A practical guide for in situ mechanical testing of musculoskeletal tissues using synchrotron tomography.

Musculoskeletal tissues are complex hierarchical materials where mechanical response is linked to structural and material properties at different dimensional levels. Therefore, high-resolution three-dimensional tomography is very useful for assessing tissue properties at different scales. In particular, Synchrotron Radiation micro-Computed Tomography (SR-microCT) has been used in several applications to analyze the structure of bone and biomaterials. In the past decade the development of digital volume correlation (DVC) algorithms applied to SR-microCT images and its combination with in situ mechanical testing (four-dimensional imaging) have allowed researchers to visualise, for the first time, the deformation of musculoskeletal tissues and their interaction with biomaterials under different loading scenarios. However, there are several experimental challenges that make these measurements difficult and at high risk of failure. Challenges relate to sample preparation, imaging parameters, loading setup, accumulated tissue damage for multiple tomographic acquisitions, reconstruction methods and data processing. Considering that access to SR-microCT facilities is usually associated with bidding processes and long waiting times, the failure of these experiments could notably slow down the advancement of this research area and reduce its impact. Many of the experimental failures can be avoided with increased experience in performing the tests and better guidelines for preparation and execution of these complex experiments; publication of negative results could help interested researchers to avoid recurring mistakes. Therefore, the goal of this article is to highlight the potential and pitfalls in the design and execution of in situ SR-microCT experiments, involving multiple scans, of musculoskeletal tissues for the assessment of their structural and/or mechanical properties. The advice and guidelines that follow should improve the success rate of this type of experiment, allowing the community to reach higher impact more efficiently.

Full-field strain of regenerated bone tissue in a femoral fracture model.

The mechanical behaviour of regenerated bone tissue during fracture healing is key in determining its ability to withstand physiological loads. However, the strain distribution in the newly formed tissue and how this influences the way a fracture heals it is still unclear. X-ray Computed Tomography (XCT) has been extensively used to assess the progress of mineralised tissues in regeneration and when combined with in situ mechanics and digital volume correlation (DVC) has been proven a powerful tool to understand the mechanical behaviour and full-field three-dimensional (3D) strain distribution in bone. The purpose of this study is therefore to use in situ XCT mechanics and DVC to investigate the strain distribution and load-bearing capacity in a regenerating fracture in the diaphyseal bone, using a rodent femoral fracture model stabilised by external fixation. Rat femurs with 1 mm and 2 mm osteotomy gaps were tested under in situ XCT step-wise compression in the apparent elastic region. High strain was present in the newly formed bone (εp1 and εp3 reaching 29 000 µÎµ and -43 000 µÎµ, respectively), with a wide variation and inhomogeneity of the 3D strain distribution in the regenerating tissues of the fracture gap, which is directly related to the presence of unmineralised tissue observed in histological images. The outcomes of this study will contribute in understanding natural regenerative ability of bone and its mechanical behaviour under loading.

X-ray computed tomography evaluations of additive manufactured multimaterial composites.

Additive Manufacturing (AM) often produces complex engineered structures by precisely distributing materials in a layer-by-layer fashion. Multimaterial AM is a particularly flexible technique able to combine a range of hard and soft materials to produce designed composites. Critically, the design of AM multimaterial structures requires the development of precise three-dimensional (3D) computed aided design (CAD) files. While such digital design is heavily used, techniques able to validate the physically manufactured composite against the digital design from which it is generated are lacking for AM, especially as any evaluations must be able to distinguish material variation across the 3D space. Nowadays, there is a growing interest in volumetric tools that can provide topological information hidden by the surface of shaped materials. So far, technologies such as Optical microscopy (OM), Scanning Electron Microscopy (SEM), and Coordinate Measuring Machine (CMM) have paved the way into the metrology field to measure the external geometry of physical objects. Currently, alongside conventional metrology tools, X-ray computed tomography (XCT) is emerging to measure the subsurface of the objects but maintaining the integrity of the probed samples. Thereby, the volumetric nature of the XCT investigations and its associated imaging techniques, ensure 3D quantitative measurements comparable to the output data from 2D metrology tools, but above all, supply the missing subsurface description for an exhaustive metrology study. The reward associated with XCT applied to multimaterial AM is a map reflecting the fabricated distribution of materials following CAD, with the benefits of better understanding the mechanical interplay within phases, hence, describing the hidden processes as well as the changes in phases due to a range of mechanical or chemical phenomena. In this study, a nondestructive approach using X-ray computed tomography (XCT) is used to fully evaluate the 3D distribution of multimaterials from an AM process. Specifically, two diverse hard and soft materials are alternatively produced in the form of a fibre embedded in a matrix via ink-jet printing. XCT coupled with imaging evaluation were able to distinguish between the differing materials and, importantly, to demonstrate a reduction in the expected fabricated volumes when compared to the respective CAD designs. LAY DESCRIPTION: Additive Manufacturing (AM) has recently become important in producing complex engineered structures. Using 3D CAD files and/or reconstructed data sets from imaging, hard and soft materials are manufactured independently or in combination, according to geometrical features and shapes in the input data. However, the evaluation of the resultant manufactured parts in comparison with the original 3D drawing is currently lacking. In this sense, X-ray computed tomography (XCT) provides an important metrology tool for mono and multimaterial AM. In this work a volumetric metrology investigation is proposed using higher resolution XCT to provide 3D information comparable to that of the 3D CAD drawings. A commercial high-resolution multijetting material printer (ProJet 5500X, 3D Systems, USA) is used to manufacture single fibre composites, through a complementary deposition of photo sensible polymers. Hard and soft plastics are produced using a UV curable step, resulting in materials of similar attenuation under an X-ray probe. A critical aim of the evaluations is the potential for XCT to distinguish between different UV curable 3D printing materials.

Hierarchical electrospun tendon-ligament bioinspired scaffolds induce changes in fibroblasts morphology under static and dynamic conditions.

The regeneration of injured tendons and ligaments is challenging because the scaffolds needs proper mechanical properties and a biomimetic morphology. In particular, the morphological arrangement of scaffolds is a key point to drive the cells growth to properly regenerate the collagen extracellular matrix. Electrospinning is a promising technique to produce hierarchically structured nanofibrous scaffolds able to guide cells in the regeneration of the injured tissue. Moreover, the dynamic stretching in bioreactors of electrospun scaffolds had demonstrated to speed up cell shape modifications in vitro. The aim of the present study was to combine different imaging techniques such as high-resolution X-ray tomography (XCT), scanning electron microscopy (SEM), fluorescence microscopy and histology to investigate if hierarchically structured poly (L-lactic acid) and collagen electrospun scaffolds can induce morphological modifications in human fibroblasts, while cultured in static and dynamic conditions. After 7 days of parallel cultures, the results assessed that fibroblasts had proliferated on the external nanofibrous sheath of the static scaffolds, elongating themselves circumferentially. The dynamic cultures revealed a preferential axial orientation of fibroblasts growth on the external sheath. The aligned nanofibre bundles inside the hierarchical scaffolds instead, allowed a physiological distribution of the fibroblasts along the nanofibre direction. Inside the dynamic scaffolds, cells appeared thinner compared with the static counterpart. This study had demonstrated that hierarchically structured electrospun scaffolds can induce different fibroblasts morphological modifications during static and dynamic conditions, modifying their shape in the direction of the applied loads. LAY DESCRIPTION: To enhance the regeneration of injured tendons and ligaments cells need to growth on dedicated structures (scaffolds) with mechanical properties and a fibrous morphology similar to the natural tissue. In particular, the morphological organisation of scaffolds is fundamental in leading cells to colonise them, regenerating the collagen extracellular matrix. Electrospinning is a promising technique to produce fibres with a similar to the human collagen fibres, suitable to design complex scaffolds able to guide cells in the reconstruction of the natural tissue. Moreover, it is well established that the cyclic stretching of these scaffolds inside dedicated systems called bioreactors, can speed up cells growth and their shape modification. The aim of the present study was to investigate how hierarchically structured electrospun scaffolds, made of resorbable material such as poly(L-lactic acid) and collagen, could induce morphological changes in human fibroblasts, while cultured during static and dynamic conditions. These scaffolds were composed by an external electrospun membrane that grouped inside it a ring-shaped bundle, made of axially aligned nanofibres, resembling the morphological arrangement of tendon and ligament tissue. After 7 days of parallel cultures, the scaffolds were investigated using the following imaging techniques: (i) high-resolution X-ray tomography (XCT); (ii) scanning electron microscopy (SEM); (iii) fluorescence microscopy and (iv) histology. The results showed that fibroblasts were able to grow on the external nanofibrous sheath of the static scaffolds, by elongating themselves along their circumference. The dynamic cultures revealed instead a preferential axial orientation of fibroblasts grown on the external sheath. The aligned nanofibre bundles inside the hierarchical scaffolds allowed an axial distribution of the fibroblasts along the nanofibres direction. This study has demonstrated that the electrospun hierarchically structured scaffolds investigated can modify the fibroblasts morphology both in static and dynamic conditions, in relation with the direction of the applied loads.

Applications of X-ray computed tomography for the evaluation of biomaterial-mediated bone regeneration in critical-sized defects.

Bone as such displays an intrinsic regenerative potential following fracture; however, this capacity is limited with large bone defects that cannot heal spontaneously. The management of critical-sized bone defects remains a major clinical and socioeconomic need with osteoregenerative biomaterials constantly under development aiming at promoting and enhancing bone healing. X-ray computed tomography (XCT) has become a standard and essential tool for quantifying structure-function relationships in bone and biomaterials, facilitating the development of novel bone tissue engineering strategies. This paper presents recent advancements in XCT analysis of biomaterial-mediated bone regeneration. As a noninvasive and nondestructive technique, XCT allows for qualitative and quantitative evaluation of three-dimensional (3D) scaffolds and biomaterial microarchitecture, bone growth into the scaffold as well as the 3D characterisation of biomaterial degradation and bone regeneration in vitro and in vivo. Furthermore, in combination with in situ mechanical testing and digital volume correlation (DVC), XCT demonstrated its potential to better understand the bone-biomaterial interactions and local mechanics of bone regeneration during the healing process in relation to the regeneration achieved in vivo, which will likely provide valuable knowledge for the development and optimisation of novel osteoregenerative biomaterials. LAY DESCRIPTION: Bone, being a dynamically adaptable material, displays excellent regenerative properties following fracture. However, the self-healing capacity of bone becomes more difficult with large bone defects. Those defects are common and occur in many clinical situations; hence, biomaterials are mostly used to restore both bone structure and function in the defect site. X-ray computed tomography (XCT) is a powerful tool to evaluate bone regeneration in critical-sized defects after the implantation of biomaterials, allowing to an improved understanding of the regeneration process following different bone tissue engineering approaches. This paper focuses on recent advancements in XCT analysis to characterise biomaterial-mediated bone regeneration in critical-sized defects. XCT supports three-dimensional (3D) analysis of biomaterials, scaffolds and regenerated bone microarchitecture, as well as bone ingrowth into the scaffold. As a nondestructive technique, XCT allows for a 3D characterisation of biomaterial degradation and bone regeneration over time. In addition, XCT combined with in situ mechanical experiments and digital volume correlation (DVC) provides a 3D evaluation and quantification of bone-biomaterial interactions and deformation mechanisms during the regeneration process. This remains essential for the development and enhancement of novel biomaterials able to produce bone that is comparable with the native tissue they aim to replace.

Optimization of digital volume correlation computation in SR-microCT images of trabecular bone and bone-biomaterial systems.

A micromechanical characterization of biomaterials for bone tissue engineering is essential to understand the quality of the newly regenerated bone, enabling the improvement of tissue regeneration strategies. A combination of microcomputed tomography in conjunction with in situ mechanical testing and digital volume correlation (DVC) has become a powerful technique to investigate the internal deformation of bone structure at a range of dimensional scales. However, in order to obtain accurate three-dimensional strain measurement at tissue level, high-resolution images must be acquired, and displacement/strain measurement uncertainties evaluated. The aim of this study was to optimize imaging parameters, image postprocessing and DVC settings to enhance computation based on 'zero-strain' repeated high-resolution synchrotron microCT scans of trabecular bone and bone-biomaterial systems. Low exposures to SR X-ray radiation were required to minimize irradiation-induced tissue damage, resulting in the need of advanced three-dimensional filters on the reconstructed images to reduce DVC-measured strain errors. Furthermore, the computation of strain values only in the hard phase (i.e. bone, biomaterial) allowed the exclusion of large artefacts localized in the bone marrow. This study demonstrated the suitability of a local DVC approach based on synchrotron microCT images to investigate the micromechanics of trabecular bone and bone-biomaterial composites at tissue level with a standard deviation of the errors in the region of 100 microstrain after a thorough optimization of DVC computation. LAY DESCRIPTION: Understanding the quality of newly regenerated bone after implantation of novel biomaterials is essential to improve bone tissue engineering strategies and formulation of biomaterials. The relationship between microstructure and mechanics of bone has been previously addressed combining microcomputed tomography with in situ mechanical testing. The addition of an image-based experimental technique such as digital volume correlation (DVC) allows to characterize the deformation of materials in a three-dimensional manner. However, in order to obtain accurate information at the micro-scale, high-resolution images, obtained for example by using synchrotron radiation microcomputed tomography, as well as optimization of the DVC computation are needed. This study presents the effect of different imaging parameters, image postprocessing and DVC settings for as accurate investigation of trabecular bone structure and bone-biomaterial interfaces. The results showed that when appropriate image postprocessing and DVC settings are used DVC computation results in very low strain errors. This is of vital importance for a correct understanding of the deformation in bone-biomaterial systems and the ability of such biomaterials in producing new bone comparable with the native tissue they are meant to replace.

Automatic diameter and orientation distribution determination of fibrous materials in micro X-ray CT imaging data.

Fibrous nanomaterials such as electrospun materials have many uses ranging from tissue engineering to biosensors. High-resolution imaging is an important component in the characterization of these materials. Important parameters required to predict and study the properties of fibre rich materials include diameter and orientation distribution as well as fibre spacing. The orientations and the relative dimensions of the fibres can be measured via specially designed imaging software. Difficulties in this measurement process can arise if fibres are distributed in close proximity to each other in relation to the resolution of the imaging modality. For example, if some automation is required in the measurement process and, particularly, if the automated processes are not designed for situations where the fibres are in close proximity to each other. This work is therefore concerned with the development of automated measurement techniques to provide estimates of the diameters of fibres and also the orientation distribution. The software automatically detects special points in the fibrous materials where fibres can be considered to have some delineation from surrounding fibres. These sparse points are considered to be points at which estimates of the fibres' properties can be quantified. Aligned and randomly distributed electrospun poly(caprolactone) nanofibres were prepared. Imaging of these materials was performed with an X-ray Computer Tomography system with an image voxel size of 0.15 × 0.15 × 0.15 µm3 . Scanning Electron Microscopy images were also obtained. Fibre diameters estimated using images from both modalities using the developed techniques were found to be in agreement. Orientation distribution was summarized with multiscale entropy and found to be consistent with visual observation across different scales. LAY DESCRIPTION: Fibres are present in many types of materials which can include, for some materials, very small fibres e.g. a few nanometres in diameter. Very small fibres are present in collagen and elastin which are common tissues of many organs in many types of living things. The sizes of these very small fibres and how they are arranged are important information that can help in the understanding of the overall properties of these materials. Materials with very small fibres can also be synthesized using specialised techniques. The properties of these synthesized fibrous materials are also important to help in understanding how the materials will perform in various different applications. Applications are many and can range from tissue engineering to drug delivery. Some properties of these materials can be shown, visually, with the aid of 3D imaging techniques such as X-ray Computer Tomography (XCT) or in 2D, with Scanning Electron Microscopy (SEM) but at a higher magnification. The work described here is centred around the development of computer algorithms to automatically determine material properties from 3D XCT images. Tests are performed with material samples, where the fibres are aligned (in semi-parallel fashion) and another where the fibres are randomly oriented (criss-crossing). The tests show that the developed algorithms are able to successfully and relatively accurately determine the diameters of the fibres. The tests also show that it is possible to quantify the relative randomness of the orientations of the fibres.

High-resolution x-ray tomographic morphological characterisation of electrospun nanofibrous bundles for tendon and ligament regeneration and replacement.

Repair of ligaments and tendons requires scaffolds mimicking the spatial organisation of collagen in the natural tissue. Electrospinning is a promising technique to produce nanofibres of both resorbable and biostable polymers with desired structural and morphological features. The aim of this study was to perform high-resolution x-ray tomography (XCT) scans of bundles of Nylon6.6, pure PLLA and PLLA-Collagen blends, where the nanofibres were meant to have a predominant direction. Characterisation was carried out via a dedicated methodology to firmly hold the specimen during the scan and a workflow to quantify the directionality of the nanofibres in the bundle. XCT scans with 0.4 and 1.0 µm voxel size were successfully collected for all bundle compositions. Better image quality was achieved for those bundles formed by thicker nanofibres (i.e. 0.59 µm for pure PLLA), whereas partial volume effect was more pronounced for thinner nanofibres (i.e. 0.26 µm for Nylon6.6). As expected, the nanofibres had a predominant orientation along the axis of the bundles (more than 20% of the nanofibres within 3° and more than 60% within 18° from the bundle axis), with a Gaussian-like dispersion in the other directions. The directionality assessment was validated by comparison against a similar analysis performed on SEM images: the XCT analysis overestimated the amount of nanofibres very close to the bundle axis, especially for the materials with thinnest nanofibres, but adequately identified the amount of nanofibres within 12°. LAY DESCRIPTION: Repair of ligaments and tendons requires dedicated materials (scaffolds) mimicking the spatial organisation of the collagen (the main material composing such natural tissue). Electrospinning is a promising technique that allows production of fibres with nanometric dimension using high voltage to stretch very tiny drops of polymeric solutions. Electrospinning allows processing both polymers that can be resorbed by the host tissue, and nonresorbable ones, to obtain the desired structural and morphological features by arranging the nanofibres in bundles. The aim of this study was to perform high-resolution x-ray computed tomography (XCT) scans of bundles, where the nanofibres were meant to have a predominant direction. The investigation included bundles of different compositions: a biostable polymer (Nylon) and bioresorbable ones (pure Poly-L-lactic acid (PLLA) and PLLA-Collagen blends). The electrospun bundles were produced using a validated method (Sensini et al 2017: https://doi.org/10.1088/1758-5090/aa6204). To this end, we developed a dedicated methodology to scan such small specimens, and a workflow to quantify the directionality of the nanofibres in the bundle. For all the compositions, XCT scans with extremely high resolution (i.e. down to 0.4 µm) were successfully collected. As expected, better images were obtained for those bundles where the nanofibres were thicker than the scanning resolution (i.e. 0.59 µm for pure PLLA). The images of the thinnest nanofibres (i.e. 0.26 µm for Nylon) were poorer because the fibre diameter was smaller than the resolution (partial volume effect). The nanofibres had a predominant orientation along the axis of the bundles (more than 60% of the nanofibres were within 18° from the bundle axis). The nanofibres had a Gaussian-like dispersion in the other directions. As this is the first time that XCT is used to quantify the directionality of this kind of bundles, the directionality assessment was further validated by comparison against a similar analysis performed on SEM images. Overall, this study has demonstrated the usefulness and reliability of using high-resolution x-ray computed tomography (XCT) scans to investigate the morphology of polymeric scaffolds made of electrospun nanofibres.

Morphological and Mechanical Biomimetic Bone Structures.

Cortical bone is an example of a mineralized tissue containing a compositional distribution of hard and soft phases in 3-dimensional space for mechanical function. X-ray computed tomography (XCT) is able to describe this compositional and morphological complexity but methods to provide a physical output with comparable mechanical function is lacking. A workflow is presented here to establish a method of using high contrast XCT to establish a virtual model of cortical bone that is manufactured using a multiple material capable 3D printer. Resultant 3D printed structures were produced based on more and less remodelled bone designs exhibiting a range of secondary osteon density. Variation in resultant mechanical properties of the 3D printed composite structures for each bone design was achieved using a combination of material components and reasonable prediction of elastic modulus provided using a Hashin-Shtrikman approach. The ability to 3D print composite structures using high contrast XCT to distinguish between compositional phases in a biological structure promises improved anatomical models as well as next-generation mechano-mimetic implants.

Novel mutations in KARS cause hypertrophic cardiomyopathy and combined mitochondrial respiratory chain defect.

Mutations in KARS, which encodes for both mitochondrial and cytoplasmic lysyl-tRNA synthetase, have been so far associated with three different phenotypes: the recessive form of Charcot-Mary-Tooth polyneuropathy, the autosomal recessive nonsyndromic hearing loss and the last recently described condition related to congenital visual impairment and progressive microcephaly. Here we report the case of a 14-year-old girl with severe cardiomyopathy associated to mild psychomotor delay and mild myopathy; moreover, a diffuse reduction of cytochrome C oxidase (COX, complex IV) and a combined enzymatic defect of complex I (CI) and complex IV (CIV) was evident in muscle biopsy. Using the TruSight One sequencing panel we identified two novel mutations in KARS. Both mutations, never reported previously, occur in a highly conserved region of the catalytic domain and displayed a dramatic effect on KARS stability. Structural analysis confirmed the pathogenic role of the identified variants. Our findings confirm and emphasize that mt-aminoacyl-tRNA synthetases (mt-ARSs) enzymes are related to a broad clinical spectrum due to their multiple and still unknown functions.

Computation of full-field displacements in a scaffold implant using digital volume correlation and finite element analysis.

Measurements of three-dimensional displacements in a scaffold implant under uniaxial compression have been obtained by two digital volume correlation (DVC) methods, and compared with those obtained from micro-finite element models. The DVC methods were based on two approaches, a local approach which registers independent small volumes and yields discontinuous displacement fields; and a global approach where the registration is performed on the whole volume of interest, leading to continuous displacement fields. A customised mini-compression device was used to perform in situ step-wise compression of the scaffold within a micro-computed tomography (µCT) chamber, and the data were collected at steps of interest. Displacement uncertainties, ranging from 0.006 to 0.02 voxel (i.e. 0.12-0.4 µm), with a strain uncertainty between 60 and 600 µÎµ, were obtained with a spatial resolution of 32 voxels using both approaches, although the global approach has lower systematic errors. Reduced displacement and strain uncertainties may be obtained using the global approach by increasing the element size; and using the local approach by increasing the number of intermediary sub-volumes. Good agreements between the results from the DVC measurements and the FE simulations were obtained in the primary loading direction as well as in the lateral directions. This study demonstrates that volumetric strain measurements can be obtained successfully using DVC, which may be a useful tool to investigate mechanical behaviour of porous implants.

Damage evolution in acetabular reconstructs under physiological testing in a saline environment.

Damage development in cemented acetabular reconstructs has been studied under a combined cyclic loading block representative of routine activities in a saline environment. A custom-made environmental chamber was designed and installed on the Portsmouth hip simulator to allow testing of acetabular reconstructs in a wet condition for the first time. Damage was monitored and detected by scanning at selected loading intervals using micro-focus computed tomography (µCT). The preliminary results show that, although, as in dry cases, debonding at the bone-cement interface defined the failure of the cement fixation, the combination of mechanical loading and saline environment significantly affected the damage initiation and development, with drastically reduced survival lives of the reconstructs. Debonding was found to be initiated at the bone-cement interface near the rim of the acetabular cup, or DeLee zone I, in wet condition, as opposed to initiation in DeLee zone II near the dome region in dry cases. The survival time of the reconstruct in wet condition is less than 10% of that in dry condition under a given applied hip contact force.

A Pluto-like radius and a high albedo for the dwarf planet Eris from an occultation.

The dwarf planet Eris is a trans-Neptunian object with an orbital eccentricity of 0.44, an inclination of 44 degrees and a surface composition very similar to that of Pluto. It resides at present at 95.7 astronomical units (1 AU is the Earth-Sun distance) from Earth, near its aphelion and more than three times farther than Pluto. Owing to this great distance, measuring its size or detecting a putative atmosphere is difficult. Here we report the observation of a multi-chord stellar occultation by Eris on 6 November 2010 UT. The event is consistent with a spherical shape for Eris, with radius 1,163 ± 6 kilometres, density 2.52 ± 0.05 grams per cm(3) and a high visible geometric albedo, Pv = 0.96(+0.09)(-0.04). No nitrogen, argon or methane atmospheres are detected with surface pressure larger than ∼1 nanobar, about 10,000 times more tenuous than Pluto's present atmosphere. As Pluto's radius is estimated to be between 1,150 and 1,200 kilometres, Eris appears as a Pluto twin, with a bright surface possibly caused by a collapsed atmosphere, owing to its cold environment. We anticipate that this atmosphere may periodically sublimate as Eris approaches its perihelion, at 37.8 astronomical units from the Sun.

The surface composition and temperature of asteroid 21 Lutetia as observed by Rosetta/VIRTIS.

The Visible, InfraRed, and Thermal Imaging Spectrometer (VIRTIS) on Rosetta obtained hyperspectral images, spectral reflectance maps, and temperature maps of the asteroid 21 Lutetia. No absorption features, of either silicates or hydrated minerals, have been detected across the observed area in the spectral range from 0.4 to 3.5 micrometers. The surface temperature reaches a maximum value of 245 kelvin and correlates well with topographic features. The thermal inertia is in the range from 20 to 30 joules meter(-2) kelvin(-1) second(-0.5), comparable to a lunarlike powdery regolith. Spectral signatures of surface alteration, resulting from space weathering, seem to be missing. Lutetia is likely a remnant of the primordial planetesimal population, unaltered by differentiation processes and composed of chondritic materials of enstatitic or carbonaceous origin, dominated by iron-poor minerals that have not suffered aqueous alteration.

Compressive behaviour of bovine cancellous bone and bone analogous materials, microCT characterisation and FE analysis.

Compressive behaviour of bovine cancellous bone and three open-cell metallic foams (AlSi7Mg (30 ppi and 45 ppi); CuSn12Ni2 (30 ppi)) has been studied using mechanical testing, micro-focus computed tomography and finite element modelling. Whilst the morphological parameters of the foams and the bone appear to be similar, the mechanical properties vary significantly between the foams and the bone. Finite element models were built from the CT images of the samples and multi-linear constitutive relations were used for modelling of the bone and the foams. The global responses of the bone and foam samples were reasonably well captured by the FE models, whilst the percentage of yielded elements as a measure of damage evolution during compression seems to be indicative of the micro-mechanical behaviour of the samples. The damage evolution and distribution patterns across the bone and the foams are broadly similar for the strain range studied, suggesting possible substitution of trabecular bones with appropriate foams for biomechanical studies.

Effect of protein glutathionylation on neuronal cytoskeleton: a potential link to neurodegeneration.

Neurons are highly susceptible to oxidative stress and oxidation of cytoskeletal proteins is considered one of the first steps of neurodegeneration. Protein glutathionylation is a key event in the redox regulation of protein function and constitutes a sensor of tissue oxidative stress in patho-physiological conditions. In this study, we analyzed for the first time tubulin glutathionylation and its relation to neurites degeneration. For this purpose, we exposed motoneuronal cells to the physiological oxidant glutathione disulfide (GSSG) and we analyzed the extent and morphology of axonal changes caused by protein glutathionylation in these cells. Then we studied the effect of glutathionylation on the distribution of stable and dynamic microtubules in the same cells. Our results indicate that oxidative stress conditions determined by an increased intracellular level of oxidized glutathione may cause an alteration of the cytoskeleton organization and function leading to axon degeneration. These findings might contribute to understand the sequence of pathogenic events involved in the axonal degeneration that characterizes many diseases of the nervous system associated with oxidative stress.

Bone-cement interfacial behaviour under mixed mode loading conditions.

Interfacial behaviour of the bone-cement interface has been studied under tensile, shear and mixed mode loading conditions. Bovine cancellous bone was used to bond with acrylic bone cement to form bone-cement interface samples, which were mechanically tested under selected tensile, shear and mixed mode loading conditions. The influence of the loading angle and the extent of the cement penetration on the interfacial behaviour were examined. The failure mechanisms with regard to loading mode were examined using micro-focus computed tomography. The measured tensile and shear responses were utilized in a cohesive zone constitutive model, from which the pre-yield linear and the post-yield exponential strain softening behaviour under mixed mode loading conditions was predicted. The implications of the work on the studies of cemented joint replacements are also discussed.

[Legislation and inspection for the health and safety of workers. Efficacy and limits]. / Legislazione e ispezione per la salute e la sicurezza dei lavoratori. Efficacia e limiti.

OBJECTIVES: To provide information regarding Occupational Health and Safety (OHS) Inspections in Europe. METHODS AND RESULTS: The dynamics that are transforming regulatory subsystems and complementary inspection services are described. Simplification initiatives, the limits and difficulties of applying the different models of Health and Safety Management Systems are discussed. Examples are given on how to evaluate legislation and technical standards during planning and enforcement. Different approaches for studying characteristics, methodologies and efficacy in practice of OHS Inspection are provided. CONCLUSIONS: Targeted inspections need to respond to the needs of enterprises and workers. Impartiality must be guaranteed and workers' participation should be facilitated.