Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles
Authors
Spasovski, VesnaRomolo, Anna
Zagorc, Urška
Arrigler, Vesna
Kisovec, Matic
Zavec, Apolonija Bedina
Arko, Matevž
Molnár, Adrienn
Schlosser, Gitta
Iglič, Aleš
Kogej, Ksenija
Kralj-Iglič, Veronika
Article (Published version)
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Show full item recordAbstract
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 n...m), 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
Keywords:
hybridosomes / liposomes / nanovesicles / extracellular particles / small cellular particles / drug deliverySource:
International Journal of Nanomedicine, 2024, 19, 1709-1721Publisher:
- Dove Press Ltd
Funding / projects:
- This research was funded by Slovenian Research Agency, grant numbers P2-0232, P3-0388, P1-0201, P1-0391, J3-3066, L3-2621, J2-4447, J2-4427 and IO-0003. Project no. SNN 138407 has been implemented with the support provided by the Ministry of Innovation and Technology of Hungary from the National Research, Development and Innovation Fund, financed under the SNN_21 funding scheme
URI
https://www.dovepress.com/characterization-of-nanohybridosomes-from-lipids-and-spruce-homogenate-peer-reviewed-fulltext-article-IJNhttps://imagine.imgge.bg.ac.rs/handle/123456789/2332
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Spasovski, Vesna AU - Romolo, Anna AU - Zagorc, Urška AU - Arrigler, Vesna AU - Kisovec, Matic AU - Zavec, Apolonija Bedina AU - Arko, Matevž AU - Molnár, Adrienn AU - Schlosser, Gitta AU - Iglič, Aleš AU - Kogej, Ksenija AU - Kralj-Iglič, Veronika PY - 2024 UR - https://www.dovepress.com/characterization-of-nanohybridosomes-from-lipids-and-spruce-homogenate-peer-reviewed-fulltext-article-IJN UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2332 AB - 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 PB - Dove Press Ltd T2 - International Journal of Nanomedicine T1 - Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles EP - 1721 SP - 1709 VL - 19 DO - 10.2147/IJN.S432836 ER -
@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" }
Spasovski, V., Romolo, A., Zagorc, U., Arrigler, V., Kisovec, M., Zavec, A. B., Arko, M., Molnár, A., Schlosser, G., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2024). Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles. in International Journal of Nanomedicine Dove Press Ltd., 19, 1709-1721. https://doi.org/10.2147/IJN.S432836
Spasovski V, Romolo A, Zagorc U, Arrigler V, Kisovec M, Zavec AB, Arko M, Molnár A, Schlosser G, Iglič A, Kogej K, Kralj-Iglič V. Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles. in International Journal of Nanomedicine. 2024;19:1709-1721. doi:10.2147/IJN.S432836 .
Spasovski, Vesna, Romolo, Anna, Zagorc, Urška, Arrigler, Vesna, Kisovec, Matic, Zavec, Apolonija Bedina, Arko, Matevž, Molnár, Adrienn, Schlosser, Gitta, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles" in International Journal of Nanomedicine, 19 (2024):1709-1721, https://doi.org/10.2147/IJN.S432836 . .