Department of Information and Communication Technologies, Universitat Pompeu Fabra, Roc Boronat, 138, 08018, Barcelona, Spain.
Department of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute and State University, 325 Stranger St, Blacksburg, VA, 24061, USA.
Ann Biomed Eng. 2020 May;48(5):1451-1462. doi: 10.1007/s10439-020-02462-8. Epub 2020 Feb 5.
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.
高频不可逆电穿孔（H-FIRE）作为传统不可逆电穿孔（IRE）的替代方法出现，以克服 IRE 治疗中出现的与神经肌肉电刺激相关的问题。在 H-FIRE 中，IRE 中常用的单极脉冲被短双极脉冲群取代。目前，对于不同波形的使用如何影响细胞死亡动力学和机制，人们知之甚少。在这项研究中，用人胰腺腺癌细胞进行了典型的 IRE 方案和各种具有相同通电时间的 H-FIRE 方案的处理。在处理后不同时间评估细胞活力、膜完整性和 Caspase 3/7 活性。在两种处理中，我们都确定了两种不同的死亡动力学（即时和延迟），并量化了导致每种动力学的电场范围。虽然在典型的 IRE 方案中，导致延迟细胞死亡的电场范围非常狭窄，但在 H-FIRE 中这个范围更宽，可以通过缩短脉冲长度来增加。在所有处理中，延迟细胞死亡的细胞的膜完整性都表现出相似的时间演变，然而，只有在 H-FIRE 处理的细胞中观察到 Caspase 3/7 的表达。
