Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (20): 3637-3640.doi: 10.3969/j.issn.1673-8225.2010.20.007

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AAV-hBMP-4 versus AAV-EGFP for osteogenic induction of rabbit bone marrow mesenchymal stem cells in vitro

Zhao Xi-chun1, Dong Zhi-yong1, Zheng Zhao-min2, Kuang Guan-ming3    

  1. 1Department of Orthopaedics, Jiaozhou Central Hospital of Qingdao City, Qingdao  266300, Shandong Province, China;
    2Department of Spinal Surgery, First Affiliated Hospital of Sun Yet-sen University, Guangzhou  510080, Guangdong Province, China;
    3Department of Orthopaedics, the University of Hong Kong, Hong Kong, China
  • Online:2010-05-14 Published:2010-05-14
  • Contact: Zheng Zhao-min, Professor, Doctor, Doctoral supervisor, Department of Spinal Surgery, First Affiliated Hospital of Sun Yet-sen University, Guangzhou 510080, Guangdong Province, China zhengzm1@163.com
  • About author:Zhao Xi-chun, Associate professor, Associate chief physician, Department of Orthopaedics, Jiaozhou Central Hospital of Qingdao City, Qingdao 266300, Shandong Province, China zhiyongzsy@163.com
  • Supported by:

    the Fok Ying-Tong Education Foundation, No.94020*

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Abstract:

BACKGROUND: The releasing speed of human bone morphogenetic protein (hBMP) is not consistent with the new bone formation, thus, it is necessary to adjust the BMP release speed by suitable vector. 

OBJECTIVE: To determine osteogenic effects of transfection of adeno-associated virus (AAV)-hBMP-4 and AAV-enhanced green fluorescent proteins (EGFP) on rabbit bone marrow mesenchymal stem cells (BMSCs) in vitro. 

METHODS: The BMSCs were cultured using total bone marrow method, and then the cells were transfected with AAV-hBMP-4 and AAV-EGFP, respectively, with 5×104 infection value. After that, the differences of osteogenic effects of 2 groups were compared by observing cell morphological changes, alkali phosphatase staining, Von Kossa staining, alizarin bordeaux staiing and alkali phosphatase content.

RESULTS AND CONCLUSION: The ossification of BMSCs was obviously observed after transfected with AAV-hBMP4, which was identified by alkali phosphatase staining, Von Kossa staining and alizarin bordeaux staining. No similar changes were observed in the AAV-EGFP group. The content of alkali phosphatase was greater in the AAV-hBMP4 group than that of the AAV-EGFP group (P < 0.01). The results demonstrated that AAV-EGFP transfected BMSCs presented notably osteogenic activity.

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