Electroporation induced changes in extracellular vesicle profile

Extracellular vesicles (EVs) are promising drug delivery systems (DDSs). Electroporation is widely applied in the loading of therapeutic payloads but has not been optimized for EV loading. Understanding the potential effects of electroporation on EV profile and integrity is important if they are to be applied therapeutically. In the present study, EVs were isolated and subjected to electroporation at different voltages (500-1000 mV), pulse numbers (1-3), and pulse widths (10-30 ms). Particle concentration, size distribution, polydispersity index (PDI), zeta potential (ZP), and protein concentration were analyzed and western blotting was performed to evaluate possible variations in EV surface markers. Suspension in electroporation buffer (EB) significantly reduced EV concentration, increased particle size, and reduced the ZP. Native EV profile could not be recovered following washing. Electroporation parameters (EPs) applied had variable effects on in EV profile, with reductions in surface protein concentration and a more neutral ZP observed. In conclusion, we identified that the electroporation protocol had a considerable impact on basic EV properties, which could impact their application as DDS. Further optimization of EBs and protocols is required to retain native EV profile following loading.