Ternary lipid bilayers containing cholesterol in a high curvature silica xerogel environment.

The phase behavior of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)/1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) (1/1 mol ratio)/cholesterol (0-60 mol %) supported lipid bilayers agreed with a DOPC/DSPC/cholesterol ternary phase diagram by Zhao et al. when a mica support was used (Zhao, J.; Wu, J.; Heberle, F. A.; Mills, T. T.; Klawitter, P.; Huang, G.; Costanza, G.; Feigenson, G. W. Biochim. Biophys. Acta, Biomembr. 2007, 1768, 2764-2776). However, when a silica xerogel support was used, the phase behavior deviated from the phase diagram. Specifically, miscibility and trend lines of DSPC-rich domain area fraction, domain shape, and domain size versus cholesterol, obtained by analysis of fluorescence and atomic force microscopy (AFM) images, were as expected for mica-supported lipid bilayers, but were substantially stretched to higher cholesterol concentrations for silica xerogel-supported lipid bilayers. In addition, this behavior was found in three other ternary lipid compositions substituting slightly shorter acyl chain lengths in comparison to DSPC or a saturated lipid versus unsaturated DOPC. Qualitative comparison of domain characteristics of DOPC/DSPC/cholesterol (0 and 15 mol %) bilayers supported by silica xerogel, mica, borosilicate glass, and quartz showed that the networked surface layer of high curvature (0.04 nm(-1)) silica beads was the dominant influence as opposed to the surface chemistry. Based upon the literature, we postulate two curvature-based mechanisms that explain our results. In the first mechanism, cholesterol was transferred from the higher curvature supported lipid bilayer to the lower curvature vesicles in the medium during the vesicle fusion and thermal cooling step, resulting in a lowered cholesterol concentration of the supported lipid bilayer. In the second mechanism, high curvature promoted sustained lipid demixing as the cholesterol concentration was increased, thus creating a new phase diagram in which coexisting phases persist to a higher cholesterol concentration. These results suggest that a surface layer of high curvature features can be used to observe and study curvature-induced intrabilayer transport or demixing over large areas and that curvature can play an important role in sorting and localization of biomembrane components.

Silica xerogel/aerogel-supported lipid bilayers: consequences of surface corrugation.

The objective of this paper was to review our recent investigations of silica xerogel and aerogel-supported lipid bilayers. These systems provide a format to observe relationships between substrate curvature and supported lipid bilayer formation, lipid dynamics, and lipid mixtures phase behavior and partitioning. Sensitive surface techniques such as quartz crystal microbalance and atomic force microscopy are readily applied to these systems. To inform current and future investigations, we review the experimental literature involving the impact of curvature on lipid dynamics, lipid and phase-separated lipid domain localization, and membrane-substrate conformations and we review our molecular dynamics simulations of supported lipid bilayers with the atomistic and molecular information they provide.

Effect of support corrugation on silica xerogel--supported phase-separated lipid bilayers.

Lipid bilayers supported by substrates with nanometer-scale surface corrugations hold interest in understanding both nanoparticle-membrane interactions and the challenges of constructing models of cell membranes on surfaces with desirable properties, e.g., porosity. Here, we successfully form a two-phase (gel-fluid) lipid bilayer supported by nanoporous silica xerogel. Surface topology, lateral diffusion coefficient, and lipid density in comparison to mica-supported lipid bilayers were characterized by atomic force microscopy, fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), and quantitative fluorescence microscopy, respectively. We found that the two-phase lipid bilayer follows the silica xerogel surface contours. The corrugation imparted on the lipid bilayer results in a lipid density that is twice that on a flat mica surface in the fluid regions. In direct agreement with the doubling of actual bilayer area in a projected area, we find that the lateral diffusion coefficient (D) of fluid lipids on silica xerogel (approximately 1.7 microm2/s) is lower than on mica (approximately 3.9 microm2/s) by both FRAP and FCS techniques. Furthermore, the gel-phase domains on silica xerogel compared to mica were larger and less numerous. Overall, our results suggest the presence of a relatively defect-free continuous two-phase lipid bilayer that penetrates approximately midway into the first layer of approximately 50 nm silica xerogel beads.

AFM for structure and dynamics of biomembranes.

We review structure and dynamic measurements of biomembranes by atomic force microscopy (AFM). We focus mainly on studies involving supported lipid bilayers (SLBs), particularly formation by vesicle rupture on flat and corrugated surfaces, nucleation and growth of domains in phase-separated systems, anesthetic-lipid interactions, and protein/peptide interactions in multicomponent systems. We show that carefully designed experiments along with real-time AFM imaging with superior lateral and z resolution (0.1 nm) have revealed quantitative details of the mechanisms and factors controlling vesicle rupture, domain shape and size, phase transformations, and some model biological interactions. The AFM tip can also be used as a mechanical transducer and incorporated in electrochemical measurements of membrane components; therefore, we touch on these important applications in both model and cell membranes.

Production and characterization of films from cotton stalk xylan.

Composite film production based on cotton stalk xylan was studied, and the mechanical and physical properties of the films formed were investigated. Xylan and lignin were separated from cellulose by alkali extraction and, then, lignin was removed using ethanol washing. Self-supporting continuous films could not be produced using pure cotton stalk xylan. However, film formation was achieved using 8-14% (w/w) xylan without complete removal of lignin during xylan isolation. Keeping about 1% lignin in xylan (w/w) was determined to be sufficient for film formation. Films were produced by casting the film-forming solutions, followed by solvent evaporation in a temperature (20 degrees C) and relative humidity (40%) controlled environment. The elastic modulus and hypothetical coating strength of the films obtained by using 8% xylan were significantly different from the ones containing 10-14% xylan. The water vapor transfer rates (WVTR) decreased with increasing xylan concentration, which made the films thicker. The glycerol addition as an additional plasticizer resulting in more stretchable films having higher WVTR and lower water solubility values. As a result, film production was successfully achieved from xylan, which was extracted from an agricultural waste (cotton stalk), and the film-forming effect of lignin on pure xylan has been demonstrated.