃描電鏡觀察細胞生長、增殖情況。結(jié)果表明MG-63細胞增殖明顯；經(jīng)過6d培養(yǎng)后MG-63細胞基本長滿材料表面，說明炭支架材料具有優(yōu)越的生物相容性。本文采用Alamar Blue法定量地測定MG-63細胞在該多孔炭支架的增殖狀況和在培養(yǎng)板平面的增殖狀況。檢測結(jié)果表明：MG-63細胞在多孔炭支架材料上的增殖速度明顯高于培養(yǎng)板上的增殖速度，說明多孔炭支架材料相比平面培養(yǎng)更利于MG-63細胞的生長和增殖。為了探索多孔炭支架材料促進MG-63細胞的生長和增殖的分子生物學機理，本文采用了RT-PCR方法來檢測與生長、分化相關(guān)基因的轉(zhuǎn)錄水平。通過與二維平面培養(yǎng)的MG-63細胞對比，在多孔炭支架上培養(yǎng)的MG-63細胞內(nèi)ColI、BGN、ON等基因的轉(zhuǎn)錄水平有明顯的不同。
[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

本文編號：2244354