Fundamentals of Pulsed Field Ablation at the Individual Cardiomyocyte Level

Background Pulsed Field Ablation (PFA) is a rapidly adopted catheter ablation method employing high-voltage pulses to treat cardiac arrhythmias by cardiomyocyte electroporation. Objective We systematically characterized cardiomyocyte responses in vitro to different PFA waveforms to better understand transient electrophysiological effects observed in clinical PFA. Methods Isolated adult rat ventricular cardiomyocytes were exposed to clinically relevant waveforms: monophasic 100 μs, biphasic 2 μs, and monophasic 200 ns pulses. Transmembrane voltage, intracellular Ca 2+ , and sarcomere shortening were monitored simultaneously. Computational modeling and machine learning were used to characterize responses. Results We observed six distinct response types ranging from no response to complete loss of excitability. All three waveforms induced the same spectrum of responses albeit at different electric field strengths. Cell orientation influenced thresholds in a pulse duration-dependent manner. Modeling demonstrated that electroporation-induced increase in membrane conductance explains the observed responses. Machine learning revealed that cells retain signatures of electroporation in their action potentials and Ca 2+ transients, differing between microsecond and submicrosecond waveforms. On a longer time scale, 12–36% of cells that initially lost excitability recovered within ∼10 minutes. Conclusion All PFA waveforms produced similar cardiomyocyte responses through electroporation-induced increase of membrane conductance. Substantial fraction of cells recover excitability within minutes, providing a plausible cellular explanation for the clinically observed transient loss of intracardiac electrograms following PFA, and a rationale for pulsed field mapping based on reversible electroporation. While fundamental effects are waveform-independent, signatures in recovered cells show waveform-dependent features. These findings advance mechanistic understanding of PFA at the cellular level.