Page 176 - Read Online

Page 18 Ribovski et al. Extracell Vesicles Circ Nucleic Acids 2023;4:283-305 https://dx.doi.org/10.20517/evcna.2023.26

development of EVs as drug delivery systems a.o. to reveal the EV population that is responsible for exerting
functional effects. In addition, further understanding of the mechanisms of EV biogenesis, cellular
internalization and intracellular fate is needed. To obtain a comprehensive understanding of EV-mediated
cargo release, it is advisable to examine relevant producer-recipient cell combinations and analyze the
stoichiometry of the delivery process. Most importantly, the delivery process needs to be studied under
[143]
conditions that mimic the in vivo situation and/or be verified in vivo, which is not common practice . As
an example, the influence of bodily fluids (blood, plasma, CSF, etc.) needs to be reflected in the experiments,
which will certainly add another layer of complexity. While exosomes are naturally present in saliva, urine,
blood, and plasma, in the majority of experiments, their mechanism of action is investigated under standard
cell culture conditions in physiological buffers, neglecting the influence of environmental factors present in
their natural extracellular surrounding. This includes the presence of specific macromolecules in the
extracellular fluid. For synthetic nanoparticles, it is well-known that a protein corona is formed upon
administration of the nanoparticles in biological fluids, e.g., blood plasma, which is known to influence the
biodistribution of these nanoparticles [144-146] . Recently, the formation of a protein corona on EVs in blood
[147]
plasma was shown . As the nanoparticle protein corona composition is, amongst others, influenced by
nanoparticle size and surface chemistry, it is easy to envision that the investigation of EV protein corona
[148]
constitutes a new challenge to the field, forming an emerging subject area for EV research .

DECLARATIONS
Authors’ Contributions
Conceived the idea for the manuscript: Zuhorn I, Ribovski L, Joshi B
Wrote, revised, and edited the manuscript: Ribovski L, Joshi B, Gao J, Zuhorn I
Prepared Figure 1: Ribovski L, Zuhorn I

Availability of Data and Materials
Not applicable.

Financial Support and Sponsorship
Zuhorn I and Ribovski L are supported by the VICI grant (18683) awarded to Zuhorn I in the Talent
Programme, which is (partly) financed by the Dutch Research Council (NWO); Gao J acknowledges
financial support from the China Scholarship Council (No. 202106210083).

Conflicts of Interest
All authors declared that there are no conflicts of interest.

Ethical approval and consent to participate
Not applicable.

Consent for publication
Not applicable.

Copyright
© The Author(s) 2023.

REFERENCES
1. Nigri J, Leca J, Tubiana SS, et al. CD9 mediates the uptake of extracellular vesicles from cancer-associated fibroblasts that promote
pancreatic cancer cell aggressiveness. Sci Signal 2022;15:eabg8191. DOI
2. Berumen Sánchez G, Bunn KE, Pua HH, Rafat M. Extracellular vesicles: mediators of intercellular communication in tissue injury
and disease. Cell Commun Signal 2021;19:104. DOI PubMed PMC