James Dean

Cell structure, stiffness and permeability of freeze-dried collagen scaffolds in dry and hydrated states

UNLABELLED Scaffolds for tissue engineering applications should be highly permeable to support mass transfer requirements while providing a 3-D template for the encapsulated biological cells. High porosity and cell interconnectivity result in highly compliant scaffolds. Overstraining occurs easily with such compliant materials and can produce misleading results. In this paper, the cell structure of freeze-dried collagen scaffolds, in both dry and hydrated states, was characterised using X-ray tomography and 2-photon confocal microscopy respectively. Measurements have been made of the scaffolds Youngs modulus using conventional mechanical testing and a customised see-saw testing configuration. Specific permeability was measured under constant pressure gradient and compared with predictions. The collagen scaffolds investigated here have a coarse cell size (∼100-150 μm) and extensive connectivity between adjacent cells (∼10-30 μm) in both dry and hydrated states. The Youngs modulus is very low, of the order of 10 kPa when dry and 1 kPa when hydrated. There is only a single previous study concerning the specific permeability of (hydrated) collagen scaffolds, despite its importance in nutrient diffusion, waste removal and cell migration. The experimentally measured value reported here (5 × 10(-)(10)m(2)) is in good agreement with predictions based on Computational Fluid Dynamics simulation and broadly consistent with the Carman-Kozeny empirical estimate. It is however about three orders of magnitude higher than the single previously-reported value and this discrepancy is attributed at least partly to the high pressure gradient imposed in the previous study. STATEMENT OF SIGNIFICANCE The high porosity and interconnectivity of tissue engineering scaffolds result in highly compliant structures (ie large deflections under low applied loads). Characterisation is essential if these scaffolds are to be systematically optimised. Scaffold overstraining during characterisation can lead to misleading results. In this study, the stiffness (in dry and hydrated states) and specific permeability of freeze-dried collagen scaffolds have been measured using techniques customised for low stiffness structures. The scaffold cell structure is investigated using X-ray computed tomography, which has been applied previously to visualise such materials, without extracting any structural parameters or simulating fluid flow. These are carried out in this work. 2-photon confocal microscopy is used for the first time to study the structure in hydrated state.

Force generation by the growth of amyloid aggregates

Significance Force generation by active biological materials, in particular through native protein polymerization, is a key feature of cellular function and motility. Protein polymerization to form amyloid fibrils is associated with a number of devastating and currently incurable protein misfolding diseases. Unlike the polymerization of actin and tubulin driving cell motility, little is known about the mechanical properties of amyloid fibril growth. Here, we present direct experimental measurements of the force generated by growing amyloid fibrils. These measurements demonstrate that amyloid growth can release comparable forces to actin and tubulin polymerization. This conclusion is remarkable as actin and tubulin, unlike proteins forming amyloid fibrils, have evolved to generate force. The generation of mechanical forces are central to a wide range of vital biological processes, including the function of the cytoskeleton. Although the forces emerging from the polymerization of native proteins have been studied in detail, the potential for force generation by aberrant protein polymerization has not yet been explored. Here, we show that the growth of amyloid fibrils, archetypical aberrant protein polymers, is capable of unleashing mechanical forces on the piconewton scale for individual filaments. We apply microfluidic techniques to measure the forces released by amyloid growth for two systems: insulin and lysozyme. The level of force measured for amyloid growth in both systems is comparable to that observed for actin and tubulin, systems that have evolved to generate force during their native functions and, unlike amyloid growth, rely on the input of external energy in the form of nucleotide hydrolysis for maximum force generation. Furthermore, we find that the power density released from growing amyloid fibrils is comparable to that of high-performance synthetic polymer actuators. These findings highlight the potential of amyloid structures as active materials and shed light on the criteria for regulation and reversibility that guide molecular evolution of functional polymers.

Limit case analysis of the “stable indenter velocity” method for obtaining creep stress exponents from constant load indentation creep tests

This study concerns a commonly-used procedure for evaluating the steady state creep stress exponent, n

Characterisation of Cell Adhesion to Substrate Materials and the Resistance to Enzymatic and Mechanical Cell-Removal

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4.6 Mechanical Properties of Metallic Fiber Network Materials

, from indentation data. The procedure involves monitoring the indenter displacement history under constant load and making the assumption that, once its velocity has stabilised, the system is in a quasi-steady state, with stage II creep dominating the behaviour. The stress and strain fields under the indenter are represented by “equivalent stress” and “equivalent strain rate” values. The estimate of n

Impact Energy Absorption in Novel, Lightweight Sandwich Panels With Metallic Fibre Cores

n

Energy absorption during projectile perforation of thin steel plates and the kinetic energy of ejected fragments

is then obtained as the gradient of a plot of the logarithm of the equivalent strain rate against the logarithm of the equivalent stress. Concerns have, however, been expressed about the reliability of this procedure, and indeed it has already been shown to be fundamentally flawed. In the present paper, it is demonstrated, using a very simple analysis, that, for a genuinely stable velocity, the procedure always leads to the same, constant value for n

Energy absorption during projectile perforation of lightweight sandwich panels with metallic fibre cores

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A procedure for extracting primary and secondary creep parameters from nanoindentation data

(either 1.0 or 0.5, depending on whether the tip shape is spherical or self-similar). This occurs irrespective of the value of the measured velocity, or indeed of any creep characteristic of the material. It is now clear that previously-measured values of n

A critical assessment of the “stable indenter velocity” method for obtaining the creep stress exponent from indentation data

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