Enhancement of ultrasound‐mediated gene transfection by membrane modification

Nozaki, Tomoyoshi; Ogawa, Ryohei; Feril, Loreto B.; Kagiya, Go; Fuse, Hideki; Kondo, Takashi

doi:10.1002/jgm.445

Cited by 55 publications

(37 citation statements)

References 32 publications

(36 reference statements)

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“…However, this also influences the stiffness of the lipid microbubble shell [31], thereby changing the ultrasound-responsiveness and behavior of the microbubbles [32]. Moreover, cell membrane fluidity is affected by temperature and this may again alter the vulnerability of cells to sonoporation [33]. A lower temperature as well as ATP depletion may hinder pore repair, since this is an energy-dependent process [19].…”

Section: Dependence Of Uptake Route On Acoustic Pressure and Moleculementioning

confidence: 99%

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

215

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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Section: Dependence Of Uptake Route On Acoustic Pressure and Moleculementioning

confidence: 99%

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

215

247

View full text Add to dashboard Cite

show abstract

“…Many studies have demonstrated that ultrasound (US) can delivery DNA, proteins, and other formulations of molecules of interest into viable cells [1][2][3][4][5][6][7][8]. It has been hypothesized that US energy, often amplified by microbubble activities (e.g.…”

Section: Introductionmentioning

confidence: 99%

“…Compelling results obtained recently have stimulated great interest in the development of sonoporation applications with exciting possibilities [1][2][3][4][5][6][7][8]. However, progress in the field is hindered by a lack of mechanistic understanding of the sonoporation process and its outcome beyond demonstrations of initial feasibility [20].…”

Section: Introductionmentioning

confidence: 99%

Effects of extracellular calcium on cell membrane resealing in sonoporation

Zhou

Shi

Cui

et al. 2008

Journal of Controlled Release

106

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Sonoporation has been exploited as a promising strategy for intracellular drug and gene delivery. The technique uses ultrasound to generate pores on the cell membrane to allow entry of extracellular agents into the cell. Resealing of these non-specific pores is a key factor determining both the uptake and post-ultrasound cell survival. This study examined the effects of extracellular Ca 2+ on membrane resealing in sonoporation, using Xenopus oocytes as a model system. The cells were exposed to tone burst ultrasound (1.06 MHz, duration 0.2 s, acoustic pressure 0.3 MPa) in the presence of 0.1% Definity ® at various extracellular [Ca 2+ ] (0-3 mM). Sonoporation inception and resealing in a single cell were monitored in real time via the transmembrane current of the cell under voltage clamp. The time-resolved measurements of transmembrane current revealed the involvement of two or more Ca 2+ related processes with different rate constants and characteristics. Rapid resealing occurred immediately after ultrasound application followed by a much slower resealing process. Complete resealing required [Ca 2+ ] above 0.54 mM. The cells resealed in 6-26 s at 1.8 mM Ca 2+ , but took longer at lower concentrations, up to 58 ~170 s at 0.54 mM Ca 2+ .

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“…The use of ultrasound in therapy (6,7) and also in gene transfection (8)(9)(10)(11)(12) has been investigated both in vitro and in vivo. Poor transfection rates remain a problem, so combinations with other methods are being used to try to improve the outcomes.…”

mentioning

confidence: 99%

Optimized ultrasound‐mediated gene transfection in cancer cells

et al. 2006

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Ultrasound-mediated gene transfection (sonotransfection) is a promising physical method for gene therapy, especially for cancer gene therapy. To investigate the optimal sonotransfection conditions and to determine whether the optimal transfection rate using sonotransfection is comparable to that of electrotransfection or liposomemediated transfection, we sonicated different cancer cell lines (U937, HeLa, PC-3, Meth A and T-24) using a 1-MHz unfocused ultrasound at different intensities, pulse repetition frequencies and exposure times. The ideal ultrasound conditions were noted to be at 1.5 Watt/cm 2 pulsed at 0.5 Hz with a duty factor of 50%. The results showed that transfection rate increased with the number of pulses, and peaked between 10 and 15 pulses before it started to decline. Using such optimal conditions, we have shown that sonotransfection is superior to electrotransfection and liposome-mediated transfection at the fixed conditions used in the present study. These findings suggest that sonotransfection could be a better alternative to other

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Enhancement of ultrasound‐mediated gene transfection by membrane modification

Abstract: This enhancement could be useful for ultrasound-mediated gene therapy in the future since both treatments for membrane modification could be directly applied to the living body.

Cited by 55 publications

References 32 publications

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

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

Effects of extracellular calcium on cell membrane resealing in sonoporation

Optimized ultrasound‐mediated gene transfection in cancer cells

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