High-frequency irreversible electroporation (H-FIRE) has emerged as an alternative to conventional irreversible electroporation (IRE) to overcome the issues associated with neuromuscular electrical stimulation that appear in IRE treatments. In H-FIRE, the monopolar pulses typically used in IRE are replaced with bursts of short bipolar pulses. Currently, very little is known regarding how the use of a different waveform affects the cell death dynamics and mechanisms. In this study, human pancreatic adenocarcinoma cells were treated with a typical IRE protocol and various H-FIRE schemes with the same energized time. Cell viability, membrane integrity and Caspase 3/7 activity were assessed at different times after the treatment. In both treatments, we identified two different death dynamics (immediate and delayed) and we quantified the electric field ranges that lead to each of them. While in the typical IRE protocol, the electric field range leading to a delayed cell death is very narrow, this range is wider in H-FIRE and can be increased by reducing the pulse length. Membrane integrity in cells suffering a delayed cell death shows a similar time evolution in all treatments, however, Caspase 3/7 expression was only observed in cells treated with H-FIRE.

中文翻译：

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常规IRE和H-FIRE处理后细胞死亡的动力学。

高频不可逆电穿孔（H-FIRE）已成为传统不可逆电穿孔（IRE）的替代产品，以克服与IRE治疗中出现的神经肌肉电刺激相关的问题。在H-FIRE中，通常在IRE中使用的单极性脉冲被短双极性脉冲的突发代替。目前，关于使用不同波形如何影响细胞死亡动力学和机制的了解甚少。在这项研究中，人类胰腺癌细胞用典型的IRE方案和各种H-FIRE方案在相同的通电时间下进行处理。在处理后的不同时间评估细胞活力，膜完整性和Caspase 3/7活性。在两种治疗中 我们确定了两种不同的死亡动态（立即和延迟），并量化了导致它们死亡的电场范围。虽然在典型的IRE协议中，导致延迟细胞死亡的电场范围非常窄，但在H-FIRE中该范围更宽，可以通过减小脉冲长度来增加。在所有处理中，遭受延迟细胞死亡的细胞的膜完整性显示相似的时间演变，但是，仅在用H-FIRE处理的细胞中观察到Caspase 3/7表达。