中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9121-.doi: 10.3969/j.issn.1673-8225.2010.49.001

• 干细胞培养与分化 •    下一篇

新型骨形态发生蛋白7多肽对骨髓基质干细胞生物学行为的影响

酆波1，2，胡冬煦1，3，张阳德1，3，武宇赤2   

  1. 1中南大学生物医学工程研究院，湖南省长沙市410008；2内蒙古医学院第三附属医院-包钢医院骨科，内蒙古自治区包头市 014010；3卫生部肝胆肠外科研究中心，湖南省长沙市
    410008
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 张阳德，博士生导师，主任医师，教授，中南大学生物医学工程研究院，卫生部肝胆肠外科研究中心，湖南省长沙市 410008
  • 作者简介:酆波☆，男，1970年生，内蒙古自治区包头市人，汉族，中南大学生物医学工程研究院在读博士，主任医师，主要从事生物医用材料的研究。 bb0472111@yahoo.com.cn

Effects of neotype bone morphogenetic protein-7 peptide on biological behavior of bone marrow stromal stem cells

Feng Bo 1,2, Hu Dong-xu 1,3, Zhang Yang-de 1,3, Wu Yu-chi2   

  1. 1 Institute of Biomedical Engineering of Central South University,  Changsha  410008, Hunan Province, China; 2 Department of Orthopaedics, Baogang Hospital, Third Affiliated Hospital of Inner Mongolia Medical College, Baotou  014010, Inner Mongolia Autonomous Region, China; 3 Research Center of Liver, Gall and Intestine Surgery, Ministry of Health, Changsha  410008, Hunan Province, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: Zhang Yang-de, Doctoral supervisor, Chief physician, Professor, Institute of Biomedical Engineering of Central South University, Changsha 410008, Hunan Province, China; Research Center of Liver, Gall and Intestine Surgery, Ministry of Health, Changsha 410008, Hunan Province, China
  • About author:Feng Bo☆, Studying for doctorate, Chief physician, Institute of Biomedical Engineering of Central South University, Changsha 410008, Hunan Province, China; Department of Orthopaedics, Baogang Hospital, Third Affiliated Hospital of Inner Mongolia Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China bb0472111@yahoo.com.cn

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402

被引次数

7

摘要：

背景：国内外对骨形态发生蛋白7的研究倾向于其在组织工程方面的应用，但是骨形态发生蛋白7除具有很强的骨诱导活性之外，还影响动物生殖系统的生长发育及生理功能，如何在充分发挥其骨诱导活性的同时，避免其他生物学作用，是骨组织工程领域面临的一个难题。
目的：观察人工合成的新型骨形态发生蛋白7多肽对骨髓基质干细胞增殖、成骨分化及矿化等生物学行为影响。
方法：分离培养大鼠骨髓基质干细胞并用流式细胞仪进行鉴定。将传至第3代的骨髓基质干细胞分为实验组和对照组，实验组在培养基加入终质量浓度为100 mg/L的骨形态发生蛋白7多肽，对照组不加多肽。分别在培养的2，4，6，8，10 d对各组细胞进行MTT检测，评价其增殖情况；培养7，14 d后，通过检测细胞内钙含量和碱性磷酸酶活性评价其成骨分化情况，并进行钙黄绿素染色观察其矿化情况。
结果与结论：原代培养大鼠骨髓基质干细胞生长良好，其纯度达到91.5%。细胞增殖实验显示骨髓基质干细胞在实验组和对照组中均增殖良好，差异无显著性意义(P > 0.05)；诱导培养7 d和14 d后，实验组碱性磷酸酶活性及钙含量显著高于对照组(P < 0.05)，实验组细胞矿化明显强于对照组。实验结果表明新型骨形态发生蛋白7多肽对骨髓基质干细胞增殖没有显著影响，但可以促进其向成骨方向分化及矿化。

关键词: 骨形态发生蛋白7, 骨髓基质干细胞, 增殖, 成骨分化, 矿化, 生物学行为

Abstract:

BACKGROUND: The study of bone morphogenetic protein-7 (BMP-7) mainly focused on its application to tissue engineering. However, BMP-7 has strong bone induction activity and affects growth, development and physiological function of animals’ reproductive system. How to fully exert its bone induction activity and to avoid other biological effects is a difficulty in bone tissue engineering field.
OBJECTIVE: To explore the effect of BMP-7 peptide on proliferation, osteogenic differentiation and mineralization of bone marrow stromal stem cells (BMSCs).
METHODS: BMSCs were harvested from rats and identified with flow cytometry. The 3rd generation of BMSCs was divided into experimental and control groups. The medium of experimental group was supplemented with 100 mg/L BMP-7 peptide. The medium of control group was not supplemented with BMP-7 peptide. After 2, 4, 6, 8, and 10 days, the proliferation of BMSCs was analyzed by MTT assay. After 7 and 14 days, the calcium contents and alkaline phosphatase activities were measured to assess the differentiation of BMSCs towards osteoblasts. The mineralization of BMSCs was determined by calcein staining.
RESULTS AND CONCLUSION: BMSCs grew well in primary culture and the purity of BMSCs was 91.5%. There was no significant difference in proliferation between experimental and control groups (P > 0.05). On days 7 and 14, calcium contents and alkaline phosphatase activities in the experimental group were significantly higher than the control group (P < 0.05). The mineralization level of BMSCs in the experimental group was significant higher than in the control group. Results suggest that BMP-7 peptide has no influence on the proliferation of BMSCs, but it can promote osteogenic differentiation and mineralization of BMSCs significantly.

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