Bone Material Properties Research Articles

Bone structure and composition in a hyperglycemic, obese, and leptin receptor-deficient rat: Microscale characterization of femur and calvarium.

Metabolic abnormalities, such as diabetes mellitus and obesity, can impact bone quantity and/or bone quality. In this work, we characterize bone material properties, in terms of structure and composition, in a novel rat model with congenic leptin receptor (LepR) deficiency, severe obesity, and hyperglycemia (type 2 diabetes-like condition). Femurs and calvaria (parietal region) from 20-week-old male rats are examined to probe bones formed both by endochondral and intramembranous ossification. Compared to the healthy controls, the LepR-deficient animals display significant alterations in femur microarchitecture and in calvarium morphology when analyzed by micro-computed X-ray tomography (micro-CT). In particular, shorter femurs with reduced bone volume, combined with thinner parietal bones and shorter sagittal suture, point towards a delay in the skeletal development of the LepR-deficient rodents. On the other hand, LepR-deficient animals and healthy controls display analogous bone matrix composition, which is assessed in terms of tissue mineral density by micro-CT, degree of mineralization by quantitative backscattered electron imaging, and various metrics extrapolated from Raman hyperspectral images. Some specific microstructural features, i.e., mineralized cartilage islands in the femurs and hyper-mineralized areas in the parietal bones, also show comparable distribution and characteristics in both groups. Overall, the altered bone microarchitecture in the LepR-deficient animals indicates compromised bone quality, despite the normal bone matrix composition. The delayed development is also consistent with observations in humans with congenic Lep/LepR deficiency, making this animal model a suitable candidate for translational research.

Effects of loading rate, age, and morphology on the material properties of human rib trabecular bone

The material and morphometric properties of trabecular bone have been studied extensively in bones bearing significant weight, such as the appendicular long bones and spine. Less attention has been devoted to the ribs, where quantification of material properties is vital to understanding thoracic injury. The objective of this study was to quantify the compressive material properties of human rib trabecular bone and assess the effects of loading rate, age, and morphology on the material properties. Material properties were quantified via uniaxial compression tests performed on trabecular bone samples at two loading rates: 0.005 s−1 and 0.5 s−1. Morphometric parameters of each sample were quantified before testing using micro-computed tomography. Rib trabecular bone material properties were lower on average compared to trabecular bone from other anatomical locations. Morphometric parameters indicated an anisotropic structure with low connectivity and a sparser density of trabeculae in the rib compared to other locations. No significant differences in material properties were observed between the tested loading rates. Material properties were only significantly correlated with age at the 0.005 s−1 loading rate, and no morphometric parameter was significantly correlated with age. Trabecular separation and thickness were most strongly correlated with the material properties, indicating the sparser trabecular matrix likely contributed to the lower material property values compared to other sites. The novel trabecular bone material properties reported in this study can be used to improve the thoracic response and injury prediction of computational models.

A Rab33b missense mouse model for Smith-McCort dysplasia shows bone resorption defects and altered protein glycosylation.

Smith McCort (SMC) dysplasia is a rare, autosomal recessive, osteochondrodysplasia that can be caused by pathogenic variants in either RAB33B or DYM genes. These genes codes for proteins that are located at the Golgi apparatus and have a role in intracellular vesicle trafficking. We generated mice that carry a Rab33b disease-causing variant, c.136A>C (p.Lys46Gln), which is identical to that of members from a consanguineous family diagnosed with SMC. In male mice at 4months of age, the Rab33b variant caused a mild increase in trabecular bone thickness in the spine and femur and in femoral mid-shaft cortical thickness with a concomitant reduction of the femoral medullary area, suggesting a bone resorption defect. In spite of the increase in trabecular and cortical thickness, bone histomorphometry showed a 4-fold increase in osteoclast parameters in homozygous Rab33b mice suggesting a putative impairment in osteoclast function, while dynamic parameters of bone formation were similar in mutant versus control mice. Femur biomechanical tests showed an increased in yield load and a progressive elevation, from WT to heterozygote to homozygous mutants, of bone intrinsic properties. These findings suggest an overall impact on bone material properties which may be caused by disturbed protein glycosylation in cells contributing to skeletal formation, supported by the altered and variable pattern of lectin staining in murine and human tissue cultured cells and in liver and bone murine tissues. The mouse model only reproduced some of the features of the human disease and was sex-specific, manifesting in male but not female mice. Our data reveal a potential novel role of RAB33B in osteoclast function and protein glycosylation and their dysregulation in SMC and lay the foundation for future studies.

Relative Effects of Radiation-Induced Changes in Bone Mass, Structure, and Tissue Material on Vertebral Strength in a Rat Model.

The observed increased risk of fracture after cancer radiation therapy is presumably due to a radiation-induced reduction in whole-bone strength. However, the mechanisms for impaired strength remain unclear, as the increased fracture risk is not fully explained by changes in bone mass. To provide insight, a small animal model was used to determine how much of this whole-bone weakening effect for the spine is attributable to changes in bone mass, structure, and material properties of the bone tissue and their relative effects. Further, because women have a greater risk of fracture after radiation therapy than men, we investigated if sex had a significant influence on bone's response to irradiation. Fractionated in vivo irradiation (10 × 3 Gy) or sham irradiation (0 Gy) was administered daily to the lumbar spine in twenty-seven 17-week-old Sprague-Dawley rats (n = 6-7/sex/group). Twelve weeks after final treatment, animals were euthanized, and lumbar vertebrae (L4 and L5 ) were isolated. Using a combination of biomechanical testing, micro-CT-based finite element analysis, and statistical regression analysis, we separated out the effect of mass, structural, and tissue material changes on vertebral strength. Compared with the sham group (mean ± SD strength = 420 ± 88 N), the mean strength of the irradiated group was lower by 28% (117 N/420 N, p < 0.0001). Overall, the response of treatment did not differ with sex. By combining results from both general linear regression and finite element analyses, we calculated that mean changes in bone mass, structure, and material properties of the bone tissue accounted for 56% (66 N/117 N), 20% (23 N/117 N), and 24% (28 N/117 N), respectively, of the overall change in strength. As such, these results provide insight into why an elevated clinical fracture risk for patients undergoing radiation therapy is not well explained by changes in bone mass alone. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Effects of Development on Bone Mineral Density and Mechanical Properties in the Aquatic Frog, Xenopus Laevis, and a Terrestrial Frog, Lithobates Catesbianus.

The limb bones of vertebrates have a critical role in supporting the weight of the body and transmitting forces that power locomotion. The loads that limb bones experience can vary in association with a range of factors, including locomotor environment or developmental stage. Limbed vertebrates that are habitually found in environments with low locomotor loads (e.g., water) might be predicted to also exhibit limb bones with less elevated mechanical properties, such as yield stiffness and yield stress. Frogs provide a distinctive case, in which these ideas can be tested as they experience changes in both locomotor style and habitat as they develop. However, while many frog taxa shift from aquatic to terrestrial habitats as they metamorphose, some lineages, such as pipids, maintain an aquatic lifestyle even after metamorphosis, providing a comparative framework for the effects of habitat shifts on developing limbs in vertebrates. This study compares the material composition and mechanical properties of the femur between frog species that are aquatic specialists (Xenopus laevis) vs generalists that spend considerable time both on land and in water (Lithobates catesbeianus) as they transition from metamorphic tadpoles to fully grown adults. MicroCT scanning was used to determine changes in bone density related to developmental stage and hindlimb use during swimming. Microindentation was then used to collect hardness values from the cortical bone of each femur, which was used to evaluate bone material properties. We found that aquatic frogs had less overall bone mineral density (BMD) than terrestrial frogs and that BMD was more elevated in the cortical region of the diaphysis than trabeculae and distal and proximal epiphyses. Despite its less elevated BMD, bone mechanical properties were not significantly different in aquatic specialist X. laevis than in more terrestrial L. catesbeianus. Our results suggest that the limb bones of aquatic frogs may experience compensatory effects through development to offset their lower BMD. Furthermore, changes in bone density and material properties across development may help to explain some of the differences in locomotor performance found between aquatic and terrestrial metamorphic frogs, providing insight into how environmental factors might correlate with bone ossification.

Breed and loading history influence in vivo skeletal strain patterns in pre-pubertal female chickens

The influence of loading history on in vivo strains within a given specie remains poorly understood, and although in vivo strains have been measured at the hindlimb bones of various species, strains engendered during modes of activity other than locomotion are lacking, particularly in non-human species. For commercial egg-laying chickens specifically, there is an interest in understanding their bones' mechanical behaviour, particularly during youth, to develop early interventions to prevent the high incidence of osteoporosis in this population. We measured in vivo mechanical strains at the tibiotarsus midshaft during steady activities (ground, uphill, downhill locomotion) and non-steady activities (perching, jumping, aerial transition landing) in 48 pre-pubescent female (egg-laying) chickens from two breeds that were reared in three different housing systems, allowing varying amounts and types of physical activity. Mechanical strain patterns differed between breeds, and were dependent on the activity performed. Mechanical strains were also affected by rearing environment: chickens that were restricted from performing dynamic load bearing activity due to caged-housing generally exhibited higher mechanical strain levels during steady, but not non-steady activities, compared to chickens with prior dynamic load-bearing activity experience. Among chickens with prior experience of dynamic load bearing activity, those reared in housing systems that allowed more frequent physical activity did not exhibit lower mechanical strains. In all groups, the tibiotarsus was subjected to a loading environment consisting of a combination of axial compression, bending, and torsion, with torsion being the predominant source of strain. Aerial transition landing produced the highest strain levels with unusual strain patterns compared to other activities, suggesting it may produce the strongest anabolic response. These results exemplify how different breeds within a given specie adapt to maintain different patterns of mechanical strains, and how benefits of physical activity in terms of resistance to strain are activity-type dependent and do not necessarily increase with increased physical activity. These findings directly inform controlled loading experiments aimed at studying the bone mechanoresponse in young female chickens and can also be associated to measures of bone morphology and material properties to understand how these features influence bone mechanical properties in vivo.

Conceptual design of compliant bone scaffolds by full-scale topology optimization

A promising new treatment for large and complex bone defects is to implant specifically designed and additively manufactured synthetic bone scaffolds. Optimizing the scaffold design can potentially improve bone in-growth and prevent under- and over-loading of the adjacent tissue. This study aims to optimize synthetic bone scaffolds over multiple-length scales using the full-scale topology optimization approach, and to assess the effectiveness of this approach as an alternative to the currently used mono- and multi-scale optimization approaches for orthopaedic applications. We present a topology optimization formulation, which is matching the scaffold's mechanical properties to the surrounding tissue in compression. The scaffold's porous structure is tuneable to achieve the desired morphological properties to enhance bone in-growth. The proposed approach is demonstrated in-silico, using PEEK, cortical bone and titanium material properties in a 2D parameter study and on 3D designs. Full-scale topology optimization indicates a design improvement of 81% compared to the multi-scale approach. Furthermore, 3D designs for PEEK and titanium are additively manufactured to test the applicability of the method. With further development, the full-scale topology optimization approach is anticipated to offer a more effective alternative for optimizing orthopaedic structures compared to the currently used multi-scale methods.

MODELLING OSTEOCHONDRAL GRAFT BEHAVIOUR IN TIBIOFEMORAL JOINTS

Autologous osteochondral grafting has demonstrated positive outcomes for treating articular cartilage defects by replacing the damaged region with a cylindrical graft consisting of bone with a layer of cartilage, taken from a non-loadbearing region of the knee. Despite positive clinical use, factors that cause graft subsidence or poor integration are relatively unknown. The aim of this study was to develop finite element (FE) models of osteochondral grafts within a tibiofemoral joint and to investigate parameters affecting osteochondral graft stability.Initial experimental tests on cadaveric femurs were performed to calibrate the bone properties and graft-bone frictional forces for use in corresponding FE models, generated from µCT scan data. The effects of cartilage defects and osteochondral graft repair were measured by examining contact pressure changes using in vitro tests on a single cadaveric human tibiofemoral joint. Six defects were created in the femoral condyles which were subsequently treated with osteochondral autografts or metal pins. Matching µCT scan-based FE models were created, and the contact patches were compared. Sensitivity to graft bone properties was investigated.The bone material properties and graft-bone frictional forces were successfully calibrated from the initial tests with good resulting levels of agreement (CCC=0.87). The tibiofemoral joint experiment provided a range of cases to model. These cases were well captured experimentally and represented accurately in the FE models. Graft properties relative to host bone had large effects on immediate graft stability despite limited changes to resultant cartilage contact pressure.Model confidence was built through extensive validation and sensitivity testing, and demonstrated that specimen-specific properties were required to accurately represent graft behaviour. The results indicate that graft bone properties affect the immediate stability, which is important for the selection of allografts and design of future synthetic grafts.AcknowledgementsSupported by the EPSRC-EP/P001076.

OPTIMIZATION OF OSTEOCHONDRAL ALLOGRAFT PREPARATION FOR TRANSPLANTATION USING FINITE ELEMENT MODELLING

Many factors have been reported to affect the functional survival of OCA transplants, including chondrocyte viability at time of transplantation, rate and extent of allograft bone integration, transplantation techniques, and postoperative rehabilitation protocols and adherence. The objective of this study was to determine the optimal subchondral bone drilling technique by evaluating the effects of hole diameter on the material properties of OCAs while also considering total surface area for potential biologic benefits for cell and vascular ingrowth.Using allograft tissues that would be otherwise discarded in combination with deidentified diagnostic imaging (MRI and CT), a model of a large shell osteochondral allograft was recreated using LS-PrePost and FEBio based on clinically relevant elastic material properties for cortical bone, trabecular bone, cartilage, and hole ingrowth tissue. The 0.8 mesh size model consisted of 4 mm trabecular bone, 4 mm cortical bone, and 3 mm cartilage sections that summed to a cross-sectional area of 1600 mm2 (40 mm x 40 mm). Holes were modeled to be 4mm deep in relation to clinical practice where holes are drilled from the deep margin of subchondral trabecular bone to the cortical subchondral bone plate. To test the biomechanic variations between drill hole sizes, models with hole sizes pertinent to standard-of-care commercially available orthopaedic drill sizes of 1.1mm, 2.4 mm, or 4.0 mm holes were loaded across the top surface over a one second duration and evaluated for effective stress, effective strain, 1st principal strain, and 3rd principal strain in compressive conditions.Results measured effective stress and strain and 1st and 3rd principal strain increased with hole depth.The results of the present FEA modeling study indicate that the larger 4.0 mm diameter holes were associated with greater stresses and strains within OCA shell graft, which may render the allograft at higher risk for mechanical failure. Based on these initial results, the smaller diameter 2.4 mm and 1.1 mm holes will be further investigated to determine optimal number, configuration, and depth of subchondral drilling for OCA preparation for transplantation

Positive medial cortical support versus anatomical reduction for trochanteric hip fractures: Finite element analysis and biomechanical testing

Background and ObjectivesThe anatomical reduction (AR) is usually considered the best option for fractures. Nevertheless, in unstable trochanteric hip fractures (UTHF), previous clinical reports found that the positive medial cortical support (PMCS, an over-reduction technique) attained higher mechanical stability, but this challenging clinical finding still needs experimental validation. MethodsThis study constructed in-silico and biomechanical PMCS and AR models, with the use of the most clinically-representative geometry design of fracture models, the multi-directional design in FE analysis, and the subject-specific (osteoporotic) bone material properties, to make the models better mimic the actual condition in clinical settings. Then multiple performance variables (von-Mises stress, strain, integral axial stiffness, displacement, structural changes, etc.) were assessed to uncover details of integral and regional stability. ResultsAmong in-silico comparison, PMCS models showed significantly lower maximum displacement than AR models, and the maximum von Mises stress of implants (MVMS-I) was significantly lower in PMCS models than in AR models (highest MVMS-I in –30°-A3-AR of 1055.80 ± 93.37 MPa). Besides, PMCS models had significantly lower maximum von Mises stress along fracture surfaces (MVMS-F) (highest MVMS-F in 30°-A2-AR of 416.40 ± 38.01 MPa). Among biomechanical testing comparison, PMCS models showed significantly lower axial displacement. Significantly lower change of neck-shaft angle (CNSA) was observed in A2-PMCS models. A fair amount of AR models converted into the obvious negative medial cortical support (NMCS) condition, whereas all PMCS models kept the PMCS condition. The results were also validated through comparison to previous clinical data. ConclusionsThe PMCS is superior to the AR in the UTHF surgery. The current study opens up the second thought of the role of over-reduction technique in bone surgery.

Influences of human thorax variability on population rib fracture risk prediction using human body models.

Rib fractures remain a common injury for vehicle occupants in crashes. The risk of a human sustaining rib fractures from thorax loading is highly variable, potentially due to a variability in individual factors such as material properties and geometry of the ribs and ribcage. Human body models (HBMs) with a detailed ribcage can be used as occupant substitutes to aid in the prediction of rib injury risk at the tissue level in crash analysis. To improve this capability, model parametrization can be used to represent human variability in simulation studies. The aim of this study was to identify the variations in the physical properties of the human thorax that have the most influence on rib fracture risk for the population of vehicle occupants. A total of 15 different geometrical and material factors, sourced from published literature, were varied in a parametrized SAFER HBM. Parametric sensitivity analyses were conducted for two crash configurations, frontal and near-side impacts. The results show that variability in rib cortical bone thickness, rib cortical bone material properties, and rib cross-sectional width had the greatest influence on the risk for an occupant to sustain two or more fractured ribs in both impacts. Therefore, it is recommended that these three parameters be included in rib fracture risk analysis with HBMs for the population of vehicle occupants.

An Overview of Selected Material Properties in Finite Element Modeling of the Human Femur

Specific finite detail modeling of the human body gives a capable primary enhancement to the prediction of damage risk through automobile impact. Currently, car crash protection countermeasure improvement is based on an aggregate of testing with installed anthropomorphic test devices (i.e., ATD or dummy) and a mixture of multibody (dummy) and finite element detail (vehicle) modeling. If an incredibly easy finite element detail version can be advanced to capture extra statistics beyond the abilities of the multi-body structures, it might allow advanced countermeasure improvement through a more targeted prediction of overall performance. Numerous research has been done on finite element analysis of broken femurs. However, there are two missing pieces of information: 1- choosing the right material properties, and 2- designing a precise model including the inner structure of the bone. In this research, most of the chosen material properties for femur bone will be discussed and evaluated.

Accuracy of osseointegrated screw-bone construct stiffness and peri-implant loading predicted by homogenized FE models relative to micro-FE models

Computational predictions of stiffness and peri-implant loading of screw-bone constructs are highly relevant to investigate and improve bone fracture fixations. Homogenized finite element (hFE) models have been used for this purpose in the past, but their accuracy has been questioned given the numerous simplifications, such as neglecting screw threads and modelling the trabecular bone structure as a continuum. This study aimed to investigate the accuracy of hFE models of an osseointegrated screw-bone construct when compared to micro-FE models considering the simplified screw geometry and different trabecular bone material models.Micro-FE and hFE models were created from 15 cylindrical bone samples with a virtually inserted, osseointegrated screw (fully bonded interface). Micro-FE models were created including the screw with threads (=reference models) and without threads to quantify the error due to screw geometry simplification. In the hFE models, the screws were modelled without threads and four different trabecular bone material models were used, including orthotropic and isotropic material derived from homogenization with kinematic uniform boundary conditions (KUBC), as well as from periodicity-compatible mixed uniform boundary conditions (PMUBC). Three load cases were simulated (pullout, shear in two directions) and errors in the construct stiffness and the volume average strain energy density (SED) in the peri-implant region were evaluated relative to the micro-FE model with a threaded screw.The pooled error caused by only omitting screw threads was low (max: 8.0%) compared to the pooled error additionally including homogenized trabecular bone material (max: 92.2%). Stiffness was predicted most accurately using PMUBC-derived orthotropic material (error: -0.7 ± 8.0%) and least accurately using KUBC-derived isotropic material (error: +23.1 ± 24.4%). Peri-implant SED averages were generally well correlated (R2 ≥ 0.76), but slightly over- or underestimated by the hFE models and SED distributions were qualitatively different between hFE and micro-FE models.This study suggests that osseointegrated screw-bone construct stiffness can be predicted accurately using hFE models when compared to micro-FE models and that volume average peri-implant SEDs are well correlated. However, the hFE models are highly sensitive to the choice of trabecular bone material properties. PMUBC-derived isotropic material properties represented the best trade-off between model accuracy and complexity in this study.

SHP-1 Protein Tyrosine Phosphatase Affects Early Postnatal Bone Development in Mice.

The Src homology region 2 domain-containing phosphatase-1 (SHP-1) is an intracellular tyrosine phosphatase that plays a negative regulatory role in immune cell signaling. Absent or diminished SHP-1 catalytic activity results in reduced bone mass with enhanced bone resorption. Here, we sought to investigate if Shp1 overexpression leads to increased bone mass and improved mechanical properties. Male and female wildtype (WT) and SHP1-transgenic (Tg) mice at 28, 56, and 84days of age were compared. We applied microcomputed tomography to assess femoral cortical bone geometry and trabecular architecture and 3-point mechanical bending to assess mid-diaphyseal structural and estimated material properties. Serum OPG, RANKL, P1NP, and CTX-1 concentrations were measured by enzyme-linked immunoassay. The majority of transgene effects were restricted to the 28-day-old mice. Trabecular bone volume per total volume, trabecular number, and connectivity density were greater in 28-day-old female SHP1-Tg mice when compared to WTs. SHP1-Tg female mice showed increased total and medullary areas, with no difference in cortical area and thickness. Cortical tissue mineral density was strongly genotype-dependent. Failure load, yield load, ultimate stress, and yield stress were all lower in 28-day-old SHP1-Tg females. In 28-day-old SHP1-Tg females, circulating levels of OPG and P1NP were higher and RANKL levels were lower than WT controls. Our study demonstrates a role for SHP-1 in early postnatal bone development; SHP-1 overexpression negatively impacted whole bone strength and material properties in females.

Effect of a new transpedicular vertebral device for the treatment or prevention of vertebral compression fractures: A finite element study

BackgroundA finite element study was performed to investigate the biomechanical performance of a novel transpedicular implant (V-STRUT©, Hyprevention, France) made of PEEK (polyetheretherketone) material in terms of strengthening the osteoporotic vertebra and the thoraco-lumbar spine.The objective was to assess numerically the efficacy of the implant to reduce the stress distribution within bone and absorb part of the stress by the implant thanks to its optimized material selection close to that of normal bone. MethodsA numerical model was generated based on a scan of an osteoporotic patient. The model is composed of three consecutive vertebrae and intervertebral discs. A heterogeneous distribution of bone material properties was assigned to the bone.In order to investigate the rationale of the device material selection, three FE models were developed (i) without the device to serve a reference model, (ii) with device made in Titanium material and (iii) with device made in PEEK material.Stiffness and stress distribution within the spine segment were computed and compared in order to assess the implants' performances. FindingsThe results obtained by the simulations indicated that the novel transpedicular implant made of PEEK material provided support to the superior vertebral endplate, restored the thoraco-lumbar spine segment stiffness and reduced the stress applied to the vertebrae under the compressive load. InterpretationImplant geometry in combination with its material properties are very important factors to restore vertebral strength and stiffness and limiting the risk of fracture at the same vertebra or adjacent ones.

Osteo-Immunomodulatory Role of Interleukin-4-Immobilized Plasma Immersion Ion Implantation Membranes for Bone Regeneration.

Barrier membranes for guided tissue regeneration are essential for bone repair and regeneration. The implanted membranes may trigger early inflammatory responses as a foreign material, which can affect the recruitment and differentiation of bone cells during tissue regeneration. The purpose of this study was to determine whether immobilizing interleukin 4 (IL4) on plasma immersion ion implantation (PIII)-activated surfaces may alter the osteo-immunoregulatory characteristics of the membranes and produce pro-osteogenic effects. In order to immobilize IL4, polycaprolactone surfaces were modified using the PIII technology. No discernible alterations were found between the morphology before and after PIII treatment or IL4 immobilization. IL4-immobilized PIII surfaces polarized macrophages to an M2 phenotype and mitigated inflammatory cytokine production under lipopolysaccharide stimulation. Interestingly, the co-culture of macrophages (on IL4-immobilized PIII surfaces) and bone marrow-derived mesenchymal stromal cells enhanced the production of angiogenic and osteogenic factors and triggered autophagy activation. Exosomes produced by PIII + IL4-stimulated macrophages were also found to play a role in osteoblast differentiation. In conclusion, the osteo-immunoregulatory properties of bone materials can be modified by PIII-assisted IL4 immobilization, creating a favorable osteoimmune milieu for bone regeneration.

Human tibial cortical bone with high porosity in type 2 diabetes mellitus is accompanied by distinctive bone material properties.

Diabetes mellitus is a metabolic disease affecting bone tissue at different length-scales. Higher fracture risk in diabetic patients is difficult to detect with common clinical fracture risk assessment due to normal or high bone mineral density in diabetic patients. The observed higher fracture risk despite normal to high areal bone mineral density in diabetic patients points towards impaired bone material quality. Here, we analyze tibial bone from individuals with type 2 diabetes mellitus using a multiscale-approach, which includes clinical and laboratory-based bone quality measures. Tibial cortical bone tissue from individuals with type 2 diabetes mellitus (T2DM) and age-matched healthy controls (n=15 each) was analyzed with in situ impact indentation, dual energy X-ray absorptiometry (DXA), high resolution peripheral microcomputed tomography (HR-pQCT), micro-computed tomography (microCT), cyclic indentation, quantitative backscattered electron microscopy (qBEI), vibrational spectroscopy (Raman), nanoindentation, and fluorescence spectroscopy. With this approach, a high cortical porosity subgroup of individuals with T2DM was discriminated from two study groups: individuals with T2DM and individuals without T2DM, while both groups were associated with similar cortical porosity quantified by means of microCT. The high porosity T2DM group, but not the T2DM group, showed compromised bone quality expressed by altered cyclic indentation properties (transversal direction) in combination with a higher carbonate-to-amide I ratio in endocortical bone. In addition, in the T2DM group with high cortical porosity group, greater cortical pore diameter was identified with HR-pQCT and lower tissue mineral density using microCT, both compared to T2DM group. Micromechanical analyses of cross-sectioned osteons (longitudinal direction) with cyclic indentation, qBEI, and nanoindentation showed no differences between the three groups. High tibial cortical porosity in T2DM can be linked to locally altered bone material composition. As the tibia is an accessible skeletal site for fracture risk assessment in the clinics (CT, indentation), our findings may contribute to further understanding the site-specific structural and compositional factors forming the basis of bone quality in diabetes mellitus. Refined diagnostic strategies are needed for a comprehensive fracture risk assessment in diabetic bone disease.

Influence of aging on mechanical properties of the femoral neck using an inverse method

Today, we are facing rapid aging of the world population, which increases the incidence of hip fractures. The gold standard of bone strength assessment in the laboratory is micro-computed finite element analysis (μFEA) based on micro-computed tomography (μCT) images. In clinics, the standard method to assess bone fracture risk is based on areal bone mineral density (aBMD), measured by dual-energy X-ray absorptiometry (DXA). In addition, homogenized finite element analysis (hFEA) constructed from quantitative computed tomography reconstructions (QCT) predicts clinical bone strength more accurately than DXA. Despite considerable evidence of degradation of bone material properties with age, in the past fifty years of finite element analysis to predict bone strength, bone material parameters remained independent of age. This study aims to assess the influence of age on apparent modulus, yield stress, and strength predictions of the human femoral neck made by laboratory-available bone volume fraction (BV/TV) and μFEA; and by clinically available DXA and hFEA. Using an inverse method, we test the hypothesis that FEA material parameters are independent of age. Eighty-six human femora were scanned with DXA (aBMD) and with QCT. The femoral necks were extracted and scanned at 16 μm resolution with μCT. The grayscale images were downscaled to 32 μm and 65 μm for linear and non-linear analyses, respectively, and segmented. The μFE solver ParOSolNL (non-linear) and a standard hFEA method were applied to the neck sections with the same material properties for all samples to compute apparent modulus, yield stress, and strength. Laboratory-available BV/TV was a good predictor of apparent modulus (R2 = 0.76), almost as good as μFEA (R2 = 0.79). However, yield stress and strength were better predicted by μFEA (R2 = 0.92, R2 = 0.86, resp.) than BV/TV (R2 = 0.76, R2 = 0.76, resp.). For clinically available variables, prediction of apparent modulus was better with hFEA than aBMD (R2 = 0.67, R2 = 0.58, resp.). hFEA outperformed aBMD for predictions of yield stress (R2 = 0.63 vs R2 = 0.34 for female and R2 = 0.55 for male) and strength (R2 = 0.48 vs R2 = 0.33 for female and R2 = 0.15 for male). The inclusion of age did not improve the multiple linear models for apparent modulus, yield stress, and strength. The resolution of the μFE meshes seems to account for most morphological changes induced by aging. The errors between the simulation and the experiment for apparent modulus, yield stress, and strength were age-independent, suggesting no rationale for correcting tissue material parameters in the current FE analysis of the aging femoral neck.

Accumulation of fluorescent advanced glycation end products and carboxymethyl-lysine in human cortical and trabecular bone

Chemical crosslinks known as advanced glycation end-products (AGEs) are associated with increased bone fracture risk and deteriorated bone mechanical properties. However, measurement of bone AGEs via ex vivo and in vitro methods has been limited to quantification of bulk fluorescent AGEs (fAGEs) and pentosidine only, which is a crosslinking fluorescent AGE. However, a non-crosslinking and non-fluorescent AGE such as carboxymethyl-lysine (CML) is found to be 40–100 times higher in quantity than pentosidine, but only one previous study has reported it in cortical bone, and one study reported it in trabecular bone. In our study, we wanted to investigate if accumulation of CML differs in cortical and trabecular compartments and if they are more strongly associated with bone mechanical properties than with fAGEs. We hypothesized that CML and fAGEs level would be higher in the trabecular compartment and show negative correlations to mechanical properties in cortical and trabecular bone. We obtained human cadaveric cortical and trabecular bone specimens, induced the formation of AGEs via the established in vitro ribosylation method, imaged specimens by microcomputed tomography to assess specimen geometry and microarchitecture, and mechanically tested cortical specimens by cyclic reference point indentation and fracture toughness tests and trabecular specimens by compression tests, followed by measurement of fAGEs and CML. fAGEs were 22 % higher in cortical bone (687 ± 44.8 ng Q/mg collagen) compared to trabecular bone (859 ± 317.1 ng Q/mg collagen), whereas CML levels were found to be 148 % higher in trabecular bone (6189.9 ± 866 ng/mg of protein) compared to cortical bone (924.6 ± 576.3 ng/mg of protein). Pooling the specimens from both the control and ribose groups, Spearman correlation analysis indicated that CML levels, but not fAGEs, are moderately associated with cortical porosity (r = +0.505, p ≤ 0.05) and mechanical properties such indentation depth (r = +0.460, p ≤ 0.05), total indentation depth (r = +0.440, p ≤ 0.05), and average energy dissipated (r = +0.465, p ≤ 0.05) in cortical bone. fAGEs showed a trend towards negative association with crack propagation toughness in cortical bone (r = −0.365, p = 0.055). No significant correlations were observed between CML and microarchitecture or mechanical properties in trabecular bone. CML levels were also associated with fAGEs in cortical bone (r = +0.596, p ≤ 0.05) but not in trabecular bone. Our preliminary findings indicate that CML, a non-crosslinking AGE, may affect bone material and mechanical properties differently than bulk fluorescent AGEs, given the higher accumulation of CML in each bone compartment. This study provides direction to future studies to quantify crosslinking and non-crosslinking AGEs separately as their effect on material and mechanical properties may be different and it would help identify better biomarkers for bone strength prediction.

Evaluation of assumptions in foot and ankle biomechanical models

BackgroundA variety of biomechanical models have been used in studies of foot and ankle disorders. Assumptions about the element types, material properties, and loading and boundary conditions are inherent in every model. It was hypothesized that the choice of these modeling assumptions could have a significant impact on the findings of the model. MethodsWe investigated the assumptions made in a number of biomechanical models of the foot and ankle and evaluated their effects on the results of the studies. Specifically, we focused on: (1) element choice for simulation of ligaments and tendons, (2) material properties of ligaments, cortical and trabecular bones, and encapsulating soft tissue, (3) loading and boundary conditions of the tibia, fibula, tendons, and ground support. FindingsOur principal findings are: (1) the use of isotropic solid elements to model ligaments and tendons is not appropriate because it allows them to transmit unrealistic bending and twisting moments and compressive forces; (2) ignoring the difference in elastic modulus between cortical and trabecular bones creates non-physiological stress distribution in the bones; (3) over-constraining tibial motion prevents anticipated deformity within the foot when simulating foot deformities, such as progressive collapsing foot deformity; (4) neglecting the Achilles tendon force affects almost all kinetic and kinematic parameters through the foot; (5) the axial force applied to the tibia and fibula is not equal to the ground reaction force due to the presence of tendon forces. InterpretationThe predicted outcomes of a foot model are highly sensitive to the model assumptions.