Shock waves: a novel method for cytoplasmic delivery of antisense oligonucleotides

Abstract

Intracytoplasmic delivery of oligonucleotides (ODN) can improve ODN-based strategies such as the antisense approach and the use of immunostimulatory CpG dinucleotide containing ODN. Shock waves are established for the treatment of nephrolithiasis and other diseases. Here we describe the use of shock waves as a new physical method for the direct transport of antisense ODN into the cytoplasm and the nucleus of cells. Human peripheral blood mononuclear cells together with antisense ODN were exposed to shock waves generated by an electrohydraulic lithotripter. ODN uptake was examined by flow cytometry and fluorescence microscopy. By optimization of physical parameters we achieved the transfer of high amounts of ODN which were detected within less than 5 min after shock wave exposure, with viability of cells higher than 95%. Transfection of human peripheral blood mononuclear cells with an antisense ODN directed against tumor necrosis factor (TNF) alpha resulted in a reduction in lipopolysaccharide-induced TNF production by 62% (n=5, P=0.006). Specificity of TNF suppression was confirmed with a four-mismatch oligonucleotide. Positive atmospheric pressure abolished antisense-mediated inhibition of TNF synthesis by blocking shock wave-induced cavitation and formation of oscillating air bubbles. Electroporation was less effective. The use of shock waves is thus an efficient physical tool for ODN delivery to cells. Shock waves may allow the evaluation of target proteins in cell types difficult to transfect with other methods and thus may improve the antisense technique for the analysis of unknown genes.

Superpulsed laser irradiation increases osteoblast activity via modulation of bone morphogenetic factors

“…The possibility that the effects of laser irradiation are in part mediated by changes in plasmamembranes may be supported by observations that shock waves, another kind of mechanical biostimulation, increase substance uptake and delivery via membrane modulation [29,32].…”

On the Destruction of Cancer Cells Using Laser-Induced Shock-Waves: A Review on Experiments and Multiscale Computer Simulations

“…38 When the energy deposited by the lithotripter is reduced below the lethal value, it is possible to permeabilize the cells without killing them. [42][43][44][45][46] In this way, molecules present in the surrounding medium can diffuse into the cells. This effect has possible applications for drug delivery and gene transfer.…”

Genetic Transformation Systems in Fungi, Volume 1

“…In vivo and in vitro studies revealed that shock waves may cause transient cell permeabilization, allowing large molecules (normally excluded by the cell membrane) to become trapped inside the cell, opening the possibility of shock wave drug delivery and gene transfection (Gambihler et al 1994 ;Lauer et al 1997 ;Bao et al 1998 ;Tschoep et al 2001 ;Schaaf et al 2003 ;Doukas and Fig. 21.1 Sketch of a pressure waveform recorded at the focus of a shock wave generator, showing the peak positive pressure p + , the peak negative pressure p − , the rise time and the full-width-half-maximum.…”