软骨细胞外基质/壳聚糖复合多孔支架和骨髓间充质干细胞构建组织工程软骨  

Constructing cartilage tissue engineering with chitosan/cartilage acellular matrix composite scaffold and bone marrow stromal cells

作　　者：王玉[1] 彭江[1] 张莉[1] 陈继凤[1] 周晓涵[1] 黄靖香[1] 许文静[1] 郭全义[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853

出　　处：《中国矫形外科杂志》2010年第20期1715-1718,共4页Orthopedic Journal of China

基　　金：全军十一五专项项目(编号:06Z057);国家自然科学基金重点项目(编号:30330570);解放军总医院科技创新基金(编号:06ZS03)

摘　　要：[目的]探讨软骨细胞外基质和壳聚糖制备复合多孔支架,同时并对小鼠骨髓间充质干细胞构建组织工程软骨的可行性进行观察。[方法]以猪关节软骨细胞外基质和壳聚糖为原料,采用冷冻干燥法制备软骨细胞外基质/壳聚糖复合多孔支架。通过扫描电镜观察材料内部结构及孔径大小,液体位移法测定材料的孔隙率,MTT方法检测支架浸提液毒性。将小鼠的骨髓间充质干细胞(BMSCs)分离培养并用TGF-β1成软骨诱导后,与材料复合培养,扫描电镜观察细胞在材料上的生长粘附情况。[结果]软骨细胞外基质/壳聚糖复合支架具有疏松多孔结构,孔径大小(159±36)μm,孔隙率为90.5%±2.3%,复合支架中的软骨细胞外基质成分甲苯胺蓝染色、番红O染色均呈阳性,MTT结果显示支架无细胞毒性。诱导的骨髓间充质干细胞在支架表面生长良好。[结论]软骨细胞外基质/壳聚糖复合材料具有合适的孔径和孔隙率,生物相容性良好,是组织工程软骨的良好支架载体。[Objective]To fabricate a nove chitosan/extracellular matrix of cartilage composite scaffold and to evaluate the possibility of application to cartilage tissue engineering.[Method]The composite scaffolds were constructed by 3%（w/v） acellular matrix of cartilage suspension and 2% chitosan solution and were poured into the cylindrical moulds.The 3-D porous scaffolds were fabricated using a simple freeze-drying method.Scanning electron microscopy was used to observe the pore size and interpores of the scaffolds.The porosity was measured by liquid displacement.MTT test was done to assess cytotoxicity of the extract fluid of the scaffolds.Bone marrow stem cells（BMSCs） of C57 mice were induced by transforming growth factor-β1and inculated into the scaffold.The condition of cell adhesion were observed by scanning electron microscopy.[Result]The 3-D porous scaffold had good pore interconnectivity with pore diameter（159±36）μm and 90.5%±2.3% porosity.The histological staining on scaffolds showed that toluidine blue,safranin O were positive.MTT test showed that the scaffolds had no cytotoxic effect on BMSCs.The adhesion of the cells on scaffold was good.[Conclusion]The 3-D porous scaffold constructed with chitosan and cartilage ECM possess good pore structure,non-toxicity and good biocompatibility,so it can be used in cartilage tissue engineering.

关 键 词：软骨细胞外基质 壳聚糖 软骨 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]