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-PRO^TM-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.

最近的研究证明，纳秒脉冲电场（nsPEF）诱导的经典生物效应可以通过传递正极性脉冲后立即产生的负极性脉冲来降低。这种现象被称为“双极抵消”，据报道，这种双极脉冲的范围很广，脉冲持续时间为2 ns至900 ns。相反，成对的脉冲，即具有相同极性的两个相同的脉冲，增加了传统的nsPEF结果。本文中，我们基于冻结波概念提出了一种新颖的鲁棒且灵活的发生器，它能够产生宽范围的脉冲，持续时间为10 ns，延迟在17到360 ns之间。进行了数值模拟和实验测量，以充分表征建议的发电机。悠宝^TM在U87-MG人胶质母细胞瘤细胞系中研究了-1摄取，作为对10 ns，11.5 MV / m nsPEF响应的膜通透性的标志。我们的结果表明，双极性消除发生在0–30 ns的延迟中，并随着相间间隔的变化而减小。此外，我们观察到，与单极性脉冲响应相比，配对nsPEF施加后的细胞响应是两倍以上，并且独立于相间间隔。