中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9126-.doi: 10.3969/j.issn.1673-8225.2010.49.002

• 干细胞培养与分化 • 上一篇    下一篇

不同接种密度骨髓间充质干细胞/骨基质明胶复合体修复大鼠桡骨缺损

田志逢1，秦书俭2，张小玲1，李占生1

 
  

  1. 1漯河医学高等专科学校解剖学教研室，河南省漯河市  462002；2辽宁医学院，辽宁省锦州市  121001
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 作者简介:田志逢★，男，1981年生，河南省濮阳市人，汉族，2008年辽宁医学院毕业，硕士，主要从事骨组织工程的研究。 zhifeng_07@sina.com

Different incubation densities of bone marrow mesenchymal stem cells and bone matrix gelatin complex in repair of radius defects in rats

Tian Zhi-feng1, Qin Shu-jian2, Zhang Xiao-ling1, Li Zhan-sheng1   

  1. 1 Department of Anatomy, Luohe Medical College, Luohe  462002, Henan Province, China; 2 Liaoning Medical University, Jinzhou  121001, Liaoning Province, China
  • Online:2010-12-03 Published:2010-12-03
  • About author:Tian Zhi-feng★, Master, Department of Anatomy, Luohe Medical College, Luohe 462002, Henan Province, China zhifeng_07@sina.com

PDF

399

被引次数

11

摘要：

背景：在构建种子细胞与载体材料的复合体时，种子细胞的接种密度是影响复合体成骨能力的一个重要因素，目前关于种子细胞的接种密度认识尚不统一。
目的：构建骨髓间充质干细胞/骨基质明胶复合体，观察构建复合体骨髓间充质干细胞的最适接种密度。
方法：体外单层培养SD大鼠骨髓间充质干细胞，以不同的细胞密度将骨髓间充质干细胞接种到异种骨基质明胶上，构建骨髓间充质干细胞/骨基质明胶复合体，通过扫描电子显微镜观察细胞在骨基质明胶中的黏附生长情况。制备SD大鼠双侧桡骨骨干5 mm节段性骨缺损模型。随机分成4组，采用骨髓间充质干细胞/骨基质明胶复合体修复骨缺损，所用的接种细胞密度分别为1×108 L-1、5×108 L-1、1×109 L-1、5×109 L-1。各组于第2，4，8，12周取材，通过大体观察大鼠的术肢活动情况，X射线放射学、组织学，免疫组织化学等检测方法，对骨缺损的修复情况进行评价。
结果与结论：4种细胞密度与骨基质明胶复合培养24 h，骨髓间充质干细胞在骨基质明胶上的黏附率随着细胞接种密度的增高而增高，当密度为1×109 L-1时，黏附率最高为(76.00±2.94)%，继续增加细胞密度其黏附率反呈下降趋势。复合培养7 d，扫描电子显微镜观察结果显示，材料上均有细胞黏附生长。X射线放射学评分显示，同一组别8，12周评分均高于2周评分(P < 0.01)；第4，8，12周1×109 L-1组与5×109 L-1组差异无显著性意义 (P > 0.05)，均高于其余两组(P < 0.01)。组织学评分结果表明，同一组别8，12周评分均高于2周评分 (P < 0.01)。结果提示，在构建骨髓间充质干细胞/骨基质明胶复合体时，骨髓间充质干细胞接种密度维持在1×109 L-1最佳。

 

关键词: 接种密度, 骨髓间充质干细胞, 骨基质明胶, 骨缺损, 大鼠

Abstract:

BACKGROUND: When the complex was made, the seed cell’s vaccination density will affect the ability of the complex, but the appropriate incubation density was not still unified.
OBJECTIVE: To construct bone marrow mesenchymal stem cells (BMSCs)/bone matrix gelatin (BMG) complex and to observe the optimal cell density of BMSCs/BMG complex.
METHODS: BMSCs from Sprague Dawley rats were cultured in vitro. BMSCs at different cell densities were incubated on the heterogenic BMG. BMSCs/BMG complex was constructed, and the adherent growth of cells on the BMG was observed using scanning electron microscope. A bone defect model (5 mm in length) was created at bilateral radius in each Sprague Dawley rat. This study was divided into four groups. BMSCs/BMG complex was used to repair bone defect. The seeding density was respectively 1×108/L, 5×108/L, 1×109/L and 5×109/L. At 2, 4, 8 and 12 weeks, the samples were obtained in each group. Gross observation on the activity of operated limb, radiographic and histological examination and immunohistochemical analysis were performed to observe the repair condition of radius defect after surgery. 
RESULTS AND CONCLUSION: After 24 hours of BMSCs/BMG complex culture, the adherent rate of cells on the BMG increased with increased cell incubation density. When the densities surpassed 1×109/L, and the maximum adhesive rate was (76.00±2.94)%. Increased cell densities showed decreased adhesive rate. At day 7, results of scanning electron microscope exhibited cells adhered on the material. X-ray radiology score showed the score was greater at 8 and 12 weeks compared with that at 2 weeks in the same group (P < 0.01). No significant difference was determined at 4, 8 and 12 weeks in the 1×109/L and 5×109/L groups (P > 0.05), and the score was greater than other two groups (P < 0.01). Histological score results have demonstrated that the score was higher at 8 and 12 weeks than that at 2 weeks in the same group (P < 0.01). Results suggest that when constructing the BMSCs/BMG complex in vitro, the optimal BMSCs seeding density was 1×109/L.

中图分类号: