A Novel 3D Microstructural Model for Trabecular Bone: II. The Relationship Between Fabric and the Yield Surface

A novel 3D microstructural model was proposed and validated in part I of this publication. In part II, the model was used to identify the yield surface of a representative volume element of human trabecular bone as a function of volume fraction and degree of anisotropy. Finite element models of open and closed cells geometries were used to calculate effective yield stresses for a variety of loading cases with periodic boundary conditions. The postyield behaviour of the trabecular tissue was assumed from data available for cortical tissue. The yield stresses defined by a 0.2% offset in the global stress-strain curve were fit to an orthotropic Hill criterion and the parameters of the surface calculated. Similarly to the previous elastic analysis, distinct but strong relationships were obtained between volume fraction, fabric and the yield surface parameters for both the open and closed cell geometries. This finding suggests that volume fraction and fabric may be used to predict the initiation of mechanical damage in human trabecular bone at the continuum level.     

Bone structural parameters, dosimetry, and relative radiation risk in the beagle skeleton

A variety of morphometric and histomorphometric parameters such as the mass of bone and marrow, bone surface areas, percentage of bone volume, percentage of the surface that is trabecular, and percentage of surfaces that are forming and resting are calculated for all major parts of the beagle skeleton. The total bone surface of the beagle is estimated at 2.9 m2 with 53.7% of the surface area being associated with trabecular bone. There are about 4.5 x 10(9) bone-lining cells and about 1 x 10(9) osteoblasts. From the fractional retention in each part of the skeleton, the initial surface concentration of 239Pu after a single injection of 592 Bq/kg body wt (0.016 microCi/kg) on resting surfaces and at sites of bone formation is calculated for various values of the affinity ratios of trabecular/cortical and forming/resting surfaces. These estimated concentrations then yield dose rates as well as cumulative and collective doses to bone-lining cells and osteoblasts in the different parts of the skeleton. On the assumption that the relative risk of tumor induction is proportional to the collective dose to either bone-lining cells or osteoblasts, the frequency of tumor occurrence is calculated and compared to observed frequencies. Both hypotheses yield approximate agreement with experimental data for different ratios of trabecular/cortical radiation sensitivity, although the differences between some bones are statistically significant.     

Some basic relationships between density values in cancellous and cortical bone

Density is a salient property of bone and plays a crucial role in determining the mechanical properties of both its cancellous and cortical structural forms. Density is defined in a number of ways at either the bone tissue (D(app), apparent) or the bone material level (D(mat), material). The concept of density is relatively simple, but measuring it in the context of bone is a complex issue. The third dimension of the problem is the concept of porosity, or BV/TV (ratio of bone material volume over tissue volume). Recent investigations from our laboratory have revealed an interdependence of D(app) and D(mat) in the cancellous bone of at least four different cohorts of human patients. To clarify the underlying causes of this behaviour, we produced here equivalent relationships from specimens originating from cortical and cancellous areas of the same bone. Plots of D(app) vs. D(mat) showed that D(mat) was not a monotonic function of increasing D(app), but instead showed a 'boomerang'-like pattern. By empirically dissecting the data in two regions for D(app) above and below a value equal to 1.3gcm(-3), we were able to objectively isolate the bone in trabecular and compact forms. Our findings may have implications not only for the segregation of bone in these two structural forms, but also for the mechanobiological and physiological processes that govern the regulation of compact and trabecular bone areas.     

A comparative analysis of the microarchitecture of cortical membranous and cortical endochondral onlay bone grafts in the craniofacial skeleton

Previous work in this laboratory established that an onlay bone graft's survival is determined primarily by its relative cortical and cancellous composition rather than its embryologic origin. A volumetric analysis of external bone graft resorption, however, does not explain the internal microarchitectural changes that may be occurring as these grafts become incorporated. To expand the knowledge of bone graft dynamics beyond volumetric parameters, a better understanding of the internal processes of bone graft remodeling is needed. In this comparative study of cortical onlay bone graft microarchitecture, the authors propose to show that cortical onlay bone grafts undergo measurable internal microarchitectural changes as they become incorporated into the surrounding craniofacial skeleton. In addition, the authors propose to further demonstrate similarities between the internal microarchitecture of cortical onlay bone grafts of different embryologic origin over time. Twenty-five adult New Zealand White rabbits were used for this study. They were divided into two groups of eight animals and one group of nine. The groups were killed at 3, 8, and 16 weeks. Cortical membranous and endochondral bone grafts were placed subperiosteally onto each rabbit's cranium. In addition, five ungrafted cortical endochondral and membranous bone specimens were used as controls. Microcomputed tomography (MCT) scanning and histomorphometric analysis were performed on all of the specimens to obtain detailed information regarding the microarchitecture of the cortical bone grafts. The parameters of bone volume fraction, bone surface area to volume, mean trabecular number, and anisotropy were used to give quantitative information about a bone's micro-organization. The results showed that there is no statistically significant difference between the cortical endochondral and the cortical membranous bone grafts for bone volume fraction, bone surface to volume, mean trabecular number, and anisotropy measurements for all time points. There were, however, statistically significant differences when comparing the control and 3-week groups to the 16-week group for all parameters. The advanced MCT technology and histomorphometric techniques proved to be effective in providing a qualitative and quantitative ultrastructural comparison of cortical endochondral and membranous onlay bone grafts over time. In this study, a statistically significant change in the internal microarchitecture of cortical onlay bone grafts of different embryologic origins was seen as they were remodeled and resorbed at all time points. Specifically, the onlay cortical bone grafts developed a less dense, more trabecular, and less organized internal ultrastructure. In addition, no difference in the three-dimensional ultrastructure of cortical endochondral and membranous bone was found. These results challenge some of the currently accepted theories of bone-graft dynamics and may eventually lead to a change in the way clinicians approach bone-graft selection for craniofacial surgery.     

Proximal tubular atrophy: Qualitative and quantitative structural changes in chronic obstructive nephropathy in the pig

Summary Kidneys of pigs with various degrees of induced chronic obstructive nephropathy were studied by light- and electron microscopy to assess the structural changes of proximal convoluted tubules with increasing degrees of atrophy. A particular aim was to evaluate the quantitative relationship between proximal tubular and interstitial changes in early tubular atrophy. The kidneys were subjected to varying degrees of ureteral obstruction and were fixed by in vivo vascular perfusion. Quantitative (morphometric) analyses were carried out on montages of electron micrographs representing randomly selected cortical areas and cross sections of individual proximal convoluted tubules. The results demonstrated that ureteral obstruction was followed by significant reductions in proximal tubular epithelium, in volume of proximal tubular mitochondria and in surface area of proximal tubular basolateral membranes. These changes were present even in the absence of any demonstrable increase in cortical interstitium or alterations in the relationships between proximal tubules and peritubular capillaries. With increase in the volume of cortical interstitium the proximal tubules were further simplified in ultra-structure with a reduced number of interdigitating lateral cell processes. Concomitantly there were significant quantitative changes in the spatial associations between tubules and capillaries due to increase in tubulo-capillary distances. The present study shows that ultrastructural changes in proximal tubules during early atrophy precede the volume increase in cortical interstitium associated with chronic obstructive nephropathy. It is suggested that the early tubular changes are due to decreased functional loads, whereas the further progression of tubular atrophy may be a result of impaired nourishment of the tubular cells due to increased interstitial tissue and altered relationships between tubules and capillaries.This work was supported by grant no 12-0727 from the Danish Medical Research Council     

The elastic moduli of human subchondral, trabecular, and cortical bone tissue and the size-dependency of cortical bone modulus

The elastic moduli of human subchondral, trabecular, and cortical bone tissue from a proximal tibia were experimentally determined using three-point bending tests on a microstructural level. The mean modulus of subchondral specimens was 1.15 GPa, and those of trabecular and cortical specimens was 4.59 GPa and 5.44 GPa respectively. Significant differences were found in the modulus values between bone tissues, which may have mainly resulted from the differences in the microstructures of each bone tissue rather than in the mineral density. Furthermore, the size-dependency of the modulus was examined using eight different sizes of cortical specimens (heights h = 100-1000 microns). While the modulus values for relatively large specimens (h greater than 500 microns) remained fairly constant (approximately 15 GPa), the values decreased as the specimens became smaller. A significant correlation was found between the modulus and specimen size. The surface area to volume ratio proved to be a key variable to explain the size-dependency.     

Plutonium worker dosimetry

Epidemiological studies of the relationship between risk and internal exposure to plutonium are clearly reliant on the dose estimates used. The International Commission on Radiological Protection (ICRP) is currently reviewing the latest scientific information available on biokinetic models and dosimetry, and it is likely that a number of changes to the existing models will be recommended. The effect of certain changes, particularly to the ICRP model of the respiratory tract, has been investigated for inhaled forms of ²³⁹Pu and uncertainties have also been assessed. Notable effects of possible changes to respiratory tract model assumptions are (1) a reduction in the absorbed dose to target cells in the airways, if changes under consideration are made to the slow clearing fraction and (2) a doubling of absorbed dose to the alveolar region for insoluble forms, if evidence of longer retention times is taken into account. An important factor influencing doses for moderately soluble forms of ²³⁹Pu is the extent of binding of dissolved plutonium to lung tissues and assumptions regarding the extent of binding in the airways. Uncertainty analyses have been performed with prior distributions chosen for application in epidemiological studies. The resulting distributions for dose per unit intake were lognormal with geometric standard deviations of 2.3 and 2.6 for nitrates and oxides, respectively. The wide ranges were due largely to consideration of results for a range of experimental data for the solubility of different forms of nitrate and oxides. The medians of these distributions were a factor of three times higher than calculated using current default ICRP parameter values. For nitrates, this was due to the assumption of a bound fraction, and for oxides due mainly to the assumption of slower alveolar clearance. This study highlights areas where more research is needed to reduce biokinetic uncertainties, including more accurate determination of particle transport rates and long-term dissolution for plutonium compounds, a re-evaluation of long-term binding of dissolved plutonium, and further consideration of modeling for plutonium absorbed to blood from the lungs.     

A comparison of the fatigue behavior of human trabecular and cortical bone tissue

The fatigue properties of trabecular bone tissue (single trabeculae) and similarly sized cortical bone specimens from human tibia were experimentally determined on a microstructural level using four-point bending cyclic tests, and they were compared based on modulus, mineral density, and microstructural characteristics. The results showed that trabecular specimens had significantly lower moduli and lower fatigue strength than cortical specimens, despite their higher mineral density values. Fracture surface and microdamage analyses illustrated different fracture and damage patterns between trabecular and cortical bone tissue, depending upon their microstructural characteristics. Based on the results from mechanical tests and qualitative observations, a possible mechanical role of the cement lines in trabecular tissue microfracture was suggested.     

Quantitative proteomics analysis of adsorbed plasma proteins classifies nanoparticles with different surface properties and size

Nanoparticle biological activity, biocompatibility and fate can be directly affected by layers of readily adsorbed host proteins in biofluids. Here, we report a study on the interactions between human blood plasma proteins and nanoparticles with a controlled systematic variation of properties using (18)O-labeling and LC-MS-based quantitative proteomics. We developed a novel protocol to both simplify isolation of nanoparticle bound proteins and improve reproducibility. LC-MS analysis identified and quantified 88 human plasma proteins associated with polystyrene nanoparticles consisting of three different surface chemistries and two sizes, as well as, for four different exposure times (for a total of 24 different samples). Quantitative comparison of relative protein abundances was achieved by spiking an (18)O-labeled "universal" reference into each individually processed unlabeled sample as an internal standard, enabling simultaneous application of both label-free and isotopic labeling quantification across the entire sample set. Clustering analysis of the quantitative proteomics data resulted in distinctive patterns that classified the nanoparticles based on their surface properties and size. In addition, temporal data indicated that the formation of the stable protein corona was at equilibrium within 5 min. The comprehensive quantitative proteomics results obtained in this study provide rich data for computational modeling and have potential implications towards predicting nanoparticle biocompatibility.     

Volume to density relation in adult human bone tissue

Uniformity of tissue mineralisation is a strongly debated issue, due to its relation with bone mechanical behaviour. Bone mineral density (BMD) is measured in the clinical practice and is applied in computational application to derive material proprieties of bone tissue. However, BMD cannot identify if the variation in bone density is related to a modification of tissue mineral density (TMD), a change in bone volume or a combination of the two. This study was aimed to investigate whether TMD can be assumed as a constant in adult human bone (trabecular and cortical).A total number of 115 cylindrical bone specimens were collected. An inter-site analysis (96 specimens, 2 donors) was performed on cortical and trabecular specimens extracted from different anatomical sites. An intra-site study (19 specimens, 19 donors) was performed on specimens extracted from femoral heads. Bone volume fraction (BV/TV) was computed by means of a micro-computed tomography. Furthermore, ash density (ρ_ash) was measured. TMD was computed as the ratio between ρ_ash and BV/TV.It was found that the TMD of trabecular (1.24±0.16 g/cm³) and cortical (1.19±0.06 g/cm³) bone were not statistically different (p=0.31). Furthermore, the linear regression between ρ_ash and BV/TV was statistically significant (r²=0.99, p<0.001). Intra- and inter-site analyses demonstrated that the mineral distribution was independent of the extraction site.The present study suggests that TMD can be assumed reasonably constant in non-pathological adult bone tissue. Consequently, it is suggested that TMD can be managed as a constant in computational models, varying only BV in relation to clinical densitometric analysis.     

Bone Strength in Growing Rats Treated with Fluoride: a Multi-dose Histomorphometric,Biomechanical and Densitometric Study

Bone deformation and fragility are common signs of skeletal fluorosis. Disorganisation of bone tissue and presence of inflammatory foci were observed after fluoride (F^?) administration. Most information about F^? effects on bone has been obtained in adult individuals. However, in fluorosis areas, children are a population very exposed to F^? and prone to develop not only dental but also skeletal fluoroses. The aim of this work was to evaluate the bone parameters responsible for the effect of different doses of F^? on fracture load of the trabecular and cortical bones using multivariate analysis in growing rats. Twenty-four 21-day-old Sprague-Dawley rats were divided into four groups: F0, F20, F40 and F80, which received orally 0, 20, 40 or 80 μmol F^?/100 g bw/day, respectively, for 30 days. After treatment, tibiae were used for measuring bone histomorphometric and connectivity parameters, bone mineral density (BMD) and bone cortical parameters. The femurs were used for biomechanical tests and bone F^? content. Trabecular bone volume was significantly decreased by F^?. Consistently, we observed a significant decrease in fracture load and Young’s modulus (YM) of the trabecular bone in F^?-treated groups. However, cortical bone parameters were not significantly affected by F^?. Moreover, there were no significant differences in cortical nor trabecular BMD. Multivariate analysis revealed a significant correlation between the trabecular fracture load and YM but not with bone volume or BMD. It is concluded that when F^? is administered as a single daily dose, it produces significant decrease in trabecular bone strength by changing the elasticity of the trabecular bone.     

Effect of239Pu on mouse hemopoietic stem cells in different types of bone marrow cavities

Summary Repopulative activity of hemopoietic stem cells of mice given i.v. 5 kBq²³⁹Pu/mouse (166.5 kBq/kg) was followed. The activity retained was measured in the whole mouse, the skeleton and the liver. Simultaneously average cumulative skeletal dose was calculated. Quantitative parameters of the stem cell compartment and the marrow cellularity were studied in variously arranged bones (femur, pelvis, lumbar vertebra) using the exocolonizing test and cytological techniques. The effects of radiation were most marked in lumbar vertebra, less serious changes were found in pelvis and only a moderate response was present in femur. The bone marrow hemopoiesis is damaged in various bone sites to different degrees and the percentage of cells at risk appears higher in trabecular than in cortical bone.     

Functional magnetic resonance imaging as a tool for investigating human cortical motor function.

Non-invasive functional magnetic resonance imaging (fMRI) mapping techniques sensitive to the local changes of blood flow, blood volume, and blood oxygenation which accompany neuronal activation have been widely used over the last few years to investigate the functional organization of human cortical motor systems, and specifically of the primary motor cortex. Validation studies have demonstrated a good correspondence between quantitative and topographic aspects of data acquired by fMRI and positron emission tomography. The spatial and temporal resolution affordable by fMRI has allowed to achieve new important information on the distributed representation of hand movements in multiple functional modules, and on the intensity and spatial extent of neural activation in the contralateral and ipsilateral primary motor cortex in relation to parametric and nonparametric aspects of movement and to the degree of handedness. Neural populations with different functional characteristics have been identified in anatomically defined regions, and the temporal aspects of the activation during voluntary movement tracked in different components of the motor system. Finally, this technique has proved useful to deepen our understanding of the neural basis of motor imagery, demonstrating increased activity in the primary motor cortex during mental representation of sequential finger movements.     

What humeri are suitable for comparative testing of suture anchors? An ultrastructural bone analysis and biomechanical study of ovine,bovine and human humeri and four different anchor types

For testing of fixation devices such as suture anchors used in rotator cuff repair often animal bones are used. They are easily obtained, inexpensive and some have been found to be similar to human bone. But can we rely on the results drawn from these studies in our daily surgical practice?The purpose of this study was to compare the trabecular bone mineral density, the trabecular bone volume fraction and the cortical layer thickness in the greater tubercle in different species to evaluate their influence on primary stability of suture anchors under a cyclic loading protocol representing the physiologic forces placed on rotator cuff repairs in vivo.Bovine and ovine humeri are not suitable for suture anchor testing. The statistical significances for pullout forces between the anchors varied from species to species. Therefore, no very applicable information can be obtained from testing suture anchors in ovine or bovine humeri with regard to ultimate failure loads in human humeri. The ultimate failure load seems to depend mainly on the cortical thickness and on the subcortical trabecular bone quality.     

Trabecular bone ontogeny in the human proximal femur

Ontogenetic changes in the human femur associated with the acquisition of bipedal locomotion, especially the development of the bicondylar angle, have been well documented. The purpose of this study is to quantify changes in the three-dimensional structure of trabecular bone in the human proximal femur in relation to changing functional and external loading patterns with age. High-resolution X-ray computed tomography scan data were collected for 15 juvenile femoral specimens ranging in age from prenatal to approximately nine years of age. Serial slices were collected for the entire proximal femur of each individual with voxel resolutions ranging from 0.017 to 0.046 mm depending on the size of the specimen. Spherical volumes of interest were defined within the proximal femur, and the bone volume fraction, trabecular thickness, trabecular number, and fabric anisotropy were calculated in three dimensions. Bone volume fraction, trabecular number, and degree of anisotropy decrease between the age of 6 months and 12 months, with the lowest values for these parameters occurring in individuals near 12 months of age. By age 2-3 years, the bone volume, thickness, and degree of anisotropy increase slightly, and regions in the femoral neck become more anisotropic corresponding to the thickening of the inferior cortical bone of the neck. These results suggest that trabecular structure in the proximal femur reflects the shift in external loading patterns associated with the initiation of unassisted walking in infants.     

The Effect of Naturally Occurring Chronic Kidney Disease on the Micro-Structural and Mechanical Properties of Bone

Chronic kidney disease (CKD) is a growing public health concern worldwide, and is associated with marked increase of bone fragility. Previous studies assessing the effect of CKD on bone quality were based on biopsies from human patients or on laboratory animal models. Such studies provide information of limited relevance due to the small size of the samples (biopsies) or the non-physiologic CKD syndrome studied (rodent models with artificially induced CKD). Furthermore, the type, architecture, structure and biology of the bone of rodents are remarkably different from human bones; therefore similar clinicopathologic circumstances may affect their bones differently. We describe the effects of naturally occurring CKD with features resembling human CKD on the skeleton of cats, whose bone biology, structure and composition are remarkably similar to those of humans. We show that CKD causes significant increase of resorption cavity density compared with healthy controls, as well as significantly lower cortical mineral density, cortical cross-sectional area and cortical cross-sectional thickness. Young''s modulus, yield stress, and ultimate stress of the cortical bone material were all significantly decreased in the skeleton of CKD cats. Cancellous bone was also affected, having significantly lower trabecular thickness and bone volume over total volume in CKD cats compared with controls. This study shows that naturally occurring CKD has deleterious effects on bone quality and strength. Since many similarities exist between human and feline CKD patients, including the clinicopathologic features of the syndrome and bone microarchitecture and biology, these results contribute to better understanding of bone abnormalities associated with CKD.     

Multiscale,Converging Defects of Macro-Porosity,Microstructure and Matrix Mineralization Impact Long Bone Fragility in NF1

Bone fragility due to osteopenia, osteoporosis or debilitating focal skeletal dysplasias is a frequent observation in the Mendelian disease Neurofibromatosis type 1 (NF1). To determine the mechanisms underlying bone fragility in NF1 we analyzed two conditional mouse models, Nf1Prx1 (limb knock-out) and Nf1Col1 (osteoblast specific knock-out), as well as cortical bone samples from individuals with NF1. We examined mouse bone tissue with micro-computed tomography, qualitative and quantitative histology, mechanical tensile analysis, small-angle X-ray scattering (SAXS), energy dispersive X-ray spectroscopy (EDX), and scanning acoustic microscopy (SAM). In cortical bone of Nf1Prx1 mice we detected ectopic blood vessels that were associated with diaphyseal mineralization defects. Defective mineral binding in the proximity of blood vessels was most likely due to impaired bone collagen formation, as these areas were completely devoid of acidic matrix proteins and contained thin collagen fibers. Additionally, we found significantly reduced mechanical strength of the bone material, which was partially caused by increased osteocyte volume. Consistent with these observations, bone samples from individuals with NF1 and tibial dysplasia showed increased osteocyte lacuna volume. Reduced mechanical properties were associated with diminished matrix stiffness, as determined by SAM. In line with these observations, bone tissue from individuals with NF1 and tibial dysplasia showed heterogeneous mineralization and reduced collagen fiber thickness and packaging. Collectively, the data indicate that bone fragility in NF1 tibial dysplasia is partly due to an increased osteocyte-related micro-porosity, hypomineralization, a generalized defect of organic matrix formation, exacerbated in the regions of tensional and bending force integration, and finally persistence of ectopic blood vessels associated with localized macro-porotic bone lesions.     

Exposure of non‐target tissues in medical diathermy

With different prevalence in different regions, radio frequency (RF) electromagnetic fields (EMF) are widely used for therapeutic tissue heating. Although short‐wave diathermy (27.12 MHz) is the most popular treatment modality, quantitative data on patient's exposure have been lacking. By numerical simulation with the numerical anatomical model NORMAN, intracorporal distributions of specific absorption rates (SAR) were investigated for different treatment scenarios and applicators. Quantitative data are provided for exposures of target treatment areas as well as for vulnerable regions such as the eye lenses, central nervous system, and testes. Different applicators and distances were investigated. Capacitive and inductive applicators exhibit quite a different heating efficiency. It could be shown that for the same output power therapeutic heat deposition can vary by almost one order of magnitude. By mimicking therapist's practice to use patient's heat perception as an indicator for output power setting, numerical data were elaborated demonstrating that muscle tissue exposures may be several times higher for inductive than for capacitive applicators. Presented quantitative data serve as a guide for power adjustment preventing relevant overexposures without compromising therapy; they also provide a basis for estimating target tissue heat load and developing therapeutic guidelines. Bioelectromagnetics 31:12–19, 2010. © 2009 Wiley‐Liss, Inc.     

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging,Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.For this purpose, we injected 10⁵ MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg¹. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula¹. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified^2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions^5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles⁷.In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.