梯度降解软骨支架材料应力刺激下与骨髓基质干细胞复合三维培养的实验研究
基质干细胞,,基质干细胞；,生物材料；,生物相容性；,组织工程,摘要,关键词,1材料与方法,2结果,3讨论,参考文献
摘 要：[目的]探讨梯度降解软骨支架材料(GDABM)的细胞生物相容性及诱导后的骨髓基质干细胞(MSCs)在三维支架上培养的生物学行为。[方法]将 MSCs与复合转化生长因子-β(TGFβ)的胶原共同混合后培养于三维软骨支架材料上，离心管内培养，10次/d，以600 r/min离心10 min。8 h、24 h、72 h、 4 d、7 d进行倒置显微镜形态学观察、扫描电镜观察。1～8 d采用MTT法检测细胞增殖活性，描绘生长曲线。通过3HProline掺入实验了解材料对MSCs胶原合成的影响。[结果]8 h后MSCs在支架材料上呈球型黏附生长，4 d后MSCs在支架材料上呈菱形或多角型复层生长，包裹于大量细胞外基质内。GDABM组细胞第3 d进入对数增长期，倍增时间为475 d，而单纯软骨细胞培养对照组倍增时间为525 d。GDABM组与单纯软骨细胞培养对照组比较胶原合成略低，3HProline掺入值有显著性差异(P<005)，说明经诱导MSCs的细胞胶原分泌功能仍低于软骨细胞。[结论]经TGFβ诱导和低转速离心应力刺激后的MSCs向软骨细胞功能分化，复合细胞因子梯度降解软骨支架材料具有良好的生物相容性，可作为MSCs的有效载体应用于组织工程化软骨的构建。
关键词：基质干细胞； 生物材料； 生物相容性； 组织工程
中图分类号 R687
Experimental studies on gradient degradation cartilage biomaterials combined with cultured marrow stroma cell in vitro
QU Yanlong, YANG Weiliang,MENG Xiangwen, et al.
Department of Orthopedic Surgery, the First Clinical Hospital, Harbin Medical University, Harbin150001, China
Abstract：[Objective]To study the biocompatibility of gradient degradation cartilage biomaterials (GDABM) and biological behavior of cultured marrow stroma cell(MSCs) combined with this threedimension scaffold in tissue engineering. [Method]MSCs(5×106) were cultured combining with GDABM in vitro in centrifuge tube, TGFβ was added to collagen and attached to GDABM. Culture tube was centrifuged in 600 r/min, 10 times/d, 10min per time. The morphological characters, cell proliferation were detected. Collagen synthesis were tracked by incorporated radioactive 3H labeled proline into culture. [Result]The MSCs attached to GDABM and grew along fibers in lozengelike or multiangularlike manner Cells in GDABM group reached logarithm increasing period at the third day. Its' doubling time was 4.75 d, While cells in control group was 5.25 d. There was difference of 3HProline value between the' two groups(P>0.05),which hint that the MSCs function was induced to cartilage in collagen synthesis, but MSCs collagen function level was lower than that of cartilage. [Conclusion]MSCs can be induced to cartilagecollagen function cell in low centrifuge stress combined with TGFβ. GDABM show good biocompatibility combined with MSCs and can provide a promising extracellular matrix scaffold for cell transplantation in cartilage tissue engineering. ......
您现在查看是摘要页，全文长 9169 字符。