Abstract
Ultrasound-mediated gene transfection in the presence of microbubbles is a recently developed, promising nonviral gene delivery method. Detailed dynamics of pore opening on the cell surface have not been clarified. In particular, pore size is one of the most essential parameters. In this study, we investigated the size effect of complexed plasmid DNA (pDNA) on transfection efficiency by packaging the pDNA within polyplex micelles. Both naked pDNA and complexed pDNA were transfected into cultured NIH3T3 cells by using ultrasound in the presence of a microbubble contrast agent, Sonazoid. The sizes of the hydrodynamic diameter of the naked and complexed pDNA, estimated by a dynamic light scattering measurement, were 600 and 120 nm, respectively. The transfection rates of the complexed pDNA, evaluated by counting the number of cells that exhibited green fluorescence, was 1.67%, while that of the naked pDNA was 0.92%. This efficiency enhancement, which depends on size reduction, showed that the pore sizes were distributed in the range of the pDNA diameters.
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