Microbubble-induced sonoporation involved in ultrasound-mediated DNA transfection in vitro at low acoustic pressures

Qiu, Yuanyuan; Zhang, Chunbing; Tu, Juan; Zhang, Dong

doi:10.1016/j.jbiomech.2012.03.011

Cited by 86 publications

(65 citation statements)

References 34 publications

(51 reference statements)

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“…A similar delivery rate was achieved at high pressures in the DNA transfection of MCF-7 cells (Qiu et al, 2010). However, high delivery rates were accomplished at the expense of cell viability, with extended cell death being observed at high pressure sonication (P pk-neg > 500kPa); in contrast, low acoustic pressures (P pk-neg < 100kPa pk-neg ) were proved to be safer but resulted in transfection efficiency below 15% 15 (Qiu et al, 2012). A common feature of most of the aforementioned studies is that experiments were conducted in the absence of flow.…”

mentioning

confidence: 78%

“…A series of pilot experiments were performed during the evaluation procedure of our flow system, using typical sonoporation ultrasound parameters (P pk-neg : 40-160kPa, PL: 10-500 cycles, N p : 10-600). Our initial selection of ultrasound parameters was based on previous studies without flow, which have shown that higher acoustic pressures increase not only the desired trans-membrane 10 delivery efficiency, but also the unwanted cellular damage (Brayman et al, 1999;Qiu et al, 2012). Studies have also shown that inertial cavitation is responsible for the decreased cell viability (Newman and Bettinger, 2007).…”

Section: Evaluation Of Ultrasound Parameters For Sonoporation During mentioning

confidence: 99%

“…Proposed routes of molecular delivery include the formation of short-lived pores in the membrane, also known as sonoporation (Yu and Xu, 2014), and endocytosis (Geers et al, 2011;van Wamel et al, 2006). In addition, increased transmembrane delivery efficiency is associated with increased damage and cell death (Qiu et al, 2012(Qiu et al, , 2010, and this efficacy-safety trade-off has limited the technology's 15 development and hence it remains unutilized in the clinic as a drug delivery tool.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Enhancement of Non-Invasive Trans-Membrane Drug Delivery Using Ultrasound and Microbubbles During Physiologically Relevant Flow

Shamout

Pouliopoulos

Lee

et al. 2015

Ultrasound in Medicine & Biology

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mentioning

confidence: 78%

Section: Evaluation Of Ultrasound Parameters For Sonoporation During mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Enhancement of Non-Invasive Trans-Membrane Drug Delivery Using Ultrasound and Microbubbles During Physiologically Relevant Flow

Shamout

Pouliopoulos

Lee

et al. 2015

Ultrasound in Medicine & Biology

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“…When microbubbles collapse, the cell membrane is perturbed and a pore is formed, allowing genetic material to enter the cell via the pore (13,14). Diagnostic US is used to introduce plasmids and short interfering RNA into cultured cells in this manner (15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Introduction of plasmids into gastric cancer cells by endoscopic ultrasound

et al. 2017

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Abstract. Short hairpin RNA of frizzled-2 (shRNA-Fz2) suppresses the cell proliferation of gastric cancer cells. Endoscopic ultrasound (EUS) is considered a suitable method for the introduction of therapeutic plasmids into cells, since the device enables the access and real-time monitoring of gastric cancer tissues. In the present study, plasmids were introduced into cells by sonoporation, as evidenced by the production of H 2 O 2 . The production of H 2 O 2 was measured by absorbance of a potassium-starch solution irradiated with EUS. Luciferase activity was analyzed in the cells irradiated with EUS after the addition of a pMetLuc2-control in the media, and cell proliferation was analyzed using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt assay after irradiation with EUS following the addition of shRNA-Fz2. Absorbance levels corresponding to free radical levels were found to be higher in the cells irradiated with EUS. Luciferase activities were found to be significantly higher in the transfected cells (plasmid with Lipofectamine LTX) than in untreated cells and were furthermore found to be higher in MKN45 cells irradiated for 0.5 min than in cells not subjected to irradiation. Luciferase activity was also found to be higher in MKN74 cells irradiated for 2 min than in cells that were not irradiated. Although the cell proliferation of the MKN45 cells tended to be suppressed by irradiation with EUS, this was non-significant suppression, while the cell proliferation of MKN74 cells was found to be suppressed by irradiation with 12 MHz for 2 min (P<0.05). In conclusion, plasmids were introduced into cultured gastric cancer cells by irradiation with EUS due to sonoporation, as evidenced by the production of H 2 O 2 ; however, the efficiency of the plasmid introduction was low compared with a traditional transfection approach.

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“…It is safer than other methods, so it may be applied in clinical application in the near future. Although the combination of US irradiation with polyethyleneimine (PEI) has already been investigated by some other groups [7,8], the transfer efficiency is still relatively low and the optimization of transfection conditions has not been studied yet. Both PEI and UTMD are promising transfection methods and novel work, but there are little reports on systematic research about these two methods.…”

Section: Introductionmentioning

confidence: 99%

The optimization of transfection in MCF-7 cells combining ultrasound irradiation with contrast agent and polyethyleneimine

Chen¹,

Qiu²,

Sun³

et al. 2012

J Cancer Ther Res

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Objective: To enhance the transfection efficiency of the human breast cancer cell lines (MCF-7) cells by plasmid DNA, the optimized condition and synergistic role of ultrasound (US) irradiation with contrast agent and polyethyleneimine (PEI) were discussed. Methods: MCF-7 cells were transfected with the compounds prepared by the plasmid DNA encoding luciferase (pCMV-luciferase-GL3) and PEI. Meanwhile, SonoVue microbubble was added to the cell suspension to serve as nucleation sites for acoustic cavitation before US irradiation. Then, the DNA expression of luciferase plasmid and viability of cells were evaluated through optimizing US irradiation. Furthermore, the influencing factor, such as the concentration of plasmid, incubation time, serum, the type of solvent and the volume of culture media, were analyzed. Results: The viability of cells and US-induced enhancement of luciferase activity were influenced by the US intensity, exposure time and duty cycle. US irradiation under an appropriate condition could accelerate the permeation of the PEI/DNA complex through the cell membrane. It could enhance the transfection efficiency of plasmid DNA. The optimal US condition for the enhancement was determined to be 1W/cm 2 , 10% DC for 3 mins. In contrast to the PEI/DNA complex alone without US irradiation or US irradiation alone, the combination of US irradiation with contrast agent and PEI had a significantly enhanced luciferase activity (P<0.01). The 2 hrs preirradiation incubation with PEI/DNA complex for MCF-7 cells exhibited a significantly enhanced luciferase activity (P<0.01). In addition, the transfection efficiency was also affected by serum, type of solvent and the volume of culture media. Conclusions: These results suggest that optimized parameters of US and transfection enhance the efficiency of gene expression in MCF-7 cells apparently. The combination of US irradiation with contrast agent and PEI has a synergy and it is not only a simple but also an applied promising method to increase plasmid DNA expression.

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Microbubble-induced sonoporation involved in ultrasound-mediated DNA transfection in vitro at low acoustic pressures

Cited by 86 publications

References 34 publications

Enhancement of Non-Invasive Trans-Membrane Drug Delivery Using Ultrasound and Microbubbles During Physiologically Relevant Flow

Enhancement of Non-Invasive Trans-Membrane Drug Delivery Using Ultrasound and Microbubbles During Physiologically Relevant Flow

Introduction of plasmids into gastric cancer cells by endoscopic ultrasound

The optimization of transfection in MCF-7 cells combining ultrasound irradiation with contrast agent and polyethyleneimine

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