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The Biomedical Imaging and Therapy facility of the Canadian Light Source comprises two beamlines, which together cover a wide X-ray energy range from 13â keV up to 140â keV. The beamlines were designed with a focus on synchrotron applications in preclinical imaging and veterinary science as well as microbeam radiation therapy. While these remain a major part of the activities of both beamlines, a number of recent upgrades have enhanced the versatility and performance of the beamlines, particularly for high-resolution microtomography experiments. As a result, the user community has been quickly expanding to include researchers in advanced materials, batteries, fuel cells, agriculture, and environmental studies. This article summarizes the beam properties, describes the endstations together with the detector pool, and presents several application cases of the various X-ray imaging techniques available to users.
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Visualization of low-density tissue scaffolds made from hydrogels is important yet challenging in tissue engineering and regenerative medicine (TERM). For this, synchrotron radiation propagation-based imaging computed tomography (SR-PBI-CT) has great potential, but is limited due to the ring artifacts commonly observed in SR-PBI-CT images. To address this issue, this study focuses on the integration of SR-PBI-CT and helical acquisition mode (i.e. SR-PBI-HCT) to visualize hydrogel scaffolds. The influence of key imaging parameters on the image quality of hydrogel scaffolds was investigated, including the helical pitch (p), photon energy (E) and the number of acquisition projections per rotation/revolution (Np), and, on this basis, those parameters were optimized to improve image quality and to reduce noise level and artifacts. The results illustrate that SR-PBI-HCT imaging shows impressive advantages in avoiding ring artifacts with p = 1.5, E = 30â keV and Np = 500 for the visualization of hydrogel scaffolds in vitro. Furthermore, the results also demonstrate that hydrogel scaffolds can be visualized using SR-PBI-HCT with good contrast while at a low radiation dose, i.e. 342â mGy (voxel size of 26â µm, suitable for in vivo imaging). This paper presents a systematic study on hydrogel scaffold imaging using SR-PBI-HCT and the results reveal that SR-PBI-HCT is a powerful tool for visualizing and characterizing low-density scaffolds with a high image quality in vitro. This work represents a significant advance toward the non-invasive in vivo visualization and characterization of hydrogel scaffolds at a suitable radiation dose.
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Síncrotrons , Alicerces Teciduais , Tomografia Computadorizada por Raios X/métodos , Engenharia Tecidual/métodos , HidrogéisRESUMO
BACKGROUND: Bone tissue engineering, based on three-dimensional (3D) printing technology, has emerged as a promising approach to treat bone defects using scaffolds. The objective of this study was to investigate the influence of porosity and internal structure on the mechanical properties of scaffolds. METHODS: We fabricated composite scaffolds (which aimed to replicate trabecular bone) from polycaprolactone (PCL) reinforced with 30% (wt.) nano-hydroxyapatite (nHAp) by extrusion printing. Scaffolds with various porosities were designed and fabricated with and without an interlayer offset, termed as staggered and lattice structure, respectively. Mechanical compressive testing was performed to determine scaffold elastic modulus and yield strength. Linear regression was used to evaluate mechanical properties as a function of scaffold porosity. RESULTS: Different relationships between mechanical properties and porosities were noted for the staggered and lattice structures. For elastic moduli, the two relationships intersected (porosity = 55%) such that the lattice structure exhibited higher moduli with porosity values greater than the intersection point; vice versa for the staggered structure. The lattice structure exhibited higher yield strength at all porosities. Mechanical testing results also indicated elastic moduli and yield strength properties comparable to trabecular bone (elastic moduli: 14-165 MPa; yield strength: 0.9-10 MPa). CONCLUSIONS: Taken together, this study demonstrates that scaffolds printed from PCL/30% (wt.) nHAp with lattice and staggered structure offer promise for treating trabecular bone defects. This study identified the effect of porosity and internal structure on scaffold mechanical properties and provided suggestions for developing scaffolds with mechanical properties for substituting trabecular bone.
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Substitutos Ósseos , Durapatita , Durapatita/química , Alicerces Teciduais/química , Poliésteres/química , Engenharia Tecidual/métodos , Porosidade , Impressão TridimensionalRESUMO
Ancestors of the Antarctic icefishes (family Channichthyidae) were benthic and had no swim bladder, making it energetically expensive to rise from the ocean floor. To exploit the water column, benthopelagic icefishes were hypothesized to have evolved a skeleton with "reduced bone," which gross anatomical data supported. Here, we tested the hypothesis that changes to icefish bones also occurred below the level of gross anatomy. Histology and micro-CT imaging of representative craniofacial bones (i.e., ceratohyal, frontal, dentary, and articular) of extant Antarctic fish species specifically evaluated two features that might cause the appearance of "reduced bone": bone microstructure (e.g., bone volume fraction and structure linear density) and bone mineral density (BMD, or mass of mineral per volume of bone). Measures of bone microstructure were not consistently different in bones from the icefishes Chaenocephalus aceratus and Champsocephalus gunnari, compared to the related benthic notothenioids Notothenia coriiceps and Gobionotothen gibberifrons. Some quantitative measures, such as bone volume fraction and structure linear density, were significantly increased in some icefish bones compared to homologous bones of non-icefish. However, such differences were rare, and no microstructural measures were consistently different in icefishes across all bones and species analyzed. Furthermore, BMD was similar among homologous bones of icefish and non-icefish Antarctic notothenioids. In summary, "reduced bone" in icefishes was not due to systemic changes in bone microstructure or BMD, raising the prospect that "reduced bone" in icefish occurs only at the gross anatomic level (i.e., smaller or fewer bones). Given that icefishes exhibit delayed skeletal development compared to non-icefish Antarctic fishes, combining these phenotypic data with genomic data might clarify genetic changes driving skeletal heterochrony.
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Densidade Óssea , Perciformes , Animais , Regiões Antárticas , Peixes/anatomia & histologia , Perciformes/anatomia & histologiaRESUMO
Simulated microgravity (SMG) inhibits osteoblast differentiation (OBD) and induces bone loss via the inhibition of the Wnt/ß-catenin pathway. However, the mechanism by which SMG alters the Wnt/ß-catenin pathway is unknown. We previously demonstrated that SMG altered the focal adhesion kinase (FAK)-regulated mTORC1, AMPK and ERK1/2 pathways, leading to the inhibition of tumor cell proliferation/metastasis and promoting cell apoptosis. To examine whether FAK similarly mediates SMG-dependent changes to Wnt/ß-catenin in osteoblasts, we characterized mouse MC3T3-E1 cells cultured under clinostat-modeled SMG (µg) conditions. Compared to cells cultured under ground (1 g) conditions, SMG reduces focal adhesions, alters cytoskeleton structures, and down-regulates FAK, Wnt/ß-catenin and Wnt/ß-catenin-regulated molecules. Consequently, protein-2 (BMP2), type-1 collagen (COL1), alkaline-phosphatase activity and matrix mineralization are all inhibited. In the mouse hindlimb unloading (HU) model, SMG-affected tibial trabecular bone loss is significantly reduced, according to histological and micro-computed tomography analyses. Interestingly, the FAK activator, cytotoxic necrotizing factor-1 (CNF1), significantly suppresses all of the SMG-induced alterations in MC3T3-E1 cells and the HU model. Therefore, our data demonstrate the critical role of FAK in the SMG-induced inhibition of OBD and bone loss via the Wnt/ß-catenin pathway, offering FAK signaling as a new therapeutic target not only for astronauts at risk of OBD inhibition and bone loss, but also osteoporotic patients.
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Proteína-Tirosina Quinases de Adesão Focal , Osteoblastos , Ausência de Peso , Via de Sinalização Wnt , beta Catenina , Células 3T3 , Animais , Ativação Enzimática , Quinase 1 de Adesão Focal/metabolismo , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Microtomografia por Raio-X , beta Catenina/metabolismoRESUMO
This paper presents a pharmacokinetic/pharmacodynamic (PK/PD) model of the action of PTH(1-34) on bone modelling and remodelling, developed for quantitatively investigating the dose- and administration pattern-dependency of the bone tissue response to this drug. Firstly, a PK model of PTH(1-34) was developed, accounting for administration via subcutaneous injections. Subsequently, the PK model was coupled to a (mechanistic) bone cell population model of bone modelling and remodelling, taking into account the effects of PTH(1-34) on the differentiation of lining cells into active osteoblasts, on the apoptosis of active osteoblasts, and on proliferation of osteoblast precursors, as well as on the key regulatory pathways of bone cell activities. Numerical simulations show that the coupled PK/PD model is able to distinguish between continuous and intermittent administration patterns of PTH(1-34), in terms of yielding both catabolic bone responses (if drug administration is carried out continuously) and anabolic bone responses (if drug administration is carried out intermittently). The model also features a non-linear relation between bone gain and drug dose (as known from experiments); doubling the dose from 80⯵g/kg/day to 160 µg/kg/day induced a 1.3-fold increase of the bone volume-to-total volume ratio. Furthermore, the model presented in this paper confirmed that bone modelling represents an essential mechanism of the anabolic response of bone to PTH(1-34) administration in rat models, and that the large amount of bone formation observed in such models cannot be explained via remodelling alone.
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Modelos Biológicos , Osteoporose/tratamento farmacológico , Hormônio Paratireóideo/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Calibragem , Modelos Animais de Doenças , Feminino , Humanos , Osteoblastos/efeitos dos fármacos , Osteoblastos/patologia , Osteoporose/sangue , Ovariectomia , Hormônio Paratireóideo/sangue , Hormônio Paratireóideo/farmacocinética , Hormônio Paratireóideo/farmacologia , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Bone acts as a reservoir for many trace elements. Understanding the extent and pattern of elemental accumulation in the skeleton is important from diagnostic, therapeutic, and toxicological perspectives. Some elements are simply adsorbed to bone surfaces by electric force and are buried under bone mineral, while others can replace calcium atoms in the hydroxyapatite structure. In this article, we investigated the extent and pattern of skeletal uptake of barium and strontium in two different age groups, growing, and skeletally mature, in healthy rats. Animals were dosed orally for 4 weeks with either strontium chloride or barium chloride or combined. The distribution of trace elements was imaged in 3D using synchrotron K-edge subtraction micro-CT at 13.5 µm resolution and 2D electron probe microanalysis (EPMA). Bulk concentration of the elements in serum and bone (tibiae) was also measured by mass spectrometry to study the extent of uptake. Toxicological evaluation did not show any cardiotoxicity or nephrotoxicity. Both elements were primarily deposited in the areas of active bone turnover such as growth plates and trabecular bone. Barium and strontium concentration in the bones of juvenile rats was 2.3 times higher, while serum levels were 1.4 and 1.5 times lower than adults. In all treatment and age groups, strontium was preferred to barium even though equal molar concentrations were dosed. This study displayed spatial co-localization of barium and strontium in bone for the first time. Barium and strontium can be used as surrogates for calcium to study the pathological changes in animal models of bone disease and to study the effects of pharmaceutical compounds on bone micro-architecture and bone remodeling in high spatial sensitivity and precision.
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Bário/metabolismo , Desenvolvimento Ósseo , Osso e Ossos/metabolismo , Estrôncio/metabolismo , Animais , Osso e Ossos/diagnóstico por imagem , Cálcio/metabolismo , Eletrocardiografia , Feminino , Imageamento Tridimensional , Ratos Sprague-Dawley , Distribuição Tecidual , Ureia/sangueRESUMO
Vascular canals in cortical bone during growth and development typically show an anisotropic pattern with canals falling into three main categories: circumferential, radial, and longitudinal. Two major hypotheses attempt to explain the preferred orientations in bone: that vascular canal orientation is optimized to resist a predominant strain direction from functional loading, or that it reflects growth requirements and velocity. We use a controlled growth experiment in broiler chickens to investigate the effect of growth rate on vascular canal orientation. Using feed restriction we set up a fast growing control group and a slow growing restricted group. We compared the microstructure in the humerus and the femur at 42 days of age using synchrotron micro-computed tomography (micro-CT), a three-dimensional (3D) method that visualizes the full canal network. We measured the 3D orientation of each canal in the whole cross-section of the bone cortex using a set of custom ImageJ scripts. Using these orientations we compute laminar, radial, and longitudinal indices that measure the proportion of circumferential, radial, and longitudinal canals, by unit of length, in the cortex. Following previous studies we hypothesized that vascular canal orientation is related to growth, with radial canals linked to a faster growth rate and related to functional loading through a high laminar index in flight bones which reflects torsional loading resulting from active flight. The control group had final body weights that were nearly twice the final weights of the restricted group and higher absolute growth rates. We found consistent patterns in the comparison between the humerus and the femur in both groups, with the humerus having higher laminar and longitudinal indices, and a lower radial index than the femur. The control group had higher radial indices and lower laminar and longitudinal indices in both the humerus and the femur than the restricted group. The higher radial indices in our control group point to a link between radial canals and faster growth, and between laminar canals and slower growth, while the higher laminar indices in the humerus point to a link between circumferential canals and torsional loading. Overall, our results indicate that the orientation of the cortical canal network in a bone is the consequence of a complex interaction between the growth rate of that bone and functional loading environment.
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Galinhas/anatomia & histologia , Galinhas/crescimento & desenvolvimento , Osso Cortical/anatomia & histologia , Osso Cortical/crescimento & desenvolvimento , AnimaisRESUMO
Cortical bone porosity and specifically the orientation of vascular canals is an area of growing interest in biomedical research and comparative/paleontological anatomy. The potential to explain microstructural adaptation is of great interest. However, the determinants of the development of canal orientation remain unclear. Previous studies of birds have shown higher proportions of circumferential canals (called laminarity) in flight bones than in hindlimb bones, and interpreted this as a sign that circumferential canals are a feature for resistance to the torsional loading created by flight. We defined the laminarity index as the percentage of circumferential canal length out of the total canal length. In this study we examined the vascular canal network in the humerus and femur of a sample of 31 bird and 24 bat species using synchrotron micro-computed tomography (micro-CT) to look for a connection between canal orientation and functional loading. The use of micro-CT provides a full three-dimensional (3D) map of the vascular canal network and provides measurements of the 3D orientation of each canal in the whole cross-section of the bone cortex. We measured several cross-sectional geometric parameters and strength indices including principal and polar area moments of inertia, principal and polar section moduli, circularity, buckling ratio, and a weighted cortical thickness index. We found that bat cortices are relatively thicker and poorly vascularized, whereas those of birds are thinner and more highly vascularized, and that according to our cross-sectional geometric parameters, bird bones have a greater resistance to torsional stress than the bats; in particular, the humerus in birds is more adapted to resist torsional stresses than the femur. Our results show that birds have a significantly (P = 0.031) higher laminarity index than bats, with birds having a mean laminarity index of 0.183 in the humerus and 0.232 in the femur, and bats having a mean laminarity index of 0.118 in the humerus and 0.119 in the femur. Counter to our expectation, the birds had a significantly higher laminarity index in the femur than in the humerus (P = 0.035). To evaluate whether this discrepancy was a consequence of methodology we conducted a comparison between our 3D method and an analogue to two-dimensional (2D) histological measurements. This comparison revealed that 2D methods significantly underestimate (P < 0.001) the amount of longitudinal canals by an average of 20% and significantly overestimate (P < 0.001) the laminarity index by an average of 7.7%, systematically mis-estimating indices of vascular canal orientations. In comparison with our 3D results, our approximated 2D measurement had the same results for comparisons between the birds and bats but found significant differences only in the longitudinal index between the humerus and the femur for both groups. The differences between our 3D and pseudo-2D results indicate that differences between our findings and the literature may be partially based in methodology. Overall, our results do not support the hypothesis that the bones of flight are more laminar, suggesting a complex relation between functional loading and microstructural adaptation.
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Aves/anatomia & histologia , Quirópteros/anatomia & histologia , Osso Cortical/anatomia & histologia , Imageamento Tridimensional/métodos , Animais , Microtomografia por Raio-XRESUMO
The nature of trapped air on submersed ultra-water-repellent interfaces has been investigated. These gaseous layers (plastrons) can last from hours to, in some examples such as the Salvinia molesta fern, months. The interface of submerged superhydrophobic surfaces with carefully controlled micropatterned surface roughness has been probed using synchrotron-based high-resolution X-ray phase tomography. This technique looks in situ, through the aqueous/gas interface in three dimensions. Long-term plastron stability appears to correlate with the appearance of scattered microdroplets <20 µm in diameter that are sandwiched within the 30 µm thick gaseous interfacial layer. These microdroplets are centered on defects or damaged sections within the substrate surface approximately 20-50 µm apart. Such irregularities represent heterogeneous micro/nano-hierarchical structures with varying surface structures and chemistry. The stability of microdroplets is governed by a combination of electrostatic repulsion, contact angle limitations, and a saturated vapor pressure, the latter of which reduces the rate of diffusion of gas out of the air layer, thus increasing underwater longevity. Homogenous surfaces exhibiting purely nano- or micro-regularity do not support such microdroplets, and, as a consequence, plastrons can disappear in <20 h compared with >160 h for surfaces with scattered microdroplets. Such behavior may be a requirement for long-term nonwetting in any system.
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OBJECTIVES: Differentiating human from nonhuman fragmented bone is often accomplished using histological methods if the observation of gross morphology proves insufficient. Linearly oriented primary and/or secondary osteonal systems, commonly referred to as osteon bands, are described as a strong indicator of nonhuman bone, particularly the occurrence of multiple bands. This phenomenon has been conventionally documented using two-dimensional (2D) histology, but such analyses are destructive and typically limited to a single cross-section. Progressive developments in high-resolution X-ray imaging, however, allow for the nondestructive three-dimensional (3D) visualization of bone microarchitecture. The primary objective of the current research was to visualize and document the occurrence of osteon banding in adult human cortical bone using high-resolution synchrotron radiation-based micro-Computed Tomography (SR micro-CT). MATERIALS AND METHODS: Synchrotron radiation-based micro-CT scanning was carried out at the Canadian Light Source (CLS) national synchrotron facility. The presence or absence of osteon banding was visualized in human skeletal elements from three adult males with representative samples from all regions of the skeleton (n = 129). If present, osteon banding was described and quantified. RESULTS: Results indicated that 23 of 129 human cortical bone specimens exhibited osteon banding, representing 18% of the sample. Linear arrangements of primary and/or secondary osteons were observed in the following skeletal elements: temporal, parietal, frontal, occipital, clavicle, mandible, femur, tibia, ulna, second metatarsal, and sacrum. DISCUSSION: The present work represents the first 3D examination of inter-element variation in osteon banding in adult human cortical bone. Findings indicate that the presence of multiple osteon bands in a single specimen is not diagnostic of nonhuman bone. As such, osteon banding categorically should not be taken as evidence of nonhuman bone in forensic and archaeological contexts.
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Osso Cortical/anatomia & histologia , Ósteon/anatomia & histologia , Adulto , Antropologia Física , Osso Cortical/diagnóstico por imagem , Ósteon/diagnóstico por imagem , Humanos , Masculino , Microtomografia por Raio-XRESUMO
This study uses synchrotron radiation-based micro-computed tomography (CT) scans to reconstruct three-dimensional networks of Haversian systems in human cortical bone in order to observe and analyse interconnectivity of Haversian systems and the development of total Haversian networks across different ages. A better knowledge of how Haversian systems interact with each other is essential to improve understanding of remodeling mechanisms and bone maintenance; however, previous methodological approaches (e.g. serial sections) did not reveal enough detail to follow the specific morphology of Haversian branching, for example. Accordingly, the aim of the present study was to identify the morphological diversity of branching patterns and transverse connections, and to understand how they change with age. Two types of branching morphologies were identified: lateral branching, resulting in small osteon branches bifurcating off of larger Haversian canals; and dichotomous branching, the formation of two new osteonal branches from one. The reconstructions in this study also suggest that Haversian systems frequently target previously existing systems as a path for their course, resulting in a cross-sectional morphology frequently referred to as 'type II osteons'. Transverse connections were diverse in their course from linear to oblique to curvy. Quantitative assessment of age-related trends indicates that while in younger human individuals transverse connections were most common, in older individuals more evidence of connections resulting from Haversian systems growing inside previously existing systems was found. Despite these changes in morphological characteristics, a relatively constant degree of overall interconnectivity is maintained throughout life. Altogether, the present study reveals important details about Haversian systems and their relation to each other that can be used towards a better understanding of cortical bone remodeling as well as a more accurate interpretation of morphological variants of osteons in cross-sectional microscopy. Permitting visibility of reversal lines, synchrotron radiation-based micro-CT is a valuable tool for the reconstruction of Haversian systems, and future analyses have the potential to further improve understanding of various important aspects of bone growth, maintenance and health.
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Envelhecimento , Ósteon/crescimento & desenvolvimento , Ósteon/ultraestrutura , Imageamento Tridimensional/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Fêmur/ultraestrutura , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Pessoa de Meia-Idade , Síncrotrons , Microtomografia por Raio-X , Adulto JovemRESUMO
The primary microstructural unit of cortical bone, the secondary osteon or Haversian system, is widely assumed to have a cylindrical shape. It is generally accepted that osteons are roughly circular in cross-section and deviations from circularity have been attributed to deviations from longitudinal orientation. To our knowledge this idealized geometric relationship, which assumes osteons are perfect cylinders, has not been rigorously explored. As such, we sought to explore two research questions: (i) Does the orientation of osteons in 3D explain variation in shapes visualized in 2D? (ii) Can differences in osteon 3D orientation explain previously reported age-related differences observed in their 2D cross-sectional shape (e.g. more circular shape and decreased area with age)? To address these questions we utilized a combination of 2D histology to identify osteon shape and superimposed micro-computed tomography data to assess osteon orientation in 3D based upon the osteonal canal. Shape was assessed by the inverse of Aspect Ratio (On.AspR(-1), based on a fitted ellipse) - which ranged from 0 (infinitely elongated shape) to 1 (perfectly circular). A sample (n = 27) of human female anterior femoral cortical bone samples from across the human lifespan (20-87 years) were included in the analysis, which involved 1418 osteons. The overall mean measure of On.AspR(-1) was 0.703 (1.42 Aspect Ratio). Mean osteon orientation was 79.1° (90° being longitudinal). While we anticipated a positive relation between orientation and On.AspR(-1), we found the opposite - a weak negative correlation (with more oblique 3D osteon alignment, the 2D shape became more circular as reflected by increased On.AspR(-1)). When analysis of covariance was performed with age and orientation as covariates, the negative relation with orientation was replaced by a significant relation with age alone. This relation with age accounted for 41% of the variation of On.AspR(-1). The results revealed that osteons, on average, are not circular in cross-section and that 3D orientation cannot account for deviation from circular shape. Osteons thus are strictly speaking not cylinders, as they tend to have elliptical cross-sections. We observed that osteons did become less elliptical in cross-section with age independent of orientation - suggesting this is a real change in morphology.
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Envelhecimento , Remodelação Óssea/fisiologia , Ósteon/anatomia & histologia , Imageamento Tridimensional , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Fêmur/crescimento & desenvolvimento , Fêmur/ultraestrutura , Técnicas Histológicas , Humanos , Masculino , Pessoa de Meia-Idade , Microtomografia por Raio-X , Adulto JovemRESUMO
Mechanical testing studies by Krauss et al. (2009) and Gupta et al. (2013) suggest that the extraordinary toughness of antler bone is primarily achieved by intrinsic/nanostructural mechanisms instead of extrinsic/microstructural mechanisms. However, this conclusion is based on data from extremely small specimens from one antler loaded only in tension, which impedes discernment of the relative importance of intrinsic vs. extrinsic mechanisms. In the present study we conducted analyses into the microstructural features of antler for details of potential additional microscale toughening characteristics, as suggested by recent mechanical testing studies of bulk specimens. The data are also considered in view of the above-mentioned studies concluding that extrinsic/microstructural toughening mechanisms are less important than nanoscale/intrinsic toughening mechanisms in antler. Mule deer antlers were evaluated using: (1) backscattered electron imaging for micro-mineralization, (2) circularly polarized light for osteonal interfacial complexity and collagen fiber orientation (CFO) heterogeneity, and (3) X-ray 3D micro-computed tomography for osteon/vessel orientation, density, and size. Results showed: (1) hyper-mineralized seams of approximately 3-4 microns thickness within relatively hypermineralized "zones" that course circuitously along osteonal interfaces, (2) highly heterogeneous CFO, including increased oblique-to-transverse CFO near/adjacent to osteon peripheries, and (3) osteons are often highly elongated in 2D. 3D reconstructions show that a considerable percentage of the vascular canals course obliquely with respect to the antler long axis. While results show multiple possible extrinsic-level histological characteristics in antler bone, it remains to be determined if microstructural characteristics become subsidiary to nanostructural characteristics in enhancing toughness during the majority of post-yield behavior of antler bone when loaded in a biologically relevant fashion.
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Chifres de Veado/ultraestrutura , Osso e Ossos/ultraestrutura , Fenômenos Mecânicos , Nanoestruturas/ultraestrutura , Animais , Calcificação Fisiológica , Cervos , Imageamento Tridimensional , Masculino , Microscopia EletrônicaRESUMO
The most abundant cell in bone, osteocytes form an interconnected system upon which the regulation of healthy bone relies. Although the complete nature of the role of osteocytes has yet to be defined, they are generally accepted to play a part in the sensing of load and the initiation of damage repair. A previous study conducted by our group identified variation of up to 30% in osteocyte lacunar density and morphological parameters between regions of a single cross-section of human femoral shaft; that study, however, was limited to a single individual. The aim of the current study was to determine whether this pattern consistently occurs in healthy young male femora. Anterior, posterior, medial and lateral blocks were prepared from the proximal femoral shaft of seven males and synchrotron radiation micro-CT imaged. Average lacunar densities (± SD) from the anterior, posterior, medial and lateral regions were 23 394 ± 1705, 30 180 ± 4860, 35 946 ± 5990 and 29 678 ± 6081 lacunae per mm(3) of bone tissue, respectively. These values were significantly different between the anterior and both the medial and posterior regions (P < 0.05). The density of the combined anterior and posterior regions was also significantly lower (P = 0.006) than the density of the combined medial and lateral regions. Although no difference was found in predominant orientation, shape differences were found; with the combined anterior-posterior regions having lacunae that were significantly more elongated and less flat than the combined medial-lateral values (P < 0.001). As expected, in this larger study, there was a dramatic difference in lacunar density between the medial and anterior region (up to ~ 54%). The study clearly demonstrates that the high variation seen in osteocyte lacunar density as well as other lacunar parameters, noted in a number of biomechanical, age and pathology studies, are well within the range of normal variation; however, the reasons for and consequences of this variation remain unclear. Lacunar parameters including abundance and shape are being increasingly incorporated into computational modeling of bone biology and this paper represents a more comprehensive description of normal healthy lacunae.
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Fêmur/citologia , Osteócitos/citologia , Adulto , Análise de Variância , Matriz Óssea/citologia , Cadáver , Contagem de Células , Fêmur/diagnóstico por imagem , Humanos , Masculino , Osteócitos/fisiologia , Tomografia Computadorizada por Raios X , Adulto JovemRESUMO
The activity of basic multicellular units (BMU) in cortical bone is classically described as a sequential order of events- resorption, reversal and formation. This simplified portrayal of the remodeling process is pervasive despite the reported variability in remodeling space morphology. These variations may reflect meaningful nuances in BMU activity but methods to quantify 3D remodeling space morphology within the context of the cellular activity are currently lacking. This study developed new techniques to define zones of BMU activity based on the 3D morphology of remodeling spaces in rabbit cortical bone and integrated morphological data with the BMU longitudinal erosion rate (LER) to elucidate the spatial-temporal coordination of BMUs and estimate mineral apposition rate (MAR). The tibiae of New Zealand white rabbits (n = 5) were imaged in vivo using synchrotron radiation and two weeks later ex vivo with desktop microCT. The in vivo and ex vivo datasets were co-registered, and 27 remodeling spaces were identified at both timepoints. A radial profile representing the 3D morphology was the platform for partitioning the remodeling spaces into resorption, reversal and formation zones. Manual, automated and semi-automated partitioning approaches were compared, and the zone-segmentations were used to calculate the length, change in radius and slope of each zone. The manual approach most accurately defined the zones of idealized remodeling spaces with known dimensions (relative error = 0.9-9.2 %) while the semi-automated method reliably defined the zones in rabbit remodeling spaces (ICC = 0.85-1.00). Combining LER and the manually derived zone dimensions indicated that a BMU passes through a cross-section in approximately 18.8 days with resorption, reversal and formation taking 4.1, 2.2, and 12.5 days, respectively. MAR estimated by the 3D analysis was not significantly different than that determined with classic histomorphometry (p = 0.48). These techniques have the potential to assess dynamic parameters of bone resorption and formation, eliminate the need for fluorochrome labeling and provide a more comprehensive perspective of the remodeling process.
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Remodelação Óssea , Reabsorção Óssea , Animais , Coelhos , Osso e Ossos , Osso Cortical/diagnóstico por imagem , Tíbia/diagnóstico por imagemRESUMO
Osteocytes are believed to be the primary agents of mechanosensing in bone. Due to this important role in the structure-function relationship of bone, osteocytes and the spaces they occupy (lacunae) are of increasing interest. Changes in lacunae with age are of particular interest in women since they are more susceptible to bone loss and fragility associated with senescent diseases including osteoporosis. This study's purpose was to test whether differences exist in lacunar density (lacunae/mm(3) of bone), orientation and morphology in the cortex of adult women spanning the human lifespan. Anterior blocks from the femoral shaft from 30 women aged 20-86years were imaged by synchrotron-radiation micro-CT. No significant relation between lacunar density and age was detected. A significant reduction in lacunar volume with age (p<0.001) was observed, alongside changes in lacunar morphology. When divided into two groups (<50 and >50years) the younger group's lacunae were â¼30% larger and were flatter (p<0.001) and less equant (spherical) (p<0.001). To our knowledge the observation that lacunar volume and morphology change over the human lifespan is novel, potentially resulting from preferential surface infilling within the extracellular space. The functional impact of this infilling is unclear but such a change in scale likely impacts the mechanosensing function of the osteocyte network. Limitations in resolution prevented us from assessing if this infilling is associated with disruption of the canaliculi. This hypothesis warrants further investigation as, if confirmed, it would represent a profound negative impact on the osteocyte network and may provide new insights into age-related bone loss.
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Fêmur/ultraestrutura , Osteócitos/fisiologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Feminino , Fêmur/diagnóstico por imagem , Humanos , Pessoa de Meia-Idade , Microtomografia por Raio-X , Adulto JovemRESUMO
OBJECTIVES: In modern bone biology, the term "remodeling" generally refers to internal bone turnover that creates secondary osteons. However, it is also widely used by skeletal biologists, including biological anthropologists as a catch-all term to refer to different skeletal changes. In this review, we investigated how "remodeling" is used across topics on skeletal biology in biological anthropology to demonstrate potential problems with such pervasive use of a generalized term. METHODS: Using PubMed and Google Scholar, we selected and reviewed 205 articles that use the term remodeling to describe skeletal processes and have anthropological implications. Nine edited volumes were also reviewed as examples of collaborative work by different experts to demonstrate the diverse and extensive use of the term remodeling. RESULTS: Four general meanings of bone "remodeling" were identified, namely, internal turnover, functional adaptation, fracture repair, and growth remodeling. Additionally, remodeling is also used to refer to a broad array of pathological skeletal changes. DISCUSSION: Although we initially identified four general meanings of bone remodeling, they are not mutually exclusive and often occur in combination. The term "remodeling" has become an extensively used catch-all term to refer to different processes and outcomes of skeletal changes, which inevitably lead to misunderstanding and a loss of information. Such ambiguity and confusion are potentially problematic as the field of biological anthropology becomes increasingly multidisciplinary. Therefore, we advocate for precise, context-specific definitions and explanations of bone remodeling as it continues to be used across disciplines within and beyond biological anthropology.
Assuntos
Antropologia , Osso e Ossos , Osso e Ossos/diagnóstico por imagem , Remodelação ÓsseaRESUMO
BACKGROUND: Scaffolds are of great importance in tissue engineering applications as they provide a mechanically supportive environment for cellular activity, which is particularly necessary for hard tissues such as bone. Notably, the mechanical properties of a scaffold vary with differing design parameters such as those related to scaffold height and internal structure. Thus, the present study aimed to explore the relationship between design parameters and mechanical properties of composite polycaprolactone (PCL) and nano-hydroxyapatite (nHAp) scaffolds fabricated by three-dimensional (3D) printing. METHODS: We designed and printed scaffolds with different internal structures (lattice and staggered) and varying heights (4, 6, 8 and 10 layers), and consistent porosity (50%) for the purpose of comparison. Then, we examined the scaffold microstructure (pore size and penetration between layers) using scanning electron microscopy (SEM) and mechanical properties (elastic modulus and yield strength) using compressive testing. RESULTS: Our results illustrated that the microstructural parameters were related to scaffold design. At higher heights, pore size increased while penetration between layers decreased; thus, mechanical properties were affected. Results of mechanical testing demonstrated that for lattice scaffolds, elastic modulus was similar for 6 vs 4, and 8 vs 4 layers but ~33% lower for 10 layers vs 4 layers. Similarly, yield strength was comparable for 6 vs 4, and 8 vs 4 layers but ~27% lower for 10 layers vs 4 layers. With staggered scaffolds, when compared to 4-layer results, elastic modulus was similar for 6 layers but was ~43% lower for 8 layers and ~38% lower for 10 layers. Staggered scaffolds had ~38%, ~51%, and ~76% lower yield strength when the number of layers were increased from 4 to 6, 8, and 10 layers, respectively. When comparing lattice and staggered scaffolds with the same layer number, elastic modulus was similar, apart from 8-layer scaffolds where the staggered design was ~42% lower than lattice. Yield strength was similar between 4-layer staggered and lattice scaffolds, while staggered scaffolds with 6, 8, and 10 number of layers showed ~43%, ~45%, ~68% lower strength, respectively, than those found in lattice scaffolds with the same layer numbers. CONCLUSIONS: Mechanical properties of 3D printed scaffolds depended on scaffold height for both lattice and staggered internal structures. Staggered scaffolds had lower mechanical properties than the lattice scaffolds with the same height and were more sensitive to the change in scaffold height. Taken together, lattice scaffolds demonstrated the advantages of more stable mechanical properties over staggered scaffolds. Also, scaffolds with lower height were more promising in terms of mechanical properties compared to scaffolds with greater height.
Assuntos
Durapatita , Alicerces Teciduais , Durapatita/química , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Osso e Ossos , Porosidade , Poliésteres/química , Impressão TridimensionalRESUMO
Cellular communication in the mechanosensory osteocyte Lacuno-Canalicular Network (LCN) regulates bone tissue remodeling throughout life. Age-associated declines in LCN size and connectivity dysregulate mechanosensitivity to localized remodeling needs of aging or damaged tissue, compromising bone quality. Synchrotron radiation-based micro-Computed Tomography (SRµCT) and Confocal Laser Scanning Microscopy (CLSM) were employed to visualize LCN and vascular canal morphometry in an age series of the anterior femur (males n = 14, females n = 11, age range = 19-101, mean age = 55). Age-associated increases in vascular porosity were driven by pore coalescence, including a significant expansion in pore diameter and a significant decline in pore density. In contrast, the LCN showed significant age-associated reductions in lacunar volume fraction, mean diameter, and density, and in canalicular volume fraction and connectivity density. Lacunar density was significantly lower in females across the lifespan, exacerbating their age-associated decline. Canalicular connectivity density was also significantly lower in females but approached comparable declining male values in older age. Our data illuminate the trajectory and potential morphometric sources of age-associated bone loss. Increased vascular porosity contributes to bone fragility with aging, while an increasingly reduced and disconnected LCN undermines the mechanosensitivity required to repair and reinforce bone. Understanding why and how this degradation occurs is essential for improving the diagnosis and treatment of age-related changes in bone quality and fragility.