Phylogenetic signal in bone microstructure of sauropsids

In spite of the fact that the potential usefulness of bone histology in systematics has been discussed for over one and a half centuries, the presence of a phylogenetic signal in the variation of histological characters has rarely been assessed. A quantitative assessment of phylogenetic signal in bone histological characters could provide a justification for performing optimizations of these traits onto independently generated phylogenetic trees (as has been done in recent years). Here we present an investigation on the quantification of the phylogenetic signal in the following bone histological, microanatomical, and morphological traits in a sample of femora of 35 species of sauropsids: vascular density, vascular orientation, index of Haversian remodeling, cortical thickness, and cross-sectional area (bone size). For this purpose, we use two methods, regressions on distance matrices tested for significance using permutations (a Mantel test) and random tree length distribution. Within sauropsids, these bone microstructural traits have an optimal systematic value in archosaurs. In this taxon, a Mantel test shows that the phylogeny explains 81.8% of the variation of bone size and 86.2% of the variation of cortical thickness. In contrast, a Mantel test suggests that the phylogenetic signal in histological traits is weak: although the phylogeny explains 18.7% of the variation of vascular density in archosaurs, the phylogenetic signal is not significant either for vascular orientation or for the index of Haversian remodeling. However, Mantel tests seem to underestimate the proportion of variance of the dependent character explained by the phylogeny, as suggested by a PVR (phylogenetic eigenvector) analysis. We also deal with some complementary questions. First, we evaluate the functional dependence of bone vascular density on bone size by using phylogenetically independent contrasts. Second, we perform a variation partitioning analysis and show that the phylogenetic signal in bone vascular density is not a by-product of phylogentic signal in bone size. Finally, we analyze the evolution of cortical thickness in diapsids by using an optimization by squared change parsimony and discuss the functional significance of this character in terms of decreased buoyancy in crocodiles and mass saving in birds. These results are placed in the framework of the constructional morphology model, according to which the variation of a character in a clade has a historical (phylogenetic) component, a functional (adaptive) component, and a structural (architectural) component.     

The significance of bone microstructure in mechanotransduction

Recent developments in modeling the relationship between bone microstructure and mechanotransduction are reviewed. The focus is on the relationship between the bone microstructure and the mechanosensation mechanism by which osteocytes sense the bone fluid motion propelled by the mechanical loading of the whole bone.     

Dependence of ultrasonic attenuation on bone mass and microstructure in bovine cortical bone

The development of the axial transmission technique now enables in vivo evaluation of cortical bone quality, which plays an important role in bone fragility. Cortical bone is a complex multiscale material, which may be made of different types of microstructure. The interaction between ultrasound and cortical bone remains unclear and most studies have been confined to wave speed analysis. The first aim of this study is to investigate the dependence of the frequency-dependent attenuation on the type of bone microstructure. The second goal is to determine whether broadband ultrasonic attenuation (BUA) is related to volumetric bone mineral density (vBMD) and mass density. Parallelepipedic samples of bovine cortical bone were cut from three specimens and tested in the axial, radial and tangential directions using an ultrasonic transmission device. BUA was evaluated over a 1-MHz wide bandwidth around 4MHz. In addition, the microstructure of each sample was determined using an optical microscope. BUA values measured in porotic microstructure are significantly higher than in Haversian microstructure. The lowest BUA values are obtained for plexiform microstructure. For all structures, BUA in the axial direction is significantly smaller than in the radial and tangential directions. Moreover, BUA is correlated with both vBMD and density (determination coefficient (R2) equal to 0.44 and 0.65, respectively, in the axial direction). BUA variations can be explained by scattering and viscoelastic mechanisms. This study suggests that BUA measurements have the potential to discriminate among different cortical bone microstructures in addition to providing material properties.     

On the relationship between the microstructure of bone and its mechanical stiffness.

A recent study of bone structure shows that the plate-shaped carbonate apatite crystals in individual lamellae are arranged in layers across the lamellae, and that the orientation of these layers are different in alternate lamellae. Based on these findings, a new micromechanical model for the Young's modulus of bone is proposed, which accounts for the anisotropy and geometrical characteristics of the material. The model incorporates the platelet-like geometry of the basic reinforcing unit, the presence of alternating thin and thick lamellae, and the orientations of the crystal platelets in the lamellae. The thin and thick lamellae are modeled as orthotropic composite layers made up of thin rectangular apatite platelets within a collagen matrix, and classical orthotropic elasticity theory is used to calculate the Young's modulus of the lamellae. Bone is viewed as an assembly of such orthotropic lamellae bent into cylindrical structures, and having a constant, alternating angle between successive lamellae. The micromechanical model employs a modified rule-of-mixtures to account for the two types of lamellae. The model provides a curve similar to the published experimental data on the angular dependence of Young's modulus, including a local maximum at an angle between 0 and 90 degrees. A rigorous testing of the model awaits additional experimental data.     

3D imaging of microstructure of spruce wood

Synchrotron radiation phase-contrast X-ray tomographic microscopy (srPCXTM) was applied to observation and identification of the features of spruce anatomy at the cellular lengthscale. The pilot experiments presented in the paper clearly revealed the features of the heartwood of Spruce (Picea abies [L.] Karst.), such as lumina and pits connecting the lumina, with a theoretical voxel size of 0.7 x 0.7 x 0.7 microm(3). The experiments were carried out on microspecimens of heartwood, measuring approximately 200 by 200 micrometers in cross-section. The technique for production and preparation of wood microsamples was developed within the framework of this investigation. The total porosity of the samples was derived and the values of the microstructural parameters, such as the diameters of tracheid, cell wall thicknesses and pit diameters were assessed non-invasively. Microstructural features as thin/small as approximately 1.5 microm were revealed and reconstructed in 3D. It is suggested that the position of sub-voxel-sized features (such as position of tori in the bordered pit pairs) can be determined indirectly using watershed segmentation. Moreover, the paper discusses the practical issues connected with a pipelined phase-contrast synchrotron-based microtomography experiment and the possible future potentials of this technique in the domain of wood science.     

The effects of freezing,boiling and degreasing on the microstructure of bone

The histology of bone has been a useful tool in research. It is commonly used to estimate the age of an individual at death, to assess if the bone is of human or non-human origin and in trauma analysis. Factors that affect the histology of bone include age, sex, population affinity and burning to name but a few. Other factors expected to affect bone histology are freezing, boiling and degreasing but very little information is available for freezing and the effect thereof, and it is unknown if boiling and degreasing affects bone histology. The aim of this study was to assess the effects of freezing, freezing and boiling, and freezing, boiling and degreasing on the histological structure of compact bone. Five cadaver tibiae were frozen at −20 °C for 21 days followed by segments being boiled in water for three days and degreased in trichloroethylene at 82 °C for three days. Anterior midshaft sections were prepared as ground sections and for Scanning Electron Microscopy (SEM). Quantitatively, there were no significant differences between freezing, boiling and degreasing; however, qualitative differences were observed using SEM. After being frozen the bone displayed cracks and after boiling the bones displayed erosion pits on the surface. It is suggested that further research, using different durations and temperatures for boiling and freezing be undertaken on bone samples representing different ages and various skeletal elements.     

Seeing the light in bone metabolism imaging

Conventional in vivo imaging of bone metabolism is dominated by gamma-ray bone scintigraphy: a technique in which gamma-ray emissions from radioactively labeled regions of metabolically active bone are mapped. More recently, however, near-infrared fluorescent probes have been developed that optically emulate these radionuclides. Although still in their infancy, techniques based on the use of such functionally targeted fluorophores might one-day offer improved resolution, sensitivity and speed in bone metabolism imaging -- without any of the health risks posed by the internalization of radioactive sources.     

Palaeobiology of Triassic procolophonids,inferred from bone microstructure

Procolophonoidea represent the most successful radiation of Parareptilia that lived during the Permo-Triassic. They are one of the few vertebrate groups that survived the end-Permian extinction and are thus important for studying the recovery of the post-extinction terrestrial ecosystem. Here, we investigate the palaeobiology of three Triassic procolophonid parareptiles, namely Sauropareion anoplus, Procolophon trigoniceps and Teratophon spinigenis, from the Karoo Basin of South Africa, inferred from histological analyses of their limb bones. Results reveal that all three taxa exhibit parallel-fibered bone tissue. Growth rings are absent in the Early Triassic Sauropareion and Procolophon whereas annuli are present in the Middle Triassic Teratophon, even during early ontogeny, suggesting a difference in life histories. Morphology and bone histology imply fossorial lifestyles for all three taxa, suggesting that burrowing may have played an important role in their survival during the harsh post-extinction Triassic environment.     

A 3D elastic micropolar model of vertebral trabecular bone from lattice homogenization of the bone microstructure

A 3D anisotropic micropolar continuum model of vertebral trabecular bone is presently developed accounting for the influence of microstructure-related scale effects on the macroscopic effective properties. Vertebral trabecular bone is modeled as a cellular material with an idealized periodic structure made of open 3D cells. The micromechanical approach relies on the discrete homogenization technique considering lattice microrotations as additional degrees of freedom at the microscale. The effective elastic properties of 3D lattices made of articulated beams taking into account axial, transverse shearing, flexural, and torsional deformations of the cell struts are derived as closed form expressions of the geometrical and mechanical microparameters. The scaling laws of the effective moduli versus density are determined in situations of low and high effective densities to assess the impact of the transverse shear deformation. The classical and micropolar effective moduli and the internal flexural and torsional lengths are identified versus the same microparameters. A finite element model of the local architecture of the trabeculae gives values of the effective moduli that are in satisfactory agreement with the homogenized moduli.     

Light-harvesting antenna composition controls the macrostructure and dynamics of thylakoid membranes in Arabidopsis

We characterized a set of Arabidopsis mutants deficient in specific light-harvesting proteins, using freeze-fracture electron microscopy to probe the organization of complexes in the membrane and confocal fluorescence recovery after photobleaching to probe the dynamics of thylakoid membranes within intact chloroplasts. The same methods were used to characterize mutants lacking or over-expressing PsbS, a protein related to light-harvesting complexes that appears to play a role in regulation of photosynthetic light harvesting. We found that changes in the complement of light-harvesting complexes and PsbS have striking effects on the photosystem II macrostructure, and that these effects correlate with changes in the mobility of chlorophyll proteins within the thylakoid membrane. The mobility of chlorophyll proteins was found to correlate with the extent of photoprotective non-photochemical quenching, consistent with the idea that non-photochemical quenching involves extensive re-organization of complexes in the membrane. We suggest that a key feature of the physiological function of PsbS is to decrease the formation of ordered semi-crystalline arrays of photosystem II in the low-light state. Thus the presence of PsbS leads to an increase in the fluidity of the membrane, accelerating the re-organization of the photosystem II macrostructure that is necessary for induction of non-photochemical quenching.     

Impact of the porous microstructure on the overall elastic properties of the osteonal cortical bone

Mechanical properties of bones are largely determined by their microstructure. The latter comprises a large number of diverse pores. The present paper analyzes a connection between structure of the porous space of the osteonal cortical bone and bone's overall anisotropic elastic moduli. The analysis is based on recent developments in the theory of porous materials that predict the anisotropic effective moduli of porous solids in terms of pores' shapes, orientations and densities. Bone's microstructure is modeled using available micrographs. The calculated anisotropic elastic constants for porous cortical bone are, mostly, in agreement with available experimental data. The influence of each of the pore types on the overall moduli is examined. The results of the analysis can also be used to estimate the extent of mineralization (hydroxyapatite content) if the overall porosity and the effective moduli are known and, vice versa, to estimate porosity from the measured moduli and the extent of mineralization.     

Specifications for machining the bovine cortical bone in relation to its microstructure

Until date, many devices have been developed for cutting human bones during orthopedic surgeries. However, bones are anisotropic material, and their machining characteristics depend on the tool feed direction. In this study, microcutting of the bovine cortical bone is performed and its structure observed under a microscope. Furthermore, the formation of cutting chips and measurement of the cutting force during bone machining are dynamically observed while considering the anisotropy of bone tissue. In particular, the fracture of secondary osteons and crack propagation in bones are observed and analyzed. The results indicate that when the cut depth exceeds 20 μm and is greater than the interval of concentric lamellae, cracks are formed together with chips. A new method for bone machining is proposed. This method is based on the characteristics of crack propagation in bones and is expected to produce low mechanical stress and realize highly efficient and precise machining of living tissues such as bones.     

Compressive strength of mandibular bone as a function of microstructure and strain rate

An investigation has been made of the compressive strength of the porcine mandible and its depedence upon microstructure and strain rate. The results are compared with the fracture behavior of bovine femoral bone. Regarding microstructural dependence, it was found that fracture behavior depends upon regularity of structure, morphology of subunits, orientation of lamellae with respect to the stress axis, amount of ground substance, density and mineral content. Fracture mode was found to be a strong function of strain rate. For both porcine mandibular and bovine femoral bone, there is a ductile-to-brittle transition which results in a change of strain rate sensitivity coefficient from 0.1 to 0.0 at the transition region. This is corroborated by a large change in work-to-fracture values at this region. Therefore, the existence of a critical velocity for bone is supported by the present data.