Especially in cardiac electrophysiology, PFA may represent a dominant future treatment 24, 25, 26.Д¬urrently, in most of the electroporation-based clinical applications, relatively long monophasic pulses of 50вЂ“100В Ојs are delivered with low repetition rates, synchronized with the heart rhythm 3, 27, 28. On the other hand, irreversible electroporation (IRE) appeared as a new medical application 3 for non-thermal tumor 18, 19, 20 and cardiac ablation (Pulse Field AblationвЂ“PFA) 21, 22, 23 providing considerable benefits over existing thermal ablation methods, such as reducing the risk of damaging the nearby critical tissue. Reversible electroporation is successfully used as combination of high-voltage pulsed electric fields with low-permeant chemotherapeutic drug or with DNA: electrochemotherapy (ECT) and gene electrotransfer (GET), respectively 11, 12, 13, 14, 15, 16, 17. Electroporation can be either reversible, when the cell recovers after the treatment and survives, or irreversible when the exposure leads to cell death 2, 3, 4.Д®lectroporation is used in multiple clinical applications 5, 6, 7 as well as in biotechnology 8, food processing 9, and environmentally relevant applications 10. Thus, plasma membrane permeability is increased and transmembrane transport of molecules is enabled which otherwise are unable to cross the membrane 1. This induces a transmembrane voltage (TMV), e.g.,В 500В mV, which far exceeds its resting TMV (typically -40В mV to -70В mV). However, further evaluation of the biphasic pulse protocols presented herein is necessary to confirm the efficiency of the newly proposed HF-EP.Д®lectroporation/electropermeabilization describes the phenomenon where the cell membrane is exposed to sufficiently strong electric field that is generated by short-duration, high-voltage pulses. The study shows that the range of the optimal pulse parameters may be increased depending on the prerequisites of the therapy. In addition, interphase and interpulse delays play a significant role in reducing the muscle contraction and/or pain sensation. Our results confirm that biphasic high-frequency pulses with a pulse width of 1 or 2В Вµs reduce muscle contraction and pain sensation as opposed to currently used longer monophasic pulses. Twenty-five healthy individuals were subjected to electrical stimulation of the tibialis anterior muscle with biphasic high-frequency pulses in the range of few microseconds and both, symmetric and asymmetric interphase and interpulse delays. This is the first study in humans examining pain sensation using biphasic high-frequency electroporation pulses. While the reduction of muscle contractions was confirmed in multiple in vivo studies, the reduction of pain sensation in humans was not confirmed yet, nor was the relationship between muscle contraction and pain sensation investigated. In cardiac tissue ablation, which utilizes irreversible electroporation, the treatment is being established as Pulsed Field Ablation. This treatment modality appeared under the term high-frequency electroporation (HF-EP), which can be potentially used for some clinical applications of electroporation such as electrochemotherapy, gene electrotransfer, and tissue ablation. To minimize neuromuscular electrical stimulation during electroporation-based treatments, the replacement of long monophasic pulses with bursts of biphasic high-frequency pulses in the range of microseconds was suggested in order to reduce muscle contraction and pain sensation due to pulse application.

Contraction intensity scale