@article{
author = "Spasovski, Vesna and Romolo, Anna and Zagorc, Urška and Arrigler, Vesna and Kisovec, Matic and Zavec, Apolonija Bedina and Arko, Matevž and Molnár, Adrienn and Schlosser, Gitta and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2024",
abstract = "Introduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized
hybridosomes or nanohybridosomes, NSHs) were considered.
Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles
from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and
assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid
chromatography–mass spectrometry.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and
interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light
scattering and interferometric light microscopy measurements agreed well on the average hydrodynamic radius of the
particles Rh (between 140 and 180 nm), while the concentrations of the particles were in the range between 1013 and
1014/mL indicating that NSHs present a considerable (more than 25%) of the sample which is much more than the yield of
natural extracellular vesicles (EVs) from spruce needle homogenate (estimated less than 1%). Spruce specific lipids and
proteins were found in hybridosomes.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that
large-scale production of NSHs from lipids and spruce needle homogenate is feasible.
Plain Language Summary: Cells shed into their exterior nanoparticles (here referred to as extracellular vesicles – EVs) that
are free to move, reach distant cells and are taken up by them. As they carry bioactive constituents, EVs may have important
impact on the recipient cells. The mechanisms of EV formation and mediation can be employed in designing therapeutic,
prophylactic and diagnostic methods for various medical issues. EVs can be harvested from biological samples; however,
their yield is small,12 and there are potential side effects. Artificial vesicles – liposomes – have high yield; however, in vivo,
they can be degraded before reaching the target and their reproducibility is yet insufficient. In order to combine advantages of
both types of nanoparticles, we have composed nanohybridosomes (NSHs) from soya lecithin, water and supernatant of
isolation of EVs from spruce needle homogenate, visualized them by cryogenic electron microscopy and characterized them
with respect to their size, concentration and protein/nucleic acid content. We have applied a recently developed interferometric light microscopy to determine the hydrodynamic radius and the concentration of EVs. We found that the majority of
composed particles are nano-sized and that they enclose more than 25% of the incoming volume of liquid, which is considerably more than about 1% that can be harvested by isolation of EVs from spruce needle homogenate by (ultra)
centrifugation",
publisher = "Dove Press Ltd",
journal = "International Journal of Nanomedicine",
title = "Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles",
pages = "1721-1709",
volume = "19",
doi = "10.2147/IJN.S432836"
}
