title: Microbubble-enhanced ultrasound exposure improves gene transfer in vascular endothelial cells.
abstract: OBJECTIVE
  To explore the effects of ultrasound exposure combined with microbubble contrast agent (SonoVue) on the permeability of the cellular membrane and on the expression of plasmid DNA encoding enhanced green fluorescent protein (pEGFP) transfer into human umbilical vein endothelial cells (HUVECs).
  
  
  METHODS
  HUVECs with fluorescein isothiocyanate-dextran (FD500) and HUVECs with pEGFP were exposed to continuous wave (1.9 MHz, 80.0 mW/cm(2)) for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytometry, respectively.
  
  
  RESULTS
  The percentage of FD500-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone (24.0% +/- 5.5% vs 66.6% +/- 4.1%, P < 0.001). Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue (16.1% +/- 1.9% vs 1.5% +/- 0.2%, P < 0.001). No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue (94.1% +/- 2.3% vs 91.1% +/- 4.1%).
  
  
  CONCLUSIONS
  The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non-invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors.
authors.affiliation: Department of Medical Ultrasonics, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Nie
authors.forename: Hui-Xiong
authors.initials: HX
authors.lastname: Xu
authors.forename: Qing
authors.initials: Q
authors.lastname: Tang
authors.forename: Ming-De
authors.initials: MD
authors.lastname: Lu
mesh_terms: D002478:Cells, Cultured; D003911:Dextrans; D042783:Endothelial Cells / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D045423:Microbubbles; D010743:Phospholipids; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D013459:Sulfur Hexafluoride; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics
chemical_list: D003911:Dextrans; D010743:Phospholipids; D011994:Recombinant Proteins; C420843:contrast agent BR1; C485184:enhanced green fluorescent protein; C015219:fluorescein isothiocyanate dextran; D049452:Green Fluorescent Proteins; D016650:Fluorescein-5-isothiocyanate; D013459:Sulfur Hexafluoride