A novel ultrasound microbubble carrying gene and Tat peptide: preparation and characterization.
OBJECTIVE
Ultrasound-targeted microbubble destruction is a promising technology for the targeted gene delivery. The purpose of the present study is to prepare a novel lipid ultrasound microbubble-carrying gene and transactivating transcriptional activator (Tat) peptide and to investigate its transfection effect in vivo.


METHODS
Lipid ultrasound microbubbles were prepared using mechanical vibration, and the appearance, distribution, concentration, diameter, and zeta potential of the lipid ultrasound microbubbles were measured. The efficiencies of the microbubble carrying gene and Tat peptide were investigated using the fluorospectrophotometer. Contrast-enhanced ultrasonography was performed on normal rabbits to observe the duration and intensity of enhancement in myocardium. Quantitative analysis was detected using the DFY Ultrasound Image Analyzer. Transfection in vivo was performed using the CGZZ ultrasound gene transfection instrument. The expression of enhanced green fluorescent protein in the organs was observed using confocal laser scanning microscope.


RESULTS
The diameter of the lipid microbubbles carrying gene and Tat was (2.3 +/- 0.4) mum, the concentration was (3.1 +/- 0.4) x10(9)/mL, and Zeta potential was (2.0 +/- 0.1) mV. The gene encapsulation efficiency of the lipid ultrasound microbubbles was 32%, and the Tat encapsulation efficiency was 35%. In vivo experiment showed that lipid ultrasound microbubbles could enhance the echo intensity and transfection efficiency.


CONCLUSIONS
Lipid microbubbles containing gene and Tat peptide can be used as a new vehicle for gene transfection.
Institute of Ultrasound Imaging, Department of Ultrasonography, the Second Affiliated Hospital of Chongqing Medical University, No. 76, Lin Jiang Road, YuZhong District, ChongQing City, 400010, China.
JianLi
J
Ren
ChuanShan
C
Xu
ZhiYi
Z
Zhou
Yong
Y
Zhang
XingSheng
X
Li
YuanYi
Y
Zheng
HaiTao
H
Ran
ZhiGang
Z
Wang
D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D042783:Endothelial Cells / Q000378:metabolism*; D015696:Gene Products, tat / Q000235:genetics* / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D045423:Microbubbles; D013010:Sonication / Q000379:methods*; D014162:Transfection / Q000379:methods*
D015696:Gene Products, tat; D004247:DNA