TY - JOUR

T1 - High-voltage 10 ns delayed paired or bipolar pulses for in vitro bioelectric experiments

AU - Orlacchio, Rosa

AU - Carr, Lynn

AU - Palego, Cristiano

AU - Arnaud-Cormos, Delia

AU - Leveque, Philippe

PY - 2021/2

Y1 - 2021/2

N2 - Recent studies proved that classical bio-effects induced by nanosecond pulsed electric field (nsPEF) can be reduced by the delivery of a negative polarity pulse generated immediately after a positive polarity pulse. This phenomenon is known as “bipolar cancellation” and it was reported for a wide range of bipolar pulses with pulse duration from 2 ns to 900 ns. On the contrary, paired pulses, i.e., two identical pulses with the same polarity, increased traditional nsPEF outcomes. Herein, we propose a novel robust and flexible generator, based on the frozen-wave concept, able to produce a broad range of pulses with the duration of 10 ns and delay between 17 and 360 ns. Numerical simulations and experimental measurements were performed to fully characterize the proposed generator. YO-PROTM-1 uptake was investigated in the U87-MG human glioblastoma cell line as a marker of membrane permeabilization in response to 10 ns, 11.5 MV/m nsPEF. Our results showed that bipolar cancellation occurred for delays of 0–30 ns and decreased as a function of the interphase interval. In addition, we observed that cellular response following the application of paired nsPEF was more than two-fold compared to the unipolar pulse response and was independent from the interphase interval.

AB - Recent studies proved that classical bio-effects induced by nanosecond pulsed electric field (nsPEF) can be reduced by the delivery of a negative polarity pulse generated immediately after a positive polarity pulse. This phenomenon is known as “bipolar cancellation” and it was reported for a wide range of bipolar pulses with pulse duration from 2 ns to 900 ns. On the contrary, paired pulses, i.e., two identical pulses with the same polarity, increased traditional nsPEF outcomes. Herein, we propose a novel robust and flexible generator, based on the frozen-wave concept, able to produce a broad range of pulses with the duration of 10 ns and delay between 17 and 360 ns. Numerical simulations and experimental measurements were performed to fully characterize the proposed generator. YO-PROTM-1 uptake was investigated in the U87-MG human glioblastoma cell line as a marker of membrane permeabilization in response to 10 ns, 11.5 MV/m nsPEF. Our results showed that bipolar cancellation occurred for delays of 0–30 ns and decreased as a function of the interphase interval. In addition, we observed that cellular response following the application of paired nsPEF was more than two-fold compared to the unipolar pulse response and was independent from the interphase interval.

KW - Bipolar cancellation

KW - Interphase pulse interval

KW - Pulsed electric field

KW - Versatile nanosecond generator

KW - YO-PRO (TM)-1 uptake

KW - nsPEF

UR - https://ars.els-cdn.com/content/image/1-s2.0-S1567539420302061-mmc1.xml

U2 - 10.1016/j.bioelechem.2020.107648

DO - 10.1016/j.bioelechem.2020.107648

M3 - Article

VL - 137

JO - Bioelectrochemistry

JF - Bioelectrochemistry

SN - 1567-5394

M1 - 107648

ER -