Ultralong recovery time in nanosecond electroporation systems enabled by orientational-disordering processes

The growing importance of applications based on molecular medicine and genetic engineering is driving the need to develop high-performance electroporation technologies. The electroporation phenomenon involves disruption of the cell for increasing membrane permeability. Although there is a multitude of research focused on exploring new electroporation techniques, the engineering of programming schemes suitable for these electroporation methods remains a challenge. Nanosecond stimulations could be promising candidates for these techniques owing to their ability to generate a wide range of biological responses. Here we control the membrane permeabilization of cancer cells using different numbers of electric-field pulses through orientational disordering effects. We then report our exploration of a few-volt nanosecond alternating-current (AC) stimulation method with an increased number of pulses for developing electroporation systems. A recovery time of ∼720 min was achieved, which is above the average of ∼76 min for existing electroporation methods using medium cell populations, as well as a previously unreported increased conductance with an increase in the number of pulses using weak bias amplitudes. All-atom molecular dynamics (MD) simulations reveal the orientation-disordering-facilitated increase in the degree of permeabilization. These findings highlight the potential of few-volt nanosecond AC-stimulation with an increased number of pulse strategies for the development of next-generation low-power electroporation systems.