Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (38): 7031-7036.doi: 10.3969/j.issn.1673-8225.2010.38.001

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Adenovirus mediated bone morphogenetic protein-2 gene transfection of bone marrow mesenchymal stem cells combined with nano-hydroxyapatite to construct bone graft material in vitro

Huang Jun-feng 1,2, Wang Da-ping2, He Chun-lei1, Yang Xin-jian2, Liu Li-jun2, Chen Hong-xian2, Peng Liang-quan2   

  1. 1 Guangzhou Medical College, Guangzhou   510182, Guangdong Province, China; 2 Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen   518035, Guangdong Province, China
  • Online:2010-09-17 Published:2010-09-17
  • Contact: Wang Da-ping, Professor, Chief physician, Doctor, Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China dapingwang@medmail.com.cn
  • About author:Huang Jun-feng★, Master, Attending physician, Guangzhou Medical College, Guangzhou 510182, Guangdong Province, China; Department of Orthopaedics, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China hjf097@163.com
  • Supported by:

    Science and Technology Projects by Health Bureau of Shenzhen, No. 200630*

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Abstract:

BACKGROUND: With advanced tissue engineering and gene engineering technologies, gene enhanced tissue engineered bone provides prosperous application for the repair of bone deficits. 
OBJECTIVE: To study the adhesion, proliferation and expression of bone marrow mesenchymal stem cells (BMSCs) on nano-hydroxyapatite (Nano-HA) bone graft material after transfection of adenovirus-mediated human bone morphogenetic protein-2 expression vector (Ad-BMP-2).  
METHODS: BMSCs were transfected using Ad-BMP-2. Immunohistochemistry and Western blot were used to detect BMP-2 expression in transfected cells. After transfected 48 hours, cells were inoculated on Nano-HA bone graft material evenly, scanning electron microscopy was used to observe the adhesion status of cells on material, and MTT test was used to assay the proliferation of transfected BMSCs. Cells adhered on material were gathered on days 3, 5, 7, Western blot was used to detect the expression of BMP-2 of the adhered cells. 
RESULTS AND CONCLUSION: After transfection, BMP-2 protein was highly expressed in BMSCs; Using scanning electron microscopy, the transfected BMSCs were seen to grow well around or in pores of the Nano-HA bone graft material, and proliferated. MTT test assay showed that, the Nano-HA bone graft material couldn’t inhibit in vitro proliferation of BMSCs. The transfection cells adhered on composite had a higher BMP-2 protein expression. Results demonstrated that Ad-BMP-2-transfected BMSCs are well biocompatible to Nano-HA bone graft material, the transfected cells in material can secrete BMP-2 stably for a long time.

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