Impact of microbubbles on shock wave-mediated DNA uptake in cells in vitro.
Gas-filled microbubbles have been successfully used as gene delivery reagents in combination with diagnostic ultrasound. Although shock wave exposure has been shown to transfect cells with naked DNA in vitro, it has not been tested whether the addition of microbubbles would augment DNA uptake under those conditions. Therefore, the aim of this study was to test the impact of microbubbles on transgene expression in vitro under shock wave exposure conditions. HEK 293 cells were treated with 60 or 120 pulses of shock waves at varying energy levels. Cells were mixed with either 100 microg/mL luciferase expressing plasmid DNA or with microbubbles that were produced with the same amount of this DNA. Cell death was evaluated after 1 h and transgene expression, after 24 h. In the presence of microbubbles, transgene expression was significantly higher (as much as 29-fold) relative to that obtained without microbubbles. Cells exposed to 120 pulses demonstrated higher transgene expression (as high as 2.7-fold) compared with cells exposed to 60 pulses. The use of microbubbles resulted in greater cell death, varying from 26% (low energy) to 78% (high energy). In conclusion, DNA-loaded microbubbles can significantly increase shock wave mediated gene transfer. However, this effect is associated with increased levels of cell destruction.
Bekeredjian
Raffi
R
Department of Internal Medicine III, University of Heidelberg, Heidelberg, Germany. raffi.bekeredjian@med.uni-heidelberg.de
Bohris
Christian
C
Hansen
Alexander
A
Katus
Hugo A
HA
Kuecherer
Helmut F
HF
Hardt
Stefan E
SE
D016923:Cell Death / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008055:Lipids / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D045423:Microbubbles; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics; D014465:Ultrasonics
D008055:Lipids; D004247:DNA; D008156:Luciferases