[Synthesis of new gene-loaded microbubbles serve as gene delivery vehicle applied in reporter gene transfer into cardiac myocytes].
To improve the stability and gene-carried capability of gene-attached microbubbles, the method for manufacture of albumin microbubbles was modified and new gene-loaded microbubbles were synthesized by incorporated gene-PEI complex into the shell of microbubbles. Agarose gel electrophoresis and bacteria transformation showed that PEI had the ability to provide the protection of plasmid DNA from ultrasonic degradation. The new gene-loaded microbubbles exhibited excellent acoustical and hemorheological properties. Moreover, they could carry more plasmid DNA than gene-attached microbubbles. beta-galactosidase plasmid transfection into cardiac myocytes was performed by using ultrasound targeted destruction of new gene-loaded microbubbles or gene-attached microbubbles. Gene expression in cardiac myocytes was detected by beta-galactosidase in situ staining and quantitive assay. It was shown that beta-galactosidase activity in cardiac myocytes was enhanced 107-fold by ultrasonic destruction of gene-loaded microbubbles compared with naked plasmid transfection and new gene-loaded microbubbles resulted in 6.85-fold increase in beta-galactosidase activity compared with optimal transfection mediated by gene-attached microbubbles. These results suggested that ultrasonic destruction of the gene-loaded microbubbles can enhance the cardiac myocytes exogenous gene transfer efficiency significantly and new gene-loaded microbubbles is an efficient and safe gene delivery vehicle.
Wang
Guozhong
G
Department of Cardiology, The First Affiliated Hospital, College of Medical Sciences, Zhejiang University, Hangzhou 310003, China.
Hu
Shenjiang
S
Zheng
Zhelan
Z
Sun
Jian
J
Zheng
Xia
X
Zhu
Zhaohui
Z
Li
Jiang
J
Yao
Yumei
Y
D000818:Animals; D002478:Cells, Cultured; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors; D007097:Imines; D045423:Microbubbles; D032383:Myocytes, Cardiac / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011095:Polyethylenes; D051381:Rats; D017208:Rats, Wistar; D013010:Sonication; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D007097:Imines; D011095:Polyethylenes; C505405:poly(ethylene imine); D001616:beta-Galactosidase