交联羧甲基甲壳素三维支架用于软骨组织工程的细胞实验  

Using cross-linked carboxymethyl chitin 3D scaffold on cartilage tissue engineering：a cell experiment

作　　者：李天石[1] 吴奕光[1] 何君君[1] 崔光辉[1] 

机构地区：[1]北京大学深圳医院整形科,广东深圳518036

出　　处：《中华生物医学工程杂志》2015年第2期138-140,共3页Chinese Journal of Biomedical Engineering

基　　金：广东省自然科学基金（s2013010012641）;深圳市科创委科技计划项目（JCYJ20130402145538710）

摘　　要：目的：制备交联羧甲基甲壳素三维支架，并用于兔耳软骨细胞的共同体外培养，讨论此三维支架用于组织工程的可行性。方法交联法制备交联羧甲基甲壳素三维支架；分离、培养的兔耳软骨细胞，接种于三维支架浸泡的培养基上，倒置显微镜观察软骨细胞形态。通过II型胶原免疫荧光染色及Ⅱ型胶原免疫组化染色来鉴定Ⅱ型胶原含量；甲苯胺兰和阿利新蓝染色进行鉴定蛋白聚糖含量。结果组织染色显示兔耳软骨细胞在交联羧甲基甲壳素三维支架膜浸泡培养基中生长良好，形态正常，增殖旺盛。结论成功制备交联羧甲基甲壳素三维支架；兔耳软骨细胞与交联羧甲基甲壳素三维支架有良好相容性，无毒性，可以用于进一步构建组织工程软骨。Objective To prepare cross-linked carboxymethyl chitin 3D scaffold for in vitro culture of rabbit ear chondrocytes,and to investigate the feasinility of using it on tissue engineering. Methods The carboxymethyl chitin 3D scaffold was prepared by cross-linked method. The separated and cultured rabbit ear chondrocytes were inoculated with the culture medium immersed in the 3D scaffold. observation of hematoxylin eosin staining of cartilage cell The morphology of the rabbit ear chondrocytes was observed under inverted microscope. The type Ⅱ collagen content was measured by type Ⅱ collagen immunofluorescence staining and immunohistochemical staining. The proteoglycan content was measured by toluidine blue and alcian blue staining. Results The tissue staining showed that the rabbit ear chondrocytes grew well in the culture medium immersed in the cross-linked carboxymethyl chitin 3D scaffold,with normal morphology and excellent proliferation. Conclusion The cross-linked carboxymethyl chitin 3D scaffold was successfully prepared,which can be further used on cartilage tissue engineering. Also,good biocompatibility is found between rabbit ear chondrocytes and cross-linked carboxymethyl chitin 3D scaffold,without toxicity.

关 键 词：羧甲基甲壳素 软骨细胞 细胞培养 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]