脱细胞基质软骨支架的制备

背景：脱细胞基质材料去除了天然材料中的细胞成分，保留了基质成分，有效降低了天然材料的免疫原性，同时能够保持材料的机械强度。 目的：拟利用洗脱方法去除兔肋软骨中的细胞基质，制备天然生物支架材料。 方法：取新西兰大白兔肋软骨，清除周围组织后随机分组处理，以未经处理的肋软骨作为正常对照组；48 h处理组以去污剂-酶化学消化48 h；96 h处理组以去污剂-酶化学消化96 h，3组均通过苏木精-伊红染色及电镜观察脱细胞效果。同时收集诱导第7天的兔骨髓间充质干细胞3×109 L-1，与同种异体肋软骨脱细胞基质体外复合培养，于第3，7天取复合物行电镜观察细胞在脱细胞基质表面的黏附生长情况。 结果与结论：新鲜肋软骨标本每个软骨陷窝内均有排列紧密的二三个软骨细胞，去污剂-酶化学消化后软骨陷窝内的细胞逐渐脱失，至消化处理96 h后，软骨陷窝内的细胞完全脱失。共培养第3天时，脱细胞基质表面有大量骨髓间充质干细胞分布，细胞为多角形，有伪足伸出，锚定在基质表面，部分区域可见细胞在基质表面增殖分裂；第7天时，脱细胞基质表面大部分均为细胞覆盖，细胞呈扁平状，有多个足突充分伸展，细胞之间互相连接，分泌大量细胞外基质沉积在基质表面，呈冰霜样改变，表明制备的脱细胞基质具有良好的细胞相容性。中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程全文链接：

Preparation of an acellular cartilage matrix scaffold

BACKGROUND: Acellular matrix which is decellularized but retains the matrix components in the nature materials can reduce the immunogenicity of natural materials effectively and keep the material mechanical strength. OBJECTIVE: To prepare the natural biological scaffold material by using elution method to remove the cells in the rabbit costicartilage matrix. METHODS: After removal of the surrounding tissues, the costicartilage samples from New Zealand white rabbits were randomly divided them into three groups: costicartilages untreated as normal control group, detergent-enzyme digestion for 48 hours as 48-hour treatment group, detergent-enzyme digestion for 96 hours as 96-hour treatment group. Hematoxylin-eosin staining and electron microscope were used to observe decellularized effects. At 7 days of induction, bone marrow mesenchymal stem cells, 3×109/L, were collected and co-cultured with allogeneic acellular costicartilage matrix in vitro. At 3 and 7 days of co-culture, the composite was  taken for observation of cell growth on the cell-free substrate surface under electron microscope. RESULTS AND CONCLUSION:Two or three cartilage cells were closely packed in each cartilage pit, began to depigment using the detergent-nuclease digestion method, and then completely depigmented at 96 hours after treatment. At 3 days of co-culture, there were a large amount of polygon-shaped adherent cells with pseudopodias observed on the acellular matrix surface in vitro, and cells could proliferate and divide in partial regions. At 7 days of co-culture, adherent cells spread mostly throughout the acellular matrix surface, these cells were flat-shaped and extended with multiple interconnected processes. Mass of secreted excellular matrixes were deposited on the matrix surface and showed frost-like changes, indicating the prepared acellular matrix has favorable cytocompatibility.