Trabecular Plate Research Articles

Individual trabecula segmentation (ITS)–based morphological analysis of microscale images of human tibial trabecular bone at limited spatial resolution

Individual trabecula segmentation (ITS), a rigorous model-independent 3D morphological analysis, has been developed to assess trabecular plate and rod microstructure separately based on micro-computed tomographic (µCT) images. We examined the influence of the limited spatial resolution, noise, and artifact of high-resolution peripheral quantitative CT (HR-pQCT) on ITS measurements of human tibial trabecular bone. In comparison with measurements from "gold standard" µCT images (25 µm), decreased spatial resolution (40, 60, and 80 µm) of µCT had minimal influence on the correlations of the scale of trabecular plates (ie, plate bone volume fraction, thickness, and surface area) and the orientation (ie, axial bone volume fraction) and structural type (ie, plate tissue fraction) of the trabecular network. ITS measurements of HR-pQCT images correlated significantly with those of µCT images at a similar voxel size (80 µm, r = 0.71-0.94); correlations were stronger for plate-related parameters, suggesting that measurements of trabecular rods are more subject to noise and artifact associated with HR-pQCT imaging technology. In comparison with measurements of "gold standard" µCT images, the percent absolute errors of HR-pQCT measurements such as axial and plate bone volume fraction, plate number and tissue fraction, and plate and rod thickness (3.5% to 10.3%) were comparable with those of bone volume fraction (9.3%). For both HR-pQCT and µCT images, measurements of the scale and junction densities of trabecular plates and orientation and structural type were strong and positive indicators of the elastic modulus of trabecular bone (r = 0.59-0.95). We conclude that ITS measurements of HR-pQCT images are highly reflective of trabecular bone microarchitecture from a biomechanical perspective.

Preventive and therapeutic effects of antler collagen on osteoporosis in ovariectomized rats

The present study was aimed at evaluating the preventive and therapeutic effects of collagen extracted from Sika deer velvet on osteoporosis in ovariectomized rats. Histomorphometric indices, serum biochemical parameters and biomechanical properties were measured in ovariectomized rats treated with/without antler collagen and in sham-operated rats. Compared with the ovariectomized group, significant elevation in the levels of bone mineral density (BMD), nitrogen monoxide (NO), NO synthase, bone-Ca, bone-P, bone morphogenetic protein (BMP), trabecular bone volume (TBV), number of trabecular (N), mean trabecular plate thickness (MTPT), and biomechanical properties, while reduction in the level of serum alkaline phosphatase (ALP), and mean trabecular plate separation (MTPS) were observed in antler collagen-treated groups. The extracted collagen was found to play a role in the prevention and treatment of osteoporosis in ovariectomized rats. Key words : Antler, collagen, osteoporosis, preventive, therapeutic

Individual trabeculae segmentation (ITS)-based morphological analysis of high-resolution peripheral quantitative computed tomography images detects abnormal trabecular plate and rod microarchitecture in premenopausal women with idiopathic osteoporosis.

Idiopathic osteoporosis (IOP) in premenopausal women is a poorly understood entity in which otherwise healthy women have low-trauma fracture or very low bone mineral density (BMD). In this study, we applied individual trabeculae segmentation (ITS)–based morphological analysis to high-resolution peripheral quantitative computed tomography (HR-pQCT) images of the distal radius and distal tibia to gain greater insight into skeletal microarchitecture in premenopausal women with IOP. HR-pQCT scans were performed for 26 normal control individuals and 31 women with IOP. A cubic subvolume was extracted from the trabecular bone compartment and subjected to ITS-based analysis. Three Young's moduli and three shear moduli were calculated by micro–finite element (µFE) analysis. ITS-based morphological analysis of HR-pQCT images detected significantly decreased trabecular plate and rod bone volume fraction and number, decreased axial bone volume fraction in the longitudinal axis, increased rod length, and decreased rod-to-rod, plate-to-rod, and plate-to-plate junction densities at the distal radius and distal tibia in women with IOP. However, trabecular plate and rod thickness did not differ. A more rod-like trabecular microstructure was found in the distal radius, but not in the distal tibia. Most ITS measurements contributed significantly to the elastic moduli of trabecular bone independent of bone volume fraction (BV/TV). At a fixed BV/TV, plate-like trabeculae contributed positively to the mechanical properties of trabecular bone. The results suggest that ITS-based morphological analysis of HR-pQCT images is a sensitive and promising clinical tool for the investigation of trabecular bone microstructure in human studies of osteoporosis. © 2010 American Society for Bone and Mineral Research.

Structural and mechanical parameters of trabecular bone estimated from in vivo high‐resolution magnetic resonance images at 3 tesla field strength

To evaluate the performance of a new 3 Tesla (T) high-resolution trabecular bone (TB) imaging technique at two resolution regimens in terms of serial reproducibility and sensitivity. The left distal tibial metaphysis of seven healthy volunteers was imaged at three time-points using a FLASE (fast large-angle spin-echo) pulse sequence at 137 x 137 x 410 mum(3) and (160 mum)(3) voxel sizes. Image artifacts, motion degradation, and serial image volume misalignments were controlled to maximize reproducibility of image-derived measures of scale, topology, orientation in terms of structural anisotropy, and finite-element derived Young's and shear moduli. Coefficients of variation (CV) and intraclass correlation coefficients (ICC) for structural and mechanical parameters were evaluated as measures of reproducibility and reliability. The ability of structural and mechanical parameters to distinguish between subjects was tested by analysis of variance. Reproducibility was generally higher in the anisotropic data (CVs 1-5% versus 1-9% for isotropic images). Anisotropic voxel size yielded greater measurement reliability (ICCs 0.75-0.99, mean = 0.92 versus 0.62-0.99, mean = 0.86) and better discrimination of the seven subjects (75% versus 50% of the possible comparisons were significantly different [P < 0.05]) except for measures of structural anisotropy and topology. Isotropic resolution improved detection of structural orientation and permitted visualization of small perforations in longitudinal trabecular plates not detected at anisotropic resolution. Improved image acquisition and processing techniques enhance reproducibility of structural and mechanical parameters derived from high-resolution MRI of metaphyseal bone in the distal tibia.

Quantitative analysis of subchondral sclerosis of the tibia by bone texture parameters in knee radiographs: site-specific relationships with joint space width

To determine the ability of radiographic bone texture (BTX) parameters to quantify subchondral tibia sclerosis and to examine clinical relevance for assessing osteoarthritis (OA) progression. We examined the relationship between BTX parameters and each of (1) location-specific joint space width (JSW) [JSW(x)] and minimum JSW (mJSW) of the affected compartment, and (2) knee alignment (KA) angle in knee radiographs of participants undergoing total knee arthroplasty (TKA). Digitized fixed-flexion knee radiographs were analyzed for run-length and topological BTX parameters in a subchondral region using an algorithm. Medial JSW(x) was computed at x=0.200, 0.225, 0.250 and 0.275 according to a coordinate system defined by anatomic landmarks. mJSW was determined for medial and lateral compartment lesions. KA angles were determined from radiographs using an anatomic landmark-guided algorithm. JSW measures and the magnitude of knee malalignment were each correlated with BTX parameters. Reproducibility of BTX parameters was measured by root-mean square coefficients of variation (RMSCV%). Run-length BTX parameters were highly reproducible (RMSCV%<1%) while topological parameters showed poorer reproducibility (>5%). In TKA participants (17 women, 13 men; age: 66+/-9 years; body mass index (BMI): 31+/-6 kg m(-2); WOMAC: 41.5+/-16.1; Kellgren-Lawrence score mode: 4), reduced trabecular spacing (Tb.Sp) and increased free ends (FE) were correlated with decreased JSW after accounting for BMI, gender and knee malalignment. These relationships were dependent on site of JSW measurement. High reproducibility in quantifying bone sclerosis using Tb.Sp and its significant relationship with JSW demonstrated potential for assessing OA progression. Increased trabecular FE and reduced porosity observed with smaller JSW suggest collapsing subchondral bone or trabecular plate perforation in advanced knee OA.

Micromechanical analyses of vertebral trabecular bone based on individual trabeculae segmentation of plates and rods

Trabecular plates play an important role in determining elastic moduli of trabecular bone. However, the relative contribution of trabecular plates and rods to strength behavior is still not clear. In this study, individual trabeculae segmentation (ITS) and nonlinear finite element (FE) analyses were used to evaluate the roles of trabecular types and orientations in the failure initiation and progression in human vertebral trabecular bone. Fifteen human vertebral trabecular bone samples were imaged using micro computed tomography (μCT), and segmented using ITS into individual plates and rods by orientation (longitudinal, oblique, and transverse). Nonlinear FE analysis was conducted to perform a compression simulation for each sample up to 1% apparent strain. The apparent and relative trabecular number and tissue fraction of failed trabecular plates and rods were recorded during loading and data were stratified by trabecular orientation. More trabecular rods (both in number and tissue fraction) failed at the initiation of compression (0.1–0.2% apparent strain) while more plates failed around the apparent yield point (>0.7% apparent strain). A significant correlation between plate bone volume fraction (pBV/TV) and apparent yield strength was found ( r 2=0.85). From 0.3% to 1% apparent strain, significantly more longitudinal trabecular plate and transverse rod failed than other types of trabeculae. While failure initiates at rods and rods fail disproportionally to their number, plates contribute significantly to the apparent yield strength because of their larger number and tissue volume. The relative failed number and tissue fraction at apparent yield point indicate homogeneous local failure in plates and rods of different orientations.

In Vivo μMRI-Based Finite Element and Morphological Analyses of Tibial Trabecular Bone in Eugonadal and Hypogonadal Men Before and After Testosterone Treatment

Osteoporosis is a major public health problem in men. Hypogonadal men have decreased BMD and deteriorated trabecular bone architecture compared with eugonadal men. Testosterone treatment improves their BMD and trabecular structure. We tested the hypothesis that testosterone replacement in hypogonadal men would also improve their bone's mechanical properties. Ten untreated severely hypogonadal and 10 eugonadal men were selected. The hypogonadal men were treated with a testosterone gel for 24 mo to maintain their serum testosterone concentrations within the normal range. Each subject was assessed before and after 6, 12, and 24 mo of testosterone treatment by microMRI of the distal tibia. A subvolume of each microMR image was converted to a microfinite element (microFE) model, and six analyses were performed, representing three compression and three shear tests. The anisotropic stiffness tensor was calculated, from which the orthotropic elastic material constants were derived. Changes in microarchitecture were also quantified using newly developed individual trabeculae segmentation (ITS)-based and standard morphological analyses. The accuracy of these techniques was examined with simulated microMR images. Significant differences in four estimated anisotropic elastic material constants and most morphological parameters were detected between the eugonadal and hypogonadal men. No significant change in estimated elastic moduli and morphological parameters was detected in the eugonadal group over 24 mo. After 24 mo of treatment, significant increases in estimated elastic moduli E(22) (9.0%), E(33) (5.1%), G(23) (7.2%), and G(12) (9.4%) of hypogonadal men were detected. These increases were accompanied by significant increases in trabecular plate thickness. These results suggest that 24 mo of testosterone treatment of hypogonadal men improves estimated elastic moduli of tibial trabecular bone by increased trabecular plate thickness.

Influence of Minor Trabecular Elements on Fast and Slow Wave Propagations through Cancellous Bone

Fast and slow longitudinal waves could propagate through cancellous bone in the direction parallel to the trabecular orientation; therefore, the major trabecular elements in the propagation direction are closely related to both the fast and slow wave propagations. To clarify in detail how the trabecular microstructure can be related to these waves, in this study, the influence of minor trabecular rods and pore spaces perpendicular to major trabecular plates was investigated using cancellous bone phantoms developed on the basis of a stratified model. In both experimental and simulated results, both waves were disturbed by trabecular rods and pore spaces. Moreover, it was investigated by simulation using realistic cancellous bone models via two erosion procedures whether the disturbance of both wave propagations due to minor trabecular elements could be induced in the real bone.

Dynamic simulation of three dimensional architectural and mechanical alterations in human trabecular bone during menopause

A three dimensional (3D) computational simulation of dynamic process of trabecular bone remodeling was developed with all the parameters derived from physiological and clinical data. Contributions of the microstructural bone formation deficits: trabecular plate perforations, trabecular rod breakages, and isolated bone fragments, to the rapid bone loss and disruption of trabecular microarchitecture during menopause were studied. Eighteen human trabecular bone samples from femoral neck (FN) and spine were scanned using a micro computed tomography (μCT) system. Bone resorption and formation were simulated as a computational cycle corresponding to 40-day resorption/160-day formation. Resorption cavities were randomly created over the bone surface according to the activation frequency, which was strictly based on clinical data. Every resorption cavity was refilled during formation unless it caused trabecular plate perforation, trabecular rod breakage or isolated fragments. A 20-year-period starting 5 years before and ending 15 years after menopause was simulated for each specimen. Elastic moduli, standard and individual trabeculae segmentation (ITS)-based morphological parameters were evaluated for each simulated 3D image. For both spine and FN groups, the time courses of predicted bone loss pattern by microstructural bone formation deficits were fairly consistent with the clinical measurements. The percentage of bone loss due to trabecular plate perforation, trabecular rod breakage, and isolated bone fragments were 73.2%, 18.9% and 7.9% at the simulated 15 years after menopause. The ITS-based plate fraction (pBV/BV), mean plate surface area (pTb.S), plate number density (pTb.N), and mean rod thickness (rTb.Th) decreased while rod fraction (rBV/BV) and rod number density (rTb.N) increased after the simulated menopause. The dynamic bone remodeling simulation based on microstructural bone formation deficits predicted the time course of menopausal bone loss pattern of spine and FN. Microstructural plate perforation could be the primary cause of menopausal trabecular bone loss. The combined effect of trabeculae perforation, breakage, and isolated fragments resulted in fewer and smaller trabecular plates and more but thinner trabecular rods.

In vivo ultra‐high‐field magnetic resonance imaging of trabecular bone microarchitecture at 7 T

To investigate the feasibility of 7 T magnetic resonance imaging (MRI) to visualize and quantify trabecular bone structure in vivo by comparison with 3T MRI and in vivo three-dimensional (3D) high-resolution peripheral quantitative computed tomography (HR-pQCT). The distal tibiae of 10 healthy volunteers were imaged. Therefore, fully balanced steady state free precession (bSSFP) and spin-echo (bSSSE) pulse sequences were implemented and optimized for 7 T. Structural bone parameters, such as apparent bone-volume over total-volume fraction (app.BV/TV), apparent trabecular plate separation (app.TbSp), apparent trabecular plate thickness (app.TbTh), and apparent trabecular plate number (app.TbN), were derived. All structural trabecular bone parameters correlated well (r > 0.6) between 7T and 3T, and between 7 T and HR-pQCT (r > 0.69), with the exception of app.TbTh, which correlated modestly (r = 0.41) between field strengths and very low with HR-pQCT (r < 0.16). Regarding absolute values, app.TbN varied only 4% between field strengths, and only 0.6% between 7 T and HR-pQCT. App.TbSp correlated best between 7 T and HR-pQCT (r = 0.89). Using bSSSE, significant smaller trabecular thickness and significant higher trabecular number were found compared to bSSFP. We concluded that imaging and quantification of the trabecular bone architecture at 7 T is feasible and preferably done using bSSSE. There exists great potential for ultra-high-field (UHF) MRI applied to trabecular bone measurements.

Complete Volumetric Decomposition of Individual Trabecular Plates and Rods and Its Morphological Correlations With Anisotropic Elastic Moduli in Human Trabecular Bone

Trabecular plates and rods are important microarchitectural features in determining mechanical properties of trabecular bone. A complete volumetric decomposition of individual trabecular plates and rods was used to assess the orientation and morphology of 71 human trabecular bone samples. The ITS-based morphological analyses better characterize microarchitecture and help predict anisotropic mechanical properties of trabecular bone. Standard morphological analyses of trabecular architecture lack explicit segmentations of individual trabecular plates and rods. In this study, a complete volumetric decomposition technique was developed to segment trabecular bone microstructure into individual plates and rods. Contributions of trabecular type-associated morphological parameters to the anisotropic elastic moduli of trabecular bone were studied. Seventy-one human trabecular bone samples from the femoral neck (FN), tibia, and vertebral body (VB) were imaged using muCT or serial milling. Complete volumetric decomposition was applied to segment trabecular bone microstructure into individual plates and rods. The orientation of each individual trabecula was determined, and the axial bone volume fractions (aBV/TV), axially aligned bone volume fraction along each orthotropic axis, were correlated with the elastic moduli. The microstructural type-associated morphological parameters were derived and compared with standard morphological parameters. Their contributions to the anisotropic elastic moduli, calculated by finite element analysis (FEA), were evaluated and compared. The distribution of trabecular orientation suggested that longitudinal plates and transverse rods dominate at all three anatomic sites. aBV/TV along each axis, in general, showed a better correlation with the axial elastic modulus (r(2) = 0.95 approximately 0.99) compared with BV/TV (r(2) = 0.93 approximately 0.94). The plate-associated morphological parameters generally showed higher correlations with the corresponding standard morphological parameters than the rod-associated parameters. Multiple linear regression models of six elastic moduli with individual trabeculae segmentation (ITS)-based morphological parameters (adjusted r(2) = 0.95 approximately 0.98) performed equally well as those with standard morphological parameters (adjusted r(2) = 0.94 approximately 0.97) but revealed specific contributions from individual trabecular plates or rods. The ITS-based morphological analyses provide a better characterization of the morphology and trabecular orientation of trabecular bone. The axial loading of trabecular bone is mainly sustained by the axially aligned trabecular bone volume. Results suggest that trabecular plates dominate the overall elastic properties of trabecular bone.

Trabecular Structure Quantified With the MRI-Based Virtual Bone Biopsy in Postmenopausal Women Contributes to Vertebral Deformity Burden Independent of Areal Vertebral BMD

In postmenopausal women with a wide range of vertebral deformities, MRI-based structural measures of topology and scale at the distal radius are shown to account for as much as 30% of vertebral deformity, independent of integral vertebral BMD. Trabecular bone architecture has been postulated to contribute to overall bone strength independent of vertebral BMD measured by DXA. However, there has thus far been only sparse in vivo evidence to support this hypothesis. Postmenopausal women, 60-80 yr of age, were screened by DXA, and those with T-scores at either the hip or spine falling within the range of -2.5 +/- 1.0 were studied with the MRI-based virtual bone biopsy, along with heel broadband ultrasound absorption and pQCT of the tibia. The data from 98 subjects meeting the enrollment criteria were subjected to microMRI at the distal tibia and radius, and measures of topology and scale of the trabecular bone network were computed. A spinal deformity index (SDI) was obtained from morphometric measurements in midline sagittal MR images of the thoracic and lumbar spine to evaluate associations between structure and deformity burden. A number of structural indices obtained at the distal radius were correlated with the SDI. Among these were the topological surface density (a measure of trabecular plates) and trabecular bone volume fraction, which were inversely correlated with SDI (p < 0.0001). Combinations of two structural parameters accounted for up to 30% of the variation in SDI (p < 0.0001) independent of spinal BMD, which was not significantly correlated. pQCT trabecular BMD was also weakly associated, whereas broadband ultrasound absorption was not. No significant association between SDI and structural indices were found at the tibia. Structural measures at the distal radius obtained in vivo by microMRI explained a significant portion of the variation in total spinal deformity burden in postmenopausal women independent of areal BMD.

Hepatitis C Virus Triggers Apoptosis of a Newly Developed Hepatoma Cell Line Through Antiviral Defense System

Hepatitis C virus (HCV) has a tendency to cause chronic viral infection. Viral evasion of host immune systems plays a key role in the pathogenesis of HCV. However, the interaction between HCV and hepatocyte innate antiviral defense systems is not understood. The aim of this study was to examine how human hepatocytes respond to HCV infection. We have established a novel human hepatoma cell line, LH86, from a well-differentiated hepatocellular carcinoma tissue. An infectious HCV isolate, JFH-1, was used to infect LH86 cells. HCV replication and apoptosis after viral infection were examined. Mechanisms of HCV-induced apoptosis were determined. Type I interferon induction and the relevant signaling molecules were examined. LH86 cells permitted JFH-1 HCV infection. The viral infection caused massive apoptosis. The apoptosis was related to viral replication, because blocking viral entry with anti-CD81 or suppressing viral replication with interferon protected cells from HCV-induced apoptosis. The HCV-induced apoptosis appeared to be triggered by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors, death receptor 4 and death receptor 5, which were up-regulated in HCV infection. HCV also activated interferon response factor 3, which induced expression of interferon and TRAIL in LH86 cells. Our study showed that a specific HCV isolate, JFH-1, is cytopathic in this new hepatoma cell line. LH86 cells mount an intact innate antiviral defense through induction of interferon and triggering apoptosis of infected cells. This study reveals a novel mechanism by which host hepatocytes respond to acute HCV infection.

Weaning Triggers a Decrease in Receptor Activator of Nuclear Factor-κB Ligand Expression, Widespread Osteoclast Apoptosis, and Rapid Recovery of Bone Mass after Lactation in Mice

A significant portion of milk calcium comes from the mother's skeleton, and lactation is characterized by rapid bone loss. The most remarkable aspect of this bone loss is its complete reversibility, and the time after weaning is the most rapid period of skeletal anabolism in adults. Despite this, little is known of the mechanisms by which the skeleton repairs itself after lactation. We examined changes in bone and calcium metabolism defining the transition from bone loss to bone recovery at weaning in mice. Bone mass decreases during lactation and recovers rapidly after weaning. Lactation causes changes in bone microarchitecture, including thinning and perforation of trabecular plates that are quickly repaired after weaning. Weaning causes a rapid decline in urinary C-telopeptide levels and stimulates an increase in circulating levels of osteocalcin. Bone histomorphometry documented a significant reduction in the numbers of osteoclasts on d 3 after weaning caused by a coordinated wave of osteoclast apoptosis beginning 48 h after pup removal. In contrast, osteoblast numbers and bone formation rates, which are elevated during lactation, remain so 3 d after weaning. The cessation of lactation stimulates an increase in circulating calcium levels and a reciprocal decrease in PTH levels. Finally, weaning is associated with a decrease in levels of receptor activator of nuclear factor-kappaB ligand mRNA in bone. In conclusion, during lactation, bone turnover is elevated, and bone loss is rapid. Weaning causes selective apoptosis of osteoclasts halting bone resorption. The sudden shift in bone turnover favoring bone formation subsequently contributes to the rapid recovery of bone mass.

Structural and functional assessment of trabecular and cortical bone by micro magnetic resonance imaging

Osteoporosis is a multifactorial disorder of bone mineral homeostasis affecting the elderly. It is a major public health issue with significant socioeconomic consequences. Recent findings suggest that bone loss-the key manifestation of the disease-is accompanied by architectural deterioration, both affecting the bone's mechanical competence and susceptibility to fracture. This article reviews the potential of quantitative micro MRI (mu-MRI), including a discussion of the technical requirements for image acquisition, processing, and analysis for assessing the architectural implications of osteoporosis and as a means to monitor the response to treatment. With current technology, the resolution achievable in clinically acceptable scan times and necessary signal-to-noise ratio (SNR) is comparable to trabecular thickness. This limited spatial resolution regime demands processing and analysis algorithms designed to operate under such limiting conditions. It is shown that three different classes of structural parameters can be distinguished, characterizing scale, topology, and orientation. There is considerable evidence that osteoporotic bone loss affects all three classes but that topological changes, resulting from conversion of trabecular plates to rods, with the latter's eventual disconnection, are particularly prominent. Clinical applications discussed can be divided into those dealing with assessment of osteoporotic fracture risk as opposed to the study of the effect of disease progression and regression in response to treatment. Current data suggest that noninvasive assessment of cortical and trabecular bone (TB) architecture by mu-MRI may provide new surrogate endpoints to assess the efficacy of intervention in osteoporosis treatment and prevention.

Protective Effects of Sodium Daidzein Sulfonate on Trabecular Bone in Ovariectomized Rats

The ovariectomized (OVX) rat, as an established animal model of human osteoporosis, was adopted in the present experiment to study the protective effects of sodium daidzein sulfonate (SDS) on trabecular bone. Six-month-oldSprague-Dawley rats were sham-operated or ovariectomized. Five days later, the OVX rats were randomly assigned to one of three experimental groups and treated for 90 days with vehicle, 17β-estradiol (E₂) or SDS. Compared with OVX rats, SDS administration (15 mg/kg) prevented OVX-induced decrease in lumbar vertebral and femoral bone mineral density (BMD), and significantly increased bone mechanical strength parameters, including ultimate stress and elastic modulus. In the OVX group, the structure of trabecular plate in the femoral head was absorbed and became progressively thinner or was removed completely, accompanied by enlargement of marrow cavities and amalgamation of two or more marrow cavities. Administration of SDS and E₂prevented the change of trabecular bone microarchitecture induced by OVX, increasing the trabecular bone area and trabecular thickness, while decreasing the trabecular separation. These results indicate that SDS administration prevents OVX-induced decrease in BMD and bone mechanical strength, and has a moderate protective effect on the microarchitecture of trabecular bone in aged Sprague-Dawley rats.

Bone microarchitecture in males with corticosteroid-induced osteoporosis

Microarchitectural changes in trabecular bone were analyzed by microcomputed tomography (microCT) and histomorphometry in 24 patients with corticosteroid-induced osteoporosis. The microCT images revealed a reduction in trabecular thickness only on frequency distribution curves, with no increase in trabecular separation. Trabecular plate thinning and perforations were easily identified. Corticosteroid-induced osteoporosis (CSIOP) is mediated by direct actions of the drug on bone cells. The result is a decrease in trabecular bone mass and a reduction in trabecular thickness, but connectivity is believed to remain rather well preserved. Twenty-four transiliac bone biopsies from patients with CSIOP were studied conjointly by histomorphometry [with two-dimensional (2D) architectural descriptors] and microCT (with 3D analysis of trabecular characteristics, including trabecular thickness and separation). The frequency distribution of thickness and separation were compared with data obtained in nine control subjects. 2D histomorphometry revealed a decrease in bone volume and trabecular thickness in the bone biopsies of the CSIOP patients when compared to those of the controls. MicroCT appeared to be able to identify the reduction in thickness only when the frequency distribution of trabecular thickness was computed. No difference for the curves of the frequency distribution of trabecular separation was evidenced between patients and controls. MicroCT and 2D histomorphometric results were correlated, but 2D analysis appeared to be more sensitive. However, microCT identified a very specific thinning of the trabecular plates in their center that corresponds to the earlier stages of perforations. Trabecular plate thinning can be observed and perforations occur on very thin plates in CSIOP patients.

Quantification of the Roles of Trabecular Microarchitecture and Trabecular Type in Determining the Elastic Modulus of Human Trabecular Bone

The roles of microarchitecture and types of trabeculae in determining elastic modulus of trabecular bone have been studied in microCT images of 29 trabecular bone samples by comparing their Young's moduli calculated by finite element analysis (FEA) with different trabecular type-specific reconstructions. The results suggest that trabecular plates play an essential role in determining elastic properties of trabecular bone. Osteoporosis is an age-related disease characterized by low bone mass and architectural deterioration. Other than bone volume fraction (BV/TV), microarchitecture of bone is also believed to be important in governing mechanical properties of trabecular bone. We quantitatively examined the role of microarchitecture and relative contribution of trabecular types of individual trabecula in determining the elastic property of trabecular bone. Twenty-nine human cadaveric trabecular bone samples were scanned at 21-mum resolution using a microCT system. Digital topological analysis (DTA) consisting of skeletonization and classification was combined with a trabecular type-specific reconstruction technique to extract the skeleton and identify topological type of trabeculae of the original trabecular bone image. Four different microCT-based finite element (FE) models were constructed for each specimen: (1) original full voxel; (2) skeletal voxel; (3) rod-reconstructed, preserving rod volume and plate skeleton; and (4) plate-reconstructed, preserving plate volume and rod skeleton. For each model, the elastic moduli were calculated under compression along each of three image-coordinate axis directions. Plate and rod tissue fractions directly measured from DTA-based topological classification were correlated with the elastic moduli computed from full voxel model. The elastic moduli of skeleton models were significantly correlated with those of full voxel models along all three coordinate axes (r(2) = 0.38 approximately 0.53). The rod-reconstructed model contained 21.3% of original bone mass and restored 1.5% of elastic moduli, whereas the plate-reconstructed model contained 90.3% of bone mass and restored 53.2% of elastic moduli. Plate tissue fraction showed a significantly positive correlation (r(2) = 0.49) with elastic modulus by a power law, whereas rod tissue fraction showed a significantly negative correlation (r(2) = 0.42). These results quantitatively show that the microarchitecture alone affects elastic moduli of trabecular bone and trabecular plates make a far greater contribution than rods to the bone's elastic behavior.

Use of Axial X-Ray Microcomputed Tomography to Assess Three-Dimensional Trabecular Microarchitecture and Bone Mineral Density in Single Comb White Leghorn Hens

Axial x-ray microcomputed tomography is a cost-effective technique with the potential to assess bone mineral density (mg/cc) in both cortical and cancellous bone in Single Comb White Leghorn hens. The technique requires little sample preparation and involves relatively simple data processing. The system described in this research is based on compact fan-beam type tomography, using a tungsten-anode x-ray tube with a relatively small focal spot (˜ 5 μm), coupled with a high-resolution x-ray detector system (˜ 10 μm). To produce a real 3-D data set using microcomputed tomography, x-ray projection views were acquired at 720 equally spaced angular positions (0.5 °) around the object of interest. These groups of views were then used to reconstruct a computed tomography image. A test grid with orthogonal test lines was used to calculate bone volume and bone surface. From these calculations, parallel plate equations were used to derive trabecular architectural parameters such as average trabecular plate thickness and average trabecular plate separation. Three-dimensional microarchitecture was evaluated using specialized stereological analysis software. Significant relationships between apparent bone mineral density (mg/cc) and 3-D structure were observed in femoral specimens from 66-wk-old Single Comb White Leghorn hens.

568. Ex Vivo Mesenchymal Stem Cell Therapy with rAAV2 Encoding BMP-2 Enhances Bone Regeneration in Ovariectomized Mouse Model

Mesenchymal stem cells (MSC) represent an ideal source for ex- vivo therapy for treatment of genetic, age-related, and segmental bone defects to increase bone mass. Recent studies have demonstrated that pluripotent MSC can home to the bone marrow and participate in new bone formation, raising the possibility that MSC-based therapies may be developed for effective intervention of bone loss. The pluripotent property of MSC necessitates selection of lineage- specific cells or restricted expression of therapeutic genes into committed progenitors with specific phenotypes for regulated gene therapy. An ideal approach to utilize them is by ex vivo gene therapy for osteopenia by modulating the MSC to function as a continuous source for providing progeny osteoblasts by retaining the capacity of self-renewal and direct the differentiated early progenitors towards osteoblast lineage, which would enhance new bone formation. Our earlier studies indicated efficient transduction of MSC by rAAV2. In the present study, we determined the efficacy of MSC transduced by rAAV-bone morphogenetic protein-2 (BMP-2) to repopulate and enhance bone density in a mouse model of osteoporosis. Low- passage MSC were transduced ex vivo with rAAV2 encoding BMP-2 under the transcriptional control of collagen type 1a promoter. Following rAAV transduction, the cells were maintained in culture for 10 days without induction of differentiation by maintaining the culture with FGF2. Approximately 107 MSC were then injected to ovariectomized female C57BL/6 mice intravenously. Prior to the injections, the cells were transfected with a plasmid encoding murine a4 integrin for bone-specific homing. Groups of mice also received rAAV2-GFP transduced MSC ectopically expressing |[alpha]|4 integrin or unmodified MSC. Dual Energy X-ray absorptiometry (DEXA) analysis was performed on all the animals prior to MSC injection and periodically after the treatments. Results indicated that the group of mice transplanted with MSC expressing BMP-2 showed statistically significant (p<0.035) increase in the bone mineral density (BMD) values compared to the rAAV2-GFP transduced MSC or unmodified MSC transplanted groups. Cohorts of mice from each group were sacrificed after 5 weeks and 10 weeks for bone histomorphometry, osteoclast/osteoblast ratio and the identification of engrafted MSC. Histomorphometric analysis using trichrome stain also confirmed the DEXA data where more osteoblasts and less osteoclasts were seen in the trabecular and growth plate area ofthe bone in BMP-2 treated group compared to controls. The long term effects of this ex vivo therapy on BMD will be discussed.