Ultrasound-mediated microbubble destruction enhances GFP gene expression in human gingival fibroblasts cells in vitro / 第三军医大学学报

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.