中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (42): 7797-7801.doi: 10.3969/j.issn.1673-8225.2010.42.003

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials • 上一篇    下一篇

纳米β-磷酸三钙/Ⅰ、Ⅱ型胶原层状支架复合骨髓间充质干细胞修复膝关节骨软骨缺损

陈竹生1，吕玉明2，张  兵1   

  1. 1广东省第二中医院骨科，广东省广州市  510095；  2广州医学院第三附属医院骨科，广东省广州市 510150
  • 出版日期:2010-10-15 发布日期:2010-10-15
  • 通讯作者: 吕玉明，博士，副主任医师，广州医学院第三附属医院骨科，广东省广州市 510150 lvyuming2008@126.com
  • 作者简介:陈竹生☆，男，1967年生，江西省南昌市人，汉族，2007年中山大学毕业，博士，副主任医师，主要从事关节方面的研究。 czs8341@163.com
  • 基金资助:

    广东省自然科学基金（7002690），课题名称“纳米β-TCP/胶原复合骨髓间充质干细胞构建骨软骨实验研究”；广东省医学科研基金（A2008128），课题名称“纳米仿生支架构建组织工程化骨软骨的实验研究”。广州医学院科研资助（0709106），课题名称“纳米β-TCP/胶原复合骨髓间充质干细胞构建骨软骨实验研究”。

Repair of articular osteochondral defects with bone marrow stromal stem cells/nano-beta-tricalcium phosphate/collagen I and II composite

Chen Zhu-sheng1, Lü Yu-ming2, Zhang Bing1   

  1. 1 Department of Orthopaedics, The Second Traditional Chinese Medicine Hospital of Guangdong Province, Guangzhou   510095, Guangdong Province, China; 2 Department of Orthopaedics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou   510150, Guangdong Province, China
  • Online:2010-10-15 Published:2010-10-15
  • Contact: Lü Yu-ming, Doctor, Associate chief physician, Department of Orthopaedics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou 510150, Guangdong Province, China lvyuming2008@126.com
  • About author:Chen Zhu-sheng☆, Doctor, Associate chief physician, Department of Orthopaedics, The Second Traditional Chinese Medicine Hospital of Guangdong Province, Guangzhou 510095, Guangdong Province, China czs8341@163.com
  • Supported by:

    Natural Science Foundation of Guangdong Province, No. 7002690*; Medical Scientific Research Fund of Guangdong Province, No. A2008128*; a grant by Guangzhou Medical College, No. 0709106*

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摘要：

背景：关节软骨损伤修复面临的难题主要表现在再生软骨的结构及生理功能距正常软骨相差较远，无法满足正常生理需要，目前修复方法很多，但效果均不理想。
目的：探讨纳米β-磷酸三钙/Ⅰ、Ⅱ型胶原层状支架-骨髓间充质干细胞复合物修复犬膝关节骨软骨缺损的可行性。
方法：12月龄杂种犬10只，随机分为实验组、缺损对照组，无菌条件下自髂后嵴处抽取骨髓分离培养骨髓间充质干细胞，传至第3代消化、收集，调整细胞浓度为2×109 L-1，与纳米β-磷酸三钙/Ⅰ、Ⅱ型胶原共培养24 h，即制成纳米β-磷酸三钙/Ⅰ、Ⅱ型胶原层状支架-骨髓间充质干细胞复合物。两组犬均制备右膝关节骨软骨缺损，实验组于缺损处植入支架-细胞复合物，缺损对照组不做任何处理。
结果与结论：第12周，实验组缺损内充填以白色半透明组织，表面光滑，触之较软，稍高出周围软骨面，软骨细胞分布较均一，排列无方向性；第24周，实验组缺损内充填白色半透明新生软骨组织，色泽与正常软骨相似，质韧，表面平整，与正常软骨界限消失，表面细胞平行于关节面，深层细胞排列紊乱，细胞呈团状，基质异染广泛，与周围正常软骨连接良好。缺损对照组缺损未修复，底部为白色纤维组织。提示骨髓间充质干细胞是修复关节软骨缺损较理想的种子细胞，在新生软骨形成的同时，纳米β-磷酸三钙/Ⅰ、Ⅱ型胶原层状支架逐渐降解吸收，是组织工程修复关节软骨缺损适宜的支架材料。

关键词: 纳米, &beta, -磷酸三钙, Ⅰ型胶原, Ⅱ型胶原, 骨髓间充质干细胞, 软骨细胞, 组织工程

Abstract:

BACKGROUND: Articular cartilage injury repair is mainly faced with the great difference in the structure and physiological functions between regenerated cartilage and normal cartilage, far away from normal physiological needs, there are many current repair methods, but the results are not satisfactory.
OBJECTIVE: To study the feasibility of repairing articular osteochondral defects in canines using nano-β-tricalcium phosphate (TCP)/collagen I and II composite loaded with bone marrow stromal stem cells (BMSCs).
METHODS: Ten 12-month-old hybrid canines were randomly divided into experimental group and defect control group, and BMSCs were sterilely obtained from iliac crest in each animal. Primary culture and subcultures were done. The third passage of BMSCs was digested and collected, cell density was adjusted to 2.0 × 106 L-1, the cells were co-cultured with nano- β-TCP/ colI/colII for 24 hours, then nano β-TCP/colI/colII/BMSCs composites were prepared. Articular osteochondral defects were made in canine right knees, those in experimental group were treated with nano β-TCP/colI/colII/BMSCs composites, while those in control group were left untreated.
RESULTS AND CONCLUSION: At 12 weeks, the defects in the experimental group were filled with white translucent tissues, which appeared smooth, soft and slightly protruded, chondrocytes distributed uniformly but aligned disorderly. At 24 weeks, the defects in the experimental group were filled with white translucent cartilage tissue, which appeared smooth and tenacious. The color and the luster were similar to that of the normal cartilage, and was ill-demarcated from the surrounding normal cartilage. The cells on the surface paralleled to joint surface. The cells in the deep layer arranged disorderly. In many areas, the cells clustered together. The matrix was extensively metachromatic and the new cartilage finely integrated with normal cartilage. In the control group, the defects showed little repair response macroscopically, only some white fibrous tissue can be seen at the bottom. BMSCs are ideal seed cells for repairing articular cartilage defects. Nano-β-TCP/colI/colII is gradually degraded and absorbed while new cartilage tissues form. It can be used as a suitable scaffold material for tissue engineered repair of articular cartilage defects.

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