Ultrasound-mediated destruction of microbubbles enhances gene expression after lipofectamine-mediated transfection of cells in vitro / 中华超声影像学杂志

ABSTRACT

Objective To study the effects of ultrasound mediated microbubbles destruction on endothelial cells transfected with lipofectamine DNA complexes.Methods One hour after addition of pIRES2 EGFP hVEGF gene and lipofectamine,endothelial cells on 6 well plates were exposed to ultrasound for 30 s,60 s,or 90 s in the presence(10 ?l,100 ?l,500 ?l) or absence of ultrasound contrast agent.The transducer (Sonos 5500,S3, 1.3 MHz,MI 1.0 ) was submerged in water and held at a distance of 1 cm.Expression of the marker gene EGFP was observed by fluorescent microscopy 2 days later.Results Ultrasound treatment of endothelial cells on plates for 30 s and 60 s yielded significant increase in transfection rates without damaging the cells,but 90 s treatment killed most of the cells.Ultrasound contrast agent was able to significantly increase endothelial cells transfection rate by enhancing cavitation effects at a concentration of 100 ?l,and damaged most cells when applied at a concentration of 500 ?l.Conclusions Ultrasound mediated destruction of microbubbles enhances gene expression after lipofectamine mediated transfection of endothelial cells in vitro; and it may represent an improved avenue for therapeutic gene delivery in vivo.