Authors:

Ana Dogan

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Advancing drug delivery method plays an important role in reducing toxic side effects and maximizing drug efficacy by (i) increasing target specificity and (ii) decreasing drug degradation. Extracellular vesicles (EVs), which are natural carriers released by all living cells, are gaining attention for usage in drug delivery due to their various advantageous properties, including (i) cargo protection from enzymatic degradation, (ii) tissue-specific targeting, and (iii) high biocompatibility. One of the leading methods for cargo transfection into EVs is electroporation, which utilizes short duration of voltage pulses to create temporary pores on the EV surface membrane, during which the cargo can enter. This process is limited by the maximum voltage EVs can withstand for pore formation and cargo encapsulation. We aim to investigate optimized cargo loading into EVs specifically through the addition of gold nanoparticles (AuNP) to the electroporation process. It has been shown that AuNP can increase the loading into cells during electroporation by allocating the electric voltage onto the cell surface, allowing more pore formation without increasing the total voltage applied. Due to similar surface properties, we hypothesized that AuNPs would provide a similar increase in transfection of EVs. To test this hypothesis, we electroporated green fluorescence protein (GFP) into EVs with the introduction of AuNPs. Fluorescence analysis through Cytation 5 revealed that the AuNPs increased the amount of GFP transfected into the EVs. Assessment with Nanoparticle Tracking Analysis and Transmission Electron Microscopy showed that the AuNPs did not significantly change the size, concentration, zeta potential, or morphology of the EVs, compared to the native EVs without electroporation, for retaining the EV originality and integrity. Overall, AuNPs have the capacity to increase drug transfection into EVs, allowing high-efficiency packaging for specific drug delivery to enhance therapeutic functions.

Source:

University of Florida / Ana Dogan, Nina Erwin, Xiaoshu Pan, Mei He* / 2023

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Co-authors:

Ana Dogan