中文摘要：

制作与鉴定自体兔骨髓间充质干细胞-海螵蛸复合支架,进行生物学安全性评估。采用密度梯度离心联合贴壁筛选法,体外培养兔骨髓间充质干细胞（BMSCs）;采用沉淀法,构建BMSCs-海螵蛸复合支架;通过电镜扫描及MTT试验,对复合支架进行检测。选取40只新西兰兔,分两组分别植入BMSCs-海螵蛸复合支架（试验组）或海螵蛸支架（对照组）,通过肌肉植入试验评估复合支架的生物学安全性。密度梯度离心联合贴壁筛选分离培养出干细胞,其免疫细胞化学检测表面抗原CD29（＋）。电镜扫描测得海螵蛸支架孔隙率为81.730%±6.770%。扫描电镜下见细胞黏附于海螵蛸支架爬行生长,分泌胞间基质。MTT检测：复合培养第1、3、5、7天之间吸光值两两比较,均有显著性差异（P〈0.05）,细胞呈逐渐增殖趋势;第7、8天比较无显著差异,表明第8天细胞无明显增殖。肌肉植入试验的血常规、血生化检测,复合支架组（试验组）与海螵蛸组（对照组）检测结果无显著差异,复合支架的各项指标达标。病理切片：第7天,试验组与对照组的组织炎症反应为III~IV级;第14天,二者的组织炎症反应为II~III级;第28天,二者的组织炎症反应为I~II级;第56天,二者的组织炎症反应为0~I级：均符合肌肉植入试验结果评定标准。自体BMSCs能与中药海螵蛸联合培养构建复合支架,支架上的细胞数量随培养时间延长而增多,第7天时达增殖高峰。研究表明,此复合支架的组织相容性良好,符合生物材料的应用要求。

英文摘要：

This study is aimed to conduct fabrication and safety evaluation for the rabbit BMSCs-Cuttlebone composite scaffold. BMSCs were separated by density gradient centrifugation and adherence screening method.The BMSCs-Cuttlebone scaffold was cultivated by precipitation method. The composite scaffold was examined by SEM observation and cell viability assay（ MTT test）. The biological safety of the composite scaffold was evaluated through muscle implantation test. A total of 40 New Zealand rabbit were randomly divided into two groups,and then BMSCs-cuttlebone scaffold（ experimental group） or cuttlebone（ control group） were implanted.BMSCs of CD29 positive were obtained by the method of density gradient centrifugation effectively. Scanning electron microscope observation showed that the porosity of cuttlefish bone scaffold was 81. 730% ± 6. 770%. The cells adhered on the cuttlebone bracket creeping growth and secreted extracellular matrix. Results obtained from MTT assay showed that the number of viable cells on scaffold increased over time. On the day 1,3,5,7 of cultivation,the cells kept proliferating; while on the day 7 and 8,the proliferation decreased and almost stopped on the day 8. Muscle implantation test showed no significant difference existed between the composite scaffold（ experimental group） and cuttlebone group（ control group） in the blood routine and blood biochemical tests. In the pathological observation,on the day 7,tissue inflammation grade III-IV for both experimental group and control group; on theday 14,the tissue inflammation grade was II-III for the two groups; on the day28,the tissue inflammation was I-II for the two groups; on the day 56,the tissue inflammation was 0-I grade for the two groups; all of which were within the limit of standard. The rabbit BMSCs could be co-cultured with cuttlebone to build complex biological scaffold. The cells embedded in the scaffold kept proliferation for 7 days. The composite scaffold exhibited good biocompatibility in the muscle implantati