Understanding basic multicellular unit activity in cortical bone through 3D morphological analysis: New methods to define zones of the remodeling space.

The activity of basic multicellular units (BMU) in cortical bone is classically described as a sequential order of events- resorption, reversal and formation. This simplified portrayal of the remodeling process is pervasive despite the reported variability in remodeling space morphology. These variations may reflect meaningful nuances in BMU activity but methods to quantify 3D remodeling space morphology within the context of the cellular activity are currently lacking. This study developed new techniques to define zones of BMU activity based on the 3D morphology of remodeling spaces in rabbit cortical bone and integrated morphological data with the BMU longitudinal erosion rate (LER) to elucidate the spatial-temporal coordination of BMUs and estimate mineral apposition rate (MAR). The tibiae of New Zealand white rabbits (n = 5) were imaged in vivo using synchrotron radiation and two weeks later ex vivo with desktop microCT. The in vivo and ex vivo datasets were co-registered, and 27 remodeling spaces were identified at both timepoints. A radial profile representing the 3D morphology was the platform for partitioning the remodeling spaces into resorption, reversal and formation zones. Manual, automated and semi-automated partitioning approaches were compared, and the zone-segmentations were used to calculate the length, change in radius and slope of each zone. The manual approach most accurately defined the zones of idealized remodeling spaces with known dimensions (relative error = 0.9-9.2 %) while the semi-automated method reliably defined the zones in rabbit remodeling spaces (ICC = 0.85-1.00). Combining LER and the manually derived zone dimensions indicated that a BMU passes through a cross-section in approximately 18.8 days with resorption, reversal and formation taking 4.1, 2.2, and 12.5 days, respectively. MAR estimated by the 3D analysis was not significantly different than that determined with classic histomorphometry (p = 0.48). These techniques have the potential to assess dynamic parameters of bone resorption and formation, eliminate the need for fluorochrome labeling and provide a more comprehensive perspective of the remodeling process.

Twelve Months of Denosumab and/or Alendronate Is Associated With Improved Bone Fatigue Life, Microarchitecture, and Density in Ovariectomized Cynomolgus Monkeys.

Prolonged use of antiresorptives such as the bisphosphonate alendronate (ALN) and the RANKL inhibitor denosumab (DMAb) are associated with rare cases of atypical femoral fracture (AFF). The etiology of AFF is unclear, but it has been hypothesized that potent osteoclast inhibitors may reduce bone fatigue resistance. The purpose of this study was to quantify the relationship between antiresorptive treatment and fatigue life (cycles to failure) in bone from ovariectomized cynomolgus monkeys. We analyzed humeral bone from 30 animals across five treatment groups. Animals were treated for 12 months with subcutaneous (sc) vehicle (VEH), sc DMAb (25 mg/kg/month), or intravenous (iv) ALN (50 µg/kg/month). Another group received 6 months VEH followed by 6 months DMAb (VEH-DMAb), and the final group received 6 months ALN followed by 6 months DMAb (ALN-DMAb). A total of 240 cortical beam samples were cyclically tested in four-point bending at 80, 100, 120, or 140 MPa peak stress. High-resolution imaging and density measurements were performed to evaluate bone microstructure and composition. Samples from the ALN (p = 0.014), ALN-DMAb (p = 0.008), and DMAb (p < 0.001) groups illustrated higher fatigue-life measurements than VEH. For example, at 140 MPa the VEH group demonstrated a median ± interquartile range (IQR) fatigue life of 1987 ± 10593 cycles, while animals in the ALN, ALN-DMAb, and DMAb groups survived 9850 ± 13648 (+395% versus VEH), 10493 ± 16796 (+428%), and 14495 ± 49299 (+629%) cycles, respectively. All antiresorptive treatment groups demonstrated lower porosity, smaller pore size, greater pore spacing, and lower number of canals versus VEH (p < 0.001). Antiresorptive treatment was also associated with greater apparent density, dry density, and ash density (p ≤ 0.03). We did not detect detrimental changes following antiresorptive treatments that would explain their association with AFF. In contrast, 12 months of treatment may have a protective effect against fatigue fractures. © 2022 American Society for Bone and Mineral Research (ASBMR).

Bringing Mechanical Context to Image-Based Measurements of Bone Integrity.

PURPOSE OF REVIEW: Image-based measurements of bone integrity are used to estimate failure properties and clinical fracture risk. This paper (1) reviews recent imaging studies that have enhanced our understanding of the mechanical pathways to bone fracture and (2) discusses the influence that inter-individual differences in image-based measurements may have on the clinical assessment of fracture risk RECENT FINDINGS: Increased tissue mineralization is associated with improved bone strength but reduced fracture toughness. Trabecular architecture that is important for fatigue resistance is less important for bone strength. The influence of porosity on bone failure properties is heavily dependent on pore location and size. The interaction of various characteristics, such as bone area and mineral content, can further complicate their influence on bone failure properties. What is beneficial for bone strength is not always beneficial for bone toughness or fatigue resistance. Additionally, given the large amount of imaging data that is clinically available, there is a need to develop effective translational strategies to better interpret non-invasive measurements of bone integrity.

Association between intracortical microarchitecture and the compressive fatigue life of human bone: A pilot study.

Many mechanical properties of cortical bone are largely governed by the underlying microarchitecture; however, the influence of microarchitecture on the fatigue life of bone is poorly understood. Furthermore, imaging-based studies investigating intracortical microarchitecture may expose bone samples to large doses of radiation that may compromise fatigue resistance. The purpose of this pilot study was to 1) investigate the relationship between intracortical microarchitecture and the fatigue life of human bone in compression and 2) examine the effects of synchrotron irradiation on fatigue life measurements. Cortical samples were prepared from the femoral and tibial shafts of three cadaveric donors. A subset of samples was imaged using synchrotron X-ray microCT to quantify microarchitecture, including porosity, canal diameter, lacunar density, lacunar volume, and lacunar orientation. A second group of control samples was not imaged and used only for mechanical testing. Fatigue life was quantified by cyclically loading both groups in zero-compression until failure. Increased porosity and larger canal diameter were both logarithmically related to a shorter fatigue life, whereas lacunar density demonstrated a positive linear relationship with fatigue life (r2 = 45-73%, depending on measure). Irradiation from microCT scanning reduced fatigue life measurements by 91%, but relationships with microarchitecture measurements remained. Additional research is needed to support the findings of this pilot study and fully establish the relationship between intracortical microarchitecture and the compressive fatigue life of bone.

Mechanical Fatigue of Bovine Cortical Bone Using Ground Reaction Force Waveforms in Running.

Stress fractures are a common overuse injury among runners associated with the mechanical fatigue of bone. Several in vivo biomechanical studies have investigated specific characteristics of the vertical ground reaction force (vGRF) in heel-toe running and have observed an association between increased loading rate during impact and individuals with a history of stress fracture. The purpose of this study was to examine the fatigue behavior of cortical bone using vGRF-like loading profiles, including those that had been decomposed into their respective impact and active phase components. Thirty-eight cylindrical cortical bone samples were extracted from bovine tibiae and femora. Hydrated samples were fatigue tested at room temperature in zero compression under load control using either a raw (n = 10), active (n = 10), low impact (n = 10), or high impact (n = 8) vGRF profile. The number of cycles to failure was quantified and the test was terminated if the sample survived 105 cycles. Fatigue life was significantly greater for both impact groups compared to the active (p < 0.001) and raw (p < 0.001) groups, with all low impact samples and 6 of 8 high impact samples surviving 105 cycles. The mean (± SD) number of cycles to failure for the active and raw groups was 12,133±11,704 and 16,552±29,612, respectively. The results suggest that loading rates associated with the impact phase of a typical vGRF in running have little influence on the mechanical fatigue behavior of bone relative to loading magnitude, warranting further investigation of the mechanism by which increased loading rates are associated with stress fracture.

Effects of footwear and stride length on metatarsal strains and failure in running.

BACKGROUND: The metatarsal bones of the foot are particularly susceptible to stress fracture owing to the high strains they experience during the stance phase of running. Shoe cushioning and stride length reduction represent two potential interventions to decrease metatarsal strain and thus stress fracture risk. METHODS: Fourteen male recreational runners ran overground at a 5-km pace while motion capture and plantar pressure data were collected during four experimental conditions: traditional shoe at preferred and 90% preferred stride length, and minimalist shoe at preferred and 90% preferred stride length. Combined musculoskeletal - finite element modeling based on motion analysis and computed tomography data were used to quantify metatarsal strains and the probability of failure was determined using stress-life predictions. FINDINGS: No significant interactions between footwear and stride length were observed. Running in minimalist shoes increased strains for all metatarsals by 28.7% (SD 6.4%; p<0.001) and probability of failure for metatarsals 2-4 by 17.3% (SD 14.3%; p≤0.005). Running at 90% preferred stride length decreased strains for metatarsal 4 by 4.2% (SD 2.0%; p≤0.007), and no differences in probability of failure were observed. INTERPRETATIONS: Significant increases in metatarsal strains and the probability of failure were observed for recreational runners acutely transitioning to minimalist shoes. Running with a 10% reduction in stride length did not appear to be a beneficial technique for reducing the risk of metatarsal stress fracture, however the increased number of loading cycles for a given distance was not detrimental either.

Experimental validation of finite element predicted bone strain in the human metatarsal.

Metatarsal stress fracture is a common injury observed in athletes and military personnel. Mechanical fatigue is believed to play an important role in the etiology of stress fracture, which is highly dependent on the resulting bone strain from the applied load. The purpose of this study was to validate a subject-specific finite element (FE) modeling routine for bone strain prediction in the human metatarsal. Strain gauge measurements were performed on 33 metatarsals from seven human cadaveric feet subject to cantilever bending, and subject-specific FE models were generated from computed tomography images. Material properties for the FE models were assigned using a published density-modulus relationship as well as density-modulus relationships developed from optimization techniques. The optimized relationships were developed with a 'training set' of metatarsals (n=17) and cross-validated with a 'test set' (n=16). The published and optimized density elasticity equations provided FE-predicted strains that were highly correlated with experimental measurements for both the training (r2≥0.95) and test (r2≥0.94) sets; however, the optimized equations reduced the maximum error by 10% to 20% relative to the published equation, and resulted in an X=Y type of relationship between experimental measurements and FE predictions. Using a separate optimized density-modulus equation for trabecular and cortical bone did not improve strain predictions when compared to a single equation that spanned the entire bone density range. We believe that the FE models with optimized material property assignment have a level of accuracy necessary to investigate potential interventions to minimize metatarsal strain in an effort to prevent the occurrence of stress fracture.