Temperature Effect on the Bilayer Stacking in Multilamellar Lipid Vesicles

Abstract

The small-angle region of the Bragg diffraction of MLV samples is a simple and powerful tool for the study of mesoscopic lipid structures either alone or in interaction with molecules of biological interest. It is also a helpful tool to obtain the much needed thermotropic phase diagrams of lipid mixtures. In the course of our work, we found that the analysis of the diffractograms obtained as a function of temperature is not as straightforward as we expected. When the aqueous medium is concentrated in univalent salts, the small-angle X-ray scattering, SAXS, develops several peaks that have been interpreted to result from regions with different thickness of the interbilayer region. However, a systematic study shows that nonzero ionic strength is by no means a necessary criterion for irregularity of bilayer stacking. We show that MLV in water are uniform and stable if made, kept, and measured at the same temperature. If not, the lamellar repeat distance is smaller than the equilibrated one, eventually developing transient multimodal SAXS diffractions. We present a detailed study of this phenomenon using SAXS and dynamic light scattering and conclude that the deviations from the equilibrium interbilayer distance is a consequence of geometric constraints created by the insufficient thermal expansion of the lipid bilayers.