Induction of cell-membrane porosity by ultrasound

Tachibana, Kunihide; Uchida, Toshiki; Ogawa, Koichi; Yamashita, Nobuya; Tamura, Kazuo

doi:10.1016/s0140-6736(99)01244-1

Cited by 424 publications

(264 citation statements)

References 4 publications

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“…Increased intracellular delivery is believed to occur due to the formation of transient pores of nanometer to micron dimensions [25,43,44]. Consistent with this expectation, our flow cytometry results show that ultrasound increased intracellular DNA delivery for both naked DNA and PEI:DNA complexes (Fig.…”

Section: Discussionsupporting

confidence: 87%

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Deshpande

Prausnitz

2007

Journal of Controlled Release

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Ultrasound and poly(ethylenimine) (PEI) have each separately been shown to increase DNA transfection efficiency. This study tested the hypothesis that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection. This in vitro study assessed transfection efficiency of two different DNA plasmids encoding green fluorescent protein and firefly luciferase in two different cells types, a primary culture of human aortic smooth muscle cells and an immortal line of human prostrate cancer cells. We found that ultrasound sonication increased transfection up to 18-fold, DNA complexation with PEI increased transfection up to 90-fold, and the combination of ultrasound and PEI synergistically increased transfection up to 200-fold, which resulted in reporter gene expression by 34% of cells. Kinetic measurements found that the effects of ultrasound alone acted quickly, whereas increased transfection by PEI either alone or in combination with ultrasound strongly benefited from a 4-h incubation with the DNA plasmid after sonication. Although serum reduced absolute expression levels, it did not affect the relative increase in transfection when ultrasound was added to PEI enhancement. Flow cytometry measurements showed that sonication increased intracellular uptake of labelled DNA complexed to PEI by 55% relative to PEI complexation alone. Electrophoresis assay showed no damage to DNA or PEI-DNA complexes after sonication. Overall, these results suggest that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection.

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Section: Discussionsupporting

confidence: 87%

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Deshpande

Prausnitz

2007

Journal of Controlled Release

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“…sonoporation, is generally accepted as main mechanism by which drugs enter cells during ultrasound application. Scanning electron microscopy images showed clear membrane disruptions after ultrasound exposure [9]. By voltage clamp techniques, an increase in transmembrane current simultaneously with ultrasound application was reported, indicating ion flux through pores [10].…”

Section: Introductionmentioning

confidence: 92%

Ultrasound and microbubble mediated drug delivery: Acoustic pressure as determinant for uptake via membrane pores or endocytosis

Cock

Zagato

Braeckmans

et al. 2015

Journal of Controlled Release

217

247

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Although promising results are achieved in ultrasound mediated drug delivery, its underlying biophysical mechanisms remain to be elucidated. Pore formation as well as endocytosis has been reported during ultrasound application. Due to the plethora of ultrasound settings used in literature, it is extremely difficult to draw conclusions on which mechanism is actually involved. To our knowledge, we are the first to show that acoustic pressure influences which route of drug uptake is addressed, by inducing different microbubble-cell interactions. To investigate this, FITC-dextrans were used as model drugs and their uptake was analyzed by flow cytometry. In fluorescence intensity plots, two subpopulations arose in cells with FITCdextran uptake after ultrasound application, corresponding to cells having either low or high uptake. Following separation of the subpopulations by FACS sorting, confocal images indicated that the low uptake population showed endocytic uptake. The high uptake population represented uptake via pores. Moreover, the distribution of the subpopulations shifted to the high uptake population with increasing acoustic pressure. Real-time confocal recordings during ultrasound revealed that membrane deformation by microbubbles may be the trigger for endocytosis via mechanostimulation of the cytoskeleton. Pore formation was shown to be caused by microbubbles propelled towards the cell. These results provide a better insight in the role of acoustic pressure in microbubble-cell interactions and the possible consequences for drug uptake. In addition, it pinpoints the need of a more rational, microbubble behavior based choice of acoustic parameters in ultrasound mediated drug delivery experiments.

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“…This observation is consistent with the previous report that ultrasound induced cell-membrane porosity. 37 Thus, we further modified ultrasound-mediated plasmid DNA transfection using echo contrast microbubble. The present study confirmed the previous observation that, ultrasound exposure in the presence of microbubble echo-contrast agents enhances acoustic, cavitation and is associated with approximately 300-fold increments in transgene expression after, naked DNA transfection in in vitro experiments.…”

Section: (B) Effect Of Transfection Of Human Hgf Plasmid On Number Ofmentioning

confidence: 99%