Study on the effect of small alcohol on lipid hydration and liposome formation_Journal of Biomedical Engineering

Authors：

JIANG Lihua ¹ ,  WANG Qiong ² , HU Ning ³ , YANG Jun ³

1. School of General Education, Chongqing City Management College, Chongqing 400055, P. R. China;
2. Department of Basic Medicine, Chongqing Medical and Pharmaceutical College, Chongqing 401331, P. R. China;
3. Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400030, P. R. China;

Corresponding author：

WANG Qiong, Email: wangqiong@cqu.edu.cn

Keywords：

Liposome; Ethanol; Hydration; Biomembrane

DOI：

10.7507/1001-5515.202105060

Video：

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Liposomes with precisely controlled composition are usually used as membrane model systems to investigate the fundamental interactions of membrane components under well-defined conditions. Hydration method is the most common method for liposome formation which is found to be influenced by composition of the medium. In this paper, the effects of small alcohol (ethanol) on the hydration of lipid molecules and the formation of liposomes were investigated, as well as its coexistence with sodium chloride. It was found that ethanol showed the opposite effect to that of sodium chloride on the hydration of lipid molecules and the formation of liposomes. The presence of ethanol promoted the formation of liposomes within a certain range of ethanol content, but that of sodium chloride suppressed the liposome formation. By investigating the fluorescence intensity and continuity of the swelled membranes as a function of contents of ethanol and sodium chloride, it was found that sodium chloride and ethanol showed the additive effect on the hydration of lipid molecules when they coexisted in the medium. The results may provide some reference for the efficient preparation of liposomes.

Citation： JIANG Lihua, WANG Qiong, HU Ning, YANG Jun. Study on the effect of small alcohol on lipid hydration and liposome formation. Journal of Biomedical Engineering, 2022, 39(1): 112-119. doi: 10.7507/1001-5515.202105060 Copy

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2.	Garten M, Mosgaard L D, Bornschlőgl T, et al. Whole-GUV patch-clamping. Proc Natl Acad Sci USA, 2017, 114(2): 328-333.
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7.	Montes L, Alonso A, Goni F M, et al. Giant unilamellar vesicles electroformed from native membranes and organic lipid mixtures under physiological conditions. Biophys J, 2007, 93(10): 3548-3554.
8.	Angelova M, Dimitrov D S. A mechanism of liposome electroformation. Prog Colloid Polym Sci, 1988, 76: 59-67.
9.	Wang Q, Li W, Hu N, et al. Ion concentration effect (Na⁺ and Cl⁻) on lipid vesicle formation. Colloid Surf B-Biointerfaces, 2017, 155: 287-293.
10.	Fan T, Wang Q, Hu N, et al. Preparation of giant lipid vesicles with controllable sizes by a modified hydrophilic polydimethylsiloxane microarray chip. J Colloid Interface Sci, 2018, 536: 53-61.
11.	Hossain M, Blanchard G J. Effects of ethanol and n-butanol on the fluidity of supported lipid bilayers. Chem Phys Lipids, 2021, 238(105091): 1-7.
12.	Ma C D, Wang C, Acevedo-Vélez C, et al. Modulation of hydrophobic interactions by proximally immobilized ions. Nature, 2015, 517(7534): 347-350.
13.	Hwang S, Shao Q, Williams H, et al. Methanol strengthens hydrogen bonds and weakens hydrophobic interactions in proteins-a combined molecular dynamics and NMR study. J Phys Chem B, 2011, 115(20): 6653-6660.
14.	Manca M L, Castangia I, Matricardi P, et al. Molecular arrangements and interconnected bilayer formation induced by alcohol or polyalcohol in phospholipid vesicles. Colloid Surf B-Biointerfaces, 2014, 117: 360-367.
15.	Feller S E, Brown C A, Nizza D T, et al. Nuclear overhauser enhancement spectroscopy cross-relaxation rates and ethanol distribution across membranes. Biophys J, 2002, 82(3): 1396-1404.
16.	Cevc G, Löbbecke L, Nagel N, et al. Phospholipid-alcohol interactions: effects of chain-length and headgroup variations. Phosphorus Sulfur, 1996, 109(1-4): 285-288.
17.	Wanderlingh U, D’angelo G, Nibali V C, et al. Interaction of alcohol with phospholipid membrane: NMR and XRD investigations on DPPC–hexanol system. Spectroscopy, 2012, 24: 375-380.
18.	Spector S, Selinger J V, Schnur J M. Thermodynamics of phospholipid tubules in alcohol/water solutions. J Am Chem Soc, 1997, 119(36): 8533-8539.
19.	Mikelj M, Praper T, Demič R, et al. Electroformation of giant unilamellar vesicles from erythrocyte membranes under low-salt conditions. Anal Biochem, 2013, 435(2): 174-180.
20.	Marcus Y. Effect of ions on the structure of water: structure making and breaking. Chem Rev, 2010, 40(22): 1346-1370.
21.	Mancinelli R, Botti A, Bruni F, et al. Hydration of sodium, potassium, and chloride ions in solution and the concept of structure maker/breaker. J Phys Chem B, 2007, 111(48): 13570-13577.
22.	Imberti S, Botti A, Bruni F, et al. Ions in water: The microscopic structure of concentrated hydroxide solutions. J Chem Phys, 2005, 122(194509): 1-9.
23.	Botti A, Bruni F, Imberti S, et al. Ions in water: The microscopic structure of concentrated NaOH solutions. J Chem Phys, 2004, 120(21): 10154-10162.
24.	Vácha R, Jurkiewicz P, Petrov M, et al. Mechanism of interaction of monovalent ions with phosphatidylcholine lipid membranes. J Phys Chem B, 2010, 114(29): 9504-9509.
25.	Vácha R, Siu S W, Petrov M, et al. Effects of alkali cations and halide anions on the DOPC lipid membrane. J Phys Chem A, 2009, 113(26): 7235-7243.
26.	Deplazes E, Tafalla B D, Cranfield C G, et al. Role of ion-phospholipid interactions in zwitterionic phospholipid bilayer ion permeation. J Phys Chem Lett, 2020, 11(15): 6353-6358.
27.	Böckmann R A, Hac A, Heimburg T, et al. Effect of sodium chloride on a lipid bilayer. Biophys J, 2003, 85(3): 1647-1655.
28.	Perttu E K, Kohli A G, Szoka F C. Inverse-phosphocholine lipids: a remix of a common phospholipid. J Am Chem Soc, 2012, 134(10): 4485-4488.
29.	Sachs J N, Woolf T B. Understanding the Hofmeister effect in interactions between chaotropic anions and lipid bilayers: molecular dynamics simulations. J Am Chem Soc, 2003, 125(29): 8742-8743.
30.	Sachs J N, Nanda H, Petrache H I, et al. Changes in phosphatidylcholine headgroup tilt and water order induced by monovalent salts: molecular dynamics simulations. Biophys J, 2004, 86(6): 3772-3782.
31.	Boström M, Parsons D F, Salis A, et al. Possible origin of the inverse and direct hofmeister series for lysozyme at low and high salt concentrations. Langmuir, 2011, 27(15): 9504-9511.
32.	Edwards S A, Williams D R M. Double layers and interparticle forces in colloid science and biology: analytic results for the effect of ionic dispersion forces. Phys Rev Lett, 2004, 92(24): 248303.
33.	Parsons D F, Ninham B W. Charge reversal of surfaces in divalent electrolytes: the role of ionic dispersion interactions. Langmuir, 2010, 26(9): 6430-6436.
34.	Parsons D F, Ninham B W. Surface charge reversal and hydration forces explained by ionic dispersion forces and surface hydration. Colloid Surf A-Physicochem Eng Asp, 2011, 383(1-3): 2-9.
35.	Santos A P D, Levin Y. Ion specificity and the theory of stability of colloidal suspensions. Phys Rev Lett, 2011, 106(16): 167801.
36.	Chong Y, Kleinhammes A, Tang P, et al. Dominant alcohol–protein interaction via hydration-enabled enthalpy-driven binding mechanism. J Phys Chem B, 2015, 119(17): 5367-5375.
37.	Kanduč M, Netz R R. From hydration repulsion to dry adhesion between asymmetric hydrophilic and hydrophobic surfaces. Proc Natl Acad Sci USA, 2015, 112(40): 12338-12343.
38.	Leekumjorn S, Sum A K. Molecular investigation of the interactions of trehalose with lipid bilayers of DPPC, DPPE and their mixture. Mol Simul, 2006, 32(3-4): 219-230.
39.	Cacela C, Hincha D K. Low amounts of sucrose are sufficient to depress the phase transition temperature of dry phosphatidylcholine, but not for lyoprotection of liposomes. Biophys J, 2006, 90(8): 2831-2842.

1. Stein H, Spindler S, Bonakdar N, et al. Production of isolated giant unilamellar vesicles under high salt concentrations. Front Physiol, 2017, 8(63): 1-16.
2. Garten M, Mosgaard L D, Bornschlőgl T, et al. Whole-GUV patch-clamping. Proc Natl Acad Sci USA, 2017, 114(2): 328-333.
3. Czogalla A, Grzybek M, Jones W, et al. Validity and applicability of membrane model systems for studying interactions of peripheral membrane proteins with lipids. Biochim Biophys Acta, 2014, 1841(8): 1049-1059.
4. Kindt J T, Szostak J W, Wang A. Bulk self-assembly of giant, unilamellar vesicles. ACS Nano, 2020, 14(11): 14627-14634.
5. Zvonimir B, Ana P, Dubravka K, et al. Effect of electrical parameters and cholesterol concentration on giant unilamellar vesicles electroformation. Cell Biochem Biophys, 2020, 78(2): 157-164.
6. Pott T, Bouvrais H, Méléard P. Giant unilamellar vesicle formation under physiologically relevant conditions. Chem Phys Lipids, 2008, 154(2): 115-119.
7. Montes L, Alonso A, Goni F M, et al. Giant unilamellar vesicles electroformed from native membranes and organic lipid mixtures under physiological conditions. Biophys J, 2007, 93(10): 3548-3554.
8. Angelova M, Dimitrov D S. A mechanism of liposome electroformation. Prog Colloid Polym Sci, 1988, 76: 59-67.
9. Wang Q, Li W, Hu N, et al. Ion concentration effect (Na⁺ and Cl⁻) on lipid vesicle formation. Colloid Surf B-Biointerfaces, 2017, 155: 287-293.
10. Fan T, Wang Q, Hu N, et al. Preparation of giant lipid vesicles with controllable sizes by a modified hydrophilic polydimethylsiloxane microarray chip. J Colloid Interface Sci, 2018, 536: 53-61.
11. Hossain M, Blanchard G J. Effects of ethanol and n-butanol on the fluidity of supported lipid bilayers. Chem Phys Lipids, 2021, 238(105091): 1-7.
12. Ma C D, Wang C, Acevedo-Vélez C, et al. Modulation of hydrophobic interactions by proximally immobilized ions. Nature, 2015, 517(7534): 347-350.
13. Hwang S, Shao Q, Williams H, et al. Methanol strengthens hydrogen bonds and weakens hydrophobic interactions in proteins-a combined molecular dynamics and NMR study. J Phys Chem B, 2011, 115(20): 6653-6660.
14. Manca M L, Castangia I, Matricardi P, et al. Molecular arrangements and interconnected bilayer formation induced by alcohol or polyalcohol in phospholipid vesicles. Colloid Surf B-Biointerfaces, 2014, 117: 360-367.
15. Feller S E, Brown C A, Nizza D T, et al. Nuclear overhauser enhancement spectroscopy cross-relaxation rates and ethanol distribution across membranes. Biophys J, 2002, 82(3): 1396-1404.
16. Cevc G, Löbbecke L, Nagel N, et al. Phospholipid-alcohol interactions: effects of chain-length and headgroup variations. Phosphorus Sulfur, 1996, 109(1-4): 285-288.
17. Wanderlingh U, D’angelo G, Nibali V C, et al. Interaction of alcohol with phospholipid membrane: NMR and XRD investigations on DPPC–hexanol system. Spectroscopy, 2012, 24: 375-380.
18. Spector S, Selinger J V, Schnur J M. Thermodynamics of phospholipid tubules in alcohol/water solutions. J Am Chem Soc, 1997, 119(36): 8533-8539.
19. Mikelj M, Praper T, Demič R, et al. Electroformation of giant unilamellar vesicles from erythrocyte membranes under low-salt conditions. Anal Biochem, 2013, 435(2): 174-180.
20. Marcus Y. Effect of ions on the structure of water: structure making and breaking. Chem Rev, 2010, 40(22): 1346-1370.
21. Mancinelli R, Botti A, Bruni F, et al. Hydration of sodium, potassium, and chloride ions in solution and the concept of structure maker/breaker. J Phys Chem B, 2007, 111(48): 13570-13577.
22. Imberti S, Botti A, Bruni F, et al. Ions in water: The microscopic structure of concentrated hydroxide solutions. J Chem Phys, 2005, 122(194509): 1-9.
23. Botti A, Bruni F, Imberti S, et al. Ions in water: The microscopic structure of concentrated NaOH solutions. J Chem Phys, 2004, 120(21): 10154-10162.
24. Vácha R, Jurkiewicz P, Petrov M, et al. Mechanism of interaction of monovalent ions with phosphatidylcholine lipid membranes. J Phys Chem B, 2010, 114(29): 9504-9509.
25. Vácha R, Siu S W, Petrov M, et al. Effects of alkali cations and halide anions on the DOPC lipid membrane. J Phys Chem A, 2009, 113(26): 7235-7243.
26. Deplazes E, Tafalla B D, Cranfield C G, et al. Role of ion-phospholipid interactions in zwitterionic phospholipid bilayer ion permeation. J Phys Chem Lett, 2020, 11(15): 6353-6358.
27. Böckmann R A, Hac A, Heimburg T, et al. Effect of sodium chloride on a lipid bilayer. Biophys J, 2003, 85(3): 1647-1655.
28. Perttu E K, Kohli A G, Szoka F C. Inverse-phosphocholine lipids: a remix of a common phospholipid. J Am Chem Soc, 2012, 134(10): 4485-4488.
29. Sachs J N, Woolf T B. Understanding the Hofmeister effect in interactions between chaotropic anions and lipid bilayers: molecular dynamics simulations. J Am Chem Soc, 2003, 125(29): 8742-8743.
30. Sachs J N, Nanda H, Petrache H I, et al. Changes in phosphatidylcholine headgroup tilt and water order induced by monovalent salts: molecular dynamics simulations. Biophys J, 2004, 86(6): 3772-3782.
31. Boström M, Parsons D F, Salis A, et al. Possible origin of the inverse and direct hofmeister series for lysozyme at low and high salt concentrations. Langmuir, 2011, 27(15): 9504-9511.
32. Edwards S A, Williams D R M. Double layers and interparticle forces in colloid science and biology: analytic results for the effect of ionic dispersion forces. Phys Rev Lett, 2004, 92(24): 248303.
33. Parsons D F, Ninham B W. Charge reversal of surfaces in divalent electrolytes: the role of ionic dispersion interactions. Langmuir, 2010, 26(9): 6430-6436.
34. Parsons D F, Ninham B W. Surface charge reversal and hydration forces explained by ionic dispersion forces and surface hydration. Colloid Surf A-Physicochem Eng Asp, 2011, 383(1-3): 2-9.
35. Santos A P D, Levin Y. Ion specificity and the theory of stability of colloidal suspensions. Phys Rev Lett, 2011, 106(16): 167801.
36. Chong Y, Kleinhammes A, Tang P, et al. Dominant alcohol–protein interaction via hydration-enabled enthalpy-driven binding mechanism. J Phys Chem B, 2015, 119(17): 5367-5375.
37. Kanduč M, Netz R R. From hydration repulsion to dry adhesion between asymmetric hydrophilic and hydrophobic surfaces. Proc Natl Acad Sci USA, 2015, 112(40): 12338-12343.
38. Leekumjorn S, Sum A K. Molecular investigation of the interactions of trehalose with lipid bilayers of DPPC, DPPE and their mixture. Mol Simul, 2006, 32(3-4): 219-230.
39. Cacela C, Hincha D K. Low amounts of sucrose are sufficient to depress the phase transition temperature of dry phosphatidylcholine, but not for lyoprotection of liposomes. Biophys J, 2006, 90(8): 2831-2842.

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