Analysis of in vitro Transfection by Sonoporation Using Cationic and Neutral Microbubbles

Abstract

The objective of the study was to examine the role of acoustic power intensity and microbubble and plasmid concentrations on transfection efficiency in HEK-293 cells using a sonoporator with a 1-MHz transducer. A green fluorescent protein (GFP) reporter plasmid was delivered in as much as 80% of treated cells, and expression of the GFP protein was observed in as much as 75% of cells, using a power intensity of 2 W/cm 2 with a 25% duty cycle. In addition, the relative transfection abilities of a lipid noncationic and cationic microbubble platform were investigated. As a positive control, cells were transfected using Lipofectamine reagent. Cell survival and transfection efficiency were inversely proportional to acoustic power and microbubble concentration. Our results further demonstrated that high-efficiency transfection could be achieved, but at the expense of cell loss. Moreover, direct conjugation of plasmid to the microbubble did not appear to significantly enhance transfection efficiency under the examined conditions, although this strategy may be important for targeted transfection in vivo.

Cationic versus Neutral Microbubbles for Ultrasound-mediated Gene Delivery in Cancer

“…Because the phosphate backbone of DNA and RNA render them highly anionic, MBs with cationic lipid shells have thus been utilized as a simple strategy to engineer MBs that spontaneously complex with genetic cargo by means of charge interactions (14,22,(26)(27)(28)(29)(30). To determine whether systemic administration of genes directly coupled onto MBs confers a transfection efficiency advantage in ultrasound-mediated gene delivery versus traditional coadministration of uncoupled genes and MBs, a direct comparison between the two approaches is necessary.…”

New chitosan nanobubbles for ultrasound-mediated gene delivery: preparation and in vitro characterization

“…The use of microbubbles as gene delivery vehicles and transfectionenhancing agents is particularly promising. 2,4,5 Various methods have been proposed for the delivery of genes using microbubbles, including: direct physical incorporation of DNA into the microbubble shell during fabrication; use of cationic lipids incorporated into the microbubble shell to bind DNA electrostatically; use of single or multiple layers of cationic polymer on the microbubble shell to bind DNA electrostatically; covalent linking of DNA-nanoparticle carriers; and use of complementary DNA strands to load nanoparticles. Most of these microbubble formulations have demonstrated high DNA-loading capacity and/or high transfection efficiency.…”

Micro- and nanobubbles: A versatile non-viral platform for gene delivery

“…Then the destruction of microbubbles may cause high-energy microstreams or microjets, that will cause shear stress on the cell membrane enhancing cell uptake, probably due to transient holes. This phenomenon has been deeply studied as a way to mediate the transfer of different molecules (Tlaxca et al 2010, Bao et al 1997, Greenleaf et al 1998, Guzman et al 2001a, Kim et al 1996, Tata et al 1997. Sonoporation has been shown to enhance the intracellular delivery of small molecules, macromolecules (Guzmán et al 2001b, Guzmán et al 2002,Karshafian et al 2009, Miller et al 1999, (Yu et al 2006) and genetic materials (Bao et al 1997).…”