Physicochemical Analysis of Extracellular Nanovesicles and Exosome-Like Nanovesicles

Extracellular vesicles (EVs) represent a promising tool for diagnostics and therapy, but high heterogeneity limits their practical use. In this work, a comparative study of the physicochemical properties of native EVs secreted by HEK-293T cells and exosome-like nanovesicles (EMNVs) obtained by extrusion of whole HEK-293T cells was carried out. Using transmission electron microscopy, dynamic and electrophoretic light scattering, and surface-enhanced Raman spectroscopy, the authors found that EMNVs form a more homogeneous population with an average diameter of ~115 nm and a high negative charge (−22.9 mV), whereas native EVs are more heterogeneous in size and have a lower zeta potential (−12.1 mV). Spectral analysis revealed the presence of nucleic acid signals in native EVs and their absence in EMNVs, indicating differences in molecular composition due to different biogenesis mechanisms. The obtained data demonstrate that EMNVs are a more standardized and colloidally stable alternative to native EVs, making them a promising platform for the development of drug-delivery systems.