Transfection effect of microbubbles on cells in superposed ultrasound waves and behavior of cavitation bubble

Kodama, Tetsuya; Tomita, Yukio; Koshiyama, Kenichiro; Blomley, Martin

doi:10.1016/j.ultrasmedbio.2006.03.004

Cited by 87 publications

(56 citation statements)

References 43 publications

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“…SonoVue microbubbles (Bracco, Milan, Italy) were used to promote cavitation and were reconstituted in saline solution at a concentration of 25-30 particles/cell, and slowly added to the cell suspension prior to ultrasound exposure (25). The UTMD experiments were performed in an exposure tank and the transducer (Accusonic, Metron Medical Australia Pty., Ltd.) was held in a positioning device at the base of the tank, as previously described (27).…”

Section: Optimization Of Survivin-shrna With the Utmd Techniquementioning

confidence: 99%

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

et al. 2012

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Abstract. Apoptosis induction by short hairpin RNA (shRNA) expression vectors may be an efficient and promising strategy for cancer gene therapy. Ultrasound-targeted microbubble destruction (UTMD) is an appealing technique; however, there few data are available to demonstrate the feasibility and to optimize the methodology for this technology. The aim of this study was to optimize this technique and to elucidate the effects on gene inhibition and apoptosis induction in vitro. Human cervical cancer cell lines were obtained and cultured. shRNA vectors were constructed, and the UTMD technique was examined to determine whether or not it was suitable for shRNA transfection into cells. Cells were then examined using flow cytometry. The results revealed that the optimal irradiation parameters obtained higher transfection efficiency and did not affect the integrity of plasmid DNA. We concluded that survivin downregulation with shRNA expression vectors, mediated by the optimal UTMD parameters, markedly induced cell apoptosis and caused cell cycle arrest, laying a foundation for further investigation of this cancer therapy.

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Section: Optimization Of Survivin-shrna With the Utmd Techniquementioning

confidence: 99%

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

et al. 2012

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“…31 Cavitation 32 has received the most attention, as many scientists have shown that the deliberate addition of echocontrast MCB 33,34 (used to enhance clinical ultrasound imaging) to the transfection medium before USE markedly increases subsequent transgene expression in vitro. 4,7,23,28,[35][36][37] Cavitation may be subdivided into two categories (see Figure 1):…”

Section: Technological Developments Have Increased the Efficiency Of mentioning

confidence: 99%

Gene therapy progress and prospects: Ultrasound for gene transfer

2007

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“…It has been hypothesized that US energy, often amplified by microbubble activities (e.g. bubble oscillation and collapse) [9][10][11][12][13][14][15][16], generates transient, non-specific pores on the cell membrane, a process termed sonoporation [14,[17][18][19]. The transient pores allow a limited time window for the otherwise non-permeable extracellular agents to enter the cell.…”

Section: Introductionmentioning

confidence: 99%

Effects of extracellular calcium on cell membrane resealing in sonoporation

Zhou

Shi

Cui

et al. 2008

Journal of Controlled Release

104

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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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Transfection effect of microbubbles on cells in superposed ultrasound waves and behavior of cavitation bubble

Cited by 87 publications

References 43 publications

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

Gene therapy progress and prospects: Ultrasound for gene transfer

Effects of extracellular calcium on cell membrane resealing in sonoporation

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