Systematic In Vitro Investigation of PEFs Pulse Parameter Specifications on HepG2 Liver Cancer Cells

Pulsed Electric Fields (PEFs) are widely investigated in cancer research, yet interpretation and optimisation of exposure protocols remain challenging due to the complex interplay between electrical parameters, thermal effects, and biological response. While voltage amplitude is often emphasised, the influence of pulse timing and structure under dose-constrained conditions is less systematically examined. This work presents an exploratory in vitro study on the HepG2 hepatocellular carcinoma cell line using a custom-built bipolar pulse delivery system. Seventy experimental conditions were tested, each varying a single pulse parameter (voltage, width, period, pulses per burst, burst count, or burst period), with temperature monitored during exposure. Cell metabolic viability was quantified 24 h post-treatment using an MTT assay. The results show that short-term viability suppression and thermal behaviour depend on pulse configuration and not on total electric dose alone. Based on comparative trends, a qualitative hierarchy of parameter influence is proposed, with total electric dosage and pulse width showing the strongest association with viability reduction, followed by voltage, pulse period, pulses per burst, and burst period. Elevated temperatures observed in some regimes indicate possibly combined electro–thermal effects rather than purely electrical responses. Overall, the study provides a parameter-sensitivity overview that highlights the importance of pulse timing and thermal management alongside voltage amplitude in PEF experiments.