MembraneDyn: simulating the dynamics of supported membrane stacks on the nanosecond timescale.

The static structure factor and the undulation dynamics of a solid-supported membrane stack have previously been calculated by Romanov and Ul'yanov [Romanov & Ul'yanov (2002). Phys. Rev. E, 66, 061701]. Based on this prior work, the calculation has been extended to cover the membrane dynamics, i.e. the intermediate scattering function as a Fourier transform of the van Hove correlation function of the membrane stack. Fortran code which calculates the intermediate scattering function for a membrane stack on a solid support is presented. It allows the static and dynamic scattering functions to be calculated according to the derivation of Romanov and Ul'yanov. The physical properties of supported phospholipid bilayers can be examined in this way and the results can be directly compared with results obtained from grazing-incidence neutron spin-echo spectroscopy experiments.

Multiple scattering and resolution effects in small-angle neutron scattering experiments calculated and corrected by the software package MuScatt.

This article deals with multiple scattering effects that are important for the method of small-angle neutron scattering (SANS). It considers three channels for the coherent elastic, the incoherent elastic and the incoherent inelastic scattering processes. The first channel contains the desired information on the experiment. Its multiple scattering effects can be desmeared, as shown in the later sections of the article. The other two channels display a nearly constant background as a function of the scattering angle. The incoherent elastic scattering is treated by the theory of Chandrasekhar, allowing for multiple scattering even at large scattering angles. The transfer to a single representative thermalized wavelength by the inelastic scattering - as a simplification - is assumed to happen by a single scattering event. Once the transition to this altered wavelength has happened, further incoherent multiple scattering is considered. The first part of the paper deals with the multiple scattering effects of light water. In the later part of the article, deconvolution algorithms for multiple scattering and instrumental resolution of the elastic coherent signal as implemented in the program MuScatt are described. All of these considerations are interesting for both reactor-based instruments with velocity selectors and time-of-flight SANS instruments and may improve the reliability of the data treatment.

Small-angle scattering for beginners.

Many experimental methods are available for the characterization of nanostructures, but most of them are limited by stringent experimental conditions. When it comes to analysing nanostructures in the bulk or in their natural environment - even as ordinary as water at room temperature - small-angle scattering (SAS) of X-rays or neutrons is often the only option. The rapid worldwide development of synchrotron and neutron facilities over recent decades has opened unprecedented possibilities for using SAS in situ and in a time-resolved way. But, in spite of its huge potential in the field of nanomaterials in general, SAS is covered far less than other characterization methods in non-specialized curricula. Presented here is a rigorous discussion of small-angle scattering, at a technical level comparable to the classical undergraduate coverage of X-ray diffraction by crystals and which contains diffraction as a particular case.

Inelastic neutron scattering analysis with time-dependent Gaussian-field models.

Converting neutron scattering data to real-space time-dependent structures can only be achieved through suitable models, which is particularly challenging for geometrically disordered structures. We address this problem by introducing time-dependent clipped Gaussian field models. General expressions are derived for all space- and time-correlation functions relevant to coherent inelastic neutron scattering for multiphase systems and arbitrary scattering contrasts. Various dynamic models are introduced that enable one to add time-dependence to any given spatial statistics, as captured, e.g., by small-angle scattering. In a first approach, the Gaussian field is decomposed into localized waves that are allowed to fluctuate in time or to move either ballistically or diffusively. In a second approach, a dispersion relation is used to make the spectral components of the field time-dependent. The various models lead to qualitatively different dynamics, which can be discriminated by neutron scattering. The methods of this paper are illustrated with oil/water microemulsion studied by small-angle scattering and neutron spin-echo. All available data-in both film and bulk contrasts, over the entire range of q and τ-are analyzed jointly with a single model. The analysis points to the static large-scale structure of the oil and water domains while the interfaces are subject to thermal fluctuations. The fluctuations have an amplitude of around 60 Å and contribute to 30% of the total interface area.

Probing the Complex Loading-Dependent Structural Changes in Ultrahigh Drug-Loaded Polymer Micelles by Small-Angle Neutron Scattering.

Drug-loaded polymer micelles or nanoparticles are being continuously explored in the fields of drug delivery and nanomedicine. Commonly, a simple core-shell structure is assumed, in which the core incorporates the drug and the corona provides steric shielding, colloidal stability, and prevents protein adsorption. Recently, the interactions of the dissolved drug with the micellar corona have received increasing attention. Here, using small-angle neutron scattering, we provide an in-depth study of the differences in polymer micelle morphology of a small selection of structurally closely related polymer micelles at different loadings with the model compound curcumin. This work supports a previous study using solid-state nuclear magnetic resonance spectroscopy and we confirm that the drug resides predominantly in the core of the micelle at low drug loading. As the drug loading increases, neutron scattering data suggests that an inner shell is formed, which we interpret as the corona also starting to incorporate the drug, whereas the outer shell mainly contains water and the polymer. The presented data clearly shows that a better understanding of the inner morphology and the impact of the hydrophilic block can be important parameters for improved drug loading in polymer micelles as well as provide insights into the structure-property relationship.

Influence of Environmental Conditions on the Fusion of Cationic Liposomes with Living Mammalian Cells.

Lipid-based nanoparticles, also called vesicles or liposomes, can be used as carriers for drugs or many types of biological macromolecules, including DNA and proteins. Efficiency and speed of cargo delivery are especially high for carrier vesicles that fuse with the cellular plasma membrane. This occurs for lipid mixture containing equal amounts of the cationic lipid DOTAP and a neutral lipid with an additional few percents of an aromatic substance. The fusion ability of such particles depends on lipid composition with phosphoethanolamine (PE) lipids favoring fusion and phosphatidyl-choline (PC) lipids endocytosis. Here, we examined the effects of temperature, ionic strength, osmolality, and pH on fusion efficiency of cationic liposomes with Chinese hamster ovary (CHO) cells. The phase state of liposomes was analyzed by small angle neutron scattering (SANS). Our results showed that PC containing lipid membranes were organized in the lamellar phase. Here, fusion efficiency depended on buffer conditions and remained vanishingly small at physiological conditions. In contrast, SANS indicated the coexistence of very small (~50 nm) objects with larger, most likely lamellar structures for PE containing lipid particles. The fusion of such particles to cell membranes occurred with very high efficiency at all buffer conditions. We hypothesize that the altered phase state resulted in a highly reduced energetic barrier against fusion.

Long-range excitations in phospholipid membranes.

We investigated the existence of long-range excitations and correlated structures in phospholipid membranes by means of grazing-incidence neutron spin echo spectroscopy, grazing-incidence small-angle neutron scattering, and corresponding theoretical calculations inspired by smectic-membrane theory. All these methods confirmed the existence of thermal excitations in the plane of the surface of the phospholipid membranes or the corresponding structures, respectively. Also, these measurements revealed a temperature dependence of these excitations. These excitations are associated with 100 nm in-plane correlations around physiological temperatures and of 75 nm at 16 °C. A single excitation has an energy around the µeV-regime. A temperature series revealed a high abundance at physiological temperatures and pronounced long-range in-plane structures, which are strongly suppressed at temperatures below 20 °C. From the length-scales and energy transfers involved we surmise that these excitations may play a role in several functions of the cell membranes such as stability and energy dissipation along the membrane. From a fundamental point of view, the observed behavior of those excitations is congruent with that of a quasi-particle (surface mode phonon, smomon) that exists in the plane of phospholipid membranes.

Nanoscale rheology at solid-complex fluid interfaces.

Here we present an approach to measure dynamic membrane properties of phospholipid membranes close to an interface. As an example we show results of the membrane dynamics of a phospholipid membrane multilayer-stack on a solid substrate (silicon). On this sample we were able to measure local interaction and friction parameters using Grazing Incidence Neutron Spin Echo Spectroscopy (GINSES), where an evanescent neutron wave probes the fluctuations close to a rigid interface. With this method it is possible to access length scales in the nano to micrometer region as well as energies in the µeV range. Using a new neutron resonator structure we achieved the required intensity gain for this experiment. During our investigations we found an excitation mode of the phospholipid membrane that has not been reported previously and only became visible using the new methodology. We speculate that the energy transported by that undulation can also serve to distribute energy over a larger area of the membrane, stabilizing it. This new methodology has the capability to probe the viscoelastic effects of biological membranes, becoming a new tool for tribology on the nanoscale and has allowed the observation of the hitherto invisible property of phospholipid membranes using neutrons.

A Thermogelling Supramolecular Hydrogel with Sponge-Like Morphology as a Cytocompatible Bioink.

Biocompatible polymers that form thermoreversible supramolecular hydrogels have gained great interest in biomaterials research and tissue engineering. When favorable rheological properties are achieved at the same time, they are particularly promising candidates as material that allow for the printing of cells, so-called bioinks. We synthesized a novel thermogelling block copolymer and investigated the rheological properties of its aqueous solution by viscosimetry and rheology. The polymers undergo thermogelation between room temperature and body temperature, form transparent hydrogels of surprisingly high strength (G' > 1000 Pa) and show rapid and complete shear recovery after stress. Small angle neutron scattering suggests an unusual bicontinuous sponge-like gel network. Excellent cytocompatibility was demonstrated with NIH 3T3 fibroblasts, which were incorporated and bioplotted into predefined 3D hydrogel structures without significant loss of viability. The developed materials fulfill all criteria for future use as bioink for biofabrication.

Influence of ibuprofen on phospholipid membranes.

A basic understanding of biological membranes is of paramount importance as these membranes comprise the very building blocks of life itself. Cells depend in their function on a range of properties of the membrane, which are important for the stability and function of the cell, information and nutrient transport, waste disposal, and finally the admission of drugs into the cell and also the deflection of bacteria and viruses. We have investigated the influence of ibuprofen on the structure and dynamics of L-α-phosphatidylcholine (SoyPC) membranes by means of grazing incidence small-angle neutron scattering, neutron reflectometry, and grazing incidence neutron spin echo spectroscopy. From the results of these experiments, we were able to determine that ibuprofen induces a two-step structuring behavior in the SoyPC films, where the structure evolves from the purely lamellar phase for pure SoyPC over a superposition of two hexagonal phases to a purely hexagonal phase at high concentrations. A relaxation, which is visible when no ibuprofen is present in the membrane, vanishes upon addition of ibuprofen. This we attribute to a stiffening of the membrane. This behavior may be instrumental in explaining the toxic behavior of ibuprofen in long-term application.

Lamellar Diblock Copolymer Thin Films during Solvent Vapor Annealing Studied by GISAXS: Different Behavior of Parallel and Perpendicular Lamellae.

The reorientation of lamellae and the dependence of the lamellar spacing, Dlam, on polymer volume fraction, ϕP, Dlam ∝ ϕP-ß, in diblock copolymer thin films during solvent vapor annealing (SVA) are examined by combining white light interferometry (WLI) and grazing-incidence small-angle X-ray scattering (GISAXS). A thin film of lamellae-forming poly(styrene-b-butadiene) prepared by spin-coating features lamellae of different orientations with the lamellar spacing depending on orientation. During annealing with ethyl acetate (EAC) vapor, it is found that perpendicular lamellae behave differently from parallel ones, which is due to the fact that their initial lamellar thicknesses differ strongly. Quantitatively, the swelling process is composed of three regimes and the drying process of two regimes. The first two regimes of swelling are associated with a significant structural rearrangement of the lamellae; i.e., the lamellae first become thicker, and then perpendicular and randomly oriented lamellae vanish, which results in a purely parallel orientation at the end of the swelling process. The rearrangement is attributed to the increase of mobility of the polymer chains imparted by the solvent and to a decrease of total free energy of the thin film. In the third regime of swelling, the scaling exponent is found to be ß = -0.32. During drying, the deswelling is nonaffine which may be a consequence of the increase of nonfavorable segmental interactions as the solvent is removed.

Complex macrophase-separated nanostructure induced by microphase separation in binary blends of lamellar diblock copolymer thin films.

The nanostructures of thin films spin-coated from binary blends of compositionally symmetric polystyrene-b-polybutadiene (PS-b-PB) diblock copolymer having different molar masses are investigated by means of atomic force microscopy (AFM) and grazing-incidence small-angle X-ray scattering (GISAXS) after spin-coating and after subsequent solvent vapor annealing (SVA). In thin films of the pure diblock copolymers having high or low molar mass, the lamellae are perpendicular or parallel to the substrate, respectively. The as-prepared binary blend thin films feature mainly perpendicular lamellae in a one-phase state, indicating that the higher molar mass diblock copolymer dominates the lamellar orientation. The lamellar thickness decreases linearly with increasing volume fraction of the low molar mass diblock copolymer. After SVA, well-defined macrophase-separated nanostructures appear, which feature parallel lamellae near the film surface and perpendicular ones in the bulk.

Drug-induced morphology switch in drug delivery systems based on poly(2-oxazoline)s.

Defined aggregates of polymers such as polymeric micelles are of great importance in the development of pharmaceutical formulations. The amount of drug that can be formulated by a drug delivery system is an important issue, and most drug delivery systems suffer from their relatively low drug-loading capacity. However, as the loading capacities increase, i.e., promoted by good drug-polymer interactions, the drug may affect the morphology and stability of the micellar system. We investigated this effect in a prominent system with very high capacity for hydrophobic drugs and found extraordinary stability as well as a profound morphology change upon incorporation of paclitaxel into micelles of amphiphilic ABA poly(2-oxazoline) triblock copolymers. The hydrophilic blocks A comprised poly(2-methyl-2-oxazoline), while the middle blocks B were either just barely hydrophobic poly(2-n-butyl-2-oxazoline) or highly hydrophobic poly(2-n-nonyl-2-oxazoline). The aggregation behavior of both polymers and their formulations with varying paclitaxel contents were investigated by means of dynamic light scattering, atomic force microscopy, (cryogenic) transmission electron microscopy, and small-angle neutron scattering. While without drug, wormlike micelles were present, after incorporation of small amounts of drugs only spherical morphologies remained. Furthermore, the much more hydrophobic poly(2-n-nonyl-2-oxazoline)-containing triblock copolymer exhibited only half the capacity for paclitaxel than the poly(2-n-butyl-2-oxazoline)-containing copolymer along with a lower stability. In the latter, contents of paclitaxel of 8 wt % or higher resulted in a raspberry-like micellar core.