Study of polyvinyl alcohols (9 and 31 kDa) aggregation in aqueous solutions by fluorescent probing
Abstract
Background: When developing low-temperature cell storage methods, a serious problem is recrystallization, which leads to cell damage during thawing. Previous studies have shown the promising use of polyvinyl alcohol (PVA) as an inhibitor of recrystallization. But the mechanisms of protective action of PVA are not finally clarified. So, it is not known what structural features contribute to implementation of PVA antirecrystallization properties in the cryoprotective concentration range.
Objectives: Establishing the peculiarities of structuring PVA molecules in aqueous solutions using the fluorescent probe.
Materials and Methods: Aqueous solutions of 0.1–5% (wt.%) PVA with molecular mass (m.m.) of 9 and 31 kDa) were studied. Fluorescence probe method, photometry, stalagmometry, and molecular modeling were used.
Results: Using the 3-hydroxy-4¢-(N,N-dimethylamino)flavones (FME) fluorescent probe it was found that in 0.1–5% of PVA (m.m. 9 and 31 kDa) aqueous solutions the structural organization of polymers changes with formation of different in size and structure of local hydrophobic regions. In PVA solutions, m.m. 9 kDa micelles with smaller cavities are formed in which FME is densely surrounded by polymer segments. In the case of PVA m.m. 31 kDa, it forms micelles with smaller cavities surrounded by polymer segments. PVA m.m. 31 kDa forms micelles with larger in size and more hydrophilic cavities. If the content is more than 3%, PVA m.m. 31 kDa aggregates are partially destroyed, which may be the result of increased water content. Under these conditions, PVA m.m. 9 kDa micelles are enlarged. as a result of aggregation. According to molecular modeling data, PVA is able to form strong hydrogen-linked complexes with the surface of ice nanocrystals. Such complex, having a hydrophobic surface, can depolarize water molecules, thus slowing down further growth of ice crystals.
Conclusions: Changes in the structural organization, which may affect the recrystallization properties, have been found in water solutions of PVA. The mechanism of implementation of polymer anticrystallization activity has been suggested. The possible role of structure and supramolecular organization of PVA in aqueous solutions in understanding the mechanisms of depressing recrystallization during freeze-thawing of cells is discussed.
Downloads
References
Mazur P. Freezing of living cells: Mechanisms and implications. Am. J. Physiol. 1984;247:125–42. https://doi.org/10.1152/ajpcell.1984.247.3.C125
Scott KL, Lecak J, Acker JP. Biopreservation of red blood cells: past, present and future. Transfus. Med. Rev. 2005;19:127–42. https://doi.org/10.1016/j.tmrv.2004.11.004
Deller RC, Vatish M, Mitchell DA, Gibson MI. Synthetic polymers enable non-vitreous cellular cryopreservation by reducing ice crystal growth during thawing. Nature Communications. 2014;5:3244–51. https://doi.org/10.1038/ncomms4244
Leclere M, Kwok BK, Wu LK, Allan DS, Ben RN. C-Linked antifreeze glycoprotein (C-AFGP) analogues as novel cryoprotectants. Bioconj.Chem. 2011;22:1804–10. https://doi.org/10.1021/bc2001837
Tam RY, Ferreira SS, Czechura P, Chaytor JL, Ben RN. Hydration index A better parameter for explaining small molecule hydration in inhibition of ice recrystallization. J. Am. Chem. Soc. 2008;130:17494–501. https://doi.org/10.1021/ja806284x
Deller RC, Congdon T, Sahid MA, Morgan M, Vatish M, Mitchell DA, et al. Ice recrystallization inhibition by polyols comparison of molecular and macromolecular inhibitors and role of hydrophobic units. Biomaterials Science. 2013;1:478–85. https://doi.org/10.1039/C3BM00194F
Gibson MI. Slowing the growth of ice with synthetic macromolecules: beyond antifreeze(glyco) proteins. Polym. Chem. 2010;1:1141–52. https://doi.org/10.1039/C0PY00089B
Congdon T, Notman R, Gibson MI. Antifreeze (glyco)protein mimetic behavior of poly(vinyl alcohol): detailed structure ice recrystallization inhibition activity study. Biomacromolecules. 2013;14:1578–86. https://doi.org/10.1021/bm400217j
Richards SJ, Jones MW, Hunaban M, Haddleton DM, MI Gibson. Probing bacterial-toxin inhibition with synthetic glycopolymers prepared by tandem post-polymerization modification: role of linker length and carbohydrate density. Angew. Chem. Int. Ed. 2012;51:7812–16. https://doi.org/10.1002/anie.201202945
Capicciotti CJ, Leclere M, Perras FA, Bryce DL, Paulin H, Harden J, et al. Potent inhibition of ice recrystallization by low molecular weight carbohydrate-based surfactants and hydrogelators. Chem. Sci. 2012;3:1408–16. https://doi.org/10.1039/C2SC00885H
Haymet ADJ, Ward LG., Harding MM. Winter flounder “antifreeze” proteins: synthesis and ice growth inhibition of analogues that probe the relative importance of hydrophobic and hydrogen-bonding interactions. J. Am. Chem. Soc., 1999;121:941–48. https://doi.org/10.1021/ja9801341
Gibson MI, Barker CA, Spain SG, Albertin L, Cameron NR. Inhibition of ice crystal growth by synthetic glycopolymers: implications for the rational design of antifreeze glycoprotein mimics. Biomacromolecules. 2009;10:328–33. https://doi.org/10.1021/bm801069x
Roshal AD, Grigorovich AV, Dorochenko AO, Pivovarenko VG, Demchenko AP. Flavonols as methal ion chelators. Complex formation with Mg2+ and Ba2+ cations in the excited state. J. Photochemistry and Photobiology A: Chemistry. 1999;127:89–100. https://doi.org/10.1016/S1010-6030(99)00105-7
Ercelen S, Klymchenko AS, Demchenko AP. Ultras ensitive fluorescent probe for the hydrophobic range of solvent polarities. Analytica Chimica Acta. 2002;464:273–87. https://doi.org/10.1016/S0003-2670(02)00493-2
Ercelen S, Klymchenko AS, Demchenko AP. Novel two-color fluorescence probe with extreme specifity to bovine serum albumin. FEBS Letters. 2003;538:25–8. https://doi.org/10.1016/S0014-5793(03)00116-9
Roshal AD, Organero JA, Douhal A. Tuning the mechanism of proton-transfer in a hydroxyflavone derivative. Chemical Physics Letters. 2003;379:53–9. https://doi.org/10.1016/j.cplett.2003.08.008
Moroz VV, Chalyi AG, Roshal AD. The properties of 4’-N,N-dimethylaminoflavonol in the ground and excited states. Russ. J. Phys. Chem. 2008;82(9):1464–69. https://doi.org/10.1134/S0036024408090100
Klymchenko AS, Demchenko AP. Multiparametric probing of intermolecular interactions with fluorescent dye exhibiting excited state intramolecular proton transfer. Phys. Chem. Chem. Phys. 2003;5:461–8. https://doi.org/10.1039/B210352D
Shynkar VV, Klymchenko AS, Piémont E, Demchenko AP, Mély Y. Dynamics of intermolecular hydrogen bonds in the excited states of 4’-dialkylamino-3-hydroxyflavones. On the pathway to an ideal fluorescent hydrogen bonding sensor. J. Phys. Chem. A. 2004;108:8151–9. https://doi.org/10.1021/jp047990l
Harkins WD, Brown FE. The determination of surface tension (free surface energy), and the weight of falling drops: the surface tension of water and benzene by the capillary height method. J. Am. Chem. Soc. 1919;41(4):499–24. https://doi.org/10.1021/ja01461a003
Abe M, editor. Measurement techniques and practices of colloid and interface phenomena. Springer Nature Singapore Pte Ltd. 2019; 145 p. https://doi.org/10.1007/978-981-13-5931-6
Smith MA, Neumann RM, Webb RA. A modification of the Algar-Flynn-Oyamada preparation of flavonols. J. Heteroc. Chem. 1968;5:425–6. https://doi.org/10.1002/jhet.5570050323
Klymchenko AS, Mely Y, Demchenko AP, Duportail G. Simultaneous probing of hydration and polarity of lipid bilayers with 3-hydroxyflavne fluorescent dyes. Biochim. Biophys. Acta. 2004;1665:6–19. https://doi.org/10.1016/j.bbamem.2004.06.004
Liu M, Cheng R, Qian R. Effect of solution concentration on the gelation of aqueous polyvinyl alcohol solution. J. Polym. Sci., Part B: Polym. Phys. 1995;33,1731–35. https://doi.org/10.1002/polb.1995.090331204
Rumyantsev MS, Gushchin AV. The effect of hexamethylphosphoramide solvent on the viscosity of polyvinyl alcohol solutions. Vestnik Nizhegorodskogo universiteta im. NI Lobachevskogo. 2013;1:91–4. Available from: http://www.unn.ru/pages/e-library/vestnik/99999999_West_2013_1(1)/20.pdf (in Russian)
Shinoda K, Yamaguchi T, Hori R. The surface tension and the critical micelle concentration in aqueous solution of β-D-alkyl glucosides and their mixtures. Bull. Chem. Soc. Jpn. 1961;34:237–41. https://doi.org/10.1246/bcsj.34.237
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
