Trabecular Bone Specimens Research Articles

Quantitative 31P NMR spectroscopy and 1H MRI measurements of bone mineral and matrix density differentiate metabolic bone diseases in rat models

In this study, bone mineral density (BMD) of normal (CON), ovariectomized (OVX), and partially nephrectomized (NFR) rats was measured by 31P NMR spectroscopy; bone matrix density was measured by 1H water- and fat-suppressed projection imaging (WASPI); and the extent of bone mineralization (EBM) was obtained by the ratio of BMD/bone matrix density. The capability of these MR methods to distinguish the bone composition of the CON, OVX, and NFR groups was evaluated against chemical analysis (gravimetry). For cortical bone specimens, BMD of the CON and OVX groups was not significantly different; BMD of the NFR group was 22.1% (by 31P NMR) and 17.5% (by gravimetry) lower than CON. For trabecular bone specimens, BMD of the OVX group was 40.5% (by 31P NMR) and 24.6% (by gravimetry) lower than CON; BMD of the NFR group was 26.8% (by 31P NMR) and 21.5% (by gravimetry) lower than CON. No significant change of cortical bone matrix density between CON and OVX was observed by WASPI or gravimetry; NFR cortical bone matrix density was 10.3% (by WASPI) and 13.9% (by gravimetry) lower than CON. OVX trabecular bone matrix density was 38.0% (by WASPI) and 30.8% (by gravimetry) lower than CON, while no significant change in NFR trabecular bone matrix density was observed by either method. The EBMs of OVX cortical and trabecular specimens were slightly higher than CON but not significantly different from CON. Importantly, EBMs of NFR cortical and trabecular specimens were 12.4% and 26.3% lower than CON by 31P NMR/WASPI, respectively, and 4.0% and 11.9% lower by gravimetry. Histopathology showed evidence of osteoporosis in the OVX group and severe secondary hyperparathyroidism (renal osteodystrophy) in the NFR group. These results demonstrate that the combined 31P NMR/WASPI method is capable of discerning the difference in EBM between animals with osteoporosis and those with impaired bone mineralization.

Diversity of bone cell activity as a histomorphometric feature of idiopathic osteoporosis in men

Although osteoporosis in men is an increasing health problem, studies on osteoporosis in males are still scarce. The aim of our study was to determine the characteristics of bone tissue and bone turnover in men with idiopathic osteoporosis. Transiliac crest bone samples were histomorphometrically analyzed after double tetracycline labeling in 32 men aged 37–65 years who were diagnosed with idiopathic osteoporosis by densitometry of the lumbar spine and hip. Bone volume, osteoid surface, osteoblast surface, eroded surface, osteoid thickness, trabecular thickness, trabecular number, trabecular separation, and mineral apposition rate (MAR) were determined in all trabecular bone specimens. Bone volume and structural parameters indicated trabecular bone loss in most patients. Cellular parameters and MAR indicated variations in bone cell actions. No age-related decrease in histomorphometric parameters was found. After the patients were grouped according to MAR values, osteoblast and eroded surfaces were found to be lower in the group with decreased MAR values and elevated in the group of patients with increased MAR parameter. Trabecular thickness was greater in patients with lower than normal MAR, due to reduced resorption and probably loss of very thin trabeculae. Our results suggest that idiopathic osteoporosis in man resembles many characteristics of postmenopausal osteoporosis in women resulting in impaired trabecular structure due to unbalanced cellular activity and bone turnover rate.

Association between collagen cross-links and trabecular microarchitecture properties of human vertebral bone

It has been suggested that age-related deterioration in trabecular microarchitecture and changes in collagen cross-link concentrations may contribute to skeletal fragility. To further explore this hypothesis, we determined the relationships among trabecular bone volume fraction (BV/TV), microarchitecture, collagen cross-link content, and bone turnover in human vertebral trabecular bone. Trabecular bone specimens from L2 vertebrae were collected from 51 recently deceased donors (54–95 years of age; 20 men and 30 women). Trabecular bone volume and microarchitecture was assessed by microCT and bone formation, reflected by osteoid surface (OS/BS, %), was measured by 2D histomorphometry. Pyridinoline (PYD), deoxypyridinoline (DPD), pentosidine (PEN) and collagen content in the cancellous bone were analysed by high-performance liquid chromatography. Associations between variables were investigated by Pearson correlations and multiple regression models, which were constructed with BV/TV and collagen cross-links as explanatory variables and microarchitecture parameters as the dependent variables. Results: Microarchitecture parameters were modestly to strongly correlated with BV/TV (r2=0.10–0.71). The amount of mature enzymatic PYD and DPD cross-links were not associated with the microarchitecture, either before or after adjustment for BV/TV. However, there was a positive correlation between PEN content and trabecular number (r=0.45, p=0.001) and connectivity density (r=0.40, p=0.004), and a negative correlation between PEN content and trabecular separation (r=−0.29, p=0.04). In the multiple regression models including BV/TV, age and PEN content was still significantly associated with several of the microarchitecture variables. In summary, this study suggests a link between trabecular microarchitecture and the collagen cross-link profile. As PEN reflects non-enzymatic glycation of collagen and generally increases with bone age, the association between PEN and trabecular architecture suggests that the preserved trabeculae may contain mainly old bone and have undergone little remodeling. Thus, vertebral fragility may not only be due to alterations in bone architecture but also to modification of collagen cross-link patterns thereby influencing bone's mechanical behavior.

Simulation of vertebral trabecular bone loss using voxel finite element analysis

Trabecular bone loss in human vertebral bone is characterised by thinning and eventual perforation of the horizontal trabeculae. Concurrently, vertical trabeculae are completely lost with no histological evidence of significant thinning. Such bone loss results in deterioration in apparent modulus and strength of the trabecular core. In this study, a voxel-based finite element program was used to model bone loss in three specimens of human vertebral trabecular bone. Three sets of analyses were completed. In Set 1, strain adaptive resorption was modelled, whereby elements which were subject to the lowest mechanical stimulus (principal strain) were removed. In Set 2, both strain adaptive and microdamage mechanisms of bone resorption were included. Perforation of vertical trabeculae occurred due to microdamage resorption of elements with strains that exceeded a damage threshold. This resulted in collapse of the trabecular network under compression loading for two of the specimens tested. In Set 3, the damage threshold strain was gradually increased as bone loss progressed, resulting in reduced levels of microdamage resorption. This mechanism resulted in trabecular architectures in which vertical trabeculae had been perforated and which exhibited similar apparent modulus properties compared to experimental values reported in the literature. Our results indicate that strain adaptive remodelling alone does not explain the deterioration in mechanical properties that have been observed experimentally. Our results also support the hypothesis that horizontal trabeculae are lost principally by strain adaptive resorption, while vertical trabeculae may be lost due to perforation from microdamage resorption followed by rapid strain adaptive resorption of the remaining unloaded trabeculae.

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

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. 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. 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. 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. 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.

Stress Deformation and Fluid Pressure of Bone Specimens under Cyclic Loading

Cyclic loading has been known to induce fluid flow and thus mechanotransduction in bones. In the past, four-point bending tests have been used exclusively in studying fluid flow in bones. In order to better understand the mechanism of deformation and fluid flow under loading, compression tests were done on trabecular bone specimens under drained and undrained conditions. In the drained tests, the volume change was observed, whereas in the undrained tests, excess pore fluid pressure was measured. Cyclic loading tests were conducted in addition to monotonic loading tests to observe the permanent volume change or excess pore fluid pressure with loading cycles. A fast loading rate gave a sharp rise in the excess fluid pressure compared to a slow loading rate. The strength and stiffness of the specimens appeared to deteriorate with an increased speed of loadings, but there was no appreciable difference between the results obtained from drained and undrained tests. The drained and undrained tests as described allowed a better understanding of bone behavior under loadings for a coupled stress-flow analysis.

Contributions of trabecular rods of various orientations in determining the elastic properties of human vertebral trabecular bone

Trabecular bone networks consist of two basic microstructural types: plates and rods. Although trabecular rods represent only a small fraction of total bone volume, their existence has important roles in failure initiation and progression. The goal of this study was to quantitatively examine the contributions of trabecular rods in various orientations to the anisotropic elastic moduli of human vertebral trabecular bone. Twenty-one human vertebral trabecular bone specimens were scanned by microcomputed tomography (μCT). A coordinate system of orthotropic axes representing the best elastic orthotropic symmetry was determined for each sample. Individual trabeculae segmentation (ITS), a 3D image analysis technique, was performed to identify each individual trabecular rod and determine its orientation in the orthotropic coordinate system. Next, three rod-removed images were created where longitudinal, oblique, or transverse trabecular rods were removed, respectively, from the original μCT images. The original and three categories of rod-removed images were then converted to finite element (FE) models for evaluation of their elastic moduli and anisotropy. Both the transverse and oblique rod-removal caused significant decreases in all six elastic moduli. However, the removal of longitudinal rods only caused significant changes in E33, G23, and G31 but not in any transverse/in-plane elastic properties (E11, E22, and G12). The analysis of covariance (ANCOVA) with repeated measures was applied to detect the moduli change in the different models caused by the effects beyond just bone volume loss. The results suggested that the loss of transverse rods induced a significant decrease in in-plane mechanical competence, which was greater than what could be explained only by the associated bone volume loss. In contrast, the reduction in the axial Young's modulus caused by the loss of transverse rods was proportional to the bone volume decrease. Furthermore, the loss of longitudinal rods affected the axial Young's modulus through both bone volume loss and architectural change. With aging, the reduction in in-plane mechanical competence would be magnified by the preferential loss of transverse rods. The predictive ability of bone mineral density, a surrogate of BV/TV in clinical measurements, may reduce more quickly for transverse mechanical properties than for the axial mechanical properties.

Density and architecture have greater effects on the toughness of trabecular bone than damage

Bone damage has been cited as an important aspect of bone quality. As such, understanding the effects of damage on the toughness of trabecular bone should provide insight into trabecular bone behavior during energy-limiting cases, such as falls. The effects of damage on the toughness of 35 bovine trabecular bone specimens were studied. Damage was induced by compressing the on-axis specimens to either 1.5% or 2.5% strain, followed by compression to 7.5% strain. The overloads resulted in significant decreases in both modulus and elastic toughness, with significantly greater decreases for the high-damage group than the low-damage group. Following damage, the elastic toughness of the high-damage group was also lower than the undamaged elastic toughness of the control group. In contrast, there was no detectable effect of damage level on toughness measured to 7.5% strain. Toughness increased linearly with BMD (R2=0.50) and by a power law relationship with volume fraction (BV/TV) (R2=0.65). Microarchitectural parameters also predicted the toughness in the absence of BV/TV or BMD. Toughness decreased with increasing slenderness ratio (Tb.Sp/Tb.Th) and structure model index (SMI) (R2=0.68, multiple regression), again independent of damage level, suggesting that failure is influenced by trabecular buckling. Taken together, the results show that normal variations in toughness due to density and architecture dominate the changes due to damage at the levels induced in this study. Moreover, measuring toughness is sensitive to the final strain, as differences found in the elastic and initial plastic regions were undetectable at higher strains. The self-limiting nature of microcracks in trabecular bone, or the trabecular architecture itself, may inhibit microcracks from propagating to macroscopic trabecular fractures, thereby limiting the effect of damage on toughness and making it difficult to detect in comparison to normal population variability.

Quantitative bone matrix density measurement by water‐ and fat‐suppressed proton projection MRI (WASPI) with polymer calibration phantoms

The density of the organic matrix of bone substance is a critical parameter necessary to clinically evaluate and distinguish structural and metabolic pathological conditions such as osteomalacia in adults and rickets in growing children. Water- and fat-suppressed proton projection MRI (WASPI) was developed as a noninvasive means to obtain this information. In this study, a density calibration phantom was developed to convert WASPI intensity to true bone matrix density. The phantom contained a specifically designed poly(ethylene oxide)/poly(methyl methacrylate) (PEO/PMMA) blend, whose MRI properties (T(1), T(2), and resonance linewidth) were similar to those of solid bone matrix (collagen, tightly bound water, and other immobile molecules), minimizing the need to correct for differences in T(1) and/or T(2) relaxation between the phantom and the subject. Cortical and trabecular porcine bone specimens were imaged using WASPI with the calibration phantom in the field of view (FOV) as a stable intensity reference. Gravimetric and amino acid analyses were carried out on the same specimens after WASPI, and the chemical results were found to be highly correlated (r(2) = 0.98 and 0.95, respectively) to the WASPI intensity. By this procedure the WASPI intensity can be used to obtain the true bone matrix mass density in g cm(-3).

Structural characterization of subchondral bone in the distal aspect of third metacarpal bones from Thoroughbred racehorses via micro–computed tomography

To characterize the microstructure of subchondral bone (SCB) plate and trabecular bone (TBB) of the distopalmar aspect of the condyles of third metacarpal bones (MC3s) from Thoroughbred racehorses at 2 different stages of SCB disease via micro-computed tomography (CT). 12 pairs of MC3s from Thoroughbred racehorses euthanized for various reasons. MC3s were collected from horses with mild (n = 6) or severe (6) SCB disease, as determined via micro-CT. Cubic (6 x 6 x 6-cm) specimens of SCB plate and TBB were cut from the palmar aspect of condyles and sagittal ridges and examined with 3-dimensional micro-CT. For each specimen, apparent bone mineral density (aBMD), true BMD (tBMD), bone volume fraction (BVF), trabecular thickness (TBT), trabecular separation (TBS), and connectivity (CN) were calculated. Condyles had higher aBMD, tBMD, BVF, and TBT and lower TBS than did the sagittal ridge. In bone specimens with mild SCB changes, SCB plate had higher aBMD, TBT, and CN and lower TBS than did TBB. In bone specimens with severe SCB disease, TBB had higher aBMD and TMD and lower TBS than it did in bone specimens with mild disease, and values were similar to those for SCB plate in bone specimens with severe disease. The microstructure of SCB of the distopalmar aspect of metacarpal condyles of horses varied according to the severity of changes identified via micro-CT. With mild SCB disease, sclerosis existed in the SCB plate of the condyles; with severe disease, sclerosis also invaded condylar TBB.

Mechanical properties of subchondral bone in the distal aspect of third metacarpal bones from Thoroughbred racehorses

To characterize the mechanical properties of subchondral bone (SCB) of the distopalmar aspect of the condyles of the third metacarpal bone (MC3) and their correlations with structural aspects of MC3s in Thoroughbred racehorses. 12 pairs of MC3s from Thoroughbred racehorses euthanized for various reasons. MC3s were collected from horses with mild (n = 6) and with severe (6) SCB changes, as determined by micro-computed tomography (CT). Specimens of SCB plate and trabecular bone were cut from the distopalmar aspect of condyles and sagittal ridge and examined with 3-dimensional micro-CT. Specimens were tested in compression, and elastic modulus, yield stress, yield strain, and toughness were calculated. Apparent and true bone mineral density, bone volume fraction, trabecular thickness, trabecular separation, and connectivity were also calculated. Differences in mechanical properties among various classifications of bone were evaluated. Correlations between structural and mechanical variables were also assessed. No differences were detected between left and right forelimbs. Specimens from condyles had higher values for elastic modulus, yield stress, and toughness than did specimens of sagittal ridge. In SCB with severe changes attributable to SCB disease, SCB plate was weaker and trabecular bone was stronger than in SCB with mild changes. Microstructural and mechanical properties were significantly correlated. A marked gradient in mechanical properties of SCB from horses, which could be involved in the pathogenesis of condylar fractures, was detected. Mechanical properties of SCB from the distal aspect of MC3s can be predicted to some extent via micro-CT.

Micro-finite element simulation of trabecular-bone post-yield behaviour – effects of material model, element size and type

Micro-finite element (micro-FE) analysis became a standard tool for the evaluation of trabecular bone mechanical properties. The accuracy of micro-FE models for linear analyses is well established. However, the accuracy of recently developed nonlinear micro-FE models for simulations of trabecular bone failure is not known. In this study, a trabecular bone specimen was compressed beyond the apparent yield point. The experiment was simulated using different micro-FE meshes with different element sizes and types, and material models based on cortical bone. The results from the simulations were compared with experimental results to study the effects of the different element and material models. It was found that a decrease in element size from 80 to 40 μm had little effect on predicted post-yield behaviour. Element type and material model had significant effects. Nevertheless, none of the established material models for cortical bone were able to predict the typical descent in the load-displacement curve seen during compression of trabecular bone.

Stimulation of osteoblast activity by homocysteine

Homocysteine (HCY) has recently been linked to fragility fractures. Moreover, HCY activates osteoclasts. Little is known about the effect of HCY on activity of human osteoblasts (OBs). We hypothesized that HCY decreases the activity of OBs. Osteoblasts obtained from tra-becular human bone specimens of eight donors were cultured with conditioned medium. Culture medium was adjusted to 0, 100, 500, 1000 and 2000 μM HCY. After 14 days alkaline phosphatase (AP) activity, pro-collagen type I N-terminal peptide (PINP) and osteocalcin (OC) secretion in the supernatant were measured. After 20 days the formation of mineralized matrix was analyzed. HCY-stimulated AP activity gradually (100 μM HCY: 118%, P= 0.006; 500 μM HCY: 125%, P < 0.001). At 1000 and 2000 μM HCY the increase of AP activity was reversible (1000 μM HCY: 106%, P= 0.317; 2000 μM HCY: 102%, P < 0.737). The PINP secretion was also stimulated by HCY reaching a maximum of 260 ± 154 μg/l at 500 μmol/l versus 205 ± 94 μ,g/l in controls. After 20 days of culture the formation of bone matrix was increased at 100 and 500 μM HCY. OC secretion was not significantly changed. The results of the present study consistently demonstrate a moderate stimulation of primary human OB activity by increasing concentrations of HCY. However, the magnitude of this effect seems to be less pronounced than recent observations on primary human osteoclasts, suggesting a dysbalance between OBs and osteoclasts in favour of osteoclasts

Quantitative Assessment of Bone Tissue Mineralization with Polychromatic Micro-Computed Tomography

Micro-computed tomography (microCT) has become an important tool for morphological characterization of cortical and trabecular bone. Quantitative assessment of bone tissue mineral density (TMD) from microCT images may be possible; however, the methods for calibration and accuracy have not been thoroughly evaluated. This study investigated hydroxyapatite (HA) phantom sampling limitations, short-term reproducibility of phantom measurements, and accuracy of TMD measurements by correlation to ash density. Additionally, the performance of a global and a local threshold for determining TMD was tested. The full length of a commercial density phantom was imaged by microCT, and mean calibration parameters were determined for a volume of interest (VOI) at 10 random positions along the longitudinal axis. Ten different VOI lengths were used (0.9-13 mm). The root mean square error (RMSE) was calculated for each scan length. Short-term reproducibility was assessed by five repeat phantom measurements for three source voltage settings. Accuracy was evaluated by imaging rat cortical bone (n = 16) and bovine trabecular bone (n = 15), followed by ash gravimetry. Phantom heterogeneity was associated with <0.5% RMSE. The coefficient of variation for five repeat measurements was generally <0.25% across all energies and phantom densities. Bone mineral content was strongly correlated to ash weight (R (2) = 1.00 for both specimen groups and both threshold methods). Ash density was well correlated for the trabecular bone specimens (R (2) > 0.80). In cortical bone specimens, the correlation was somewhat weaker when a global threshold was applied (R (2) = 0.67) compared to the local threshold method (R (2) = 0.78).

Influence of density, elasticity, and structure on ultrasound transmission through trabecular bone cylinders

The aim of this in vitro study is to evaluate the potentiality of quantitative ultrasound (QUS) to separate information on density, elasticity, and structure on specimens of trabecular bone. Fifteen cylinders of spongy bone extracted from equine vertebrae were progressively demineralized and subjected to QUS, micro computed tomography (muCT), Dual energy X-ray absorptiometry (DXA) at various mineralization levels. Eventually all cylinders underwent a compression test to calculate the Young's modulus. Correlation analysis shows that speed of sound (SOS) is strictly associated to bone mineral density (BMD), Young's modulus, and all muCT parameters except for degree of anisotropy (DA). Fast wave amplitude (FWA) is directly correlated with bone surface and total volume ratio (BS/TV) and trabecular separation (Tb Sp), and inversely correlated with trabecular number (Tb N). Because muCT parameters were strictly correlated to BMD and Young's modulus data, partial correlation analysis was performed between SOS, FWA, and structural and elastic data in order to eliminate the effect of density. SOS was significantly correlated to bone volume and total volume ratio (BV/TV), BS/TV, and Young's modulus, and FWA was significantly correlated to Tb Sp only. These results show that SOS is strongly influenced by volumetric mineral bone density and elastic modulus of the specimen, and FWA is mainly affected by trabecular separation independently on density. Therefore, SOS and FWA are able to provide different and complementary information, at least on trabecular bone samples.

Dependencies of Ultrasonic Properties on Apparent Bone Density in Trabecular Bone

The present study aims to provide insight into the dependencies of ultrasonic properties, such as speed of sound (SOS), broadband ultrasonic attenuation (BUA), and integrated reflection coefficient (IRC), on apparent bone density in trabecular bone. SOS, BUA, and IRC were measured in 19 bovine trabecular bone specimens with apparent densities from 0.411 to 0.928 g/cm3, using a matched pair of transducers with a diameter of 12.7 mm and a center frequency of 1.0 MHz. They were also compared with the predictions obtained from the modified Biot–Attenborough (MBA) model for propagation in fluid-saturated porous media.

Wave propagation in cancellous bone in terms of Biot's theory

The paper is focused on modelling of wave propagation in cancellous bones using Biot's theory. Almost all required input mechanical and structural parameters for 31 pure femoral trabecular bone specimens were measured individually. Then frequency dependent wave parameters predicted by the model were compared with the results of ultrasonic tests performed on the same specimens. To compare the predictions to measurements, additional interactions of the plane harmonic wave with the slab of cancellous was considered. The most important finding is the significant contribution of the fluid/bone and bone/fluid boundaries on the global attenuation loss. The corrected values of attenuation coefficient are of the same order of magnitude compared to measured values. The theoretical results exhibit higher attenuation of fast wave compared to that of the slow wave in good agreement with experimental observations. Moreover the amplitude ratio of simulated time domain signals of both longitudinal waves (accordingly to the Biot's model with boundary corrections), are of the same order of magnitude compared to the amplitude ratio of experimental time records. However, an analysis in the frequency domain shows that the frequency content of the simulated pulses of the fast and slow wave differs from that observed in the experiments.

Ultrasonic Propagation Through Trabecular Bone Modeled as a Random Medium

A method based on Gaussian random fields was used to generate three media having identical first and second order properties as three original trabecular bone specimens. Attenuation coefficients in both sets of specimens were computed after numerical simulations of ultrasonic wave propagation, using a finite difference time domain (FDTD) code without absorption. In generated media, they exhibited a linear trend for frequency dependence and their values were close to those obtained in the original specimens. These results suggest that combining a simplified description of trabecular bone micro-architecture with a multi-order scattering model could be relevant to predict attenuation due to scattering in trabecular bone.

The effects of clindamycin on human osteoblasts in vitro

Clindamycin is an antibiotic frequently used in different local application forms for the treatment of prosthetic joint infections, chronic osteomyelitis or as infection prophylaxis in bone cement. No information is available regarding its direct effects on bone cells, although very high local effective antibiotic concentrations can be achieved. We cultured pooled osteoblasts, previously derived from human trabecular bone specimens of four healthy donors, with different concentrations of clindamycin (0-500 microg/ml) for 24, 48 and 72 h. Cell proliferation (MTT), cytotoxicity [lactate dehydrogenase (LDH)-activity], cell metabolism [alkaline phosphatase (ALP)-activity] and extracellular matrix calcification (Alizarin staining) were assessed after antibiotic treatment. Proliferation significantly decreased in a dose-dependent manner and reached 3.5% of control samples at 500 microg/ml at 72 h. LDH-activity was unaffected at lower concentrations but significantly increased at 500 microg/ml at 48 and 72 h. ALP-activity significantly increased at 10 microg/ml at 24 and 48 h and then decreased in a time- and dose-dependent manner. Calcification increased at 10 and 25 microg/ml, while it decreased or no calcification was found at concentrations of 50 microg/ml and above. We could demonstrate that clindamycin at lower concentrations stimulated the cell metabolism of human osteoblasts and that higher clindamycin levels of 500 microg/ml had cytotoxic effects. The observed effects of high clindamycin levels on human osteoblasts highlight a potential alteration of bone metabolism in vivo and have to be taken into account in local antibiotic administration, e.g., in clindamycin-impregnated bone cement, where such high antibiotic concentrations can be achieved.

Indirect determination of trabecular bone effective tissue failure properties using micro-finite element simulations

Trabecular bone strength is marked not only by the onset of local yielding, but also by post-yield behavior. To study and predict trabecular bone elastic and yield properties, micro-finite element (micro-FE) models were successfully applied. However, trabecular bone strength predictions require micro-FE models incorporating post-yield behavior of trabecular bone tissue. Due to experimental difficulties, such data is currently not available. Here we used micro-FE modeling to determine failure behavior of trabecular bone tissue indirectly, by iteratively fitting FE simulation to experimental results. Failure parameters were fitted to an isotropic plasticity model based on Hill's yield function, using materially and geometrically nonlinear micro-FE models of seven bovine trabecular bone specimens. The predictive value of the averaged effective tissue properties was subsequently tested. The results showed that compression softening had to be included on the tissue level in order to accurately describe the apparent-level behavior of the bone specimens. A sensitivity study revealed that the simulated response was less sensitive to variations in the post-yield properties of the bone tissue than variations in the elastic and yield properties. Due to fitting of the tissue properties, apparent-level behavior could be accurately reproduced for each specimen separately. Predictions based on the averaged and fixed tissue properties were less accurate, due to inter-specimen variations in the tissue properties.