The pathophysiological basis of alterations in trabecular bone of patients with osteonecrosis of the femoral head (ONFH) remains unclear. ONFH has classically been considered a vascular disease with secondary changes in the subchondral bone. However, there is increasing evidence suggesting that ONFH could be a bone disease, since alterations in the functionality of bone tissue distant from the necrotic lesion have been observed. We comparatively studied the transcriptomic profile of trabecular bone obtained from the intertrochanteric region of patients with ONFH without an obvious aetiological factor, and patients with osteoarthritis (OA) undergoing total hip replacement in our Institution. To explore the biological processes that could be affected by ONFH, we compared the transcriptomic profile of trabecular bone from the intertrochanteric region and the femoral head of patients affected by this condition. Differential gene expression was studied using an Affymetrix microarray platform. Transcriptome analysis showed a differential signature in trabecular bone from the intertrochanteric region between patients with ONFH and those with OA. The gene ontology analyses of the genes overexpressed in bone tissue of patients with ONFH revealed a range of enriched biological processes related to cell adhesion and migration and angiogenesis. In contrast, most downregulated transcripts were involved in cell division. Trabecular bone in the intertrochanteric region and in the femoral head also exhibited a differential expression profile. Among the genes differentially expressed, we highlighted those related with cytokine production and immune response. This study identified a set of differently expressed genes in trabecular bone of patients with idiopathic ONFH, which might underlie the pathophysiology of this condition. Acknowledgements: This work was supported by grants PI18/00643 and PI22/00939 from ISCIII-FEDER, Ministerio de Ciencia, Innovación y Universidades (MICINN)-AES

Abstract. Several experimental studies derived relationships between density and macroscale material properties of trabecular bone, taking the form E=αρ. β. , where E is Young's modulus, ρ is density, and α and β are constants. Classical structural mechanics demonstrates β can vary between 1 (behaviour of the trabecular lattice is dominated by the axial stiffness of individual trabeculae) and 3 (behaviour is dominated by the bending stiffness of individual trabeculae). The ratio between rods (round trabeculae characterised by radius) and plates (flat trabeculae characterised by thickness) is also believed to govern the macroscale material properties of trabecular bone. To assess feasible ranges of α and β for trabecular bone, and their dependence on rod to plate ratio, 25 virtual samples of trabecular bone were generated as Voronoi lattices. Each 8×8×8mm sample was composed of 320 randomly generated Voronoi cells forming a foam like structure. Edges formed the rod network. Faces formed the plate network. Tissue level Young's modulus was set to 18,000MPa. Relative density was varied: 0.05, 0.1, 0.15, 0.2, 0.25. Rod to plate ratio was varied: 100:0, 75:25, 50:50, 25:75, 0:100. Macroscale Young's modulus was averaged in three orthotropic directions and used to find α and β. Around 14,000 3-noded quadratic beam elements represented rods, with average length of 0.63mm, and around 42,000 8-noded quadratic shell elements represented plates, with average area of 0.10mm. 2. Results for α and β were 3274 and 1.463 for 100% rods, 3646 and 1.067 for 50:50 rods and plates, and 4981 and 1.062 for 100% plates, showing the presence of plates improves the stiffness characteristics of trabecular bone. Work investigating the impact of element based geometry optimisation is ongoing. The work has important implications for the onset of conditions including osteoporosis and osteoarthritis, as well as those designing 3D printed scaffolds and implants

Abstract. 1.0 Objectives. Predictive structural models resulting in a trabecular bone topology closely resembling real bone would be a step toward 3D printing of sympathetic prosthetics. This study modifies an established trabecular bone structural adaptation approach, with the objective of achieving an improved adapted topology, specifically connectivity, compared to CT imaging studies; whilst retaining continuum level mechanical properties consistent with those reported in experimental studies. Strain driven structural adaptation models successfully identify trabecular trajectories, although tend to overpredict connectivity and skew trabecular radii distribution towards the smallest radius included in the adaptation. Radius adaptation of each trabecula is driven by a mechanostat approach with a target strain (1250 µɛ) below which radius is decreased (resorption), and above which radius is increased (apposition). Simulations include a lazy zone, in which neither resorption nor apposition takes place (1000 to 1500 µɛ); and a dead zone (<250 µɛ) in which complete resorption of trabeculae with the smallest included radius takes place. This study assesses the impact of increasing the dead zone threshold from <250 µɛ to <1000 µɛ, the lower limit of the lazy zone. 2.0 Methods. In-silico structural models with an initial connectivity (number of trabeculae connecting at each joint) of 14 were generated using a nearest neighbour approach applied to a random cloud of points. Trabeculae were modelled using circular beams whose radii were adapted in response to normal strains caused by the axial force and bending moments due to a vertical pressure of 1 MPa applied to the top of the lattice, with the bottom of the lattice fixed in the vertical direction. Lattices in which nodes are either able (rigid jointed) or unable (pin jointed) to transmit bending moments were considered. Five virtual samples of each lattice type were used, and each simulation repeated twice: with a dead zone of either <250 µɛ or <1000 µɛ. 3.0 Results. In pin jointed lattices the increase in dead zone threshold resulted in reduction of predicted Young's Modulus from 580 MPa (95% CI [577 MPa, 583 MPa]) to 408 MPa (95% CI [397 MPa, 419 MPa]) whilst in rigid jointed lattices it increased form 839MPa (95% CI [832 MPa, 846 MPa]) to 933 MPa (95% CI [931 MPa, 936 MPa]). Mean connectivity decreased from 10.2 to 5.8 in pin jointed simulations and from 9.6 to 3.8 in fixed joined simulations. Topological studies of trabecular bone CT images report a mean connectivity of around 3.4. Pin jointed lattice mean radius increased from 33mm to 45mm, and rigid jointed lattice mean radius increased from 33mm to 64mm. Prevalence of smallest included radius beams decreased in both. 4.0 Conclusion. Improved in-silico representations of trabecular bone can be achieved in structural adaptions by increasing the dead zone threshold and adopting a bending dominated (rigid jointed) lattice structure. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Introduction. The fixation of press-fit orthopaedic devices depends on the mechanical properties of the bone that is in contact with the implants. During the press-fit implantation, bone is compacted and permanently deformed, finally resulting in the mechanical interlock between implant and bone. For the development and design of new devices, it is imperative to understand these non-linear interactions. One way to investigate primary fixation is by using computational models based on Finite Element (FE) analysis. However, for a successful simulation, a proper material model is necessary that accurately captures the non-linear response of the bone. In the current study, we combined experimental testing with FE modeling to establish a Crushable Foam model (CFM) to represent the non-linear bone biomechanics that influences implant fixation. Methods. Mechanical testing of human tibial trabecular bone was done under uniaxial and confined compression configurations. We examined 62 human trabecular bone samples taken from 8 different cadaveric tibiae to obtain all the required parameters defining the CFM, dependent on local bone mineral density (BMD). The derived constitutive rule was subsequently applied using an in-house subroutine to the FE models of the bone specimens, to compare the model predictions against the experimental results. Results. The crushable foam model provided an accurate simulation of the experimental compression test, and was able to replicate the ultimate compression strength measured in the experiments [Figure 1]. The CFM was able to simulate the post-failure behavior that was observed in the experimental specimens up to strain levels of 50% [Figure 2]. Also, the distribution of yield strains and permanent displacement was qualitatively very similar to the experimental deformation of the bone specimens [Figure 3]. Conclusion. The crushable foam model developed in the current study was able to accurately replicate the mechanical behavior of the human trabecular bone under compression loading beyond the yield point. This advanced bone model enables realistic simulations of the primary fixation of orthopaedic devices, allowing for the analysis of the influence of interference fit and frictional properties on implant stability. In addition, the model is suitable for failure analysis of reconstructions, such as the tibial collapse of total knee arthroplasty. For any figures or tables, please contact the authors directly

3D printing can be used for the regeneration of complex tissues with intricate 3D microarchitecture. Trabecular bone is a complex and porous structure with a high degree of anisotropy. Changes in bone microarchitecture are associated with pathologies such as osteoporosis [1]. The objective of this study is to determine the viability of using 3D printing to replicate trabecular bone structures with a good control over the microarchitecture and mechanical properties. Cylindrical samples of bovine trabecular bone were used in this study. Micro-computed tomography (microCT) was carried out and an isotropic voxel size of 22 µm was obtained (Xradia Versa 520, Zeiss, USA). After 3D reconstruction the main microstructure characteristics were analysed using ImageJ (NIH, US). The 3D printed bone replicas were created by segmenting the microCT imaged bone tissue and then converted into a STL file using Avizo (FEI, US). The 3D printer used for this study was the ProJet 5500X (3D Systems, US), which allows a number of different materials to be printed in the same built with a resolution of 25 µm. Preliminary results were obtained using one single material (VisiJet CR-WT, Tensile Modulus: 1–1.6 GPa, Tensile Strength: 37–47 MPa). The 3D printed bone replicas followed a critical cleaning step to remove any remaining support material in the pores. MicroCT was then carried out for the bone replicas obtaining the same isotropic voxel size as for their biological counterparts. ImageJ was used to obtain the main microstructure characteristics. The values of bone volume fraction (BV/TV), mean trabecular thickness (Tb.Th), mean trabecular spacing (Tb.Sp), and degree of anisotropy (DA) were measured for bone samples and their 3D printed replicas [2]. Preliminary results on the first bone sample with its 3D printed replica showed similar apparent trabecular structures. Their respective BV/TV was found to be 0.24 (bone) and 0.43 (replica). The Tb.Th and Tb.Sp were 0.222 mm and 0.750 mm respectively for the bone and 0.376 mm and 0.575 mm for the replica. Finally, their respective DA was found to be 0.68 (bone) and 0.66 (replica). The main microstructure characteristics analyzed showed some differences between the bone sample and the 3D printed replica. In particular, the 3D microstructures resulted over-dimensioned mainly due to factors such as microCT voxel size, resolution of the 3D printer and supporting material removal. However this is a preliminary investigation. Further analysis will focus on optimizing the microCT imaging as well as the 3D printing process to achieve more accurate bone replicas. In addition, multi-material printing will be employed to optimize some of the mechanical properties obtained through in situ microCT testing and FE subject-specific modelling

Aims. This study investigated the effects of β-caryophyllene (BCP) on protecting bone from vitamin D deficiency in mice fed on a diet either lacking (D-) or containing (D+) vitamin D. Methods. A total of 40 female mice were assigned to four treatment groups (n = 10/group): D+ diet with propylene glycol control, D+ diet with BCP, D-deficient diet with control, and D-deficient diet with BCP. The D+ diet is a commercial basal diet, while the D-deficient diet contains 0.47% calcium, 0.3% phosphorus, and no vitamin D. All the mice were housed in conditions without ultraviolet light. Bone properties were evaluated by X-ray micro-CT. Serum levels of klotho were measured by enzyme-linked immunosorbent assay. Results. Under these conditions, the D-deficient diet enhanced the length of femur and tibia bones (p < 0.050), and increased bone volume (BV; p < 0.010) and trabecular bone volume fraction (BV/TV; p < 0.010) compared to D+ diet. With a diet containing BCP, the mice exhibited higher BV and bone mineral density (BMD; p < 0.050) than control group. The trabecular and cortical bone were also affected by vitamin D and BCP. In addition, inclusion of dietary BCP improved the serum concentrations of klotho (p < 0.050). In mice, klotho regulates the expression level of cannabinoid type 2 receptor (Cnr2) and fibroblast growth factor 23 (Fgf23) through CD300a. In humans, data suggest that klotho is connected to BMD. The expression of klotho is also associated with bone markers. Conclusion. These data indicate that BCP enhances the serum level of klotho, leading to improved bone properties and mineralization in an experimental mouse model. Cite this article: Bone Joint Res 2022;11(8):528–540

Osteoporosis is a common disorder characterized by low bone mass and reduced bone quality that affects the bone strength negatively and leads to increased risk of fracture. Bone mineral density (BMD) has been the standard instrument for the diagnosis of osteoporosis and the determination of fracture risk. Despite the approximation of the bone mass, BMD does not provide information about the bone structure. Trabecular bone score (TBS), which provides an indirect evaluation of skeletal microarchitecture, is calculated from dual X-ray absorptiometry and a simple and noninvasive method that may contribute to the prediction of osteoporotic fractures in addition to the measure of bone density. The goal of this study was to determine the mean TBS values in healthy postmenopausal women and the overall association between TBS and demographic features, bone mineral density of the lumbar spine and femoral neck and bone mineral density to body mass index ratio (BMD/BMI) of the lumbar spine. Fifty-three postmenopausal healthy women participated. The bone mineral density of the lumbar spine and femoral neck were measured dual X-ray absorptiometry. Anteroposterior lumbar spine acquisitions were used to calculate TBS for L1-L4. Age, height, weight, BMI and the ratio of BMD to BMI, which was considered to be a simple tool for assessing fracture risk in especially obese individuals, were calculated. The relationship between TBS and other variables was examined using Spearman's rank correlation coefficients. Mean BMD of the lumbar spine and the femoral neck were 0.945 ± 0.133 and 0.785 ± 0.112 g/cm2, respectively (Table 1). Mean TBS was 1.354 ± 0.107. There was a significant positive moderate correlation between TBS and total lumbar BMD/BMI ratio (r=0.595, pTBS values of postmenopausal women were negatively correlated with age and BMI and positively with bone mineral density and BMD/BMI ratio. The ratio between lumbar BMD and BMI presented a stronger correlation with TBS than that of BMD with TBS. Because of the better correlation, the BMD/BMI ratio may be used as a simple tool for the assessment of the risk of fractures. Further investigation may be needed to evaluate the factors influencing exercise intervention on TBS on this population of patients

Type-2 Diabetic (T2D) patients experience up to a 3-fold increase in bone fracture risk[1]. Paradoxically, T2D-patients have a normal or increased bone mineral density when compared to non-diabetic patients. This implies that T2D has a deleterious effect on bone quality, whereby the intrinsic material properties of the bone matrix are altered. Creating clinical challenges as current diagnostic techniques are unable to accurately predict the fracture probability in T2D-patients. To date, the relationship between cyclic fatigue loading, mechanical properties and microdamage accumulation of T2D-bone tissue has not yet been examined and thus our objective is to investigate this relationship. Ethically approved femoral heads were obtained from patients, with (n=8) and without (n=8) T2D. To obtain the mechanical properties of the sample, one core underwent a monotonic compression test to 10% strain, the other core underwent a cyclic compression test at a normalized stress ratio between 0.0035mm/mm and 0.016mm/mm to a maximum strain of 3%. Microdamage was evaluated by staining the tissue with barium sulfate precipitate [2] and conducting microcomputed tomography scanning with a voxel size of 10μm. The monotonically tested T2D-group showed no statistical difference in mechanical properties to the non-T2D-group, even when normalised against BV/TV. There was also no difference in BV/TV. For the cyclic test, the T2D-group had a significantly higher initial modulus (p<0.01) and final modulus (p<0.05). There was no difference in microdamage accumulation. Previous population-level studies have found that T2D-patients have been shown to have an increased fracture risk when compared to non-T2D-patients. This research indicates that T2D does not impair the mechanical properties of trabecular bone from the femoral heads of T2D-patients, suggesting that other mechanisms may be responsible for the increased fracture risk seen in T2D-patients

Orthopaedic reconstruction procedures to combat osteoarthritis, inflammatory arthritis, metabolic bone disease and other musculoskeletal disorders have increased dramatically, resulting in high demand on the advancement of bone implant technology. In the past, joint replacement operations were commonly performed primarily on elderly patients, in view of the prosthesis survivorship. With the advances in surgical techniques and prosthesis technology, younger patients are undergoing surgeries for both local tissue defects and joint replacements. This patient group is now more active and functionally more demanding after surgery. Today, implanted prostheses need to be more durable (load-bearing), they need to better match the patient's original biomechanics and be able to survive longer. Additive manufacturing (AM) provides new possibilities to further combat the problem of stress-shielding and promote better bone remodelling/ingrowth and thus long term fixation. This can be accomplished by matching the varying strain response (stiffness) of trabecular or subchondral bone locally at joints. The purpose of this research is therefore to determine whether a porous structure can be produced that can match the required behaviour and properties of trabecular bone regardless of skeletal location and can it be incorporated into a long-term implant. A stochastic structure visually similar to trabecular bone was designed and optimised for AM (Figure 1) and produced over a range of porosities in multiple materials, Stainless Steel 316, Titanium (Grade 23 – Ti6Al4V ELI) and Commercially Pure Titanium (Grade 2) using a Renishaw AM250 metal additive manufacturing system. Over 150 cylindrical specimens were produced per material and subjected to a compression test to determine the specimens' Elastic Modulus (Stiffness) and Compressive Yield Strength. Micro-CT scans and gravimetric analysis were also performed to determine and validate the specimens' porosity. Results were then graphed on a Strength vs. Stiffness Ashby plot (Figure 2) comparing the values to those of trabecular bone in the tibia and femur. It was found that AM can produce porous structures with an elastic modulus as low as 100 MPa up to 2.7 GPa (the highest stiffness investigated in this study). Titanium structures with a stiffness <500MPa had compressive strengths towards the bottom range of similar stiffness trabecular bone. Between 500 MPa − 1 GPa Titanium AM porous structures match the compressive strength of equivalent stiffness trabecular bone and from 1 GPa − 2 GPa the Ti structures exceed the strength of equivalent stiffness trabecular bone up to ∼2.5 times and consequently increase by a power law. These results show that AM can produce structures with similar stiffness to trabecular bone over a range of skeletal locations whilst matching or exceeding the compressive strength of bone. The results have not yet taken into account fatigue life with the fatigue life of these types of structures tending to be between 0.1 – 0.4 of their compressive strength. This means that a titanium porous structure would need to be 2.5 – 10 times stiffer or stronger than the portion of trabecular bone it is replacing. This data is highly encouraging for AM manufactured, bone stiffness matched implant technology

Objectives. Bisphosphonates (BP) are the first-line treatment for preventing fragility fractures. However, concern regarding their efficacy is growing because bisphosphonate is associated with over-suppression of remodelling and accumulation of microcracks. While dual-energy X-ray absorptiometry (DXA) scanning may show a gain in bone density, the impact of this class of drug on mechanical properties remains unclear. We therefore sought to quantify the mechanical strength of bone treated with BP (oral alendronate), and correlate data with the microarchitecture and density of microcracks in comparison with untreated controls. Methods. Trabecular bone from hip fracture patients treated with BP (n = 10) was compared with naïve fractured (n = 14) and non-fractured controls (n = 6). Trabecular cores were synchrotron scanned and micro-CT scanned for microstructural analysis, including quantification of bone volume fraction, microarchitecture and microcracks. The specimens were then mechanically tested in compression. Results. BP bone was 28% lower in strength than untreated hip fracture bone, and 48% lower in strength than non-fractured control bone (4.6 MPa vs 6.4 MPa vs 8.9 MPa). BP-treated bone had 24% more microcracks than naïve fractured bone and 51% more than non-fractured control (8.12/cm. 2. vs 6.55/cm. 2. vs 5.25/cm. 2. ). BP and naïve fracture bone exhibited similar trabecular microarchitecture, with significantly lower bone volume fraction and connectivity than non-fractured controls. Conclusion. BP therapy had no detectable mechanical benefit in the specimens examined. Instead, its use was associated with substantially reduced bone strength. This low strength may be due to the greater accumulation of microcracks and a lack of any discernible improvement in bone volume or microarchitecture. This preliminary study suggests that the clinical impact of BP-induced microcrack accumulation may be significant. Cite this article: A. Jin, J. Cobb, U. Hansen, R. Bhattacharya, C. Reinhard, N. Vo, R. Atwood, J. Li, A. Karunaratne, C. Wiles, R. Abel. The effect of long-term bisphosphonate therapy on trabecular bone strength and microcrack density. Bone Joint Res 2017;6:602–609. DOI: 10.1302/2046-3758.610.BJR-2016-0321.R1

Trabecular bone is a multiscale hierarchical composite material that is known to display time-dependant properties. However, most biomechanical models treat this material as time independent. Time-dependant properties, such as creep and relaxation, are thought to play an important role in many clinically relevant orthopaedic issues: implant loosening, vertebral collapse, and non-traumatic fractures. In this study compressive multiple-load-creep-unload-recovery (MLCUR) tests were applied to human trabecular bone specimens. 15 female femoral heads were harvested, with full ethical approval and patient consent, at the time of total hip replacement. Central cores were extracted and cut parallel under constant irrigation. Specimens were embedded in end caps using surgical cement, an epoxy tube was secured around the end caps and filled with phosphate buffered saline (PBS) to ensure the specimens remained hydrated throughout. Embedded samples were scanned by microCT (SkyScan 1172, Bruker) at a resolution of 17µm to determine microarchitecture. Bone volume fraction (BVF) was used to represent microarchitecture. Specimens had an effective length of 16.37mm (±1.90SD) with diameter of 8.08mm (±0.05SD), and BVF of 19.22% (±5.61SD). The compressive MLCUR tests were conducted at 5 strain levels, 2000µε, 4000µε, 6000µε, 8000µε and 10000µε. At each strain level, the load required to maintain each strain was held for 200s (creep) then unloaded to 1N for 600s (recovery). The instantaneous, creep, unloading and recovered strains can be easily obtained from the strain-time curves. Stress-strain plots revealed the Young's modulus. Data was modelled using line of best fit with appropriate curve fitting. R2 values were used to indicate association. Mechanical testing demonstrated the expected time independent relationship between BVF and stiffness: higher stiffness was found for specimen with higher BVF and this was consistent for all strain levels. Creep strain was found to depend on instantaneous strain and BVF. At low levels of instantaneous strain, there was a greater amount of creep strain in low BVF samples (R2 = 0.524). This relationship was no longer apparent at higher strain levels (R2 = 0.058). Residual strain also depended on the applied instantaneous strain and BVF: at low levels of strain, residual strain was similar with all BVF (R2 = 0.108) and at high levels of strain, residual strain was greater in low BVF samples (R2 = 0.319). The amount of instantaneous strain applied to each sample is constant, variations in stiffness result in different applied loads. In low BVF bone, the stiffness is also low, therefore the stress required to reach designed strain is also lower: yet, there is more creep and less recovery. We have demonstrated that even at loads below recognised yield levels, time-dependence affects the mechanical response and residual strain is present. In cases of low BVF, deflection due to creep, and increased irrecoverable strain could have clinically relevant consequences, such as implant loosening and vertebral collapse. The role of time-dependant properties of bone is seldom considered. This data could be developed into a constitutive model allowing these time-dependant behaviours to be incorporated in finite element modelling, leading to better predictions of implant loosening, especially for lower quality bone

Summary Statement. OA knee with subchondral cyst formation presented differential microstructure and mechanical competence of trabecular bone. This finding sheds light on the pivot role of subchondral cyst in OA bone pathophysiology. Introduction. Subchondral bone cyst (SBC) is a major radiological finding in knee osteoarthritis (OA), together with joint space narrowing, osteophyte and sclerotic bone formation. There is mounting evidence showing that SBC originates in the same region as bone marrow lesions (BMLs). The presence of subchondral bone cyst (SBCs), in conjunction with BMLs, was associated with the severity of pain, and was able to predict tibial cartilage lolume loss and risk of joint replacement surgery in knee OA patient. It is speculated that the presence of SBCs might increase intraosseous pressure of subchondral bone, and trigger active remodeling and high turnover of surrounding trabecular bone. Yet the exact effect of SBC on the structural and mechanical properties trabecular bone, which provides the support to overlying articular cartilage, remains to be elucidated. Therefore, this study aimed to investiate the microstructure and mechanical competence of trabecular bone of knee OA in presence or absence of SBC. Patients & Methods. A total of 20 postmenopausal women (54–87 years old) with the late-stage of primary knee OA were recruited in this study. Tibial plateau specimens were collected during joint replacement surgery. The samples were grouped for comparison according to presence or absences of SBC in micro-CT images. For micro-CT examination, a cylindrical volume of region of interest (VOI) of 10mm in diameter and 1mm in height was used to cover the trabecular bone region surrounding SBC, and then a cubic VOI of 3.5×3.5×3.5mm. 3. was applied in different anatomic locations of tibial plateau, such as medial, intermediate and lateral part, for the analyses of trabecular bone microstructure. Subsequently, two cylinders of subchondral bone specimens were drilled for each sample with micro-CT guidance from lateral portion of cystic wall along the direction of physiological loading of knee joint. The specimens were processed for micro-CT and mechanical testing using MTS 858 Mini Bionix sequentially. Each specimen was compressed in a longitudinal direction at a speed of 1mm/minute; the ultimate strength and modulus of the specimens were generated. Comparisons of microstructure and mechanical properties of trabecular bone were performed between two groups using student t test. The structure-mechanics relationship was also investigated using Pearson correlation. Results. The bone volume fraction (BV/TV, %) was significantly higher in knee OA specimens in presence of SBC (32±7%) in comparison with those in absence of SBC (16±5%, p<0.001). Meanwhile there were more plate-like trabecular bone surrounding SBC (0.78±0.61) than those without SBC (1.81±0.28, p<0.001), which was indicated by structure model index (0∼3). Furthermore, the trend in conversion of rod-like (close to 3) towards plate-like trabeculae was noticed in different locations of knee OA specimens with SBC formation. Trabecular bone around SBC presented higher modulus (73±22MPa) compared with those without SBC (45±29MPa, p=0.034). The stiffer trabecular bone in presence of SBC correlated with its plate-like morphology (r=0.696, p<0.001) as well as bone volume fraction (r=0.578, p=0.004). Conclusion. Presence of SBC was associated with conversion of trabeculae towards plate-like morphology together with the increase of mechanical competence in advanced knee OA

Strain is a robust indicator of bone failure initiation. Previous work has demonstrated the measurement of vertebral trabecular bone strain by Digital Volume Correlation (DVC) of µCT scan in both a loaded and an unloaded configuration. This project aims to improve previous strain measurement methods relying on image registration, improving resolution to resolve trabecula level strain and to improve accuracy by applying feature based registration algorithms to µCT images of vertebral trabecular bone to quantify strain. It is hypothesised that extracting reliable corresponding feature points from loaded and unloaded µCT scans can be used to produce higher resolution strain fields compared to DVC techniques. The feature based strain calculation algorithm has two steps: 1) a displacement field is calculated by finding corresponding feature points identified in both the loaded and unloaded µCT scans 2) strain fields are calculated from the displacement fields. Two methods of feature point extraction, Scale Invariant Feature Transform (SIFT) and Skeletonisation, were applied to unloaded (fixed) and loaded (moving) µCT images of a rat tail vertebra. Spatially non-uniform displacement fields were generated by automatically matching corresponding feature points in the unloaded and loaded scans. The Thin Plate Spline method and a Moving Least Squares Meshless Method were both tested for calculating strain from the displacement fields. Verification of the algorithms was performed by testing against known artificial strain/displacement fields. A uniform and a linearly varying 2% compressive strain field were applied separately to an unloaded 2D sagittal µCT slice to simulate the moving image. SIFT was unable to reliably match identified feature points leading to large errors in displacement. Skeletonisation generated a more accurate and precise displacement field. TPS was not tolerant to small displacement field errors, which resulted in inaccurate strain fields. The Meshless Methods proved much more resilient to displacement field errors. The combination of Skeletonisation with the Meshless Method resulted in best performance with an accuracy of −405µstrain and a detection limit of 1210µstrain at a strain resolution of 221.5µm. The DVC algorithm verified using the same validation test yielded a similar detection limit (1190µstrain), but with a lower accuracy for the same test (2370µstrain) for a lower resolution strain field (770µm) (Hardisty, 2009). The Skeletonisation algorithm combined with the Meshless Method calculated strain at a higher resolution, but with a similar detection limit, to that of traditional DVC methods. Future improvements to this method include the implementation of subpixel feature point identification and adapting this method of strain measurement into a 3D domain. Ultimately, a hybrid DVC/feature registration algorithm may further improve the ability to measure trabecular bone strain using µCT based image registration

We tested in compression specimens of human proximal tibial trabecular bone from 31 normal donors aged from 16 to 83 years and determined the mechanical properties, density and mineral and collagen content. Young’s modulus and ultimate stress were highest between 40 and 50 years, whereas ultimate strain and failure energy showed maxima at younger ages. These age-related variations (except for failure energy) were non-linear. Tissue density and mineral concentration were constant throughout life, whereas apparent density (the amount of bone) varied with ultimate stress. Collagen density (the amount of collagen) varied with failure energy. Collagen concentration was maximal at younger ages but varied little with age. Our results suggest that the decrease in mechanical properties of trabecular bone such as Young’s modulus and ultimate stress is mainly a consequence of the loss of trabecular bone substance, rather than a decrease in the quality of the substance itself. Linear regression analysis showed that collagen density was consistently the single best predictor of failure energy, and collagen concentration was the only predictor of ultimate strain

Introduction. Mechanical property relationships used in the computational modeling of bones are most often derived using mechanical testing of normal cadaveric bone. However, a significant percentage of patients undergoing joint arthroplasties exhibit some form of pathologic bone disease, such as osteoarthritis. As such, the objective of this study was to compare the micro-architecture and apparent modulus (E. app. ) of humeral trabecular bone in normal cadaveric specimens and bone extracted from patients undergoing total shoulder arthroplasty. Methods. Micro-CT scans were acquired at 20 µm spatial resolution for humeral heads from non-pathologic cadavers (n=12) and patients undergoing total shoulder arthroplasty (n=10). Virtual cylindrical cores were extracted along the medial-lateral direction. Custom-code was used to generate micro finite element models (µFEMs) with hexahedral elements. Each µFEM was assigned either a homogeneous tissue modulus of 20 GPa or a heterogeneous tissue modulus scaled by CT- intensity. Simulated compression to 0.5% apparent strain was performed in the medial-lateral direction. Morphometric parameters and apparent modulus-bone volume fraction relationships were compared between groups. Results. Comparing morphometric parameters, arthroplasty patients had significantly larger bone volume fractions (p = .023) and mean trabecular separation (p = .031), but no significant differences in mean trabecular thickness (p = .060) or trabecular number (p = .178). Variations were observed in the fit curves between normal and arthroplasty cases, with normal bone being best fit by power relationships, and arthroplasty bone exhibiting a more linear relationship. There was no significant difference in mean apparent modulus for homogeneous tissue moduli (p = .060) but was a significant difference for heterogeneous tissue moduli (p = .038). DISCUSSION. Consistent with previously developed relationships that map apparent mechanical properties, normal cadaveric bone was best fit by a power relationship with an exponential coefficient over 2. However, the apparent modulus- volume fraction relationship in the arthroplasty patient bone exhibited a more linear relationship. These results suggest that the architectural and mechanical properties of normal cadaveric and arthroplasty patient trabecular bone are not equal. Since these relationships are used to map apparent mechanical properties to computational models, these preliminary results suggest that relationships derived from cadaveric normal bone may map the apparent mechanical properties differently than patients who undergo arthroplasty. Additional samples added to this dataset will allow for mechanical property relationships to be developed that account for these bone mechanical property variations. This has the potential to greatly improve the computational modeling of patients undergoing arthroplasty procedures and computational models that are used to design and improve shoulder arthroplasty components

While reverse shoulder arthroplasty (RSA) is a reliable treatment option for patients with rotator cuff deficiency, loss of glenoid baseplate fixation often occurs due to screw loosening. We questioned whether an analysis of the trabecular bone density distribution in the scapula would indicate more optimal sites for screw placement. As such, the purpose of this study was to determine the anatomic distribution of trabecular bone density in regions of the scapula available for screw placement in RSA. Seven cadaveric shoulders were computed tomography (CT) scanned, and then voxels of the scapulae were isolated from the CT volume (Mimics 15.0 Materialise, Leuven, Belgium). Analyses were conducted in a common, 3D coordinate system. Volumetric regions of interest (ROI) within the scapula were identified based on potential baseplate screw sites. ROIs included areas at the base of the coracoid process lateral and inferior to the suprascapular notch, in the posterior and anterior lateral spine and in the anterosuperior and posteroinferior lateral border. Hounsfield Units (HU) were extracted from voxels corresponding to trabecular bone within each ROI. Overall bone density was summarised as the frequency of HU values above 80% of the ROI's maximum density value. Paired, two-tailed t-tests assuming unequal variance were used for pairwise comparisons (P≤0.05). Intra-region analyses compared two ROIs within the same broad anatomical structure; inter-region analyses compared ROIs between anatomical structures. Areas of the spine and lateral border of the scapula appeared to be denser than the coracoid process. Intra-region comparisons indicated no significant differences within ROI: coracoid P=0.43, spine P=0.95, lateral border P=0.41. ROI inferior to the suprascapular notch were on average 3.78% (P=0.08) and 6% (P=0.04) less dense than the anterosuperior and posteroinferior lateral border and 7.59% (P=0.006) and 7.72% (P=0.01) less dense than the anterior and posterior lateral spine. ROI lateral to the suprascapular notch were 6% (P=0.05) and 8.21% (P=0.02) less dense than the anterosuperior and posteroinferior lateral border and 9.8% (P=0.006) and 9.94% (P=0.008) less dense than the anterior and posterior lateral spine. There was no significant difference between the anterior spine and anterosuperior and posteroinferior lateral border (P=0.12, P=0.58), nor between the posterior spine and anterosuperior and posteroinferior lateral border (P=0.14, P=0.57). Results from this study indicate that the spine and lateral border of the scapula contain denser trabecular bone relative to regions in the coracoid. The higher quality bone of the spine and lateral border should be favoured over the coracoid process when fixing the glenoid baseplate in RSA. Further research may support the redesign of the glenoid baseplate geometry to better integrate the anatomy of the scapula and improve implant survival

One of the mechanisms which controls bone growth, repair remodeling and absorption is mechanical loading. There exists no long-term in vitro model to study bone cells together with their matrix, nor a model that can apply quantitative mechanical forces of physiological amplitudes and frequencies. The analysis of the mechanical properties of bone (Young’s modulus and visco-elastic moduli) on small pieces of bone is also difficult with present devices. We have built a device that can maintain full viability and physiological response of bone for a period of several weeks and integrates all three functions. 10mm diameter bone cores 5 mm thick were obtained from the trabecular bone of the distal ulna of a 24 months old cow by precision cutting with diamond saws and keyhole cutters (our pattern) in sterile 7–10°C phosphate buffered saline (PBS) and cultured in a variation of DMEM containing fructose HI GEM. Results: The results of these studies have shown that perfusion of trabecular bone can maintain all cells and maintain bone structure for at least 72 days. In conventional methods for bone organ cultures, small bones, such as rat calvaria, quickly start to resorb bone and degenerate. In our perfusion system we see no evidence of change.. Initial experiments have indicated that there are 2 visco-elastic moduli of bone with different time constants, that the elastic modulus of trabecular bone varies is site dependant and that loading to 0.4% compression raises prostaglandin E2 and insulin-like growth factor 1 within a few hours. Mechanical stiffness of bone is increased by 35% when loaded for 20 days at 4,000μ, and decreases by 25% when not loaded. PTH at 10-10M increases stiffness over the load effect and 10-6M PTH decreases stiffness even in the presence of loading. Active osteoclasts are seen during the whole culture period indicating that the stem cells are present and functional. We gratefully acknowledge support by the German Arthrose Foundation (DAH) and the AO in Davos, CH

Using the trabecular bone bioreactor (ZETOS) developed in our laboratories we have investigated the formation of bone using the fluorescent bone seeking markers calcein and alizarin red. And the association of bone formation with the increase in stiffness with mechanical loading. 10 mm diameter bone cores 5 mm thick were obtained from the distal radius /ulna of cows obtained at the slaughter house. by precision cutting with diamond saws and keyhole cutters (our pattern) in sterile 7–10°C phosphate buffered saline (PBS) and cultured in a variation of DMEM containing fructose HI GEM. Results: Loading the bone 30x 4,000μ per day resulted in an increase of stiffness of 35%, by day 30 while the non loaded controls decreased in stiffness. Calcein was added at day 27 to the circulating medium for 4 hours and then fresh medium was circulated. On day 30 alazarin red was circulated through the trabecular bone. The bones were subsequently fixed and embedded in resin and sectioned by classical histological techniques. The difference in distance between the two dyes indicated the amount of bone formation. The mechanically loaded bones showed significant evidence of formation and also significant numbers of active osteoclasts indicating high bone turnover. No evidence of necrosis or cartilage formation was found. Formation in unloaded bones was much reduced and on many areas no active osteoblasts could be observed. This is the first demonstration of bone formation ex vivo after 30 days of culture. We gratefully acknowledge support by the German Arthrose Foundation (DAH) and the AO in Davos, CH. DJ is a recipient of a Fork award from the AO

Introduction Sheep are being used increasingly for spinal and other skeletal-related research. However, there is still limited information about the molecular pathways of bone remodelling in this species compared to rats or mice. It has been demonstrated in other animal models and in the human that the receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) play major regulatory roles in controlling osteoclast activity and their differentiation. We investigated the expression of RANKL and OPG in trabecular bone of an ovariectomised steroid-treated osteopaenic sheep model. Methods Trabecular bone from the lumbar spine (LS) and proximal femur (PF) of ten osteopaenic ewes and four normal ewes were collected [. 1. ]. Total RNA was isolated and complementary DNA (cDNA) was synthesised. DNA encoding RANKL and OPG were sequenced and ovine specific primers were designed to amplify the cDNA by real time RT-PCR to generate products corresponding to mRNA encoding RANKL and OPG. The results were normalised to 18S RNA. Results Total OPG expression (in trabecular bone) from the PF region was over two fold higher than the LS (P< 0.0001). The relative expression of OPG in the both LS and PF regions were significantly higher in the treated animals (steroid & oophorectomy) compared to controls (p< 0.05). The relative expression of RANK-L in the PF was significantly higher than in the LS (P< 0.0001). However, the relative RANK-L expression in the treated animals was not significantly different from the control animals in either region. The ratio of RANK-L:OPG in the PF and the LS was not significantly different but it was significantly reduced in the osteopaenic animals. Discussion Based on this gene expression study and previous histomorphological data, it appears that trabecular bone loss is not due to increased osteoclastic activity but may rather due to lack of osteoblastic activity and function. Higher expression of OPG and RANK-L and greater bone loss compared to LS suggest that the rate of bone turnover is greater in the PF. Further investigation of the molecular pathways of bone loss in this animal model will increase its utility for osteoporosis research

Introduction. Trabecular bone score (TBS) is a parameter of bone microarchitecture that is determined by the level analysis of DXA images. TBS is associated with fractures in the preliminary case-control and prospective studies. The aim of this study was to assess the TBS role in the traumatology and orthopedics. Materials and methods. We've examined 176 healthy women aged 40–79 years (mean age – 53.4±0.6 yrs) and 117 men aged 40–79 years (mean age – 59.8±0.9 yrs). Bone mineral density (BMD) of whole body, PA lumbar spine and proximal femur were measured by DXA method (Prodigy, GEHC Lunar, Madison, WI, USA) and PA spine TBS were assessed by TBS iNsight® software package installed on the available DXA machine (Med-Imaps, Pessac, France). Results. We have observed a significant decrease of TBS as a function of age (F=6.56; p=0.0003) whereas PA spine BMD was significantly increasing with age (F=4.04; p=0.008) in the examined women. This contradiction can be traced to the spinal osteoarthritis and degenerative diseases progressing with age in the elderly patients. TBS was significantly lower in women with duration of PMP over 4 yrs (p=0.003) in comparison with women without menopause; BMD of spine significantly decreased in women with duration of PMP over 7–9 yrs (p=0.02). So, the TBS can detect changes in the state of bone tissue at the earlier stage than BMD. We have observed a significant decrease of TBS in men with ageing (F=2.44; p=0.05). Overall TBS values in men are lower than the age matched TBS values in women. Conclusion. TBS is an independent parameter which has a potential diagnostic value of its own, without taking into account the BMD results. The study concerning patients with osteoporosis and fractures is underway