[Synthesis of new gene-loaded microbubbles serve as gene delivery vehicle applied in reporter gene transfer into cardiac myocytes].

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi

Department of Cardiology, The First Affiliated Hospital, College of Medical Sciences, Zhejiang University, Hangzhou 310003, China.

Published: August 2006

AI Article Synopsis

  • The method for creating gene-loaded microbubbles was enhanced by incorporating gene-PEI complexes into their structure, improving stability and gene delivery capacity.
  • Agarose gel electrophoresis and bacterial transformation demonstrated that PEI protects plasmid DNA during ultrasonic exposure, allowing better gene transfer.
  • Ultrasound-targeted destruction of these new microbubbles significantly increased gene expression in cardiac myocytes, showing a 107-fold increase in beta-galactosidase activity compared to traditional methods, highlighting their effectiveness as a gene delivery vehicle.

Article Abstract

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.

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