Progress in the development of ultrasound-mediated gene delivery systems utilizing nano- and microbubbles.
ABSTRACT Recently, ultrasound-mediated gene delivery with nano- and microbubbles was developed as a novel non-viral vector system. In this gene delivery system, microstreams and microjets, which are induced by disruption of nano/microbubbles exposed to ultrasound, are used as the driving force to transfer genes into cells by opening transient pores in the cell membrane. This system can directly deliver plasmid DNA and siRNA into cytosol without endocytosis pathway. Therefore, these genes are able to escape from degradation in lysosome and result in enhancing the efficiency of gene expression. In addition, it is expected that ultrasound-mediated gene delivery using nano/microbubbles would be a system to establish non-invasive and tissue specific gene expression because ultrasound can transdermally expose to target tissues and organs. This review focuses on the current ultrasound-mediated gene delivery system using nano/microbubbles. We discuss about the feasibility of this gene delivery system as novel non-viral vector system.
Article: New chitosan nanobubbles for ultrasound-mediated gene delivery: preparation and in vitro characterization.[show abstract] [hide abstract]
ABSTRACT: The development of nonviral gene delivery systems is one of the most intriguing topics in nanomedicine. However, despite the advances made in recent years, several key issues remain unsettled. One of the main problems relates to the difficulty in designing nanodevices for targeted delivery of genes and other drugs to specific anatomic sites. In this study, we describe the development of a novel chitosan nanobubble-based gene delivery system for ultrasound-triggered release. Chitosan was selected for the nanobubble shell because of its low toxicity, low immunogenicity, and excellent biocompatibility, while the core consisted of perfluoropentane. DNA-loaded chitosan nanobubbles were formed with a mean diameter of less than 300 nm and a positive surface charge. Transmission electron microscopic analysis confirmed composition of the core-shell structure. The ability of the chitosan nanobubbles to complex with and protect DNA was confirmed by agarose gel assay. Chitosan nanobubbles were found to be stable following insonation (2.5 MHz) for up to 3 minutes at 37°C. DNA release was evaluated in vitro in both the presence and absence of ultrasound. The release of chitosan nanobubble-bound plasmid DNA occurred after just one minute of insonation. In vitro transfection experiments were performed by exposing adherent COS7 cells to ultrasound in the presence of different concentrations of plasmid DNA-loaded nanobubbles. In the absence of ultrasound, nanobubbles failed to trigger transfection at all concentrations tested. In contrast, 30 seconds of ultrasound promoted a moderate degree of transfection. Cell viability experiments demonstrated that neither ultrasound nor the nanobubbles affected cell viability under these experimental conditions. Based on these results, chitosan nanobubbles have the potential to be promising tools for ultrasound-mediated DNA delivery.International Journal of Nanomedicine 01/2012; 7:3309-18. · 3.13 Impact Factor
[show abstract] [hide abstract]
ABSTRACT: This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.Proceedings of the National Academy of Sciences 09/2012; 109(41):16486-91. · 9.68 Impact Factor
Dataset: Racz ActaPhysHun 2011 98