Robert Wallace

Do bisphosphonates inhibit direct fracture healing?: A laboratory investigation using an animal model

Fracture repair occurs by two broad mechanisms: direct healing, and indirect healing with callus formation. The effects of bisphosphonates on fracture repair have been assessed only in models of indirect fracture healing. A rodent model of rigid compression plate fixation of a standardised tibial osteotomy was used. Ten skeletally mature Sprague-Dawley rats received daily subcutaneous injections of 1 µg/kg ibandronate (IBAN) and ten control rats received saline (control). Three weeks later a tibial osteotomy was rigidly fixed with compression plating. Six weeks later the animals were killed. Fracture repair was assessed with mechanical testing, radiographs and histology. The mean stress at failure in a four-point bending test was significantly lower in the IBAN group compared with controls (8.69 Nmm(-2) (sd 7.63) vs 24.65 Nmm(-2) (sd 6.15); p = 0.017). On contact radiographs of the extricated tibiae the mean bone density assessment at the osteotomy site was lower in the IBAN group than in controls (3.7 mmAl (sd 0.75) vs 4.6 mmAl (sd 0.57); p = 0.01). In addition, histological analysis revealed progression to fracture union in the controls but impaired fracture healing in the IBAN group, with predominantly cartilage-like and undifferentiated mesenchymal tissue (p = 0.007). Bisphosphonate treatment in a therapeutic dose, as used for risk reduction in fragility fractures, had an inhibitory effect on direct fracture healing. We propose that bisphosphonate therapy not be commenced until after the fracture has united if the fracture has been rigidly fixed and is undergoing direct osteonal healing.

Micro-CT based finite element models of cancellous bone predict accurately displacement once the boundary condition is well replicated: A validation study

Non-destructive 3D micro-computed tomography (microCT) based finite element (microFE) models are used to estimate bone mechanical properties at tissue level. However, their validation remains challenging. Recent improvements in the quantification of displacements in bone tissue biopsies subjected to staged compression, using refined Digital Volume Correlation (DVC) techniques, now provide a full field displacement information accurate enough to be used for microFE validation. In this study, three specimens (two humans and one bovine) were tested with two different experimental set-ups, and the resulting data processed with the same DVC algorithm. The resulting displacement vector field was compared to that predicted by microFE models solved with three different boundary conditions (BC): nominal force resultant, nominal displacement resultant, distributed displacement. The first two conditions were obtained directly from the measurements provided by the experimental jigs, whereas in the third case the displacement field measured by the DVC in the top and bottom layer of the specimen was applied. Results show excellent relationship between the numerical predictions (x) and the experiments (y) when using BC derived from the DVC measurements (UX: y=1.07x-0.002, RMSE: 0.001mm; UY: y=1.03x-0.001, RMSE: 0.001mm; UZ: y=x+0.0002, RMSE: 0.001 mm for bovine specimen), whereas only poor correlation was found using BCs according to experiment set-ups. In conclusion, microFE models were found to predict accurately the vectorial displacement field using interpolated displacement boundary condition from DVC measurement.

A micro-architectural evaluation of osteoporotic human femoral heads to guide implant placement in proximal femoral fractures

Background and purpose The micro-architecture of bone has been increasingly recognized as an important determinant of bone strength. Successful operative stabilization of fractures depends on bone strength. We evaluated the osseous micro-architecture and strength of the osteoporotic human femoral head. Material and methods 6 femoral heads, obtained during arthroplasty surgery for femoral neck fracture, underwent micro-computed tomography (microCT) scanning at 30 μm, and bone volume ratio (BV/TV), trabecular thickness, structural model index, connection density, and degree of anisotropy for volumes of interest throughout the head were derived. A further 15 femoral heads underwent mechanical testing of compressive failure stress of cubes of trabecular bone from different regions of the head. Results The greatest density and trabecular thickness was found in the central core that extended from the medial calcar to the physeal scar. This region also correlated with the greatest degree of anisotropy and proportion of plate-like trabeculae. In the epiphyseal region, the trabeculae were organized radially from the physeal scar. The weakest area was found at the apex and peripheral areas of the head. The strongest region was at the center of the head. Interpretation The center of the femoral head contained the strongest trabecular bone, with the thickest, most dense trabeculae. The apical region was weaker. From an anatomical and mechanical point of view, implants that achieve fixation in or below this central core may achieve the most stable fixation during fracture healing.

Major source of error when calculating bone mechanical properties.

Mechanical testing of bone and fracture callus is performed to assess the functional properties of the tissue. In particular, 3‐ point bending is a commonly performed technique when experimentally evaluating mechanical properties. The Euler‐ Bernoulli equation used to calculate the bending stiffness assumes that the beam is long and slender. In practice, beams that have a span to depth ratio (aspect ratio) of greater than 20:1 are considered to be “slender.” (1) If this method is used on non‐ slender bones an error results as the contribution of the bending due to shear deformation is not taken into account. A review of articles published in the last 2 years in this journal indicated that in the 14 papers that used 3‐point bending to determine elastic modulus none accounted for the effect on deformation due to shear. The error from ignoring the contribution of shear deformation on bones with these properties can be as high as 38%. If the analysis does not take shear deformation into account, large errors in the evaluated material properties will result in an estimated 95% of bones. Three‐point bending is often carried out using long bones from animals such as the mouse, rat, or rabbit that are too small to allow sectioning into smaller samples and must therefore be tested whole. Tests performed on these bones will therefore always be subject to inaccuracies if they assumed to be slender beams. The problem is compounded by the fact that the test length of the specimen is not the length of the bone, but the distance between the supports. In order to provide a suitable location, the supports are generally placed at the metaphyses of the bone. Therefore, the span is less than the total length of the bone, further reducing the aspect ratio, often by 25%, resulting in an increase to the potential error. The problem of not accounting for deflection due to shear is further exacerbated as mechanical testing is often used to determine the influence of medical treatments or physical diseases. In order to study these, animals (usuallymice or rats) are bred with genetic deficiencies. In some cases, such as is found in the FGFR‐3 deficient mouse, this results in musculoskeletal changes. These changes can result in significantly different aspect ratio of limbs in comparison to the wild and the modified specimen as a limb may be longer or thicker without a corresponding change in the complimentary dimension. Assumptions made about the cross‐sectional shape, (ie, whether it is approximately circular as with the shaft of the femur or approximately triangular as for the shaft of the tibia) can also result in errors. However, for whole bones, themajor determinant of this error is the aspect ratio rather than the cross‐sectional geometry. It is recognized that small errors may arise due to non‐ prismatic geometry of the bone, ie, changes in cross‐section from proximal to distal, and inhomogeneous material properties. These are best considered through the use of numerical simulation (eg, finite element analysis), which require a full 3D geometry construction via a CT scan and assignment of variable material properties from CT attenuation data. As a consequence it requires considerably more resources: scanning; conversion of images to numerical models; and analysis of models and interpretation of results. As a result, this type of analysis is rarely performed on small animal studies such as those conducted using rat or mouse limbs. Numerical analysis, due to the additional work and resources required, is usually restricted limited to human bones, as the limited supply of these bones can often warrant the use of additional resources. The deflection due to bending and that attributed to shear can be derived following the methods set out in Wang.

Hybrid electrolytes with 3D bicontinuous ordered ceramic and polymer microchannels for all-solid-state batteries

Hybrid solid electrolytes, composed of 3D ordered bicontinuous conducting ceramic and insulating polymer microchannels are reported. The ceramic channels provide continuous, uninterrupted pathways, maintaining high ionic conductivity between the electrodes, while the polymer channels permit improvement of the mechanical properties from that of the ceramic alone, in particular mitigation of the ceramic brittleness. The conductivity of a ceramic electrolyte is usually limited by resistance at the grain boundaries, necessitating dense ceramics. The conductivity of the 3D ordered hybrid is reduced by only the volume fraction occupied by the ceramic, demonstrating that the ceramic channels can be sintered to high density similar to a dense ceramic disk. The hybrid electrolytes are demonstrated using the ceramic lithium ion conductor Li1.4Al0.4Ge1.6(PO4)3 (LAGP). Structured LAGP 3D scaffolds with empty channels were prepared by negative replication of a 3D printed polymer template. Filling the empty channels with non-conducting polypropylene (PP) or epoxy polymer (epoxy) creates the structured hybrid electrolytes with 3D bicontinuous ceramic and polymer microchannels. Printed templating permits precise control of the ceramic to polymer ratio and the microarchitecture; as demonstrated by the formation of cubic, gyroidal, diamond and spinodal (bijel) structures. The electrical and mechanical properties depend on the microarchitecture, the gyroid filled with epoxy giving the best combination of conductivity and mechanical properties. An ionic conductivity of 1.6 × 10−4 S cm−1 at room temperature was obtained, reduced from the conductivity of a sintered LAGP pellet only by the volume fraction occupied by the ceramic. The mechanical properties of the gyroid LAGP–epoxy electrolyte demonstrate up to 28% higher compressive failure strain and up to five times the flexural failure strain of a LAGP pellet before rupture. Notably, this demonstrates that ordered ceramic and polymer hybrid electrolytes can have superior mechanical properties without significantly compromising ionic conductivity, which addresses one of the key challenges for all-solid-state batteries.

Increased Circulating Adiponectin in Response to Thiazolidinediones: Investigating the Role of Bone Marrow Adipose Tissue

Background Bone marrow adipose tissue (MAT) contributes to increased circulating adiponectin, an insulin-sensitizing hormone, during caloric restriction (CR), but whether this occurs in other contexts remains unknown. The antidiabetic thiazolidinediones (TZDs) also promote MAT expansion and hyperadiponectinemia, even without increasing adiponectin expression in white adipose tissue (WAT). Objectives To test the hypothesis that MAT expansion contributes to TZD-associated hyperadiponectinemia, we investigated the effects of rosiglitazone, a prototypical TZD, in wild-type (WT) or Ocn-Wnt10b mice. The latter resist MAT expansion during CR, leading us to postulate that they would also resist this effect of rosiglitazone. Design Male and female WT or Ocn-Wnt10b mice (C57BL/6J) were treated with or without rosiglitazone for 2, 4, or 8 weeks, up to 30 weeks of age. MAT content was assessed by osmium tetroxide staining and adipocyte marker expression. Circulating adiponectin was determined by ELISA. Results In WT mice, rosiglitazone caused hyperadiponectinemia and MAT expansion. Compared to WT mice, Ocn-Wnt10b mice had significantly less MAT in distal tibiae and sometimes in proximal tibiae; however, interpretation was complicated by the leakage of osmium tetroxide from ruptures in some tibiae, highlighting an important technical consideration for osmium-based MAT analysis. Despite decreased MAT in Ocn-Wnt10b mice, circulating adiponectin was generally similar between WT and Ocn-Wnt10b mice; however, in females receiving rosiglitazone for 4 weeks, hyperadiponectinemia was significantly blunted in Ocn-Wnt10b compared to WT mice. Notably, this was also the only group in which tibial adiponectin expression was lower than in WT mice, suggesting a close association between MAT adiponectin production and circulating adiponectin. However, rosiglitazone significantly increased adiponectin protein expression in WAT, suggesting that WAT contributes to hyperadiponectinemia in this context. Finally, rosiglitazone upregulated uncoupling protein 1 in brown adipose tissue (BAT), but this protein was undetectable in tibiae, suggesting that MAT is unlikely to share thermogenic properties of BAT. Conclusion TZD-induced hyperadiponectinemia is closely associated with increased adiponectin production in MAT but is not prevented by the partial loss of MAT that occurs in Ocn-Wnt10b mice. Thus, more robust loss-of-MAT models are required for future studies to better establish MAT’s elusive functions, both on an endocrine level and beyond.

Sertoli Cells Modulate Testicular Vascular Network Development, Structure, and Function to Influence Circulating Testosterone Concentrations in Adult Male Mice

The testicular vasculature forms a complex network, providing oxygenation, micronutrients, and waste clearance from the testis. The vasculature is also instrumental to testis function because it is both the route by which gonadotropins are delivered to the testis and by which T is transported away to target organs. Whether Sertoli cells play a role in regulating the testicular vasculature in postnatal life has never been unequivocally demonstrated. In this study we used models of acute Sertoli cell ablation and acute germ cell ablation to address whether Sertoli cells actively influence vascular structure and function in the adult testis. Our findings suggest that Sertoli cells play a key role in supporting the structure of the testicular vasculature. Ablating Sertoli cells (and germ cells) or germ cells alone results in a similar reduction in testis size, yet only the specific loss of Sertoli cells leads to a reduction in total intratesticular vascular volume, the number of vascular branches, and the numbers of small microvessels; loss of germ cells alone has no effect on the testicular vasculature. These perturbations to the testicular vasculature leads to a reduction in fluid exchange between the vasculature and testicular interstitium, which reduces gonadotropin-stimulated circulating T concentrations, indicative of reduced Leydig cell stimulation and/or reduced secretion of T into the vasculature. These findings describe a new paradigm by which the transport of hormones and other factors into and out of the testis may be influenced by Sertoli cells and highlights these cells as potential targets for enhancing this endocrine relationship.

Nonlinear viscoelastic characterization of bovine trabecular bone

The time-independent elastic properties of trabecular bone have been extensively investigated, and several stiffness–density relations have been proposed. Although it is recognized that trabecular bone exhibits time-dependent mechanical behaviour, a property of viscoelastic materials, the characterization of this behaviour has received limited attention. The objective of the present study was to investigate the time-dependent behaviour of bovine trabecular bone through a series of compressive creep–recovery experiments and to identify its nonlinear constitutive viscoelastic material parameters. Uniaxial compressive creep and recovery experiments at multiple loads were performed on cylindrical bovine trabecular bone samples (

Tensioning of Prolene reduces creep under cyclical load: relevance to a simple pre-operative manoeuvre

n = 19