High-intensity irreversible electroporation targeting intracellular structures enhance tumor ablation in lung cancer models

Abstract

This study aimed to optimize irreversible electroporation (IRE) parameters to enhance intracellular injury, specifically targeting nuclear and mitochondrial structures that are insufficiently affected by conventional protocols. To address limitations of standard 1000 similar to 2500 V/cm clinical settings, we experimentally and computationally evaluated both low- and high-electric-field conditions and identified pulse parameters capable of safely achieving electric field strengths exceeding 4,000 V/cm, values that remain below the predicted arcing threshold for our electrode configuration while permitting effective intracellular electroporation. In vitro studies using A549 lung cancer cells demonstrated that high-field IRE markedly intensified oxidative stress, resulting in a 30-fold increase in hydrogen peroxide production and pronounced disruption of mitochondrial membrane potential. Transmission electron microscopy further confirmed severe ultrastructural injury, including plasma membrane rupture, nuclear membrane deformation, and complete loss of mitochondrial cristae, culminating in irreversible cell death. In vivo experiments corroborated these findings: high-field IRE produced extensive and uniform tumor ablation, whereas conventional lower field strengths generated only localized and partial damage. These results indicate that elevating electric-field intensity in IRE protocols can overcome the inherent limitations of traditional approaches by reliably inducing intracellular organelle damage, suppressing cellular repair pathways, and enhancing overall ablation completeness. Further studies are warranted to evaluate long-term safety and therapeutic durability of high-field IRE in vivo.