Pore-Opening Dynamics of Single Nanometer Biovesicles at an Electrified Interface

Release from nanobiovesicles via a pore generated by membrane electroporation at an electrified interface can be monitored by vesicle impact electrochemical cytometry (VIEC) and provides rich information about the various vesicular content transfer processes, including content homeostasis, intraphase content transfer, or the transient fusion of vesicles. These processes are primarily influenced by the vesicular pore-opening dynamics at the electrified interface which has not been disclosed at the single nanobiovesicle level yet. In this work, after simultaneously measuring the size and release dynamics of individual vesicles, we employed a moving mesh-finite element simulation algorithm to reconstruct the accurate pore-opening dynamics of individual vesicles with different sizes during VIEC. We investigated the expansion times and maximal pore sizes as two characteristics of different vesicles. The pore expansion times between nanobiovesicles and pure lipid liposomes were compared, and that of the nanobiovesicles is much longer than that for the liposomes, 2.1 ms vs 0.18 ms, respectively, which reflects the membrane proteins limiting the electroporation process. For the vesicles with different sizes, a positive relationship of pore size (R_p,max) with the vesicle size (R_ves) and also their ratio (R_p,max/R_ves) versus the vesicle sizes is observed. The mechanism of the pore size determination is discussed and related to the membrane proteins and the vesicle size. This work accurately describes the dynamic pore-opening process of individual vesicles which discloses the heterogeneity in electroporation of different sized vesicles. This should allow us to examine the more complicated vesicular content transfer process between intravesicular compartments.