[In vivo endochondral bone formation by implanting human bone morphogenetic protein-2-producing fibroblasts into nude mouse muscle].
OBJECTIVE
To determine whether fibroblasts can be used to promote endochondral bone formation in vivo by transfer of human bone morphogenetic protein-2(hBMP-2) into fibroblasts.


METHODS
pcDNA3-hBMP-2 was constructed by use of gene clone and recombined technique. NIH3T3 fibroblasts were transfected with pcDNA3-hBMP-2. The positive cell clones were selected with G418. In NIH3T3 fibroblasts transferred with pcDNA3-hBMP-2, the expression of hBMP-2 was determined by in situ hybridization and immunohistochemical analysis; alkaline phosphatase activity was measured. hBMP-2-producing fibroblasts were implanted into nude mouse muscle to observe endochondral bone formation in vivo.


RESULTS
pcDNA3-hBMP-2 was successfully constructed. In NIH3T3 fibroblasts transfected with pcDNA3-hBMP-2, the BMP-2 expression was stable; alkaline phophatase activity was much higher than that in non-transfected NIH3T3 cells. Endochondral bone formation in vivo was observed at the site of implantation 4 weeks later.


CONCLUSIONS
Fibroblasts transfected by hBMP-2 gene can be used to promote endochondral bone formation in vivo.
Li
Xiang-dong
XD
Institute of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an Shaanxi, P. R. China 710032.
Hu
Yun-yu
YY
D016475:3T3 Cells; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts / Q000502:physiology*; D005822:Genetic Vectors; D051379:Mice; D008819:Mice, Nude; D010012:Osteogenesis; D014161:Transduction, Genetic; D014162:Transfection; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta