Acute bone loss following SARS-CoV-2 infection in mice.

The novel coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and has infected more than 650 million people worldwide. Approximately 23% of these patients developed lasting "long-haul" COVID symptoms, including fatigue, joint pain, and systemic hyperinflammation. However, the direct clinical impact of SARS-CoV-2 infection on the skeletal system including bone and joint health has not been determined. Utilizing a humanized mouse model of COVID-19, this study provides the first direct evidence that SARS-CoV-2 infection leads to acute bone loss, increased osteoclast number, and thinner growth plates. This bone loss could decrease whole-bone mechanical strength and increase the risk of fragility fractures, particularly in older patients, while thinner growth plates may create growth disturbances in younger patients. Evaluating skeletal health in patients that have recovered from COVID-19 will be crucial to identify at-risk populations and develop effective countermeasures.

Training drives turnover rates in racehorse proximal sesamoid bones.

Focal bone lesions are often found prior to clinically relevant stress-fractures. Lesions are characterized by low bone volume fraction, low mineral density, and high levels of microdamage and are hypothesized to develop when bone tissue cannot sufficiently respond to damaging loading. It is difficult to determine how exercise drives the formation of these lesions because bone responds to mechanical loading and repairs damage. In this study, we derive steady-state rate constants for a compartment model of bone turnover using morphometric data from fractured and non-fractured racehorse proximal sesamoid bones (PSBs) and relate rate constants to racing-speed exercise data. Fractured PSBs had a subchondral focus of bone turnover and microdamage typical of lesions that develop prior to fracture. We determined steady-state model rate constants at the lesion site and an internal region without microdamage using bone volume fraction, tissue mineral density, and microdamage area fraction measurements. The derived undamaged bone resorption rate, damage formation rate, and osteoid formation rate had significant robust regression relationships to exercise intensity (rate) variables, layup (time out of exercise), and exercise 2-10 months before death. However, the direction of these relationships varied between the damaged (lesion) and non-damaged regions, reflecting that the biological response to damaging-loading differs from the response to non-damaging loading.

Exercise history predicts focal differences in bone volume fraction, mineral density and microdamage in the proximal sesamoid bones of Thoroughbred racehorses.

Medial proximal sesamoid bones (PSBs) from Thoroughbred racehorses that did (Case) or did not (Control) experience unilateral biaxial PSB fracture were evaluated for bone volume fraction (BVF), apparent mineral density (AMD), tissue mineral density (TMD), and microdamage in Case fractured, Case contralateral limb intact, and Control bones. A majority of Case bones had a subchondral lesion with high microdamage density, and low BVF, AMD, and TMD. Lesion microdamage and densitometric measures were associated with training history by robust linear regression. Exercise intensity was negatively related to BVF (0.07 ≤ R2 ≤ 0.12) and positively related to microcrack areal density (0.21 ≤ R2 ≤ 0.29) in the lesion; however, in an undamaged site, the relationships were opposite in direction. Regardless of location, TMD decreased with event frequency for both Case and Control, suggesting increased bone remodeling with exercise. Measures of how often animals were removed from active training (layups) predicted a decrease in TMD, AMD, BVF, and microdamage at regions away from the lesion site. A steady-state compartment model was used to organize the differences in the correlations between variables within the data set. The overall conclusions are that at the osteopenic lesion site, repair of microdamage by remodeling was not successful (e.g., lower bone mass, increased damage, and lower mineralization) but that in regions away from the lesion remodeling successfully controlled damage (e.g., higher bone mass, less microdamage, and lower mineralization).

In vitro motions of the medial and lateral proximal sesamoid bones under mid-stance load conditions are consistent with racehorse fracture configurations.

Proximal sesamoid bone (PSB) fractures in racehorses are likely fatigue fractures that occur due to repetitive loads and stress remodeling. The loading circumstances that may induce damage in the PSBs are not well understood. The goal of this study was to determine in three-dimensions, PSB motions relative to the opposing metacarpal condyle during simulated mid-stance loads. Seven equine cadaveric forelimbs were axially loaded in a material testing system to simulate standing and mid-stance walk, trot, and gallop load conditions (1.8-10.5 kN). Joint angles were determined by tracking the positions of bone-fixed kinematic markers. Internal-external rotation, abduction-adduction, and flexion-extension of each PSB relative to the third metacarpal condyle were compared between loads and between PSBs using an ANOVA with Tukey-Kramer post hoc tests for pairwise comparisons. The medial PSB rotated externally and the lateral PSB apex abducted during limb loading. Medial PSB external rotation was significantly greater at the gallop load condition than at the walk or trot load conditions. The medial and lateral PSB motions observed in this study are consistent with location of fatigue damage and fracture configurations frequently seen in medial and lateral PSBs from Thoroughbred racehorses. Specifically, medial PSB external rotation is consistent with the development of an abaxial subchondral medial PSB lesion that is reported in association with medial PSB transverse fracture and lateral PSB abduction is consistent with axial longitudinal fracture of the lateral PSB.

In vitro assessment of the motion of equine proximal sesamoid bones relative to the third metacarpal bone under physiologic midstance loads.

OBJECTIVE: To assess the motion of the proximal sesamoid bones (PSBs) relative to the third metacarpal bone (MC3) of equine forelimbs during physiologic midstance loads. SAMPLE: 8 musculoskeletally normal forelimbs (7 right and 1 left) from 8 adult equine cadavers. PROCEDURES: Each forelimb was harvested at the mid-radius level and mounted in a material testing system so the hoof could be moved in a dorsal direction while the radius and MC3 remained vertical. The PSBs were instrumented with 2 linear variable differential transformers to record movement between the 2 bones. The limb was sequentially loaded at a displacement rate of 5 mm/s from 500 N to each of 4 loads (1.8 [standing], 3.6 [walking], 4.5 [trotting], and 10.5 [galloping] kN), held at the designated load for 30 seconds while lateromedial radiographs were obtained, and then unloaded back to 500 N. The position of the PSBs relative to the transverse ridge of the MC3 condyle and angle of the metacarpophalangeal (fetlock) joint were measured on each radiograph. RESULTS: The distal edge of the PSBs moved distal to the transverse ridge of the MC3 condyle at 10.5 kN (gallop) but not at lower loads. The palmar surfaces of the PSBs rotated away from each other during fetlock joint extension, and the amount of rotation increased with load. CONCLUSIONS AND CLINICAL RELEVANCE: At loads consistent with a high-speed gallop, PSB translations may create an articular incongruity and abnormal bone stress distribution that contribute to focal subchondral bone lesions and PSB fracture in racehorses.

Subchondral focal osteopenia associated with proximal sesamoid bone fracture in Thoroughbred racehorses.

BACKGROUND: Proximal sesamoid bone (PSB) fracture is the most common fatal injury in Thoroughbred (TB) racehorses in the United States. Epidemiological and pathological evidence indicates PSB fracture is likely the acute culmination of a chronic stress-related process. However, the aetiopathogenesis of PSB fracture is poorly understood. OBJECTIVE: To characterise bone abnormalities that precede PSB fracture. STUDY DESIGN: Two retrospective case-control groups of PSBs from TB racehorses with, and without, unilateral biaxial PSB fracture. METHODS: Proximal sesamoid bones were harvested post-mortem from TB racehorses subjected to euthanasia for unilateral biaxial PSB fracture (cases) or causes unrelated to PSB fracture (controls) while racing or training. The fractured medial PSB (FX-PSB) and contralateral intact medial PSB (CLI-PSB) from racehorses that sustained PSB fracture, and an intact medial PSB (CTRL-PSB) from racehorses that did not have a PSB fracture were collected as case and control specimens. Study 1 distributions of morphological features were compared among case and control groups using visual examination, photographs, radiographs and histology of whole PSBs and serial sagittal sections (10 FX-PSB, 10 CLI-PSB and 10 CTRL-PSB). Study 2 local bone volume fraction and mineral densities were compared among case and control PSBs using microcomputed tomography (9 FX-PSB, 9 CLI-PSB and 9 CTRL-PSB). RESULTS: A focal subchondral lesion characterised by colocalised focal discoloration, radiolucency, osteopenia, low tissue mineral density and a surrounding region of dense cancellous bone was identified in most case horses but not in controls. This subchondral lesion was found in a slightly abaxial mid-body location and was bilaterally present in most case horses. MAIN LIMITATIONS: The post-mortem samples may not represent the spectrum of abnormalities that occur throughout the development of the subchondral lesion. Lateral PSBs were not examined, so their contribution to biaxial PSB fracture pathogenesis is unknown. CONCLUSION: Abaxial subchondral lesions are consistent with pre-existing injury and likely associated with PSB fracture.

Age Dependence of Systemic Bone Loss and Recovery Following Femur Fracture in Mice.

The most reliable predictor of future fracture risk is a previous fracture of any kind. The etiology of this increased fracture risk is not fully known, but it is possible that fracture initiates systemic bone loss, leading to greater fracture risk at all skeletal sites. In this study, we investigated systemic bone loss and recovery after femoral fracture in young (3-month-old) and middle-aged (12-month-old) mice. Transverse femur fractures were created using a controlled impact, and whole-body bone mineral density (BMD), trabecular and cortical microstructure, bone mechanical properties, bone formation and resorption rates, mouse voluntary movement, and systemic inflammation were quantified at multiple time points post-fracture. We found that fracture led to decreased whole-body BMD in both young and middle-aged mice 2 weeks post-fracture; this bone loss was recovered by 6 weeks in young but not middle-aged mice. Similarly, trabecular bone volume fraction (BV/TV) of the L5 vertebral body was significantly reduced in fractured mice relative to control mice 2 weeks post-fracture (-11% for young mice, -18% for middle-aged mice); no significant differences were observed 6 weeks post-fracture. At 3 days post-fracture, we observed significant increases in serum levels of interleukin-6 and significant decreases in voluntary movement in fractured mice compared with control mice, with considerably greater changes in middle-aged mice than in young mice. At this time point, we also observed increased osteoclast number on L5 vertebral body trabecular bone of fractured mice compared with control mice. These data show that systemic bone loss occurs after fracture in both young and middle-aged mice, and recovery from this bone loss may vary with age. This systemic response could contribute to increased future fracture risk after fracture; these data may inform clinical treatment of fractures with respect to improving long-term skeletal health. © 2018 American Society for Bone and Mineral Research.

Hitting the ground running: Evaluating an integrated racehorse limb and race surface computational model.

Race surface mechanics contribute to musculoskeletal injury in racehorses. These mechanics affect ground reaction forces applied to the hoof, and thus limb motions during stance that can contribute to musculoskeletal pathologies. Race surface design has been largely empirical within the industry, with little uniform consensus for injury prevention and performance. Furthermore, race surface installations are too expensive to install experimentally. The objective of this research was to develop and evaluate an integrated racehorse limb and race surface computational model. Combined forward/inverse dynamic simulations of distal leading forelimb motions of a galloping horse during stance were compared to 2D distal leading forelimb kinematics of actual galloping racehorses on race surfaces with measured mechanics. Model predicted angular and translational kinematic profiles had similar qualitative shapes as experimental data, with comparable peak magnitudes. Model predictions of peak metacarpophalangeal position and timing were within 11° and 8ms of mean experimental data. The model overestimated peak fetlock angular velocity on consolidated surfaces (up to 1390°/s), and hoof displacements (up to 4cm) during stance. The model's ability to produce comparable qualitative kinematic profiles to experimental data and biologically reasonable fetlock and hoof motions support the future use of this model to explore the effect of race surface parameters on increasing or decreasing distal limb motions and provide supportive evidence for potential mechanisms of injury.

Bone Material Properties and Skeletal Fragility.

Deformations of vertebrae and sudden fractures of long bones caused by essentially normal loading are a characteristic problem in osteoporosis. If the loading is normal, then the explanation for and prediction of unexpected bone failure lies in understanding the mechanical properties of the whole bone-which come from its internal and external geometry, the mechanical properties of the hard tissue, and from how well the tissue repairs damage. Modern QCT and MRI imaging systems can measure the geometry of the mineralized tissue quite well in vivo-leaving the mechanical properties of the hard tissue and the ability of bone to repair damage as important unknown factors in predicting fractures. This review explains which material properties must be measured to understand why some bones fail unexpectedly despite our current ability to determine bone geometry and bone mineral content in vivo. Examples of how to measure the important mechanical properties are presented along with some analysis of potential drawbacks of each method. Particular attention is given to methods useful to characterize the loss of bone toughness caused by mechanical fatigue, drug side effects, and damage to the bone matrix.

Modeling equine race surface vertical mechanical behaviors in a musculoskeletal modeling environment.

Race surfaces have been associated with the incidence of racehorse musculoskeletal injury, the leading cause of racehorse attrition. Optimal race surface mechanical behaviors that minimize injury risk are unknown. Computational models are an economical method to determine optimal mechanical behaviors. Previously developed equine musculoskeletal models utilized ground reaction floor models designed to simulate a stiff, smooth floor appropriate for a human gait laboratory. Our objective was to develop a computational race surface model (two force-displacement functions, one linear and one nonlinear) that reproduced experimental race surface mechanical behaviors for incorporation in equine musculoskeletal models. Soil impact tests were simulated in a musculoskeletal modeling environment and compared to experimental force and displacement data collected during initial and repeat impacts at two racetracks with differing race surfaces - (i) dirt and (ii) synthetic. Best-fit model coefficients (7 total) were compared between surface types and initial and repeat impacts using a mixed model ANCOVA. Model simulation results closely matched empirical force, displacement and velocity data (Mean R(2)=0.930-0.997). Many model coefficients were statistically different between surface types and impacts. Principal component analysis of model coefficients showed systematic differences based on surface type and impact. In the future, the race surface model may be used in conjunction with previously developed the equine musculoskeletal models to understand the effects of race surface mechanical behaviors on limb dynamics, and determine race surface mechanical behaviors that reduce the incidence of racehorse musculoskeletal injury through modulation of limb dynamics.

Directional tortuosity as a predictor of modulus damage for vertebral cancellous bone.

There are many methods used to estimate the undamaged effective (apparent) moduli of cancellous bone as a function of bone volume fraction (BV/TV), mean intercept length(MIL), and other image based average microstructural measures. The MIL and BV/TV are both only functions of the cancellous microstructure and, therefore, cannot directly account for damage induced changes in the intrinsic trabecular hard tissue mechanical properties. Using a nonlinear finite element (FE) approximation for the degradation of effective modulus as a function of applied effective compressive strain, we demonstrate that a measurement of the directional tortuosity of undamaged trabecular hard tissue strongly predicts directional effective modulus (r2>0.90) and directional effective modulus degradation (r2>0.65). This novel measure of cancellous bone directional tortuosity has the potential for development into an anisotropic approach for calculating effective mechanical properties as a function of trabecular level material damage applicable to understanding how tissue microstructure and intrinsic hard tissue moduli interact to determine cancellous bone quality.

Trabecular bone loss at a distant skeletal site following noninvasive knee injury in mice.

Traumatic injuries can have systemic consequences, as the early inflammatory response after trauma can lead to tissue destruction at sites not affected by the initial injury. This systemic catabolism may occur in the skeleton following traumatic injuries such as anterior cruciate ligament (ACL) rupture. However, bone loss following injury at distant,unrelated skeletal sites has not yet been established. In the current study, we utilized a mouse knee injury model to determine whether acute knee injury causes a mechanically significant trabecular bone loss at a distant, unrelated skeletal site (L5 vertebral body).Knee injury was noninvasively induced using either high-speed (HS; 500 mm/s) or lowspeed(LS; 1 mm/s) tibial compression overload. HS injury creates an ACL rupture by midsubstance tear, while LS injury creates an ACL rupture with an associated avulsion bone fracture. At 10 days post-injury, vertebral trabecular bone structure was quantified using high-resolution microcomputed tomography (lCT), and differences in mechanical properties were determined using finite element modeling (FEM) and compressive mechanical testing. We hypothesized that knee injury would initiate a loss of trabecular bone structure and strength at the L5 vertebral body. Consistent with our hypothesis, we found significant decreases in trabecular bone volume fraction (BV/TV) and trabecular number at the L5 vertebral body in LS injured mice compared to sham (8.8% and 5.0%, respectively), while HS injured mice exhibited a similar, but lower magnitude response (5.1% and 2.5%, respectively). Contrary to our hypothesis, this decrease intrabecular bone structure did not translate to a significant deficit in compressive stiffness or ultimate load of the full trabecular body assessed by mechanical testing or FEM. However,we were able to detect significant decreases in compressive stiffness in both HS and LS injured specimens when FE models were loaded directly through the trabecular bone region (9.9% and 8.1%, and 3, respectively). This finding may be particularly important for osteoporotic fracture risk, as damage within vertebral bodies has been shown to initiate within the trabecular bone compartment. Altogether, these data point to a systemic trabecular bone loss as a consequence of fracture or traumatic musculoskeletal injury, which may be an underlying mechanism contributing to increased risk of refracture following an initial injury. This finding may have consequences for treatment of acute musculoskeletal injuries and the prevention of future bone fragility.

Torsional properties of distal femoral cortical defects.

The optimal management of pathologic long bone lesions remains a challenge in orthopedic surgery. The goal of the current study was to investigate the effect of defect depth on the torsional properties of the distal femur. A laterally placed distal metaphyseal cylindrical defect was milled in the cortex of the distal femur in 20 composite models. The proximal extent of the defects was constant. By decreasing the radius of the cylinder that intersected this predefined cord, 4 different radii defining 4 different depths of resection of the distal femur were created for testing: 17%, 33%, 50%, and 67% cortical defects, when normalized to the width of the femur at the level of resection. Each femur was mounted into a hydraulic axial/torsion materials testing machine and each specimen underwent torsional stiffness testing and torsional failure in external rotation. The specimens with less than a 33% cortical loss consistently demonstrated a superiorly oriented spiral fracture pattern, while the specimens with greater than a 50% cortical loss consistently demonstrated an inferiorly oriented transverse fracture pattern. The cortical defects were all statistically (P<.05) less stiff in torsion as the defect grew larger. There was a strong linear correlation between the mean torsional stiffness and cortical defect size (r(2)=0.977). This observation is supported by finite element analysis. The amount of femur remaining is crucial to stability. This biomechanical analysis predicts a critical loss of torsional integrity when a cortical defect approaches 50% of the width of the femur.

Strain-induced optical changes in demineralized bone.

Bone "stress-whitens," becoming visibly white during mechanical loading, immediately prior to failure. Stress-whitening is known to make materials tougher by dissipating mechanical energy. A greater understanding of stress-whitening, both an optical and mechanical phenomenon, may help explain age-related increases in fracture risk that occur without changes in bone mineralization. In this work, we directly measure the optical properties of demineralized bone as a function of deformation and immersing fluid (with different hydrogen-bonding potentials, water, and ethanol). The change in refractive index of demineralized bone was linear: with deformation and not applied force. Changes in refractive index were likely due to pushing low-refractive-index fluid out of specimens and secondarily due to changes in the refractive index of the collagenous phase. Results were consistent with stress-whitening of demineralized bone previously observed. In ethanol, the refractive index values were lower and less sensitive to deformation compared with deionized water, corroborating the sensitivity to fluid hydration. Differences in refractive index were consistent with structural changes in the collagenous phase such as densification that may also occur under mechanical loading. Understanding bone quality, particularly stress-whitening investigated here, may lead to new therapeutic targets and noninvasive methods to assess bone quality.

Remineralized bone matrix as a scaffold for bone tissue engineering.

There is a need for improved biomaterials for use in treating non-healing bone defects. A number of natural and synthetic biomaterials have been used for the regeneration of bone tissue with mixed results. One approach is to modify native tissue via decellularization or other treatment for use as natural scaffolding for tissue repair. In this study, our goal was to improve on our previously published alternating solution immersion (ASI) method to fabricate a robust, biocompatible, and mechanically competent biomaterial from natural demineralized bone matrix (DBM). The improved method includes an antigen removal (AR) treatment step which improves mineralization and stiffness while removing unwanted proteins. The chemistry of the mineral in the remineralized bone matrix (RBM) was consistent with dicalcium phosphate dihydrate (brushite), a material used clinically in bone healing applications. Mass spectrometry identified proteins removed from the matrix with AR treatment to include α-2 HS-glycoprotein and osteopontin, noncollagenous proteins (NCPs) and known inhibitors of biomineralization. Additionally, the RBM supported the survival, proliferation, and differentiation of human mesenchymal stromal cells (MSCs) in vitro as well or better than other widely used biomaterials including DBM and PLG scaffolds. DNA content increased more than 10-fold on RBM compared to DBM and PLG; likewise, osteogenic gene expression was significantly increased after 1 and 2 weeks. We demonstrated that ASI remineralization has the capacity to fabricate mechanically stiff and biocompatible RBM, a suitable biomaterial for cell culture applications.

Remineralization of demineralized bone matrix (DBM) via alternating solution immersion (ASI).

In order to achieve successful clinical outcomes, biomaterials used for bone grafts must possess a number of traits including biocompatibility and osteoconductivity. These materials must also demonstrate appropriate mechanical stability to withstand handling as well as support potentially significant stresses at the implant site. Synthetic and natural polymer scaffolds used for bone tissue engineering (BTE) often lack necessary mechanical properties. Our goal was to internally mineralize natural collagenous matrix, thereby increasing mechanical properties of the material to useful levels. Published methods for intrafibrillar collagen mineralization were applied to clinically relevant-sized constructs but did not successfully deposit mineral in the interior of the constructs. To address this limitation, we developed a new technique for the remineralization of demineralized bone matrix (DBM) based on alternating solution immersion, or ASI. Mineral was removed from equine bone specimens, leaving behind a demineralized bone matrix (DBM). This matrix provides a framework for the nucleation and growth of a replacement mineral phase. Plain film radiography and microcomputed tomography (microCT) indicated accumulation of mineral within the DBM, and mechanical testing (3 point bending and compression) revealed a significant increase in stiffness between the DBM and the remineralized bone matrix (RBM). We believe this remineralization process will be useful in the preparation of stiff and strong allografts for clinical application.

A comparison of cartilage stress-relaxation models in unconfined compression: QLV and stretched exponential in combination with fluid flow.

Cartilage exhibits nonlinear viscoelastic behaviour. Various models have been proposed to explain cartilage stress relaxation, but it is unclear whether explicit modelling of fluid flow in unconfined compression is needed. This study compared Fung's quasi-linear viscoelastic (QLV) model with a stretched-exponential model of cartilage stress relaxation and examined each of these models both alone and in combination with a fluid-flow model in unconfined compression. Cartilage explants were harvested from bovine calf patellofemoral joints and equilibrated in tissue culture for 5 days before stress-relaxation testing in unconfined compression at 5% nominal strain. The stretched exponential models fit as well as the QLV models. Furthermore, the average stretched exponential relaxation time determined by this model lies within the range of experimentally measured relaxation times for extracted proteoglycan aggregates, consistent with the hypothesis that the stretched exponential model represents polymeric mechanisms of cartilage viscoelasticity.

A Novel Method for Curvefitting the Stretched Exponential Function to Experimental Data.

The stretched exponential function has many applications in modeling numerous types of experimental relaxation data. However, problems arise when using standard algorithms to fit this function: we have observed that different initializations result in distinct fitted parameters. To avoid this problem, we developed a novel algorithm for fitting the stretched exponential model to relaxation data. This method is advantageous both because it requires only a single adjustable parameter and because it does not require initialization in the solution space. We tested this method on simulated data and experimental stress-relaxation data from bone and cartilage and found favorable results compared to a commonly-used Quasi-Newton method. For the simulated data, strong correlations were found between the simulated and fitted parameters suggesting that this method can accurately determine stretched exponential parameters. When this method was tested on experimental data, high quality fits were observed for both bone and cartilage stress-relaxation data that were significantly better than those determined with the Quasi-Newton algorithm.

Mechanical and morphological properties of trabecular bone samples obtained from third metacarpal bones of cadavers of horses with a bone fragility syndrome and horses unaffected by that syndrome.

OBJECTIVE: To determine morphological and mechanical properties of trabecular bone of horses with a bone fragility syndrome (BFS; including silicate-associated osteoporosis). SAMPLE: Cylindrical trabecular bone samples from the distal aspects of cadaveric third metacarpal bones of 39 horses (19 horses with a BFS [BFS bone samples] and 20 horses without a BFS [control bone samples]). PROCEDURES: Bone samples were imaged via micro-CT for determination of bone volume fraction; apparent and mean mineralized bone densities; and trabecular number, thickness, and separation. Bone samples were compressed to failure for determination of apparent elastic modulus and stresses, strains, and strain energy densities for yield, ultimate, and failure loads. Effects of BFS and age of horses on variables were determined. RESULTS: BFS bone samples had 25% lower bone volume fraction, 28% lower apparent density, 18% lower trabecular number and thickness, and 16% greater trabecular separation versus control bone samples. The BFS bone samples had 22% lower apparent modulus and 32% to 33% lower stresses, 10% to 18% lower strains, and 41 % to 52% lower strain energy densities at yield, ultimate, and failure loads, compared with control bone samples. Differences between groups of bone samples were not detected for mean mineral density and trabecular anisotropy. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that horses with a BFS had osteopenia and compromised trabecular bone function, consistent with bone deformation and pathological fractures that develop in affected horses. Effects of this BFS may be systemic, and bones other than those that are clinically affected had changes in morphological and mechanical properties.

Relating micromechanical properties and mineral densities in severely suppressed bone turnover patients, osteoporotic patients, and normal subjects.

Mineralization of bone, from the tissue level to whole bones, is associated with mechanical properties. The relationship between bone tissue mineralization and micromechanical properties may be affected by age, disease, and drug treatment. Patients with severely suppressed bone turnover (SSBT) suffered atypical fractures while on bisphosphonate treatment. The role of tissue level mineralization in predicting material level properties of SSBT bone may be different from that of other osteoporotic patients and of normal subjects. The aim of this study was to compare the relationships between mineralization and micromechanical properties of bone biopsies from patients with SSBT, bisphosphonate-naive osteoporotic patients with typical vertebral fracture, and normal young and age-matched subjects. We used nanoindentation and quantitative backscattered electron microscopy to characterize the elastic modulus, contact hardness, plastic deformation resistance, and tissue mineralization of the biopsies at site-matched locations within each biopsy. The linear mineralization-mechanical property relationships were different among the groups with respect to the intercepts for only cortical bone tissue but not the slopes for cortical and trabecular bone tissues. For a given mineral density, there was a trend of greater plastic deformation resistance in SSBT cortical bone compared to young normal bone. Similarly, there was a trend of greater plastic deformation resistance in osteoporotic trabecular bone compared to young normal bone for a given mineral density. The age-matched normal group had higher elastic modulus and a trend of higher contact hardness compared to the young normal group for a given mineral density. However, the mechanical property-mineralization relationships within an individual were weak, and only 21 of 53 biopsies that were analyzed had at least one significant association between mineralization and a mechanical property measurement for either cortical or trabecular bone tissues. The average properties of microstructural regions (deep and superficial remodeling packets in trabecular bone; osteonal and interstitial regions in cortical bone) were consistent with mineral accumulation with tissue age, with the exception of the SSBT group. SSBT trabecular bone deep packets had higher hardness and plastic deformation resistance than superficial packets, but mineralization levels and tissue modulus were not different between packet types. We conclude that relationships between mineral and mechanical properties were different between fracture and normal groups and between young and old normal groups, and that atypical fracture may be associated with changed microstructural material properties and tissue level mineralization compared to osteoporotic patients with vertebral fracture and normal subjects. We hypothesize that tissue level bone quality may be an important determinant in fracture risk, such that tissue mineral density may predict different material properties in different patient groups.