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Stability of erythrocyte-derived nanovesicles assessed by light scattering and electron microscopy
ID
Božič, Darja
(
Author
),
ID
Hočevar, Matej
(
Author
),
ID
Kisovec, Matic
(
Author
),
ID
Pajnič, Manca
(
Author
),
ID
Pađen, Ljubiša
(
Author
),
ID
Jeran, Marko
(
Author
),
ID
Bedina Zavec, Apolonija
(
Author
),
ID
Podobnik, Marjetka
(
Author
),
ID
Kogej, Ksenija
(
Author
),
ID
Iglič, Aleš
(
Author
),
ID
Kralj-Iglič, Veronika
(
Author
)
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https://www.mdpi.com/1422-0067/22/23/12772
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Abstract
Extracellular vesicles (EVs) are gaining increasing amounts of attention due to their potential use in diagnostics and therapy, but the poor reproducibility of the studies that have been conducted on these structures hinders their breakthrough into routine practice. We believe that a better understanding of EVs stability and methods to control their integrity are the key to resolving this issue. In this work, erythrocyte EVs (hbEVs) were isolated by centrifugation from suspensions of human erythrocytes that had been aged in vitro. The isolate was characterised by scanning (SEM) and cryo-transmission electron microscopy (cryo-TEM), flow cytometry (FCM), dynamic/static light scattering (LS), protein electrophoresis, and UV-V spectrometry. The hbEVs were exposed to various conditions (pH (4–10), osmolarity (50–1000 mOsm/L), temperature (15–60 °C), and surfactant Triton X-100 (10–500 μM)). Their stability was evaluated by LS by considering the hydrodynamic radius (R$_h$), intensity of scattered light (I), and the shape parameter (ρ). The morphology of the hbEVs that had been stored in phosphate-buffered saline with citrate (PBS–citrate) at 4 °C remained consistent for more than 6 months. A change in the media properties (50–1000 mOsm/L, pH 4–10) had no significant effect on the R$_h$ (=100–130 nm). At pH values below 6 and above 8, at temperatures above 45 °C, and in the presence of Triton X-100, hbEVs degradation was indicated by a decrease in I of more than 20%. Due to the simple preparation, homogeneous morphology, and stability of hbEVs under a wide range of conditions, they are considered to be a suitable option for EV reference material.
Language:
English
Keywords:
light scattering
,
vesicle characterization
,
vesicle stability
,
extracellular vesicle reference material
,
nanovesicles
,
cellular vesicles
,
vesicle shape
,
scanning electron microscopy of extracellular vesicles
,
cryo-electron microscopy of extracellular vesicles
Work type:
Article
Typology:
1.01 - Original Scientific Article
Organization:
ZF - Faculty of Health Sciences
FE - Faculty of Electrical Engineering
FKKT - Faculty of Chemistry and Chemical Technology
MF - Faculty of Medicine
Publication status:
Published
Publication version:
Version of Record
Year:
2021
Number of pages:
28 str.
Numbering:
Vol. 22, iss. 23, art. 12772
PID:
20.500.12556/RUL-133687
UDC:
577
ISSN on article:
1422-0067
DOI:
10.3390/ijms222312772
COBISS.SI-ID:
88130563
Publication date in RUL:
09.12.2021
Views:
1565
Downloads:
183
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Record is a part of a journal
Title:
International journal of molecular sciences
Shortened title:
Int. j. mol. sci.
Publisher:
MDPI
ISSN:
1422-0067
COBISS.SI-ID:
2779162
Licences
License:
CC BY 4.0, Creative Commons Attribution 4.0 International
Link:
http://creativecommons.org/licenses/by/4.0/
Description:
This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:
01.12.2021
Secondary language
Language:
Slovenian
Keywords:
sipanje svetlobe
,
karakterizacija veziklov
,
stabilnost veziklov
,
referenčni material zunajceličnih veziklov
,
nanovezikli
,
celični vezikli
,
oblika veziklov
,
vrstična elektronska mikroskopija zunajceličnih veziklov
,
krioelektronska mikroskopija zunajceličnih veziklov
Projects
Funder:
EC - European Commission
Funding programme:
H2020
Project number:
801338
Name:
Extracellular vesicles from a natural source for tailor-made nanomaterials
Acronym:
VES4US
Funder:
ARRS - Slovenian Research Agency
Project number:
P2-0232
Name:
Funkcije in tehnologije kompleksnih sistemov
Funder:
ARRS - Slovenian Research Agency
Project number:
P3-0388
Name:
Mehanizmi varovanja zdravja
Funder:
ARRS - Slovenian Research Agency
Project number:
P1-0201
Name:
Fizikalna kemija
Funder:
ARRS - Slovenian Research Agency
Project number:
P1-0391
Name:
Molekulske interakcije
Funder:
ARRS - Slovenian Research Agency
Project number:
P2-0132
Name:
Fizika in kemija površin kovinskih materialov
Funder:
ARRS - Slovenian Research Agency
Project number:
L3-2621
Name:
Mehanizmi omajanja endoproteze kolka
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