Ultrasound-mediated microbubble destruction enhances GFP gene expression in human gingival fibroblasts cells in vitro
- VernacularTitle:超声微泡造影剂介导pEGFP-N_1质粒转染HGFs的实验研究
- Author:
Shumei LUO
;
Yunyan ZHANG
;
Xiaobo ZHONG
;
Lin ZHONG
- Publication Type:Journal Article
- Keywords:
utrasonics;
contrast media;
human gingival fibroblasts;
gene transfection
- From:Journal of Third Military Medical University
2003;0(24):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether ultrasound-mediated microbubble destruction can effectively deliver pEGFP-N1 plasmid into human gingival fibroblasts (HGFs).Methods The primary cultured HGFs were divided into 4 groups,that is,plasmid,microbubble+plasmid,ultrasound+plasmid,and ultrasound+microbubble+plasmid groups.pEGFP-N1 plasmid was used to transfect to HGFs as a gene marker with ultrasound-mediated microbubble destruction.The last group was further divided into subgroups to optimize the transfection conditions.After 48 h,phase-contrast fluorescent microscopy was employed to evaluate the expression of green fluorescent protein (GFP).The cell vitality was measured by the MTT assay.Results The transfection efficiency of the ultrasound+microbubble+plasmid group was higher than other experiment groups.Optimal gene expression was found when ultrasound was radiated at 1.5 W/cm2 for 60 s,microbubble was at a concentration of 10% and plasmid was at a concentration of 6.67 ?g/ml,and the transfection efficiency was highest under this condition.Conclusion Under specific conditions,ultrasound mediated microbubble destruction enhances the reporter gene transfection and expression in HGFs.
