Ultrasound-mediated microbubble destruction enhances exogenous gene expression in NIH3T3 cells in vitro.
- Author:
Qin-feng SUN
1
;
Yu LIU
;
Pi-shan YANG
;
Fang DU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bone Morphogenetic Protein 2; Genetic Therapy; Humans; In Vitro Techniques; Mice; Microbubbles; NIH 3T3 Cells; Plasmids; Recombinant Proteins; Transfection; Transforming Growth Factor beta; Ultrasonics
- From: West China Journal of Stomatology 2008;26(2):198-205
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the transfection efficiency of the recombinant human bone morphogenetic protein-2 (hBMP-2) gene in targeted cells by ultrasound-mediated microbubble destruction.
METHODSNIH3T3 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.
RESULTSThe 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).
CONCLUSIONUltrasound-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.