A. J. Bushby

Nanomechanical properties and mineral concentration in articular calcified cartilage and subchondral bone

We studied articular calcified cartilage (ACC) and the immediately subchondral bone (SCB) in normal and osteoarthritic human femoral heads. Thick slices of human normal reference post mortem (PM) and osteoarthritic (OA) femoral heads (age 55–89 years) were embedded in PMMA, micromilled, carbon coated and studied using quantitative backscattered electron (qBSE) imaging to determine variations in degree of mineralization. With exact microanatomical location, nanoindentation was performed on the same block faces in representative superior (more highly loaded) and medial regions of the joint surface. Using a partial unloading method, elastic modulus as a function of indenter penetration depth was determined using a spherical tipped diamond indenter. A pointed indenter was used to determine the tissue hardness in selected locations. The relationship between mineralization and indentation modulus was more distinct in ACC than in SCB, the latter having a higher matrix concentration with variable collagen orientation. In OA, the bulk of the measurements were coincident with those in the PM samples, although there was a greater range in the levels of mineralization and modulus in ACC. In OA, extremely hypermineralized ACC was found in ACC proper, especially in superior regions, and translocated into SCB and hyaline cartilage. The very highly mineralized cartilage fragments may function as a hard grinding abrasive, accelerating wear rates whether attached to or fragmented from the eburnated surfaces of OA ACC. Highly mineralized regions would also alter loading patterns and thereby contribute to further destruction of the joint tissues.

Viscoelastic properties of bone as a function of hydration state determined by nanoindentation

Spherical indentation creep testing was used to examine the effect of hydration state on bone mechanical properties. Analysis of creep data was based on the elastic–viscoelastic correspondence principle and utilized a direct solution for the finite loading-rate experimental conditions. The zero-time shear modulus was computed from the creep compliance function and compared to the indentation modulus obtained via conventional indentation analysis, based on an elastic unloading response. The method was validated using a well-known polymer material under three different loading conditions. The method was applied to bone samples prepared with different water content by partial exchange with ethanol, where 70% ethanol was considered as the baseline condition. A hydration increase was associated with a 43% decrease in stiffness, while a hydration decrease resulted in a 20% increase in bone tissue stiffness.

High phosphate content significantly increases apatite formation of fluoride-containing bioactive glasses

Bioactive glass-containing toothpastes for treating dentine hypersensitivity work by precipitating hydroxycarbonate apatite (HCA) onto the tooth surface, but concerns exist over the long-term durability of HCA in the mouth. Fluoride-containing bioactive glasses form fluorapatite (FAp) in physiological solutions, which is more chemically stable against acid attack. The influence of phosphate content on apatite formation was investigated by producing a low-phosphate (about 1 mol% P(2)O(5)) and a high-phosphate (about 6 mol%) series of melt-derived bioactive glasses in the system SiO(2)P(2)O(5)CaONa(2)O; increasing amounts of CaF(2) were added by keeping the ratio of all other components constant. pH change, ion release and apatite formation during immersion in Tris buffer at 37°C over up to 7 days were investigated. Crystal phases formed in Tris buffer were characterized using infrared spectroscopy, X-ray diffraction and solid-state nuclear magnetic resonance (NMR) spectroscopy. An increase in phosphate or fluoride content allowed for apatite formation at lower pH; fluoride enhanced apatite formation due to lower solubility of FAp compared to hydroxyapatite or HCA. High phosphate content glasses formed apatite significantly faster (within 6h) than low phosphate content glasses (within 3 days). In addition, an increase in phosphate content favoured apatite formation rather than fluorite (CaF(2)). (19)F magic angle spinning NMR showed the apatite formed by fluoride-containing glasses to be FAp, which makes these glasses of particular interest for dental applications. This study shows that by varying the phosphate content, the reactivity and apatite formation of bioactive glasses can be controlled successfully.

A preliminary comparison of the mechanical properties of chemically cured and ultrasonically cured glass ionomer cements, using nano-indentation techniques.

There is a requirement for a dental cement with properties comparable or superior to conventional glass ionomer cements (GICs) but with the command set properties of the resin-modified GICs. The objective of this work was to show that the application of ultrasound to conventional Fuji IX commercial glass ionomer cement imparts a command set, whilst improving the short-term surface mechanical properties. Nano-indentation techniques were employed to highlight the improvements in hardness and creep resistance imparted to the cement through the application of ultrasound. The instant set imparted by the application of ultrasound provides improved surface hardness and creep, particularly within the first 24 h after setting. The surface hardness of the chemically cured Fuji IX (176 M Pa) increased by an order of magnitude when set ultrasonically (2620 M Pa), whilst creep reduced to a negligible amount. Rapid setting allows for shorter chair time and an improved clinical technique, making restorations more convenient for both the patient and clinician.

Composite bounds on the elastic modulus of bone

Advances in diagnosis and treatment of some bone disorders can be made by understanding the linkage between mineral content and mechanical function. Bone is approximately half by volume a hydrated protein network, and the remainder is a biomineral analogue of hydroxyapatite. In the current work, paired measurements of mechanical properties, using nanoindentation, and of bone mineral volume fraction, computed from quantitative back-scattered electron imaging, were made on six different types of normal and outlier bone samples. Local elastic modulus was plotted against mineral fraction and compared with predictions of engineering bounds for a two-phase composite material. Experimental data spanning the composite bounds showed no one-to-one relationship between mechanical stiffness and bone composition, excluding the possibility of any single, simple composites model for bone at nanometer length-scales.

Control of starch granule numbers in Arabidopsis chloroplasts

The aim of this work was to investigate starch granule numbers in Arabidopsis (Arabidopsis thaliana) leaves. Lack of quantitative information on the extent of genetic, temporal, developmental, and environmental variation in granule numbers is an important limitation in understanding control of starch degradation and the mechanism of granule initiation. Two methods were developed for reliable estimation of numbers of granules per chloroplast. First, direct measurements were made on large series of consecutive sections of mesophyll tissue obtained by focused ion beam-scanning electron microscopy. Second, average numbers were calculated from the starch contents of leaves and chloroplasts and estimates of granule mass based on granule dimensions. Examination of wild-type plants and accumulation and regulation of chloroplast (arc) mutants with few, large chloroplasts provided the following new insights. There is wide variation in chloroplast volumes in cells of wild-type leaves. Granule numbers per chloroplast are correlated with chloroplast volume, i.e. large chloroplasts have more granules than small chloroplasts. Mature leaves of wild-type plants and arc mutants have approximately the same number of granules per unit volume of stroma, regardless of the size and number of chloroplasts per cell. Granule numbers per unit volume of stroma are also relatively constant in immature leaves but are greater than in mature leaves. Granule initiation occurs as chloroplasts divide in immature leaves, but relatively little initiation occurs in mature leaves. Changes in leaf starch content over the diurnal cycle are largely brought about by changes in the volume of a fixed number of granules.

Grain size and sample size interact to determine strength in a soft metal

Understanding the strengthening of small-scale materials and structures is one of the key issues in nanotechnology. Many theories exist, each addressing a small domain of experimentally observed size effects and invoking different mechanisms. Measurements of the stress–strain relationship of nickel foils in flexure by the load–unload method provide strikingly accurate data from the elastic region through the yield point and to high plastic strain. The data show that the effects on the rate of work-hardening due to crystallite size and sample size interact, whereas in existing theories they should be independent. Existing theories cannot be complete. The symmetry of the dependence of flow stress on grain size and structure size suggests that strengthening effects are due to a finite strained volume, however this is delimited.

Materials mechanical size effects: a review

Abstract Knowledge of the mechanical behaviour of materials is essential in materials science and applied mechanics. At small sizes, this behaviour shows significant departures from the classical elastic–plastic model. The strength of a material increases either when the structure is small or when only a small volume is under strain: the so called size effect. Intrinsic size effects arise due to microstructural constraints, such as grain size or second phase particle precipitation. Extrinsic size effects are caused by dimensional constraints. These extrinsic constraints might be due to small sample size, where dislocation motion and other physical mechanisms are affected by the presence of a surface or interface, or due to small strained volume, where the dimensional constraint arises from the testing system. Generally, both sample size and strained volume constrain deformation. Interactions between intrinsic and extrinsic size effects are particularly interesting, though current understanding of this topic is limited. The purpose of the present review is to survey small scale strengthening phenomena and to assess the merits of classical and current mechanisms proposed to explain these effects. Practical applications of size effects are briefly reviewed.

Nanoscale-Targeted Patch-Clamp Recordings of Functional Presynaptic Ion Channels

Summary Direct electrical access to presynaptic ion channels has hitherto been limited to large specialized terminals such as the calyx of Held or hippocampal mossy fiber bouton. The electrophysiology and ion-channel complement of far more abundant small synaptic terminals (≤1 μm) remain poorly understood. Here we report a method based on superresolution scanning ion conductance imaging of small synapses in culture at approximately 100–150 nm 3D resolution, which allows presynaptic patch-clamp recordings in all four configurations (cell-attached, inside-out, outside-out, and whole-cell). Using this technique, we report presynaptic recordings of K+, Na+, Cl−, and Ca2+ channels. This semiautomated approach allows direct investigation of the distribution and properties of presynaptic ion channels at small central synapses. Video Abstract

Study of the interaction between the indentation size effect and Hall–Petch effect with spherical indenters on annealed polycrystalline copper

Methods to obtain tensile stress–strain properties of materials from a practically non-destructive indentation test are of great industrial interest. Nanoindentation is a good candidate. However, to do this successfully, indentation size effects must be accounted for. An indentation size effect with spherical indenters has been shown for a range of fcc metals with relatively large grain size (Spary et al 2006 Phil. Mag. 86 5581–93); the increase in yield stress being proportional to the inverse cube root of indenter radius. Here, we investigate these differences further and present results for the indentation size effect with spherical indenters on Cu samples with a range of different grain sizes from 1 µm to single crystal. The important experimental control parameter, of the relative size of the indentation compared with the grain size, is also explored by using indenters of different radii on the different grain sized samples. When the grain size, d, is less than 6 times the radius of the projected contact area, a, a Hall–Petch-like behaviour is observed superimposed on the indentation size effect. For d > 6a the indentation size effect dominates. The two effects may be combined by addition in quadrature. This new parametric function is able to predict the indentation pressure in annealed copper given input values of indenter radius and grain size.