兔骨髓间充质干细胞与异体脱细胞耳软骨支架的体外复合

背景：耳软骨作为脱细胞基质可选择的支架,进行脱细胞处理可去除了软骨细胞的抗原性,从而与种子细胞具有良好的相容性。目的：体外提取、培养兔骨髓基质干细胞,并与异体脱细胞耳软骨支架复合,观察其生物相容性。方法：提取兔骨髓间充质干细胞行体外培养,诱导为软骨细胞,以胰蛋白酶-曲拉通联合法获得脱细胞软骨支架,将两者于体外复合,10d后复合支架固定行组织学染色及扫描电镜观察。结果与结论：兔骨髓间充质干细胞体外诱导14d可形成软骨细胞,Ⅱ型胶原免疫细胞化学示胞浆呈棕黄色;兔耳软骨脱细胞基质呈乳白色,组织学染色及扫描电镜观察示经脱细胞后支架孔隙均匀,结构完整,仍保存大量酸性黏多糖及胶原成分。其孔径长度（33.70±4.33）μm,孔隙率（65.23±7.35）%。复合支架组织学染色及扫描电镜示两者黏附良好,并伴有多量基质分泌。说明兔骨髓间充质干细胞诱导为软骨细胞与异体脱细胞耳软骨有良好的生物相容性。

In vitro composite of rabbit bone marrow mesenchymal stem cells and xenogenous acellular ear cartilage framework

BACKGROUND： Ear cartilage, an optional framework for acellular matrix, loses its antigenicity to chondrocytes after acellular treatment; therefore it has a good compatibility with seed cells. OBJECTIVE： To extract and culture rabbit bone marrow mesenchymal stem cells; to composite the stem cells with xenogenous acellular ear cartilage framework; and to explore the biocompatibility between them both. METHODS： Rabbit bone marrow mesenchymal stem cells were isolated, cultured in vitro and induced into cartilage cells. The acellular cartilage framework was obtained by trypsin-Triton combined method; then the two were composited in vitro. The composited framework was fixed for histological staining and scanning electron microscopic observation on 10 day after composite. RESULTS AND CONCLUSION： Rabbit bone marrow mesenchymal stem cells were induced into cartilage cells on 14 day after in vitro induction. Type Ⅱ collagen immunohistochemistry showed the cytoplasm was brownish yellow; rabbit ear acellular cartilage matrix was ivory white. Histological staining and scanning electron microscopy indicated that the pores were evenly distributed on the framework after acellular treatment; the framework has complete structure with massive acid mucopolysaccharide and collagen components. The pore size was （33.70±4.33） μm; the porosity was （65.23±7.35）%. Histological staining and scanning electron microscopy of the composite framework showed good adhesion between the two, accompanied by large amounts of matrix secretion. These findings indicate that the chondrocytes induced from rabbit bone marrow mesenchymal stem cells have a good biocompatibility with allogenic acellular ear cartilage.