续断皂苷Ⅵ诱导的脂肪间充质干细胞复合纳米羟基磷灰石/壳聚糖支架治疗大鼠桡骨骨缺损  

Treatment of rat radial bone defect by n-HA/CS combined with adipose-derived mesenchymal stem cells induced by the dipsacus saponin Ⅵ

作　　者：赵刚[1] 刘学元[1] 姚素艳[2] 郑德宇[1] 

机构地区：[1]辽宁医学院解剖学教研室,锦州121000 [2]辽宁医学院病理生理学教研室,锦州121000

出　　处：《解剖学杂志》2016年第4期416-421,共6页Chinese Journal of Anatomy

基　　金：辽宁省高等学校创新团队项目(LT2012016)

摘　　要：摘要目的：探讨续断皂苷Ⅵ诱导的大鼠脂肪间充质干细胞（ADMSCs）联合纳米羟基磷灰石／壳聚糖（n-HA／CS）人工骨材料修复大鼠桡骨骨缺损的效果。方法：应用细胞表面抗原鉴定ADMSCs。取传至第3代的ADMSCs，培养基中加入续断皂苷Ⅵ诱导15d。诱导后的ADMSCs和n-HA／CS复合培养获得的植骨材料修复大鼠桡骨骨缺损模型，于术后第4、12周骨缺损区行X光拍片并按照改良Gray标准进行结果分析。H—E染色，并按改良Nilsson的组织学评分标准进行成骨效应评估。结果：ADMSCs呈成纤维细胞样贴壁生长，高表达CD44。续断皂苷Ⅵ诱导后的细胞呈三角形、矮方形及多边形。茜素红染色可见散在的红色阳性结节，骨钙素表达阳性，阳性细胞率85．46％±5．22％。骨缺损修复术后4周移植材料已部分被吸收。术后12周实验组部分新生骨皮质恢复连续，开始出现骨髓腔。改良Gray评分显示，在各时间点实验组均高于各对照组，差异有统计学意义。第4周行H-E染色可见植入支架部分被吸收，实验组支架间隙和周围均充满细胞。第12周实验组出现类似骨单位样结构，骨基质中为淡粉色。改良Nilsson评分显示实验组在术后的4周和12周均高于各个对照组，有统计学意义。结论：续断皂苷Ⅵ能够诱导大鼠ADMSCs分化为成骨细胞，复合n-HA／CS支架能很好修复骨缺损。Objective： To explore the recovery effect of radial bone defects in rats by dipsacus saponin Ⅵ induced adipose- derived mesenchymal stem cells（ADMSCs） in combination with artificial bone graft composed of n-hydroxyapatite/chitosan （n-HA/CS）. Methods： The surface antigens were used to detect ADMSCs. ADMSCs induced by dipsacus saponin VI after 15 days. The artificial graft, formed by the induced ADMSCs, osteoblast, cocultured with the n-HA/CS binary vector, were used to repair the radial bone defect of rat models. X-ray photo for bone defect regions were taken 4 and 12 weeks respectively after the operation and the results were analyzed according to the improved Gray＇s criteria. The new formation was detected by H-E staining. And the osteogenic effects were assessed based on the improved Nilsson histologic score criteria. Results： The primary ADMSCs were in the shape of fibroblasts and grew adherently. And the CD44, mesenchymal stem cell-related markers were highly expressed. The phenotype of induced ADMSCs was triangle, square or polygonal shape and connected each other in some place. The sporadic red particles, formed by the calcium deposits, were found by alizarin red staining. The positive ratio of osteocalcin was 85 % ~ 5.22 %. The artificial grafts of binary were partially absorbed postoperation 4 weeks. Partial compact bone of two ends with bone defect was connected 12 weeks after the operation. Bone marrow cavities of bone defect were found 12 weeks later. Gray scores of experimental group were higher than that of other control groups in each time points respectively. For the H-E staining, the major part of the implanted artificial graft was absorbed after 4 weeks. The new cells could be found in the space in the artificial graft or around the graft. Osteon-liked structures could be found 12 weeks after the operation. And the matrix was in the pink color, indicating that the matrix was not mature yet. Nilsson scores of experimental group were significantly higher than that of other 4 and 1

关 键 词：续断皂苷Ⅵ 脂肪间充质干细胞 纳米羟基磷灰石/壳聚糖支架 骨缺损 

分 类 号：R318.08[医药卫生—生物医学工程] R68[医药卫生—基础医学]