Irreversible Electroporation of AC16 Cardiomyocytes Induced by Nanosecond High Voltage Pulse In Vitro

ABSTRACT Background Cardiomyocyte lines are widely used in cardiovascular research. For cardiac ablation, the factors affecting cardiomyocytes by pulsed electrical field (PEF) ablation are being investigated due to the sensitivity of cardiomyocytes to electrical pulses. Previous studies have shown that AC16 death is independent of the number and frequency of PEF. We aim to investigate the relationship between the depth, size, and electric field strength of irreversible (reversible) electroporation formed by nanosecond PEF. Methods and Results This study used different detection methods to investigate the effects on AC16 cardiomyocytes under different electric fields strength of nanosecond PEF. Macroscopically, the impact of nanosecond PEF on the survival rate of AC16 cardiomyocytes was preliminarily determined through trypan blue staining and CCK‐8. Then, apoptosis, protein expression, and cell morphology were observed by mitochondrial membrane potential, western blot analysis, and electron microscopy to determine the electric field threshold of irreversible (reversible) electroporation in AC16 cardiomyocytes and to study the relationship between pore formation and electric field strength changes. Conclusions For AC16 cardiomyocytes, increasing the electric field strength of nanosecond PEF induces progressive deepening of electroporation. Specifically, at 1500 V/cm, the pores reached an average depth of approximately 30 nm, while at 2200 V/cm, the depth extended to approximately 80 nm with localized membrane rupture. Pore diameter and density also increased with higher field strengths, and a clear electric field threshold distinguished reversible from irreversible electroporation.