Patrick Hill

Bi-directional exchange of membrane components occurs during co-culture of mesenchymal stem cells and nucleus pulposus cells.

Mesenchymal stem cell (MSC)-based therapies have been proposed as novel treatments for intervertebral disc (IVD) degeneration. We have previously demonstrated that when MSCs are co-cultured with nucleus pulposus (NP) cells with direct cell-cell contact, they differentiate along the NP lineage and simultaneously stimulate the degenerate NP cell population to regain a normal (non-degenerate) phenotype, an effect which requires cell-cell communication. However, the mechanisms by which NP cells and MSCs interact in this system are currently unclear. Thus, in this study we investigated a range of potential mechanisms for exchange of cellular components or information that may direct these changes, including cell fusion, gap-junctional communication and exchange of membrane components by direct transfer or via microvesicle formation. Flow cytometry of fluorescently labeled MSCs and NP cells revealed evidence of some cell fusion and formation of gapjunctions, although at the three timepoints studied these phenomena were detectable only in a small proportion of cells. While these mechanisms may play a role in cell-cell communication, the data suggests they are not the predominant mechanism of interaction. However, flow cytometry of fluorescently dual-labeled cells showed that extensive bi-directional transfer of membrane components is operational during direct co-culture of MSCs and NP cells. Furthermore, there was also evidence for secretion and internalization of membrane-bound microvesicles by both cell types. Thus, this study highlights bi-directional intercellular transfer of membrane components as a possible mechanism of cellular communication between MSC and NP cells.

Tenocyte contraction induces crimp formation in tendon-like tissue

Tendons are composed of longitudinally aligned collagen fibrils arranged in bundles with an undulating pattern, called crimp. The crimp structure is established during embryonic development and plays a vital role in the mechanical behaviour of tendon, acting as a shock-absorber during loading. However, the mechanism of crimp formation is unknown, partly because of the difficulties of studying tendon development in vivo. Here, we used a 3D cell culture system in which embryonic tendon fibroblasts synthesise a tendon-like construct comprised of collagen fibrils arranged in parallel bundles. Investigations using polarised light microscopy, scanning electron microscopy and fluorescence microscopy showed that tendon constructs contained a regular pattern of wavy collagen fibrils. Tensile testing indicated that this superstructure was a form of embryonic crimp producing a characteristic toe region in the stress–strain curves. Furthermore, contraction of tendon fibroblasts was the critical factor in the buckling of collagen fibrils during the formation of the crimp structure. Using these biological data, a finite element model was built that mimics the contraction of the tendon fibroblasts and monitors the response of the Extracellular matrix. The results show that the contraction of the fibroblasts is a sufficient mechanical impulse to build a planar wavy pattern. Furthermore, the value of crimp wavelength was determined by the mechanical properties of the collagen fibrils and inter-fibrillar matrix. Increasing fibril stiffness combined with constant matrix stiffness led to an increase in crimp wavelength. The data suggest a novel mechanism of crimp formation, and the finite element model indicates the minimum requirements to generate a crimp structure in embryonic tendon.

One-step synthesis, characterisation and properties of core/shell silicogermanate zeotype crystals

The core/shell composite silicogermanate zeotype material, Si-ASU-7, has been prepared in a one-step solvothermal synthesis process from a reactant mixture of GeO2 and SiO2 in a dimethylamine/H2O/HF solvent mixture. The core/shell nature of the product material has been determined using a combination of X-ray diffraction on powder and single crystal samples, and energy dispersive X-ray spectroscopy. The crystal structure of Si-ASU-7, ([Si0.221Ge0.779O2]10·(Me2NH)(H2O), tetragonal, space group P4/mnc, a = 12.3484(3) A, c = 14.3979(5) A, V = 2195.43(11) A3) determined using single crystal X-ray diffraction data indicates that incorporation of silicon into the structure introduces a degree of disorder to the orientation of the double 4-ring secondary building units from which the framework is constructed. The thermal stability of the as-synthesised composite Si-ASU-7 material is enhanced compared to the pure germanate material ASU-7.

Growth of Hierarchically Structured High-Surface Area Alumina on FeCrAl Alloy Wires

The formation of metastable alumina phases due to the oxidation of commercial FeCrAl alloy wires (0.5 mm thickness) at various temperatures and time periods has been examined. Samples were isothermally oxidised in air using a thermogravimetric analyzer (TGA). The morphology of the oxidised samples was analyzed using an Electronic Scanning Electron Microscope (ESEM) and X-ray on the surface analysis was done using an Energy Dispersive X-Ray (EDX) analyzer. The technique of X-Ray Diffraction (XRD) was used to characterize the phase of the oxide growth. The entire study showed that it was possible to grow high-surface area gamma alumina on the FeCrAl alloy wire surfaces when isothermally oxidised above 800°C over several hours.