[Adenovirus-mediated human bone morphogenetic protein 2 gene transferred to rabbit intervertebral disc cells in vitro].
OBJECTIVE
To study the adenovirus-mediated human bone morphogenetic protein-2 gene (Ad-hBMP-2)transferred to the intervertebral disc cells of the New Zealand rabbit in vitro.


METHODS
The cells of New Zealand white rabbits were isolated from their lumbar discs. The cells were grown in the monolayer and treated with an adenovirus encoding the LacZ gene (Ad-LacZ) and Ad-hBMP-2 (50, 100, 150 MOI, multiplicity of infection) in the Dulbecco's Modified Eagle Medium and the Ham's F-12 Medium in vitro. Three days after the Ad-hBMP-2 treatment, the expression of hBMP-2 in the cells that had been infected by different doses of MCI was determined by immunofluorescence and the Western blot analysis, and the expression was determined in the cells with the Ad-LacZ treatment in a dose of 150 MDI. Six days after the Ad-hBMP-2 treatment, mRNA was extracted fur the reverse transcription polymerase chain reaction (RT-PCR) and the difference was detected between the control group and the culture group that was treated with Ad-hBMP-2 in doses of 50, 100 and 150 MCI so that the expressions of aggrecan and collagen II mRNA could be observed.


RESULTS
The expression of hBMP-2 in the cells was gradually increased after the transfection in an increasing dose, which was observed by immunofluorescence and the Western blot analysis. At 6 days the aggrecan and collagen type II mRNA expressions were up-regulated by Ad-hBMP-2 after the transfection at an increasing viral concentration in the dose-dependent manner.


CONCLUSIONS
The results show that Ad-hBMP-2 can transfect the rabbit intervertebral disc cells in vitro with a high efficiency rate and the expression of hBMP-2 after the infection is dose-dependent in the manner. Ad-hBMP-2 after transfection can up-regulate the expression of aggrecan and collagen IL mRNA at an increasing viral concentration.
Gu
Hailun
H
Department of Orthopedics, First Affiliated Hospital, China Medical University, Shenyang Liaoning, 110001, PR China.
Lv
Gang
G
Liu
Li
L
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D002478:Cells, Cultured; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology*; D008159:Lumbar Vertebrae; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D014162:Transfection
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D012333:RNA, Messenger