TY  - 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 . .

TY  - CONF
AU  - Spasovski, Vesna
AU  - Romolo, Anna
AU  - Kisovec, Matic
AU  - Zagorc, Urška
AU  - Arrigler, Vesna
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Iglič, Aleš
AU  - Kogej, Ksenija
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2265
AB  - Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
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 with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
C3  - Small New World 2.0
T1  - Hybridosomes from spruce needle homogenate
EP  - 85
SP  - 85
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2265
ER  - 

@conference{
author = "Spasovski, Vesna and Romolo, Anna and Kisovec, Matic and Zagorc, Urška and Arrigler, Vesna and Arko, Matevž and Bedina Zavec, Apolonija and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
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 with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.",
publisher = "Serbian Society for Extracellular Vesicles (SrbEVs), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Serbian Society for Extracellular Vesicles (SrbEVs)",
journal = "Small New World 2.0",
title = "Hybridosomes from spruce needle homogenate",
pages = "85-85",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2265"
}

Spasovski, V., Romolo, A., Kisovec, M., Zagorc, U., Arrigler, V., Arko, M., Bedina Zavec, A., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2023). Hybridosomes from spruce needle homogenate. in Small New World 2.0
Serbian Society for Extracellular Vesicles (SrbEVs)., 85-85.
https://hdl.handle.net/21.15107/rcub_imagine_2265

Spasovski V, Romolo A, Kisovec M, Zagorc U, Arrigler V, Arko M, Bedina Zavec A, Iglič A, Kogej K, Kralj-Iglič V. Hybridosomes from spruce needle homogenate. in Small New World 2.0. 2023;:85-85.
https://hdl.handle.net/21.15107/rcub_imagine_2265 .

Spasovski, Vesna, Romolo, Anna, Kisovec, Matic, Zagorc, Urška, Arrigler, Vesna, Arko, Matevž, Bedina Zavec, Apolonija, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Hybridosomes from spruce needle homogenate" in Small New World 2.0 (2023):85-85,
https://hdl.handle.net/21.15107/rcub_imagine_2265 .

TY  - CONF
AU  - Spasovski, Vesna
AU  - Romolo, Anna
AU  - Kisovec, Matic
AU  - Zagorc, Urška
AU  - Arrigler, Vesna
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Iglič, Aleš
AU  - Kogej, Ksenija
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2264
AB  - Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
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 with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Slovenian Network for Extracellular Vesicles (SiN-EV)
C3  - Small New World 2.0
T1  - Hybridosomes from spruce needle homogenate
EP  - 27
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2264
ER  - 

@conference{
author = "Spasovski, Vesna and Romolo, Anna and Kisovec, Matic and Zagorc, Urška and Arrigler, Vesna and Arko, Matevž and Bedina Zavec, Apolonija and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
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 with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.",
publisher = "Serbian Society for Extracellular Vesicles (SrbEVs), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Slovenian Network for Extracellular Vesicles (SiN-EV)",
journal = "Small New World 2.0",
title = "Hybridosomes from spruce needle homogenate",
pages = "27-27",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2264"
}

Spasovski, V., Romolo, A., Kisovec, M., Zagorc, U., Arrigler, V., Arko, M., Bedina Zavec, A., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2023). Hybridosomes from spruce needle homogenate. in Small New World 2.0
Serbian Society for Extracellular Vesicles (SrbEVs)., 27-27.
https://hdl.handle.net/21.15107/rcub_imagine_2264

Spasovski V, Romolo A, Kisovec M, Zagorc U, Arrigler V, Arko M, Bedina Zavec A, Iglič A, Kogej K, Kralj-Iglič V. Hybridosomes from spruce needle homogenate. in Small New World 2.0. 2023;:27-27.
https://hdl.handle.net/21.15107/rcub_imagine_2264 .

Spasovski, Vesna, Romolo, Anna, Kisovec, Matic, Zagorc, Urška, Arrigler, Vesna, Arko, Matevž, Bedina Zavec, Apolonija, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Hybridosomes from spruce needle homogenate" in Small New World 2.0 (2023):27-27,
https://hdl.handle.net/21.15107/rcub_imagine_2264 .

TY  - JOUR
AU  - Troha, Kaja
AU  - Vozel, Domen
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Dolinar, Drago
AU  - Hočevar, Matej
AU  - Jan, Zala
AU  - Kisovec, Matic
AU  - Kocjančič, Boštjan
AU  - Pađen, Ljubiša
AU  - Pajnič, Manca
AU  - Penič, Samo
AU  - Romolo, Anna
AU  - Repar, Neža
AU  - Spasovski, Vesna
AU  - Steiner, Nejc
AU  - Šuštar, Vid
AU  - Iglič, Aleš
AU  - Drobne, Damjana
AU  - Kogej, Ksenija
AU  - Battelino, Saba
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2087
AB  - The preparation of autologous platelet and extracellular vesicle-rich plasma (PVRP) has been explored in many medical fields with the aim to benefit from its healing potential. In parallel, efforts are being invested to understand the function and dynamics of PVRP that is complex in its composition and interactions. Some clinical evidence reveals beneficial effects of PVRP, while some report that there were no effects. To optimize the preparation methods, functions and mechanisms of PVRP, its constituents should be better understood. With the intention to promote further studies of autologous therapeutic PVRP, we performed a review on some topics regarding PVRP composition, harvesting, assessment and preservation, and also on clinical experience following PVRP application in humans and animals. Besides the acknowledged actions of platelets, leukocytes and different molecules, we focus on extracellular vesicles that were found abundant in PVRP.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review
IS  - 4
SP  - 3420
VL  - 24
DO  - 10.3390/ijms24043420
ER  - 

@article{
author = "Troha, Kaja and Vozel, Domen and Arko, Matevž and Bedina Zavec, Apolonija and Dolinar, Drago and Hočevar, Matej and Jan, Zala and Kisovec, Matic and Kocjančič, Boštjan and Pađen, Ljubiša and Pajnič, Manca and Penič, Samo and Romolo, Anna and Repar, Neža and Spasovski, Vesna and Steiner, Nejc and Šuštar, Vid and Iglič, Aleš and Drobne, Damjana and Kogej, Ksenija and Battelino, Saba and Kralj-Iglič, Veronika",
year = "2023",
abstract = "The preparation of autologous platelet and extracellular vesicle-rich plasma (PVRP) has been explored in many medical fields with the aim to benefit from its healing potential. In parallel, efforts are being invested to understand the function and dynamics of PVRP that is complex in its composition and interactions. Some clinical evidence reveals beneficial effects of PVRP, while some report that there were no effects. To optimize the preparation methods, functions and mechanisms of PVRP, its constituents should be better understood. With the intention to promote further studies of autologous therapeutic PVRP, we performed a review on some topics regarding PVRP composition, harvesting, assessment and preservation, and also on clinical experience following PVRP application in humans and animals. Besides the acknowledged actions of platelets, leukocytes and different molecules, we focus on extracellular vesicles that were found abundant in PVRP.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review",
number = "4",
pages = "3420",
volume = "24",
doi = "10.3390/ijms24043420"
}

Troha, K., Vozel, D., Arko, M., Bedina Zavec, A., Dolinar, D., Hočevar, M., Jan, Z., Kisovec, M., Kocjančič, B., Pađen, L., Pajnič, M., Penič, S., Romolo, A., Repar, N., Spasovski, V., Steiner, N., Šuštar, V., Iglič, A., Drobne, D., Kogej, K., Battelino, S.,& Kralj-Iglič, V.. (2023). Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review. in International Journal of Molecular Sciences
MDPI., 24(4), 3420.
https://doi.org/10.3390/ijms24043420

Troha K, Vozel D, Arko M, Bedina Zavec A, Dolinar D, Hočevar M, Jan Z, Kisovec M, Kocjančič B, Pađen L, Pajnič M, Penič S, Romolo A, Repar N, Spasovski V, Steiner N, Šuštar V, Iglič A, Drobne D, Kogej K, Battelino S, Kralj-Iglič V. Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review. in International Journal of Molecular Sciences. 2023;24(4):3420.
doi:10.3390/ijms24043420 .

Troha, Kaja, Vozel, Domen, Arko, Matevž, Bedina Zavec, Apolonija, Dolinar, Drago, Hočevar, Matej, Jan, Zala, Kisovec, Matic, Kocjančič, Boštjan, Pađen, Ljubiša, Pajnič, Manca, Penič, Samo, Romolo, Anna, Repar, Neža, Spasovski, Vesna, Steiner, Nejc, Šuštar, Vid, Iglič, Aleš, Drobne, Damjana, Kogej, Ksenija, Battelino, Saba, Kralj-Iglič, Veronika, "Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review" in International Journal of Molecular Sciences, 24, no. 4 (2023):3420,
https://doi.org/10.3390/ijms24043420 . .