Electroporation induced changes in extracellular vesicle profile.

Area of Science:

• Biotechnology
• Nanomedicine
• Materials Science

Background:

• Extracellular vesicles (EVs) show potential as drug delivery systems (DDSs).
• Electroporation is a common method for loading therapeutic payloads but requires optimization for EV applications.
• Understanding electroporation's impact on EV integrity and characteristics is vital for their therapeutic use.

Purpose of the Study:

• To investigate the effects of electroporation on the physical and protein profile of extracellular vesicles (EVs).
• To evaluate how varying electroporation parameters influence EV characteristics for potential drug delivery applications.

Main Methods:

• EVs were isolated and subjected to electroporation with varied voltages, pulse numbers, and pulse widths.
• Analysis included particle concentration, size distribution, polydispersity index (PDI), zeta potential (ZP), and protein concentration.
• Western blotting was used to assess changes in EV surface markers.

Main Results:

• Suspension in electroporation buffer (EB) reduced EV concentration, increased particle size, and decreased zeta potential (ZP).
• Electroporation parameters (EPs) variably affected EV profiles, reducing surface protein concentration and leading to more neutral ZP.
• Native EV characteristics were not fully recovered after washing post-electroporation.

Conclusions:

• Electroporation protocols significantly impact fundamental EV properties, potentially affecting their efficacy as drug delivery systems (DDSs).
• Further optimization of electroporation buffers (EBs) and protocols is necessary to preserve the native EV profile after payload loading.