Probability and kinetics of rupture and electrofusion in giant unilamellar vesicles under various frequencies of direct current pulses

Abstract

Irreversible electroporation induces permanent permeabilization of lipid membranes of vesicles, resulting in vesicle rupture upon the application of a pulsed electric field. Electrofusion is a phenomenon wherein neighboring vesicles can be induced to fuse by exposing them to a pulsed electric field. We focus how the frequency of direct current (DC) pulses of electric field impacts rupture and electrofusion in cell-sized giant unilamellar vesicles (GUVs) prepared in a physiological buffer. The average time, probability, and kinetics of rupture and electrofusion in GUVs have been explored at frequency 500, 800, 1050, and 1250 Hz. The average time of rupture of many ‘single GUVs’ decreases with the increase in frequency, whereas electrofusion shows the opposite trend. At 500 Hz, the rupture probability stands at 0.45 ± 0.02, while the electrofusion probability is 0.71 ± 0.01. However, at 1250 Hz, the rupture probability increases to 0.69 ± 0.03, whereas the electrofusion probability decreases to 0.46 ± 0.03. Furthermore, when considering kinetics, at 500 Hz, the rate constant of rupture is (0.8 ± 0.1)×10−2 s-1, and the rate constant of fusion is (2.4 ± 0.1)×10−2 s-1. In contrast, at 1250 Hz, the rate constant of rupture is (2.3 ± 0.8)×10−2 s-1, and the rate constant of electrofusion is (1.0 ± 0.1)×10−2 s-1. These results are discussed by considering the electrical model of the lipid bilayer and the energy barrier of a prepore.