Optimizing Electroporation Pulses by Balancing Ablation and pH-Induced Damage: A Trajectory-Based Framework

Background: Electroporation (EP), in which pulsed electric fields permeabilize cell membranes, is widely used in biomedical and food-processing applications, including electrochemotherapy, irreversible electroporation (IRE), and gene electrotransfer (GET). Optimizing EP protocols is critical to maximize therapeutic efficacy while minimizing unintended tissue damage. Methods: We introduce a theoretical framework for EP protocol optimization based on the spatiotemporal trajectories of ablation size, electroporation threshold, and pH-induced tissue damage. The framework analyzes how the time gradient of the electric field, derived from first principles, governs the evolution and interaction of these trajectories. The approach is evaluated using in silico and in vitro potato models and applied in vivo to GET protocols using a dorsal skinfold chamber model. Results: The time gradient of the electric field was found to govern all three trajectories, and their interaction determined protocol efficiency. Within the proposed framework, the entire ablation-size trajectory could be reconstructed from a single measurement obtained at the final pulse, eliminating the need to experimentally track the EP threshold trajectory. In the absence of pH effects, ablation size increased logarithmically with pulse number, while the EP threshold decayed exponentially. When pH-induced damage was included, ablation size increased logarithmically, whereas damage increased linearly. Combining these trends enabled the identification of a critical pulse number that maximizes the difference between ablation size and pH-induced damage. Conclusion: This trajectory-based framework provides a unified description of EP dynamics and a first–principles–based method for optimizing pulse number across EP-based protocols. Joint consideration of ablation, threshold, and damage trajectories can guide the design of safer and more effective EP treatments.