不同支架材料体外构建组织工程软骨的研究

目的:体外构建组织工程软骨,筛选更为适合组织工程软骨构建的支架材料。方法:体外获取SD大鼠肋软骨细胞。采用第一代软骨细胞作为种子细胞,接种于壳聚糖/明胶和BMG/生物蛋白胶支架,体外培养的不同时间对其进行HE、甲苯胺蓝染色、Mas s on染色、免疫学检测、扫描电镜观察。结果:在培养2周时,BMG/生物蛋白胶各种染色结果显示软骨细胞在其表面以及内部分布均匀,蛋白多糖和Ⅱ型胶原染色阳性;壳聚糖/明胶表面细胞稍多于前者,但内部细胞数量极少且分布不均,染色结果不如前者明显。随着培养时间的延长各种检测均显示有大量的软骨细胞特异性的蛋白多糖和Ⅱ型胶原的表达,壳聚糖/明胶凸显出明显的优势。结论:体外成功构建组织工程软骨,软骨细胞在BMG/生物蛋白胶上的生长、增殖和分泌基质情况优于壳聚糖/明胶支架。

Study on constructing tissue-engineered cartilage in different scaffolds in vitro

Objective Try to construct the tissue-engineered cartilage and selection a better scaffold for the constructing.Methods Treating the first generation chondrocytes as the seeds which was separated from the SD rat.Through the adjusting the concentration of chondrocytes,we can make the complex which is composed by cancellous bone matrix(BMG) / biological fibrin glue,chitosan/gelatin(Cs-gel) and chondrocytes.In different culture time,HE,toluidine blue(TB),Massons triad colour,immunohistochemistry,immunofluorescence and scanning electron microscope were used to test the complex.Results After co-culturing two weeks,we can certify the BMG/ biological fibrin glue is precendecer than Cs/gel.Concretly,the chondrocytes is uniform in BMG/biological fibrin glue,but not adqulis in Cs/gel.aggrecan and collagenⅡis obviously in testing.But the longer the co-culturing,all tests display that special aggrean and collagen Ⅱabout chondrocytes are obvious at eight weeks the latter is better than the former.Conclusion Succeeding in constructing the tissue-engineered cartilage.In early culture,the growth,proliferation and secretion of chondrocytes in BMG/ biological fibrin glue was better.But Cs/gel have significallly advantage in longer co-culturing in vitro.