Journal for Veterinary Medicine, Biotechnology and Biosafety
Volume
10, Issue 4, October 2024, Pages 24–27
ISSN 2411-3174 (print version) ISSN 2411-0388
(online version)
EVALUATION
OF MECHANICAL STABILITY OF DOG ERYTHROCYTES UNDER THE INFLUENCE OF CRYOPROTECTANTS
Denysova O. M.
State Biotechnological
University, Kharkiv, Ukraine, e-mail: denysova78@gmail.com
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PDF (print version)
Citation for print version: Denysova, O. M.
(2024) ‘Evaluation of mechanical
stability of dog erythrocytes under the influence of cryoprotectants’,
Journal for Veterinary Medicine,
Biotechnology and Biosafety, 10(4), pp. 24–27.
Download
PDF (online version)
Citation for online version: Denysova, O. M.
(2024) ‘Evaluation of mechanical
stability of dog erythrocytes under the influence of cryoprotectants’,
Journal for Veterinary Medicine,
Biotechnology and Biosafety, Journal for
Veterinary Medicine, Biotechnology and Biosafety. [Online] 10(4), pp. 24–27.
DOI: 10.36016/JVMBBS-2024-10-4-3.
Summary. The mechanical stability of erythrocytes is a critical
factor in ensuring their effective functioning during storage, transportation,
and cryopreservation. The objective of this study was to ascertain the impact
of diverse cryoprotectants, including glycerol,
sucrose, dimethyl sulfoxide (DMSO),
polyethylene glycol-1500 (PEG-1500), and hydroxyethyl
starch (HES), on hemolytic damage to dog erythrocytes
subjected to mechanical stress. For this purpose, dog erythrocytes were incubated in varying concentrations of cryoprotectants and NaCl. The
cells were subjected to mechanical stress by stirring
the suspension in a container filled with plastic beads at room temperature.
The resulting hemolysis was evaluated spectrophotometrically. The results demonstrated that the
most pronounced stabilization of erythrocyte membranes was
observed during incubation with PEG-1500 and HES,
while high glycerol concentrations caused membrane destabilization. Sucrose
demonstrated a dual effect: at low concentrations, it exhibited protective properties
for cellular membranes, while at higher concentrations, enhanced membrane
vulnerability to stress. The results demonstrated that DMSO
at all studied concentrations did not significantly change the mechanical
stability of erythrocytes compared to the control group. Our findings indicate
that an increase in salt concentration in the extracellular medium is
associated with a reduction in the mechanical stability of dog erythrocytes.
The effect of cryoprotectants on the mechanical
stability of erythrocytes was found to be closely related
to their physicochemical properties. This highlights the importance of precise
selection of cryoprotectant concentrations to improve
the results of red blood cell storage and transportation. The conclusions of
this study are important for further optimization of technologies for the
long-term storage of canine erythrocytes, in particular in cryobanks
Keywords: mechanical stress,
cryopreservation
References:
Alvarez, J. G.
and Storey, B. T. (1992) ‘Evidence
for increased lipid peroxidative damage and loss of
superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation’, Journal
of Andrology, 13(3), pp. 232–241. doi:
10.1002/j.1939-4640.1992.tb00306.x.
Baskurt, O. K. and Meiselman, H. J.
(2013) ‘Red blood cell mechanical stability test’, Clinical Hemorheology and Microcirculation, 55(1),
pp. 55–62. doi: 10.3233/CH-131689.
CE (The Council
of Europe). (1986) European Convention
for the Protection of Vertebrate Animals Used for Experimental and Other
Scientific Purposes. (European Treaty Series,
No. 123). Strasbourg: The Council of Europe.
Available at: https://conventions.coe.int/treaty/en/treaties/html/123.htm.
CEC (The Council of the European Communities)
(2010) ‘Directive 2010/63/EU of the European Parliament and of the
Council of 22 September 2010 on the protection of animals used for
scientific purposes’, The Official
Journal of the European Communities, L 276, pp. 33–79. Available at: http://data.europa.eu/eli/dir/2010/63/oj.
Denysova, О.
and Zhegunov, G. (2021) ‘Cryopreservation
of canine erythrocytes using dimethyl sulfoxide,
polyethylene glycol and sucrose’, Problems of Cryobiology and Cryomedicine, 31(1), pp. 38–50. doi:
10.15407/cryo31.01.038.
Elliott, G. D., Wang, S. and
Fuller, B. J. (2017) ‘Cryoprotectants:
A review of the actions and applications of cryoprotective
solutes that modulate cell recovery from ultra-low temperatures’, Cryobiology,
76, pp. 74–91. doi:
10.1016/j.cryobiol.2017.04.004.
Gao, D. and Critser, J. K.
(2000) ‘Mechanisms of cryoinjury in living
cells’, ILAR Journal, 41(4),
pp. 187–196. doi:
10.1093/ilar.41.4.187.
Ishiguro, H.
and Rubinsky, B. (1994) ‘Mechanical
interactions between ice crystals and red blood cells during directional
solidification’, Cryobiology, 31(5), pp. 483–500. doi:
10.1006/cryo.1994.1059.
Kameneva, M. V., Repko, B. M.,
Krasik, E. F., Perricelli, B. C.
and Borovetz, H. S. (2003)
‘Polyethylene glycol additives reduce hemolysis in red blood cell
suspensions exposed to mechanical stress’, ASAIO
Journal, 49(5), pp. 537–542. doi: 10.1097/01.MAT.0000084176.30221.CF.
Saragusty, J., Gacitua, H.,
Rozenboim, I. and Arav, A.
(2009) ‘Do physical forces contribute to cryodamage?’, Biotechnology and Bioengineering, 104(4),
pp. 719–728. doi:
10.1002/bit.22435.
Shpakova, N. M.,
Orlovа, N. V. and Alexandrova, D. І.
(2010) Method for Destruction of
Erythrocytes [Sposib destruktsii
erytrotsytiv]. Patent no. UA 52701. Available at: https://sis.nipo.gov.ua/uk/search/detail/257082.
[in Ukrainian].
Shpakova, N. M., Orlova, N. V.,
Nipot, E. E. and Aleksandrova, D. I.
(2015) ‘Comparative study of mechanical stress effect on human and animal
erythrocytes’ [Porivnialne vyvchennia
dii mekhanichnoho stresu na erytrotsyty liudyny i tvaryn], Fiziologichnyi Zhurnal,
61(3), pp. 75–80. doi:
10.15407/fz61.03.075.
[in Ukrainian].
Simmonds, R. C. (2017)
‘Chapter 4. Bioethics and animal use in programs of research,
teaching, and testing’, in Weichbrod, R. H.,
Thompson, G. A. and Norton, J. N. (eds.) Management of
Animal Care and Use Programs in Research, Education, and Testing. 2nd ed. Boca Raton: CRC
Press, pp. 35–62. doi:
10.1201/9781315152189-4.
Tarasev, M., Chakraborty, S. and Alfano, K.
(2015) ‘RBC mechanical fragility as a direct blood quality metric to
supplement storage time’, Military Medicine, 180(3S),
pp. 150–157. doi: 10.7205/MILMED-D-14-00404.
Ugurel, E., Kucuksumer, Z.,
Eglenen, B. and Yalcin, O.
(2017) ‘Blood storage alters mechanical stress responses of
erythrocytes’, Clinical Hemorheology and
Microcirculation, 66(2), pp. 143–155. doi: 10.3233/CH-160219.
VRU (Verkhovna Rada Ukrainy) (2006) ‘Law
of Ukraine No. 3447-IV of
Zemlianskykh, N. G.
(2018) ‘The effects of cryoprotective
substances on the mechanical stability and geometric parameters of human
erythrocytes’, Biophysics, 63(1), pp. 66–76. doi:
10.1134/S0006350918010219.
Zemlyanskikh, N. G.
and Denisova, O. N. (2009) ‘Changes
in the erythrocyte membrane-cytoskeleton complex induced by dimethyl sulfoxide, polyethylene glycol, and low temperature’,
Biophysics, 54(4), pp. 490–496. doi:
10.1134/S0006350909040162.
Zhegunov, G. F.
and Denisova, O. N. (2010)
‘Permeability of mammalian erythrocytes for the molecules of glycerol and
DMSO and the level of cellular viability after
freezing-thawingʼ [Pronitsaemost’
eritrotsitov mlekopitayushchikh
dlya molekul glitserina i DMSO i stepen’ sokhrannosti kletok posle zamorazhivaniya-ottaivaniya],
Reports of the National Academy of
Sciences of Ukraine [Dopovidi Natsionalnoi
akademii nauk Ukrainy], 12, pp. 139–143. Available at: http://dspace.nbuv.gov.ua/handle/123456789/31118. [in Russian].
Zhegunov, G., Denysova, O.
and Zhegunova, G. (2022) ‘Blood
hypothermic storage and erythrocyte cryopreservation in dogs’, Problems
of Cryobiology and Cryomedicine, 32(4),
pp. 245–255. doi:
10.15407/cryo32.04.245.
