Membrane permeabilization and cell damage by ultrashort electric field shocks

Pakhomov, Andrei G.; Shevin, Rachael; White, Jeremy; Kolb, Juergen F.; Pakhomova, Olga N.; Joshi, R. P.; Schoenbach, K.H.

doi:10.1016/j.abb.2007.05.003

Cited by 179 publications

(152 citation statements)

References 46 publications

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“…Nanopore formation, in contrast to the larger pores formed by longer pulses (electroporation), are less likely to pass large ions such as propidium, but freely allow passage of small ions. 7,8 Various studies using diverse techniques such as electrophysiology, [9][10][11] fluorescence microscopy, 7,11 and direct ion measurement in bulk solution for longer pulse widths 12 have supported the hypothesis that such "nanopores" exist. Interestingly, when exposed cells are under a whole cell patch clamp, nanopore activity appears to have unique electrical characteristics.…”

Section: Introductionmentioning

confidence: 94%

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Nanosecond pulsed electric field thresholds for nanopore formation in neural cells

Roth¹,

Tolstykh

Payne

et al. 2013

J. Biomed. Opt

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Abstract. The persistent influx of ions through nanopores created upon cellular exposure to nanosecond pulse electric fields (nsPEF) could be used to modulate neuronal function. One ion, calcium (Ca 2þ ), is important to action potential firing and regulates many ion channels. However, uncontrolled hyper-excitability of neurons leads to Ca 2þ overload and neurodegeneration. Thus, to prevent unintended consequences of nsPEF-induced neural stimulation, knowledge of optimum exposure parameters is required. We determined the relationship between nsPEF exposure parameters (pulse width and amplitude) and nanopore formation in two cell types: rodent neuroblastoma (NG108) and mouse primary hippocampal neurons (PHN). We identified thresholds for nanoporation using Annexin V and FM1-43, to detect changes in membrane asymmetry, and through Ca 2þ influx using Calcium Green. The ED50 for a single 600 ns pulse, necessary to cause uptake of extracellular Ca 2þ , was 1.76 kV∕cm for NG108 and 0.84 kV∕cm for PHN. At 16.2 kV∕cm, the ED50 for pulse width was 95 ns for both cell lines. Cadmium, a nonspecific Ca 2þ channel blocker, failed to prevent Ca 2þ uptake suggesting that observed influx is likely due to nanoporation. These data demonstrate that moderate amplitude single nsPEF exposures result in rapid Ca 2þ influx that may be capable of controllably modulating neurological function. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Section: Introductionmentioning

confidence: 94%

“…1(b)]. The exposure configuration has been described in depth in previous publications 9 [ Fig. 1(c)].…”

Section: Exposure and Imagingmentioning

confidence: 99%

Nanosecond pulsed electric field thresholds for nanopore formation in neural cells

Roth¹,

Tolstykh

Payne

et al. 2013

J. Biomed. Opt

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“…However by using smaller fluorescent probes and more sensitive analytical methods, it has become clear that nsPEFs produce unique pores or aqueous channels that are distinct from conventional electroporation pores in that they are much smaller and are thus called nanopores. Pakhomov and coworkers have provided significant detail to the behavior of nanopores induced by nsPEFs [Bowman et al, 2010;Pakhomov et al, 2007aPakhomov et al, , b, 2009] but have only begun to fully characterize them. Pulses with durations of 60ns and electric fields of 12 kV/cm showed long-lasting (minutes) reduction of the cell membrane resistance and a corresponding loss of the plasma membrane potential.…”

Section: Effects Of Ultra-high Pulse Power On Cell Plasma Membranesmentioning

confidence: 99%

Pulse Power Ablation of Melanoma with Nanosecond Pulsed Electric Fields

Beebe¹,

Ford²,

Ren³

et al. 2011

Treatment of Metastatic Melanoma

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“…Large endocytic vacuoles were selectively permeabilized with little effect on the integrity of the outer cell membrane [14]. However, recent studies show that by applying high-voltage nanosecond pulses, the cell plasma membrane is also permeabilized [15], [16]. Researchers have demonstrated that nanosecond pulsed electric fields can induce apoptosis and inhibit tumor growth [17], [18] and so may be further developed as a possible treatment for basal cell carcinoma [19].…”

Section: Introductionmentioning

confidence: 99%

Blumlein Configuration for High-Repetition-Rate Pulse Generation of Variable Duration and Polarity Using Synchronized Switch Control

Reberšek

Kranjc

Pavliha

et al. 2009

IEEE Trans. Biomed. Eng.

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Abstract-Blumlein generators are used in different applications such as radars, lasers, and also recently in various biomedical studies, where the effects of high-voltage nanosecond pulses on biological cells are evaluated. In these studies, it was demonstrated that by applying high-voltage nanosecond pulses to cells, plasma membrane and cell organelles are permeabilized. As suggested in a recent publication, the repetition rate and polarity of nanosecond high-voltage pulses could have an important effect on the electropermeabilization process, and consequently, on the observed phenomena. Therefore, we designed a new Blumlein configuration that enables a higher repetition rate of variable duration of either bipolar or unipolar high-voltage pulses. We achieved a maximal pulse repetition rate of 1.1 MHz. However, theoretically, this rate could be even higher. We labeled endocytotic vesicles with lucifer yellow and added propidium iodide to a cell suspension for testing the cell plasma membrane integrity, so we were able to observe the permeabilization of endocytotic vesicles and the cell plasma membrane at the same time. The new design of pulse generator was built, verified, and also tested in experiments. The resulting flexibility and variability allow further in vitro experiments to determine the importance of the pulse repetition rate and pulse polarity on membrane permeabilization-both of the cell plasma membrane as well as of cell organelle membranes.

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Membrane permeabilization and cell damage by ultrashort electric field shocks

Cited by 179 publications

References 46 publications

Nanosecond pulsed electric field thresholds for nanopore formation in neural cells

Nanosecond pulsed electric field thresholds for nanopore formation in neural cells

Pulse Power Ablation of Melanoma with Nanosecond Pulsed Electric Fields

Blumlein Configuration for High-Repetition-Rate Pulse Generation of Variable Duration and Polarity Using Synchronized Switch Control

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