Molecular dynamics study of surface-tethered S(CH2CH2O)6CH3: helix formation and thermal disorder.

We present the results of a molecular dynamics study of a set of surface-tethered S(CH2CH2O)6CH3 chains. In this study, we analyze helix formation, in addition to thermal disorder, and find that spontaneous helix formation and details of helix morphology depend on charge partitioning ascribed to oxygen and the methylene groups. The effects of varying surface coverage as well as chain-surface interaction strength indicate that a set of approximately 7/2 helical structures oriented predominantly normal to the surface are formed at near full coverage. This occurs even though thermal disorder clearly precludes a description based on the concept of a perfect crystalline monolayer. Thermal fluctuations in chain morphology in the vicinity of the terminal methyl groups lead to the exposure of oxygen to the external environment. We also find that the persistence of compact helix-containing domains at partial surface coverage results in the formation of well-defined cavities or void regions that expose the bare surface, even in the presence of strong chain-surface attractive interactions.

Osteoblast cell membrane hybrid bilayers for studying cell-cell interactions.

Osteoblast-like cells were grown on a surface that presents cell membrane components to the cells in culture. The culture surface was a bimolecular layer formed by the interaction of osteoblast plasma membrane vesicles with an alkanethiol monolayer. The potential of these osteoblast-membrane hybrid bilayers for promoting osteoblast adhesion, growth and differentiation was examined. UMR-106 osteoblast-like cells cultured on these surfaces are normal in appearance, and in the presence of serum, proliferate as well or better than on control surfaces. The level of alkaline phosphatase production in the presence and absence of serum suggests that the osteoblast-like cells retain their differentiated phenotype, and appear to respond to the cell surface ligands presented by the osteoblast-membrane biomimetic surface. These observations suggest that biomimetic membrane films prepared from osteoblast cell membranes support osteoblast cell growth, allow the cells to maintain their differentiation state and may be suitable as a model system to probe cell-cell interactions.

Raman and FTIR spectroscopies of fluorescein in solutions.

Raman and Fourier transform-infra red (FT-IR) spectroscopies of fluorescein in aqueous solutions have been investigated in the pH range from 9.1 to 5.4. At pH 9.1 fluorescein is in the dianion form. At pH 5.4, fluorescein is a mixture of monoanion (approximately 85%), dianion and neutral forms (together approximately 15%). The fluorescence quantum yield drops from 0.93 for the dianion form to 0.37 for the monoanion form. The Raman and FT-IR studies focused on the frequency range from 1000 to 1800 cm(-1) which contains the skeletal vibrational modes of the xanthene moiety of fluorescein. At pH 9.1, the spectroscopic feature of fluorescein dianion are consistent with a picture of an electron delocalized among the xanthene moiety and two identical oxygens attached to opposite ends of the xanthene moiety, forming a very symmetric structure. The characteristic of fluorescein dianion is the presence of the phenoxide-like stretch at 1310 cm(-1). At pH 5.4, fluorescein monoanion has lost the symmetric structure characteristic of the dianion. The spectra of the monoanion have distinctive contributions from the phenolic bend at 1184 cm(-1). The assignments of the vibrational bands shown in Raman and FT-IR spectra are given based on both literature and the ab initio calculations at the Hartree-Fock level with HF/6-31 + +G* basis set. Excellent correlation is found between the experimental and calculated spectra.

First-principles determination of hybrid bilayer membrane structure by phase-sensitive neutron reflectometry.

The application of a new, phase-sensitive neutron reflectometry method to reveal the compositional depth profiles of biomimetic membranes is reported. Determination of the complex reflection amplitude allows the related scattering length density (SLD) profile to be obtained by a first-principles inversion without the need for fitting or adjustable parameters. The SLD profile so obtained is unique for most membranes and can therefore be directly compared with the SLD profile corresponding to the chemical compositional profile of the film, as predicted, for example, by a molecular dynamics simulation. Knowledge of the real part of the reflection amplitude, in addition to enabling the inversion, makes it possible to assign a spatial resolution to the profile for a given range of wavevector transfer over which the reflectivity data are collected. Furthermore, the imaginary part of the reflection amplitude can be used as a sensitive diagnostic tool for recognizing the existence of certain in-plane inhomogeneities in the sample. Measurements demonstrating the practical realization of this phase-sensitive technique were performed on a hybrid bilayer membrane (self-assembled monolayer of thiahexa (ethylene oxide) alkane on gold and a phospholipid layer) in intimate contact with an aqueous reservoir. Analysis of the experimental results shows that accurate compositional depth profiles can now be obtained with a spatial resolution in the subnanometer range, primarily limited by the background originating from the reservoir and the roughness of the film's supporting substrate.

Hybrid bilayer membranes in air and water: infrared spectroscopy and neutron reflectivity studies.

In this report we describe the fabrication and characterization of a phospholipid/alkanethiol hybrid bilayer membrane in air. The bilayer is formed by the interaction of phospholipid with the hydrophobic surface of a self-assembled alkanethiol monolayer on gold. We have characterized the resulting hybrid bilayer membrane in air using atomic force microscopy, spectroscopic ellipsometry, and reflection-absorption infrared spectroscopy. These analyses indicate that the phospholipid added is one monolayer thick, is continuous, and exhibits molecular order which is similar to that observed for phospholipid/phospholipid model membranes. The hybrid bilayer prepared in air has also been re-introduced to water and characterized using neutron reflectivity and impedance spectroscopy. Impedance data indicate that when moved from air to water, hybrid bilayers exhibit a dielectric constant and thickness that is essentially equivalent to hybrid bilayers prepared in situ by adding phospholipid vesicles to alkanethiol monolayers in water. Neutron scattering from these samples was collected out to a wave vector transfer of 0.25 A(-1), and provided a sensitivity to changes in total layer thickness on the order of 1-2 A. The data confirm that the acyl chain region of the phospholipid layer is consistent with that observed for phospholipid-phospholipid bilayers, but suggest greater hydration of the phospholipid headgroups of HBMs than has been reported in studies of lipid multilayers.