Single-cell electroporation with high-frequency nanosecond pulse bursts: Simulation considering the irreversible electroporation effect and experimental validation

Abstract

To study the electroporation characteristics of cells under high-frequency nanosecond pulse bursts (HFnsPBs), the original electroporation mathematical model was improved. By setting a threshold value for irreversible electroporation (IRE) and considering the effect of an electric field on the surface tension of a cell membrane, a mathematical model of electroporation considering the effect of IRE is proposed for the first time. A typical two-dimensional cell system was discretized into nodes using MATLAB, and a mesh transport network method (MTNM) model was established for simulation. The dynamic processes of single-cell electroporation and molecular transport under the application of 50 unipolar HFnsPBs with field intensities of 9 kV cm−1 and different frequencies (10 kHz, 100 kHz and 500 kHz) to the target system was simulated with a 300 s simulation time. The IRE characteristics and molecular transport were evaluated. In addition, a PI fluorescent dye assay was designed to verify the correctness of the model by providing time-domain and spatial results that were compared with the simulation results. The simulation achieved IRE and demonstrated the cumulative effects of multipulse bursts and intraburst frequency on irreversible pores. The model can also reflect the cumulative effect of multipulse bursts on reversible pores by introducing an assumption of stable reversible pores. The experimental results agreed qualitatively with the simulation results. A relative calibration of the fluorescence data gave time-domain molecular transport results that were quantitatively similar to the simulation results. This article reveals the cell electroporation characteristics under HFnsPBs from a mechanism perspective and has important guidance for fields involving the IRE of cells.