Incorporation of short-chain alcohols into fluid bilayers and its effect on membrane dynamic properties as seen by neutron scattering

In this work, we investigate the effect of concentrated alcoholic solutions (up to 40%w) on extruded 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) liposomes using scattering techniques. Extrusion in alcoholic aqueous solutions (methanol, ethanol, or butanol) reduces liposome size, evidenced from the decrease in hydrodynamic radius (RH) obtained from dynamic light scattering. Short-chain alcohols such as ethanol and butanol soften the lipid membranes, as observed by the decrease of the unrelaxed bending modulus (κ̃ ) obtained by neutron spin-echo spectroscopy. Thus, softer membranes are easily ruptured during extrusion, leading to a reduction in vesicle radius corroborated by a reduction in both RH and radius of gyration as seen by dynamic and static light scattering. Moreover, model-free analysis of small-angle neutron scattering (SANS) curves suggests that solvent molecules become incorporated into the lipid membrane. Analysing deviations in the scattering invariant from values predicted by mass balance, we find that the volume fraction of liposomes increases, modifying the scattering length density of the assemblies. By quantifying these changes, we estimate that 12% to 18% of the liposome membrane is composed of solvent molecules (alcohol + water), depending on the type of alcohol and its concentration present in solution, with the exception of glycerol where next to no incorporation was observed. Finally, structural parameters obtained through light scattering and changes in contrast profiles calculated from analysis of the invariant were corroborated through modelling of the SANS curves. Modelling suggests that a reduction of bilayer thickness takes place for liposomes dispersed in ethanolic and butanolic solutions, but not in the presence of methanol or glycerol.