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[Study of rat osteoblasts transfected by transforming growth factor-beta 1 gene]
Yong Liu1, Jing-Yuan Du, Qi-Xin Zheng
1Department of Orthopedic Surgery, Xiehe Hospital, Tongji Medical School, Huazhong University of Science and Technology, Wuhan, Hubei, P. R. China 430022.
Objective:
To investigate the effect of transforming growth factor-beta 1 (TGF-beta 1) gene transfer on the biological characteristics of osteoblasts.
Methods:
The expression of TGF-beta 1 in the transfected osteoblasts was detected by in situ hybridization and assay of TGF-beta 1 activity in the supernatant (mink lung epithelium cell growth-inhibition test). The effects of gene transfer and supernatant of the transfected osteoblasts on the proliferation and alkaline phosphatase(ALP) activity of osteoblasts were detected by 3H-TdR and MTT.
Results:
The results of in situ hybridization analysis suggested that the osteoblasts transfected by TGF-beta 1 gene could express TGF-beta 1 obviously. The complex medium, which was the mixture of serum-free DMEM and the activated supernatant according to 1:1, 1:2, 1:4, could inhibit growth of Mv-1-Lu evidently and the ratios of inhibition were 16.3%, 22.7%, 28.2% respectively. TGF-beta 1 gene transfer had no effect on the biological characteristics of osteoblasts, but the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts.
Conclusion:
TGF-beta 1 gene transfer promotes the expression of TGF-beta 1 and the biological characteristics of transfected osteoblasts are stable, which is helpful for gene therapy of bone defects in vivo.