Computed tomography porosity and spherical indentation for determining cortical bone millimetre-scale mechanical properties.

The cortex of the femoral neck is a key structural element of the human body, yet there is not a reliable metric for predicting the mechanical properties of the bone in this critical region. This study explored the use of a range of non-destructive metrics to measure femoral neck cortical bone stiffness at the millimetre length scale. A range of testing methods and imaging techniques were assessed for their ability to measure or predict the mechanical properties of cortical bone samples obtained from the femoral neck of hip replacement patients. Techniques that can potentially be applied in vivo to measure bone stiffness, including computed tomography (CT), bulk wave ultrasound (BWUS) and indentation, were compared against in vitro techniques, including compression testing, density measurements and resonant ultrasound spectroscopy. Porosity, as measured by micro-CT, correlated with femoral neck cortical bone's elastic modulus and ultimate compressive strength at the millimetre length scale. Large-tip spherical indentation also correlated with bone mechanical properties at this length scale but to a lesser extent. As the elastic mechanical properties of cortical bone correlated with porosity, we would recommend further development of technologies that can safely measure cortical porosity in vivo.

Assessing bone quality through mechanical properties in postmenopausal trabecular bone.

BACKGROUND: The inner structure of trabecular bone is a result of structural optimization provided by remodeling processes. Changes in hormonal status related to menopause cause bone tissue loss and micro-architectural deterioration with a consequent susceptibility to fracture. Accumulation of micro-damage in bone, as a function of the rate of production and rate of repair, underlies the development of stress fractures, increasing fragility associated to age and osteoporosis, especially in transmenopausal women. PATIENTS AND METHODS: Quasi-static and nano-dynamic mechanical characterization were undertaken in trabecular bone from femoral neck biopsies of postmenopausal women. AFM (Atomic Force Microscopy) complementary studies were performed to determine nano-roughness (SRa) and the fibrils width of collagen. Nanoindentations were used to quantify transmenopausal changes in intrinsic mechanical properties of trabecular bone: hardness (Hi), modulus of Young (Ei), complex modulus (E*), tan delta (δ), storage modulus (E') and loss modulus (E"). RESULTS: As result of the quasi-static measurements, 0.149 (0.036) GPa and 2.95 (0.73) GPa of Hi and Ei were obtained, respectively. As result of the nano-dynamic measurements, 17.94 (3.15), 0.62 (0.10), 13.79 (3.21 and 6.39 (1.28) GPa of E*, tan (δ), E' and E" were achieved, respectively. 101.07 SRa and 831.28 nm of fibrils width were additionally obtained. CONCLUSIONS: This study poses a first approach to the measurement of bone quality in postmenopausal trabecular bone by combining quasi-static, nano-DMA analysis and tribology of dentin surface through AFM characterization.

Biochemical assessment of nanostructures in human trabecular bone: Proposal of a Raman microspectroscopy based measurements protocol.

BACKGROUND: Improvements to the understating of the compositional contributions of bone mineral and organic components to the competence of trabecular bone are crucial. The purpose of this study was to propose a protocol to study biochemical composition of trabecular bone, based on two combined Raman analysis methodologies. MATERIAL AND METHODS: Both cluster and single point Raman mappings were obtained, in order to assess bone degeneration associated with aging, disease, or injury, and to help in the evaluation and development of successful therapies. In this study, human trabecular bone has been analysed throughout a) Raman cluster analysis: bone mineral content, carbonate-to-phosphate ratio (both from the mineral components), the crosslinking and nature/secondary structure of collagen (both from the organic components); and b) Single point Raman spectra, where Raman points related to the minerals and organic components were also obtained, both techniques were employed in spectra attained at 400 to 1700 cm-1. RESULTS: Multivariate analysis confirmed: 1) the different spectral composition, 2) the existence of centroids grouped by chemical affinity of the various components of the trabecular bone, and 3) the several traces of centroids and distribution of chemical compositional clusters. CONCLUSIONS: This study is important, because it delivers a study protocol that provides molecular variations information in both mineral and collagen structure of trabecular bone tissue. This will enable clinicians to benefit knowing the microstructural differences in the bone subjected to degeneration of their patients.

Commonality in the microarchitecture of trabecular bone: A preliminary study.

Understanding the relationship between the microstructure and mechanical function of trabecular bone is critical for prediction and prevention of bone fragility fractures. However, a detailed understanding of the structural design of trabecular microarchitecture is still missing. This study hypothesized that there exists a commonality in the underlying probabilistic distributions of microstructural features of trabecular bones, whereas the microstructural differences among individuals are primarily describe by a set of scalar parameters. To test the hypothesis, twenty-three trabecular bone specimens were obtained from two anatomic locations (i.e., femoral neck and vertebral body) and a diverse group of seventeen donors of different age and sex. The number, size, spatial location, and orientation of individual plates and rods in the trabecular bone specimens were determined via volumetric decomposition of 3D µCT images using the Individual Trabecula Segmentation (ITS) technique. Then, m/n bootstrap Kolmogorov-Smirnov tests were performed to compare the normalized distributions of size, orientation, and spatial arrangement of trabecular plates and rods in the specimens. The results showed that 100% of the twenty-three normalized distributions of each microstructural feature were statistically equivalent irrespective of individual differences among the bone specimens, except the distributions of rod spatial arrangement (<100%). On the other hand, nonparametric Mann-Whitney U tests showed that a set of scalar parameters (i.e., the number, average size, and average nearest neighbor distance of trabecular plates and rods) were statistically different among the individual specimens (p<0.05). Due to the commonality of the underlying distributions, the individual differences in the trabecular microstructure among the specimens seemed to be reflected primarily by changes in the scalar parameters. The above results strongly support the hypothesis of this study and may shed more light on understanding the natural design of trabecular bone microstructures.

Porotic paradox: distribution of cortical bone pore sizes at nano- and micro-levels in healthy vs. fragile human bone.

Bone is a remarkable biological nanocomposite material showing peculiar hierarchical organization from smaller (nano, micro) to larger (macro) length scales. Increased material porosity is considered as the main feature of fragile bone at larger length-scales. However, there is a shortage of quantitative information on bone porosity at smaller length-scales, as well as on the distribution of pore sizes in healthy vs. fragile bone. Therefore, here we investigated how healthy and fragile bones differ in pore volume and pore size distribution patterns, considering a wide range of mostly neglected pore sizes from nano to micron-length scales (7.5 to 15000 nm). Cortical bone specimens from four young healthy women (age: 35 ± 6 years) and five women with bone fracture (age: 82 ± 5 years) were analyzed by mercury porosimetry. Our findings showed that, surprisingly, fragile bone demonstrated lower pore volume at the measured scales. Furtnermore, pore size distribution showed differential patterns between healthy and fragile bones, where healthy bone showed especially high proportion of pores between 200 and 15000 nm. Therefore, although fragile bones are known for increased porosity at macroscopic level and level of tens or hundreds of microns as firmly established in the literature, our study with a unique assessment range of nano-to micron-sized pores reveal that osteoporosis does not imply increased porosity at all length scales. Our thorough assessment of bone porosity reveals a specific distribution of porosities at smaller length-scales and contributes to proper understanding of bone structure which is important for designing new biomimetic bone substitute materials.

Creating High-Resolution Multiscale Maps of Human Tissue Using Multi-beam SEM.

Multi-beam scanning electron microscopy (mSEM) enables high-throughput, nano-resolution imaging of macroscopic tissue samples, providing an unprecedented means for structure-function characterization of biological tissues and their cellular inhabitants, seamlessly across multiple length scales. Here we describe computational methods to reconstruct and navigate a multitude of high-resolution mSEM images of the human hip. We calculated cross-correlation shift vectors between overlapping images and used a mass-spring-damper model for optimal global registration. We utilized the Google Maps API to create an interactive map and provide open access to our reconstructed mSEM datasets to both the public and scientific communities via our website www.mechbio.org. The nano- to macro-scale map reveals the tissue's biological and material constituents. Living inhabitants of the hip bone (e.g. osteocytes) are visible in their local extracellular matrix milieu (comprising collagen and mineral) and embedded in bone's structural tissue architecture, i.e. the osteonal structures in which layers of mineralized tissue are organized in lamellae around a central blood vessel. Multi-beam SEM and our presented methodology enable an unprecedented, comprehensive understanding of health and disease from the molecular to organ length scale.

Influence of segmentation on micro-CT images of trabecular bone.

Segmentation of biomedical images is of great importance in various studies aiming to both the identification of regions of interests within the image and the performance of quantified measurements. Nevertheless, the segmentation of the biomedical images represents a wide range of medical cases and there is not a unique technique applicable to all kinds of medical images. In this study, three popular techniques for segmenting micro-CT images of bone microstructures are evaluated. Fixed threshold, Otsu's algorithm and a modified version of the Chan-Vese segmentation technique have been applied on micro-CT images and have been compared to higher resolution golden standard, that is histological images. The modification of the Chan-Vese technique is based on the novel implementation of a new initialization process called the Branch Point Initialization. Stereological measurements were performed on all the segmented images and statistically compared to the golden standard. Fixed threshold and the modified Chan-Vese technique have shown comparable results, with a maximum significant error of about 10%. However, Chan-Vese showed an easier, faster and more reliable segmentation procedure for optimal settings identification. The Otsu's method showed a maximum error larger than 20%. Given the limits and advantages of the known segmentation techniques, the proposed modified Chan-Vese active contour technique shows high potential for use in the segmentation of micro-CT images as well as in other high-resolution X-ray images. This potential is augmented by the recent introduction of high-resolution clinical technologies for which standard techniques have already shown to be insufficient.

Nano-structural, compositional and micro-architectural signs of cortical bone fragility at the superolateral femoral neck in elderly hip fracture patients vs. healthy aged controls.

To unravel the origins of decreased bone strength in the superolateral femoral neck, we assessed bone structural features across multiple length scales at this cortical fracture initiating region in postmenopausal women with hip fracture and in aged-matched controls. Our combined methodological approach encompassed atomic force microscopy (AFM) characterization of cortical bone nano-structure, assessment of mineral content/distribution via quantitative backscattered electron imaging (qBEI), measurement of bone material properties by reference point indentation, as well as evaluation of cortical micro-architecture and osteocyte lacunar density. Our findings revealed a wide range of differences between the fracture group and the controls, suggesting a number of detrimental changes at various levels of cortical bone hierarchical organization that may render bone fragile. Namely, mineral crystals at external cortical bone surfaces of the fracture group were larger (65.22nm±41.21nm vs. 36.75nm±18.49nm, p<0.001), and a shift to a higher mineral content and more homogenous mineralization profile as revealed via qBEI were found in the bone matrix of the fracture group. Fracture cases showed nearly 35% higher cortical porosity and showed significantly reduced osteocyte lacunar density compared to controls (226±27 vs. 247±32#/mm(2), p=0.05). Along with increased crystal size, a shift towards higher mineralization and a tendency to increased cortical porosity and reduced osteocyte lacunar number delineate that cortical bone of the superolateral femoral neck bears distinct signs of fragility at various levels of its structural organization. These results contribute to the understanding of hierarchical bone structure changes in age-related fragility.

Influence of porosity, pore size, and cortical thickness on the propagation of ultrasonic waves guided through the femoral neck cortex: a simulation study.

The femoral neck is a common fracture site in elderly people. The cortical shell is thought to be the major contributor to the mechanical competence of the femoral neck, but its microstructural parameters are not sufficiently accessible under in vivo conditions with current X-ray-based methods. To systematically investigate the influences of pore size, porosity, and thickness of the femoral neck cortex on the propagation of ultrasound, we developed 96 different bone models (combining 6 different pore sizes with 4 different porosities and 4 different thicknesses) and simulated the ultrasound propagation using a finite-difference time-domain algorithm. The simulated single-element emitter and receiver array consisting of 16 elements (8 inferior and 8 superior) were placed at anterior and posterior sides of the bone, respectively (transverse transmission). From each simulation, we analyzed the waveform collected by each of the inferior receiver elements for the one with the shortest time of flight. The first arriving signal of this waveform, which is associated with the wave traveling through the cortical shell, was then evaluated for its three different waveform characteristics (TOF: time point of the first point of inflection of the received signal, Δt: difference between the time point at which the signal first crosses the zero baseline and TOF, and A: amplitude of the first extreme of the first arriving signal). From the analyses of these waveform characteristics, we were able to develop multivariate models to predict pore size, porosity, and cortical thickness, corresponding to the 96 different bone models, with remaining errors in the range of 50 µm for pore size, 1.5% for porosity, and 0.17 mm for cortical thickness.

Dependence of mechanical properties of trabecular bone on plate-rod microstructure determined by individual trabecula segmentation (ITS).

Individual trabecula segmentation (ITS) technique can decompose the trabecular bone network into individual trabecular plates and rods and is capable of quantifying the plate/rod-related microstructural characteristics of trabecular bone. This novel technique has been shown to be able to provide in-depth insights into micromechanics and failure mechanisms of human trabecular bone, as well as to distinguish the fracture status independent of area bone mineral density in clinical applications. However, the plate/rod microstructural parameters from ITS have never been correlated to experimentally determined mechanical properties of human trabecular bone. In this study, on-axis cylindrical trabecular bone samples from human proximal tibia (n=22), vertebral body (n=10), and proximal femur (n=21) were harvested, prepared, scanned using micro computed-tomography (µCT), analyzed with ITS and mechanically tested. Regression analyses showed that the plate bone volume fraction (pBV/TV) and axial bone volume fraction (aBV/TV) calculated by ITS analysis correlated the best with elastic modulus (R(2)=0.96-0.97) and yield strength (R(2)=0.95-0.96). Trabecular plate-related microstructural parameters correlated highly with elastic modulus and yield strength, while most rod-related parameters were found inversely and only moderately correlated with the mechanical properties. In addition, ITS analysis also identified that trabecular bone at human femoral neck had the highest trabecular plate-related parameters while the other sites were similar with each other in terms of plate-rod microstructure.

Bone mineralization is elevated and less heterogeneous in adults with type 2 diabetes and osteoarthritis compared to controls with osteoarthritis alone.

PURPOSE: The purpose of this study was to determine whether trabecular bone mineralization differed in adults with type 2 diabetes compared to adults without type 2 diabetes. METHODS: Proximal femur specimens were obtained following a total hip replacement procedure from men and women ≥65 years of age with and without type 2 diabetes. A scanning electron microscope was used for quantitative backscattered electron imaging (qBEI) analysis of trabecular bone samples from the femoral neck. Gray scale images (pixel size=5.6 µm(2)) were uploaded to ImageJ software and gray level (GL) values were converted to calcium concentrations (weight [wt] % calcium [Ca]) using data obtained with energy dispersive X-ray spectrometry. The following bone mineralization density distribution (BMDD) outcomes were collected: the weighted mean bone calcium concentration (CaMEAN), the most frequently occurring bone calcium concentration (CaPEAK) and mineralization heterogeneity (CaWIDTH). Differences between groups were assessed using the Student's t-test for normally distributed data and Mann-Whitney U-test for non-normally distributed data. An alpha value of <0.05 was considered significant. RESULTS: Thirty-five Caucasian participants were recruited (mean [standard deviation, SD] age, 75.5 [6.5]years): 14 adults with type 2 diabetes (years since type 2 diabetes diagnosis, 13.5 [7.4]years) and 21 adults without type 2 diabetes. In the adults with type 2 diabetes, bone CaMEAN was 4.9% greater (20.36 [0.98]wt.% Ca versus 19.40 [1.07]wt.% Ca, p=0.015) and CaWIDTH was 9.4% lower (median [interquartile range] 3.55 [2.99-4.12]wt.% Ca versus 3.95 [0.71]wt.% Ca, p<0.001) compared to controls. There was no between-group difference in CaPEAK (21.12 [0.97]wt.% Ca for type 2 diabetes versus 20.44 [1.30]wt.% Ca for controls, p=0.121). CONCLUSION: The combination of elevated mean calcium concentration in bone and lower mineralization heterogeneity in adults with type 2 diabetes may have deleterious effects on the biomechanical properties of bone. These microscopic alterations in bone mineralization, which may be mediated by suppressed bone remodeling, further elucidate higher fracture risk in adults with type 2 diabetes.

Age-dependence of power spectral density and fractal dimension of bone mineralized matrix in atomic force microscope topography images: potential correlates of bone tissue age and bone fragility in female femoral neck trabeculae.

There is an increasing interest in bone nano-structure, the ultimate goal being to reveal the basis of age-related bone fragility. In this study, power spectral density (PSD) data and fractal dimensions of the mineralized bone matrix were extracted from atomic force microscope topography images of the femoral neck trabeculae. The aim was to evaluate age-dependent differences in the mineralized matrix of human bone and to consider whether these advanced nano-descriptors might be linked to decreased bone remodeling observed by some authors and age-related decline in bone mechanical competence. The investigated bone specimens belonged to a group of young adult women (n = 5, age: 20-40 years) and a group of elderly women (n = 5, age: 70-95 years) without bone diseases. PSD graphs showed the roughness density distribution in relation to spatial frequency. In all cases, there was a fairly linear decrease in magnitude of the power spectra with increasing spatial frequencies. The PSD slope was steeper in elderly individuals (-2.374 vs. -2.066), suggesting the dominance of larger surface morphological features. Fractal dimension of the mineralized bone matrix showed a significant negative trend with advanced age, declining from 2.467 in young individuals to 2.313 in the elderly (r = 0.65, P = 0.04). Higher fractal dimension in young women reflects domination of smaller mineral grains, which is compatible with the more freshly remodeled structure. In contrast, the surface patterns in elderly individuals were indicative of older tissue age. Lower roughness and reduced structural complexity (decreased fractal dimension) of the interfibrillar bone matrix in the elderly suggest a decline in bone toughness, which explains why aged bone is more brittle and prone to fractures.

Proximal femoral epiphysis anatomy in chilean population: orthopedic and forensic aspects / Anatomía del epífisis proximal del fémur en la población chilena. Aspectos traumatológicos y forenses

From a biomechanical standpoint, bone geometry and density are factors correlated to the bone resistance of the femur when supporting body weight, with geometric parameters like the diameter of the femoral head and neck, the length of the femoral neck, and the femoral neck angle as determinant factors in the incidence of hip fractures, which increase in frequency and seriousness in osteoporotic patients. In Chile, morphometric data that contributes to relating the anatomy of the proximal epiphysis of the femur as an associated factor in hip fractures does not exist; likewise, there are no anthropometric indexes that may contribute to the forensic sciences. The purpose of this study is to establish average measurements of the proximal epiphysis of the femur in the adult Chilean population. Descriptive Study. The proximal epiphyses of 81 dry adult femurs were analyzed (44 right and 37 left bones), measuring the following parameters: length of the femoral neck (LN), femoral neck angle (FNA), circumference of the femoral head (CH) and circumference of the femoral neck (CN). The statistical relationship between the measurements and the side of each sample was analyzed (t-test p=0.05). The average lengths were LN= 3.59cm (+/- 0.43 cm); FNA= 124.17 (+/- 6.37), CH= 14.34 cm (+/- 1.27 cm) and CN= 9.7 cm (+/- 0.87 cm). No significant differences between the left and right sides were found. Average numbers were obtained for the anatomy of the proximal femoral epiphysis from a sample in the Chilean population. With the data obtained, we propose to carry out anatomo-clinical, epidemiologic and forensic studies in this population.

Desde un punto de vista biomecánico, la geometría y la densidad ósea son factores correlacionados con la resistencia del hueso del fémur al apoyar el peso corporal, con los parámetros geométricos, como el diámetro de la cabeza femoral y el cuello, la longitud del cuello del fémur, y el ángulo del cuello femoral factores determinantes en la incidencia de fracturas de cadera, que aumentan en frecuencia y gravedad en los pacientes con osteoporosis. En Chile, no existen datos morfométricos que relacionen la anatomía de la epífisis proximal del fémur como un factor asociado a las fracturas de cadera ni índices antropométricos que pueden contribuir a las ciencias forenses. El propósito de este estudio es establecer las medidas promedio de la epífisis proximal de fémur en población adulta chilena. Estudio Descriptivo. Se analizaron la epífisis proximal de 81 fémures adultos secos (44 derechos y 37 izquierdos), midiendo los siguientes parámetros: longitud del cuello femoral (LC) , ángulo cérvico-diafisiario femoral (ACD), circunferencia de la cabeza femoral (CCa) y circunferencia del cuello femoral (CCu). Se analizó la relación estadística de las medidas con el lado de cada muestra (test Chi cuadrado p:0,05) Las longitudes promedios fueron LC: 3,59 cm (+/- 0,43 cm); ACD: 124,17 (+/- 6,37 cm); CCa: 14,34 cm (+/- 1,27 cm) y CCu: 9,7 cm (+/- 0,87 cm). No se encontraron diferencias significativas entre el lado derecho e izquierdo. Los resultados proponen la necesidad de realizar estudios anatomo-clínicos y epidemiológicos actualizados en población chilena donde la geometría de la epífisis proximal del fémur se incluya dentro del análisis.

Micro-structural basis for particular vulnerability of the superolateral neck trabecular bone in the postmenopausal women with hip fractures.

In this study we analyzed the trabecular bone micro-architecture in the inferomedial and superolateral subregions of the femoral neck in a group with hip fractures and a control group of elderly women, with aim to clarify the micro-structural basis of bone fragility. Proximal femora from 29 Caucasian female cadavers were collected at Institute of Forensic Medicine in Belgrade (15 women with hip fracture: age 79.5±8.5 yrs.; and 14 women without hip fractures: age 74.1±9.3 yrs.). The femoral neck section was scanned in dry conditions using a micro-computed tomography (Scanco µCT 40), at 70 kV, 114 µA, 300 ms integration time, 36 µm resolution, isotropic, 1024×1024 pixels per slice, automatically evaluating trabecular micro-architecture using the built-in program of the micro-CT with direct 3D morphometry. The samples were foam padded to avoid any movement artifacts during scanning. Analysis of the neck section in the fracture group compared to the control cases demonstrated significantly lower bone volume fraction (mean: 6.3% vs. 11.2%, p=0.002), lower connectivity density (0.33/mm(3) vs. 0.74/mm(3), p=0.019) and higher trabecular separation (0.87 mm vs. 0.83 mm, p=0.030). Division into the superolateral and inferomedial regions of interest revealed that the superolateral neck displayed even more differences in micro-architectural properties between the fracture and non-fracture groups. Namely, while in the inferomedial neck only bone volume fraction and degree of anisotropy displayed significant inter-group variability (lower BV/TV with higher degree of anisotropy in the fracture group), in the superolateral neck almost all parameters were different between the fracture cases and the controls, where the fracture group showed a lower trabecular bone volume fraction (3.6% vs. 8.2%, p=0.001), lower connectivity (0.21 vs. 0.63/mm(3), p=0.008), more rod like trabecular structure (SMI: 2.94 vs. 2.62, p=0.049), higher separation and the thinned trabeculae (Tb.Sp: 0.89 vs. 0.85 mm, p=0.013; Tb.Th: 0.17 vs. 0.20 mm, p=0.05). In addition, after adjusting for the effects of BV/TV, the majority of differences disappeared, demonstrating that the bone loss manifests itself via the changes in micro-architectural parameters: trabecular thinning, rising the spacing between individual trabeculae, reducing trabecular connectivity and accentuating trabecular perforations leading to predominance of rod-like trabecular elements. Preferential impairment of the superolateral neck trabecular structure and organization in women with hip fracture reveals the region-dependent micro-structural basis of bone fragility in elderly women.

Age- and gender-dependent changes in three-dimensional microstructure of cortical and trabecular bone at the human femoral neck.

UNLABELLED: We investigated age- and gender-related variation of both cortical and trabecular microstructure in human femoral neck. We found that age-related change of cortical porosity is more noticeable than that of trabecular parameter. Our data may help to gain more insight into the potential mechanism of osteoporotic femoral neck fractures. INTRODUCTION: Variations in the microstructure of cortical and trabecular bone contribute to decreased bone strength. Age- and gender-related changes in cortical and trabecular microstructure of femoral neck is unclear. The aim of this study was to identify three-dimensional (3D) microstructural changes of both cortical and trabecular bone simultaneously in human femoral neck with age and gender, using micro-computed tomography (micro-CT). We hypothesized that there would be differences in age-related changes of cortical and trabecular bone for both women and men. METHODS: We used 56 femoral necks of 28 women and men (57-98 years of age) from a Japanese population. The subjects were chosen to give an even age and gender distribution. Both women and men were divided into three age groups: middle (57-68 years), old (72-82 years), and elderly (87-98 years) groups. We examined cortical bone specimen from the inferior sector of femoral neck and trabecular bone specimen from the middle of femoral neck using micro-CT and 3D bone analysis software. RESULTS: Cortical thickness (Ct.Th) decreased by 10-15%, cortical porosity (Ca.V/TV) almost doubled, and canal diameter (Ca.Dm) increased by 65-77% between the middle-aged and elderly groups for both women and men. The trabecular bone volume fraction (BV/TV) decreased by around 20%; trabecular thickness (Tb.Th), trabecular number (Tb.N), and connectivity density (Conn.D) decreased; and trabecular separation (Tb.Sp) and structure model index (SMI) increased with age for both women and men. As compared with women, men had higher Ct.Th and BV/TV and lower Ca.V/TV and Ca.Dm among three age groups. There was a significant inverse correlation between Ca.V.TV and BV/TV for both women and men. CONCLUSION: Our findings indicate that Ct.Th and BV/TV decreased, and Ca.V/TV and Ca.Dm increased in femoral neck with age for both women and men. The most obvious age-related change is the increase of Ca.V/TV. The decrease of BV/TV with age is more noticeable than that of Ct.Th. This is the first study that has provided both cortical and trabecular microstructural data simultaneously in a Japanese sample. These data may help us to gain more insight into the potential mechanism of osteoporotic femoral neck fractures.

Age-and region-dependent changes in three-dimensional microstructural properties of proximal femoral trabeculae.

UNLABELLED: This study investigated regional variations in the 3D microstructure of trabecular bone in human proximal femur, with respect to aging. The results demonstrate that age-related changes in trabecular microstructure significantly varied from different sub-regions of the proximal femur. INTRODUCTION: We hypothesize that the age-related changes in trabecular bone microstructure appear to be varied from specific anatomic sub-regions of the proximal femur followed by non-uniform bone loss. The purpose of this study was therefore to explore regional variations in the 3D microstructure of trabecular bone in human proximal femur, with respect to aging. METHODS: A total of 162 trabecular bone cores from six regions of 27 femora of male cadaver donors were scanned using micro-computed tomography (micro-CT). The following microstructural parameters were calculated: bone volume fraction (BV/TV), trabecular number (Tb.N), thickness (Tb.Th) and separation (Tb.Sp), structure model index (SMI), and degree of anisotropy (DOA). RESULTS: Age-related changes in trabecular microstructure varied from different regions of the proximal femur. There was a significant decrease in bone volume fraction and an almost identical decrease in trabecular thickness associated with aging at any region. Regional analysis demonstrated a significant difference in BV/TV, Tb.Th, Tb.Sp, Tb.N and DOA between superior and inferior neck, as well as a significant difference in BV/TV, Tb.Sp, Tb.N, SMI and DOA between superior and inferior trochanter. CONCLUSIONS: Age-related changes in bone loss and trabecular microstructure within the male proximal femur are not uniform in this cadaveric population.

Cortical vascular canals in human mandible and other bones.

The human mandible is highly mineralized. We hypothesized that this is related to the local vascularity of the bone. This could not be examined directly, but, as a surrogate, intracortical vascular canal spaces of the human mandible were studied so that we could determine possible relationships with age, gender, location, dental status, and tissue mineralization. Canal numbers, area, and volume fraction were calculated from quantitative backscattered electron images of human mandibles aged 16-96 years. Data were compared with calvaria, maxilla, lumbar vertebra, femoral neck, and iliac crest. In the mandible, the buccal aspect of the midline was the most porous, the canals being larger and more numerous. The cortical porosity in the posterior of partially dentate mandibles was significantly greater than that of either dentate or edentate mandibles, and there was a significant increase in the size of canals in the mandible with increasing age. Female mandibles had more porous cortices. No relationship was found between cortical porosity and the degree of bone mineralization.

Structural and remodeling indices in the cancellous bone of the proximal femur across adulthood.

Fragility fractures, including fractures of the femoral neck, result from reductions in the amount, quality and architecture of bone. However, investigations of the underlying structural changes that might predispose to fracture have been largely limited to skeletal sites that do not fracture, such as the iliac crest (IC). The aim of this study was to use histomorphometry to map changes in the architecture and the static remodeling indices of cancellous bone, as a function of age and sex, in bone samples taken from the intertrochanteric (IT) region of the proximal femur at routine autopsy (18-88 years of age). Bone samples for histology were processed from 10-mm cubes of IT cancellous bone. Histomorphometry was performed using an ocular-mounted 10 x 10 graticule at a magnification of x100. An age-dependent decrease in trabecular bone volume was observed in both females and males, as expected (r=-0.75 and r=-0.63, p<0.001, respectively). The underlying mechanisms for bone turnover appeared to be different between males and females. Thus, while the static index of bone resorption (ES/BV) was positively age-dependent in males and females (p<0.001, p<0.03, respectively), the index of bone formation (OS/BV) correlated positively with age in the female group only (p<0.001 vs. NS). Perhaps reflecting an increase in bone formation in older females, the OS/ES ratio was greater in older females than younger females or males. Surprisingly, while resorption indices increased in older males compared with their younger counterparts, bone formation indices increased only in the older female cohort. The IT region in the proximal femur is adjacent to the site commonly involved in fragility fracture. With the limitation that these results describe cross-sectional data, they provide useful insights into changes in the cancellous bone structure and at the bone surface of females and males over the age range of 20-90 years, at a clinically relevant skeletal site.

Collagen fibril diameter in relation to bone site. a quantitative ultrastructural study.

The collagen fibril diameter was measured in cortical bone samples from the femoral neck, rear and front tibia of rats using electron microscopy analysis. Statistically significant differences (0.001

Femoral neck trabecular microstructure in ovariectomized ewes treated with calcitonin: MRI microscopic evaluation.

UNLABELLED: Ovariectomy induces deterioration of the trabecular structure in the femoral neck of ewes, as depicted by MR microscopic imaging. This structural deterioration is prevented by salmon calcitonin treatment. INTRODUCTION: This study evaluated the trabecular (Tb) microarchitecture of an ovariectomy (OVX)-induced osteoporotic model in ewes and determined the effects of salmon calcitonin (sCT), an osteoclast inhibitor, on the Tb structure. This is the first report of OVX-induced changes in the Tb structure in the femoral neck in the ewes and effect of sCT on the microarchitecture. MATERIALS AND METHODS: Ewes (5-8 years old, n = 28) were equally allocated into sham (Sham), OVX injected with vehicle, or OVX injected with sCT at 50 or 100 IU, three injections per week. They were killed 6 months after OVX. The femoral neck was examined with an MR imager at 9.4 T in axial, coronal, and sagittal planes. An internal calibration procedure as a means of standardizing image analysis was used to adjust the segmentation threshold. Data from all three planes were averaged. RESULTS AND CONCLUSIONS: Compared with Sham, OVX induced significant changes (p < 0.0125) in the MRI-derived femoral neck Tb structure: Tb bone volume fraction (BV/TV), -18%; Tb number, -20%; Tb separation, +23%; number of free ends, +28%; number of nodes, -39%; number of Tb branches, -23%; mean length of Tb branches, -19%. Compared with OVX, treatment of sCT at 100 IU significantly improved all the Tb structural parameters to the Sham level (p < 0.0001 approximately p = 0.0281), whereas 50 IU significantly increased the Tb number and the mean length of the Tb branches. BV/TV explained 74% of the variation of compressive stress of the trabecular cylinder cores of the femoral neck. Combining all structural parameters in a multivariate regression analysis significantly improved the explanation to 84%, and adding BMD further improved the predictive ability of the model to 92%. We conclude that OVX induces deterioration of the MRI-derived Tb microstructure in the femoral neck of ewes. sCT treatment prevents OVX-induced changes. The femoral neck microarchitecture significantly correlates with its biomechanical properties. Combining microstructural parameters with BMD further improves the prediction of bone biomechanical properties. The effects of sCT on OVX ewes may help explain reduced fracture risk in postmenopausal osteoporotic women treated with sCT.