Friction, lubrication, and in situ mechanics of poroelastic cellulose hydrogels.

The tribology between biphasic materials is challenging to predict and interpret due to the interrelationship between mechanical properties, microstructure and movement of the fluid phase contained within. A new approach is presented to deconvolute these effects for cellulose hydrogels, which have a fibrous network that is akin to the microstructure of articular cartilage and plant cell walls. This is achieved by developing a tribo-rheological technique that uniquely incorporates in situ mechanical characterisation (compression-relaxation and small amplitude oscillatory shear) immediately prior to measuring the tribological response between pairs of hydrogels. A radial pressure gradient is generated upon compression-relaxation of the poroelastic hydrogels that results in a non-uniform film thickness at the interface between them. Simulations of this process show that contact between gels occurs in an outer annulus region. Accounting for the predicted contact area between hydrogels varying in cellulose density and pectin solution viscosity causes measured tribology data to collapse onto a single curve; the apparent static friction between hydrogel tribopairs increases with the storage modulus of the hydrogels according to a power law with exponent 0.67. The method is used to compare the influence of plant cell wall polysaccharides, xyloglucan and arabinoxylan, on the interactive forces between cellulose fibres; xyloglucan is found to reduce the static friction between the hydrogels while arabinoxylan had no significant effect. The methodologies presented should provide a new framework for studying the friction between gels and other biphasic soft materials and polymeric surface films.

Surface rearrangement of adsorbed EGCG-mucin complexes on hydrophilic surfaces.

The kinetic adsorption-desorption behaviour of porcine gastric mucin in the presence of physiologically relevant concentrations of the polyphenol epigallocatechin gallate (EGCG) was investigated using high-resolution kinetic optical waveguide lightmode spectroscopy (OWLS) and atomic force microscopy (AFM). Comparison with dynamic light scattering results from EGCG-mucin mixtures indicates that discrete particles are formed whose size increases with increasing EGCG:mucin ratio. These particles are deduced to be the adsorbing entities, which fuse on the surface to form complex surface layers. At low molar EGCG:mucin ratios (<∼1000), aggregates fuse on the surface to form a monolayer similar to one of pure mucin. With increasing EGCG concentration, the surface assembly of aggregates becomes consistent with their rearrangement and spreading in the shape of a spherical segment. At the highest molar ratios investigated (>12,000) the particles begin to destabilize. The presence of EGCG leads to birefringence hysteresis during adsorption-desorption, indicating structural rearrangement, even at molar ratios ∼1000. The intensification of the phenomenon with increasing EGCG:mucin ratio mimics what was previously observed with the increase of mucin concentration in an EGCG-free system.

Interpreting atomic force microscopy nanoindentation of hierarchical biological materials using multi-regime analysis.

We present a novel Multi-Regime Analysis (MRA) routine for interpreting force indentation measurements of soft materials using atomic force microscopy. The MRA approach combines both well established and semi-empirical theories of contact mechanics within a single framework to deconvolute highly complex and non-linear force-indentation curves. The fundamental assumption in the present form of the model is that each structural contribution to the mechanical response acts in series with other 'mechanical resistors'. This simplification enables interpretation of the micromechanical properties of materials with hierarchical structures and it allows automated processing of large data sets, which is particularly indispensable for biological systems. We validate the algorithm by demonstrating for the first time that the elastic modulus of polydimethylsiloxane (PDMS) films is accurately predicted from both approach and retraction branches of force-indentation curves. For biological systems with complex hierarchical structures, we show the unique capability of MRA to map the micromechanics of live plant cells, revealing an intricate sequence of mechanical deformations resolved with precision that is unattainable using conventional methods of analysis. We recommend the routine use of MRA to interpret AFM force-indentation measurements for other complex soft materials including mammalian cells, bacteria and nanomaterials.

Concentration of salivary protective proteins within the bound oral mucosal pellicle.

OBJECTIVES: To study which salivary proteins form the protective bound mucosal pellicle and to determine the role of transglutaminase in pellicle development. MATERIALS AND METHODS: Oral epithelial cells were collected and underwent washes of different strengths, followed by homogenisation. SDS-PAGE, western blotting, IgA ELISAs and amylase activity assays were completed on cell homogenates and compared to saliva samples to confirm which salivary proteins were bound to cell surfaces. RESULTS: Salivary mucins, MUC5B and MUC7, were strongly retained on the oral epithelial cell surface. Other bound proteins including cystatin S, carbonic anhydrase VI, secretory component and IgA could be washed off. IgA was present in concentrated levels in the bound mucosal pellicle compared to amounts in saliva. Amylase, one of the most abundant proteins present in saliva, showed minimal levels of binding. Transglutaminase 3 presence was confirmed, but proteins that it catalyses cross-links between, statherin and proline-rich proteins, showed minimal presence. CONCLUSION: Some protective salivary proteins including mucins and IgA become concentrated on oral surfaces in the bound mucosal pellicle, through specific interactions. Concentration of mucins would contribute to lubrication to prevent abrasion damage to soft tissues, whilst increased IgA could create an 'immune reservoir' against mucosal infection.

Understanding glycoprotein behaviours using Raman and Raman optical activity spectroscopies: characterising the entanglement induced conformational changes in oligosaccharide chains of mucin.

We illustrate the great potential of Raman and ROA spectroscopies for investigating the structure and organisation of glycoproteins and the complex matrices they can form. In combination these spectroscopic techniques are sensitive to changes in conformation revealing details of secondary and tertiary structures, probing hydrogen bonding interactions, as well as resolving side chain orientation and the absolute configuration of chiral substructures. To demonstrate this potential we have characterised the structural changes in a complex glycoprotein, mucin. Spectral changes were observed during the entanglement transition as the mucin concentration was increased. By applying two-dimensional correlation analysis (2DCos) to the ROA and Raman concentration-dependent spectral sets delicate transitions in mucin conformation could also be determined. From ~20-40 mg/ml conformational transitions assigned mainly to the sugar N-acetyl-d-galactosamine (GalNAc), which is the linking saccharide unit to the protein backbone, were monitored. Further changes in local oligosaccharide conformation above 40 mg/ml were also monitored, together with other structural transitions observed in the protein core, particularly ß-structure formation. Consequently, these spectral techniques were shown to monitor the formation of transient entanglements formed by brush-brush interactions between oligosaccharide combs of mucin molecules identifying changes in both carbohydrate and protein moieties. This work clearly shows how these methods can be used to elucidate fresh insights into the complex behaviour of these large complex molecules.

Collective dynamics of an end-grafted polymer brush in solvents of varying quality.

The dynamic structure of a chemically end-grafted polystyrene brush bathed in solvents of varying interactions was studied by evanescent wave dynamic light scattering. It reveals distinct behavior under good and poor solvent conditions. The cooperative diffusion is a generic feature of a good solvent environment, whereas a second slow relaxation mode appears in the theta solvent regime. Its characteristics resemble self-diffusion of clusters in a gel while weak concentration fluctuations in the polymer brush decay similarly to a semidilute polymer solution.