中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (21): 3806-3810.doi: 10.3969/j.issn.1673-8225.2010.21.002

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

自制人骨形态发生蛋白2复合骨修复材料诱导骨再生

汪  炬1，肖  刚1，陈  阳1，周智优1，林永亮2，吴  昊3，查振刚3，周天鸿1   

  1. 1暨南大学生命科学技术学院，广东省广州市  510632；2广东冠昊生物科技有限公司，广东省广州市  510663；3暨南大学第一附属医院骨科，广东省广州市  510632
  • 出版日期:2010-05-21 发布日期:2010-05-21
  • 通讯作者: 查振刚，博士，教授，暨南大学第一附属医院骨科，广东省广州市 510632
  • 作者简介:汪 炬★，女，1968年生，安徽省绩溪县人，汉族，暨南大学毕业，硕士，副研究员，主要从事蛋白质工程研究。 wangju1688@163.com
  • 基金资助:

    省部产学研项目(2007A090302083)“新型rhBMP2人工骨复合材料的产业化”；省科技攻关项目(2009B030801237) ”新型细胞因子仿生复合骨生物材料的研制及产业化”；广东省教育部科技部企业科技特派员行动计划(2009B090600134)资助项目“rhBMP2 复合生物型人工骨新一代骨诱导材料及制品的商品化研究”。

Osteoinductive activity of self-made human bone morphogenetic protein-2 composite bone in bone repairing

Wang Ju1, Xiao Gang1, Chen Yang1, Zhou Zhi-you1, Lin Yong-liang2, Wu Hao3, Zha Zhen-gang3, Zhou Tian-hong1   

  1. 1 College of Life Science and Technology, Jinan University, Guangzhou   510632, Guangdong Province, China; 2 Grandhope Biotechnology Limited Company, Guangzhou   510663, Guangdong Province, China; 3 Department of Orthopaedics, Jinan University Affiliated First Hospital, Guangzhou   510632, Guangdong Province, China
  • Online:2010-05-21 Published:2010-05-21
  • Contact: Zha Zhen-gang, Doctor, Professor, Department of Orthopaedics, Jinan University Affiliated First Hospital, Guangzhou 510632, Guangdong Province, China
  • About author:Wang Ju★, Master, Associate investigator, College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China wangju1688@163.com
  • Supported by:

    Province and Ministry Project of Industry, University and Research, No. 2007A090302083*; Provincial Scientific and Technological Project, No. 2009B030801237*; Enterprise Scientific Special Agents by Guangdong Province, Ministry of Education and Ministry of Science and Technology, No. 2009B090600134*

PDF

472

被引次数

12

摘要：

背景：骨形态发生蛋白是目前发现的一类惟一可独立诱导骨再生的细胞因子，临床应用前景广阔。但它们价格昂贵，限制了在中国的应用。
目的：探讨自主生产的重组人骨形态发生蛋白2的骨诱导活性及对骨缺损的修复作用。
方法：通过电泳及质谱的方法检测其复性后重组人骨形态发生蛋白2的纯度，通过C2C12细胞在重组人骨形态发生蛋白2诱导下向成骨细胞分化检测其骨诱导活性。将重组人骨形态发生蛋白2与以动物骨为原料加工制成的新型骨修复材料复合，植入新西兰兔桡骨缺损实验模型观察其对骨缺损的修复作用。
结果与结论：自主生产的重组人骨形态发生蛋白2纯度≥95%，能明显诱导C2C12细胞向成骨细胞分化，分化早期大量表达碱性磷酸酶，分化末期形成大量的矿化小节，诱骨活性良好。在兔桡骨修复试验中，X射线影像学及苏木精-伊红染色组织学检查结果表明，重组人骨形态发生蛋白2复合材料组在2个月时形成大量骨痂，4~6个月时载体与自体骨融合，材料大量降解，修复效果良好；单纯骨填充材料组4~6个月时只有少量新生骨桥与自体骨连接，降解量极少；空白对照组损伤部位为纤维化组织所填充。结果提示自主生产的重组人骨形态发生蛋白2促进骨修复作用明显，适合应用于临床。

关键词: 自主生产, 人骨形态发生蛋白, 生物型骨填充材料, 桡骨缺损, 骨修复

Abstract:

BACKGROUND: Bone morphogenetic protein is a kind of growth factors only which can induce bone regeneration independently that we found until now. The prospect of clinical application of bone morphogenetic proteins is very bright. But their high cost limits their use in China.
OBJECTIVE: To evaluate the osteoinductive activity and the bone-defect repairing ability of self-made recombinant human bone morphogenetic protein (rhBMP-2).
METHODS: The purity of rhBMP-2 was assayed by SDS-PAGE and mass spectrography. The osteoblastic to differentiation markers of C2C12 cells after induction of rhBMP-2 were assayed to identify the osteoinductive activity of rhBMP-2. The bone-defect repairing ability of rhBMP-2 was verified by New Zealand rabbit radial bone repair trial after the composition of material and rhBMP-2.
RESULTS AND CONCLUSION: The purity of rhBMP-2 was ≥95%, rhBMP-2 could greatly induce C2C12 cells to differentiate into osteoblasts, alkaline phosphatase largely expressed at early stage of the differentiation, and many mineralized nodules formed at end stage, indicating good osteoinductive activity of the self-produced rhBMP-2. In the New Zealand rabbit radial bone repair trials, the results of X-ray assay and hematoyline-eosin staining showed that, massive callus formed on the surface of rhBMP-2 composite material at 2 months, and fused with autologous bone tissue at 4 months, a large amount of materials degraded. In the pure material group, a small amount of bone bridges produced newly linked with the planted material in which the degradation was little. In blank group, the defect area was filled with fibrosis tissue. The results suggested that self-produced rhBMP-2 could markedly promote the bone repairing and could be used for clinical application.

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