[Ultrasound-mediated microbubble destruction enhances exogenous gene expression in NIH3T3 cells in vitro].
OBJECTIVE
To investigate the transfection efficiency of the recombinant human bone morphogenetic protein-2 (hBMP-2) gene in targeted cells by ultrasound-mediated microbubble destruction.


METHODS
NIH3T3 cells' anabiosis was completed and went down to the 3rd or 4th generation, and cultured in 6 well plates. The cells were divided into 2 groups: Plasmid DNA and Lipofectamine 2000 group (liposome group), plasmid DNA and ultrasound and microbubble group (ultrasound-mediated microbubble destruction group). Plasmid DNA was transfected into cells with liposome or ultrasound and microbubble. 24-48 hours later, the transfection efficiency and the concentrations of hBMP-2 were measured with fluoresence microscope and enzyme-linked immunosorbent assay (ELISA) respectively. The data were analyzed by curve fitting and t-test of SPSS 11.5.


RESULTS
The transfection efficiency rate was (7.30 +/- 1.58)% in liposome group, compared with (11.77 +/- 3.16)% in ultrasound-mediated microbubble destruction group (P< 0.05). The concentration of hBMP-2 after transfection was (1164.35 +/- 724.67) pg/mL in liposome group, versus (2932.70 +/- 656.27) pg/mL in ultrasound-mediated microbubble destruction group (P<0.05).


CONCLUSIONS
Ultrasound-mediated microbubble destruction could significantly improve the transfection efficiency and expression of hBMP-2 gene in NIH3T3 cells. It may provide a new and effective gene delivery system for gene therapy in periodontal regeneration.
Sun
Qin-Feng
QF
Department of Periodontology, School of Stomatology, Shandong University, Jinan 250012, China.
Liu
Yu
Y
Yang
Pi-Shan
PS
Du
Fang
F
D000818:Animals; D055396:Bone Morphogenetic Protein 2; D015316:Genetic Therapy; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D045423:Microbubbles; D041681:NIH 3T3 Cells; D010957:Plasmids; D011994:Recombinant Proteins; D014162:Transfection; D016212:Transforming Growth Factor beta; D014465:Ultrasonics
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2