Bone Mechanical Properties Research Articles

PRG4 deficiency in mice alters skeletal structure, mechanics, and calvarial osteoclastogenesis, and rhPRG4 inhibits in vitro osteoclastogenesis.

Osteoporosis is a chronic disease characterized by reduced bone mass and increased fracture risk, estimated to affect over 10 million people in the United States alone. Drugs used to treat bone loss often come with significant limitations and/or long-term safety concerns. Proteoglycan-4 (PRG4, also known as lubricin) is a mucin-like glycoprotein best known for its boundary lubricating function of articular cartilage. In more recent years, it has been shown that PRG4 has anti-inflammatory properties, contributes to the maintenance of subchondral bone integrity, and patients with PRG4 mutations are osteopenic. However, it remains unknown how PRG4 impacts mechanical and material properties of bone. Therefore, our objective was to perform a phenotyping study of bone in a Prg4 gene trap (GT) mouse (PRG4 deficient). We found that femurs of Prg4 GT mice have altered mechanical, structural, and material properties relative to wildtype littermates. Additionally, Prg4 GT mice have a greater number of calvarial osteoclasts than wildtype mice, but do not have a notable inflammatory serum profile. Finally, Prg4 GT mice do not have an altered rate of bone formation, and exogenous recombinant human PRG4 (rhPRG4) administration inhibited osteoclastogenesis in vitro, suggesting that the skeletal phenotype may be due to changes in bone resorption. Overall, this work demonstrates that PRG4 deficiency affects several integral properties of bone structure, mechanics, and skeletal cell activity, and provides the foundation and insight toward future work evaluating PRG4 as a potential therapeutic target in treating bone loss.

Evaluation of bone-related mechanical properties in female patients with long-term remission of Cushing's syndrome using quantitative computed tomography-based finite element analysis.

Hypercortisolism in Cushing's syndrome (CS) is associated with bone loss, skeletal fragility, and altered bone quality. No studies evaluated bone geometric and strain-stress values in CS patients after remission thus far. Thirty-two women with CS in remission (mean age [±SD] 51 ± 11; body mass index [BMI], 27 ± 4 kg/m2; mean time of remission, 120 ± 90 months) and 32 age-, BMI-, and gonadal status-matched female controls. Quantitative computed tomography (QCT) was used to assess volumetric bone mineral density (vBMD) and buckling ratio, cross-sectional area, and average cortical thickness at the level of the proximal femur. Finite element (FE) models were generated from QCT to calculate strain and stress values (maximum principal strain [MPE], maximum strain energy density [SED], maximum Von Mises [VM], and maximum principal stress [MPS]). Areal BMD (aBMD) and trabecular bone score (TBS) were assessed by dual-energy X-ray absorptiometry (2D DXA). Trabecular vBMD at total hip and trochanter were lower in CS as compared with controls (P < .05). Average cortical thickness was lower, and buckling ratio was greater in CS vs controls (P < .01). All strain and stress values were higher in CS patients vs controls (P < .05). 2D DXA-derived measures were similar between patients and controls (P > .05). Prior hypercortisolism predicted both VM (β .30, P = .014) and MPS (β .30, P = .015), after adjusting for age, BMI, menopause, delay to diagnosis, and duration of remission. Women with prior hypercortisolism have reduced trabecular vBMD and impaired bone geometrical and mechanical properties, which may contribute to an elevated fracture risk despite long-term remission.

Calcium supplementation attenuates fluoride-induced bone injury via PINK1/Parkin-mediated mitophagy and mitochondrial apoptosis in mice

Excessive consumption of fluoride can cause skeletal fluorosis. Mitophagy has been identified as a novel target for bone disorders. Meanwhile, calcium supplementation has shown great potential for mitigating fluoride-related bone damage. Hence, this study aimed to elucidate the association between mitophagy and skeletal fluorosis and the precise mechanisms through which calcium alleviates these injuries. A 100 mg/L sodium fluoride (NaF) exposure model in Parkin knockout (Parkin-/-) mice and a 100 mg/L NaF exposure mouse model with 1% calcium carbonate (CaCO3) intervention were established in the current study. Fluoride exposure caused the impairment of mitochondria and activation of PTEN-induced putative kinase1 (PINK1)/E3 ubiquitin ligase Park2 (Parkin)-mediated mitophagy and mitochondrial apoptosis in the bones, which were restored after blocking Parkin. Additionally, the intervention model showed fluoride-exposed mice exhibited abnormal bone trabecula and mechanical properties. Still, these bone injuries could be effectively attenuated by adding 1% calcium to their diet, which reversed fluoride-activated mitophagy and apoptosis. To summarize, fluoride can activate bone mitophagy through the PINK1/Parkin pathway and mitochondrial apoptosis. Parkin-/- and 1% calcium provide protection against fluoride-induced bone damage. Notably, this study provides theoretical bases for the prevention and therapy of animal and human health and safety caused by environmental fluoride contamination.

Strukturne determinante povećane koštane fragilnosti kod starijih osoba - višestruke perspektive

Numerous epidemiological studies have shown that increased bone fragility and a higher risk of fractures are present in the aged, which reduces their quality of life and represents a significant socio-economic burden for the healthcare system. However, morphological and structural determinants underlying increased bone fragility have yet to be fully explained. This paper aimed to provide an overview of modern studies that dealt with determinants of increased bone fragility, analyzing different hierarchical levels of bone tissue organization (macro-, micro-, and nano-levels) in aged individuals and individuals with chronic comorbidities (mainly in individuals with chronic liver disease, renal disorders, and type 2 diabetes mellitus). Also, variable frequency of fractures at different skeletal sites in aged persons and individuals with chronic diseases was shown, indicating that aging-related bone loss is not a uniform process. A complete understanding of the spatial pattern of impaired bone quality can aid in the targeted evaluation of individualized fracture risk. Establishing a firm connection between the results of the clinical assessment of bone status and the analysis of numerous structural and mechanical bone properties (on various hierarchical levels) can represent a solid base for developing adequate guidelines and algorithms for prevention and treatment of increased bone fragility in aged individuals and individuals with chronic diseases.

Development and Structural Evaluation of Dog Bone Particle–Reinforced Epoxy Composites for Biomedical Applications

There has been great interest in developing high‐performance, sustainable and eco‐friendly materials. This study presents an innovative development and research of the mechanical properties of dog bone–reinforced epoxy composites to determine their suitability for biomedical applications. Good surface compatibility for the matrix and reinforcement was the target, complementing the structural compatibility needed for such applications. Hand‐lay open mould technique was used to fabricate the composites with varying weight percentage of dog bone particles (DBPs), and mechanical properties (tensile strength, Young’s modulus, percentage elongation and flexural strength) were determined. The results show that the developed composites’ mechanical properties were enhanced, and the tensile strength of 21.10 MPa–27.97 MPa, Young’s modulus of 1531.00 MPa–1732.90 MPa, ductility of 2.10%–2.60% and flexural strength of 27.53 MPa–35.67 MPa were recorded for the biocomposites. Energy‐dispersive X‐ray spectroscopy (EDS) revealed the main elements’ contents of the DBPs as calcium (50.3%), carbon (20.77%) and oxygen (11.76%). Scanning electron microscopy (SEM) examination of the fractured surfaces revealed the homogeneous distribution of the dog bone particulates within the epoxy matrix, which caused the property enhancement. The findings of this research underline the potency of processed animal waste as filler materials.

Elucidating the role of diverse mineralisation paradigms on bone biomechanics - a coarse-grained molecular dynamics investigation.

Bone as a hierarchical composite structure plays a myriad of roles in vertebrate skeletons including providing the structural stability of the body. Despite this critical role, the mechanical behaviour at the sub-micron levels of bone's hierarchy remains poorly understood. At this scale, bone is composed of Mineralised Collagen Fibrils (MCF) embedded within an extra-fibrillar matrix that consists of hydroxyapatite minerals and non-collagenous proteins. Recent experimental studies hint at the significance of the extra-fibrillar matrix in providing the bone with the stiffness and ductility needed to serve its structural roles. However, due to limited resolution of experimental tools, it is not clear how the arrangement of minerals, and in particular their relative distribution between the intra- and extra-fibrillar space contribute to bone's remarkable mechanical properties. In this study, a Coarse Grained Molecular Dynamics (CGMD) framework was developed to study the mechanical properties of MCFs embedded within an extra-fibrillar mineral matrix and the precise roles extra- and intra-fibrillar mineralisation on the load-deformation response was investigated. It was found that the presence of extra-fibrillar mineral resulted in the development of substantial residual stress in the system, by limiting MCF shortening that took place during intra-fibrillar mineralisation, resulting in substantial compressive residual stresses in the extra-fibrillar mineral phase. The simulation results also revealed the crucial role of extra-fibrillar mineralisation in determining the elastic response of the Extrafibrillar mineralised MCF (EFM-MCF) system up to the yield point, while the fibrillar collagen affected the post-yield behaviour. When physiological levels of mineralisation were considered, the mechanical response of the EFM-MCF systems was characterised by high ductility and toughness, with micro-cracks being distributed across the extra-fibrillar matrix, and MCFs effectively bridging these cracks leading to an excellent combination of strength and toughness. Together, these results provide novel insight into the deformation mechanisms of an EFM-MCF system and highlight that this universal building block, which forms the basis for lamellar bone, can provide an excellent balance of stiffness, strength and toughness, achieving mechanical properties that are far beyond the capabilities of the individual constituents acting alone.

The osteocytic actions of glucocorticoids on bone mass, mechanical properties, or perilacunar remodeling outcomes are not rescued by PTH(1-34).

Glucocorticoids (GC) and parathyroid hormone (PTH) are widely used therapeutic endocrine hormones where their effects on bone and joint arise from actions on multiple skeletal cell types. In osteocytes, GC and PTH exert opposing effects on perilacunar canalicular remodeling (PLR). Suppressed PLR can impair bone quality and joint homeostasis, including in GC-induced osteonecrosis. However, combined effects of GC and PTH on PLR are unknown. Given the untapped potential to target osteocytes to improve skeletal health, this study sought to test the feasibility of therapeutically mitigating PLR suppression. Focusing on subchondral bone and joint homeostasis, we hypothesize that PTH(1-34), a PLR agonist, could rescue GC-suppressed PLR. The skeletal effects of GC and PTH(1-34), alone or combined, were examined in male and female mice by micro-computed tomography, mechanical testing, histology, and gene expression analysis. For each outcome, females were more responsive to GC and PTH(1-34) than males. GC and PTH(1-34) exerted regional differences, with GC increasing trabecular bone volume but reducing cortical bone thickness, stiffness, and ultimate force. Despite PTH(1-34)'s anabolic effects on trabecular bone, it did not rescue GC's catabolic effects on cortical bone. Likewise, cartilage integrity and subchondral bone apoptosis, tartrate-resistant acid phosphatase (TRAP) activity, and osteocyte lacunocanalicular networks showed no evidence that PTH(1-34) could offset GC-dependent effects. Rather, GC and PTH(1-34) each increased cortical bone gene expression implicated in bone resorption by osteoclasts and osteocytes, including Acp5, Mmp13, Atp6v0d2, Ctsk, differences maintained when GC and PTH(1-34) were combined. Since PTH(1-34) is insufficient to rescue GC's effects on young female mouse bone, future studies are needed to determine if osteocyte PLR suppression, due to GC, aging, or other factors, can be offset by a PLR agonist.

Osteoblast-specific down-regulation of NLRP3 inflammasome by aptamer-functionalized liposome nanoparticles improves bone quality in postmenopausal osteoporosis rats.

Rationale: NLRP3 inflammasome is critical in the development and progression of many metabolic diseases driven by chronic inflammation, but its effect on the pathology of postmenopausal osteoporosis (PMOP) remains poorly understood. Methods: We here firstly examined the levels of NLRP3 inflammasome in PMOP patients by ELISA. Then we investigated the possible mechanisms underlying the effect of NLRP3 inflammasome on PMOP by RNA sequencing of osteoblasts treated with NLRP3 siRNA and qPCR. Lastly, we accessed the effect of decreased NLRP3 levels on ovariectomized (OVX) rats. To specifically deliver NLRP3 siRNA to osteoblasts, we constructed NLRP3 siRNA wrapping osteoblast-specific aptamer (CH6)-functionalized lipid nanoparticles (termed as CH6-LNPs-siNLRP3). Results: We found that the levels of NLRP3 inflammasome were significantly increased in patients with PMOP, and were negatively correlated with estradiol levels. NLRP3 knock-down influenced signal pathways including immune system process, interferon signal pathway. Notably, of the top ten up-regulated genes in NLRP3-reduced osteoblasts, nine genes (except Mx2) were enriched in immune system process, and five genes were related to interferon signal pathway. The in vitro results showed that CH6-LNPs-siNLRP3 was relatively uniform with a dimeter of 96.64 ± 16.83 nm and zeta potential of 38.37 ± 1.86 mV. CH6-LNPs-siNLRP3 did not show obvious cytotoxicity and selectively delivered siRNA to bone tissue. Moreover, CH6-LNPs-siNLRP3 stimulated osteoblast differentiation by activating ALP and enhancing osteoblast matrix mineralization. When administrated to OVX rats, CH6-LNPs-siNLRP3 promoted bone formation and bone mass, improved bone microarchitecture and mechanical properties by decreasing the levels of NLRP3, IL-1β and IL-18 and increasing the levels of OCN and Runx2. Conclusion: NLRP3 inflammasome may be a new biomarker for PMOP diagnosis and plays a key role in the pathology of PMOP. CH6-LNPs-siNLRP3 has potential application for the treatment of PMOP.

Effects of spray-dried plasma on performance, carcass parameters, tibia quality and Newcastle disease vaccine efficacy in broiler chicken fed corn-soy diets with two varying levels of digestible amino acids and AMEn density.

This study aimed to determine the effects of spray dried plasma (SDP) on growth performance, carcass traits, tibia quality, and hemagglutination inhibition titers in broilers fed two nutritional strategies with high or low nutrient density. In the study, 816 one-day-old Ross 308 male broiler chickens were divided into a 2 × 2 factorial arrangements consisting of four treatment groups with 12 replicates (17 birds/replicate) based on diets with high nutrient density (HND) or low nutrient density (LND) from d 0 to 42 and receiving either control or 1% SDP diets during d 0 to 10. The results showed that feed intake (FI) and body weight gain (BWG) were increased (P < 0.05) and feed conversion ratio (FCR) was significantly reduced (P = 0.003) for broilers fed HND diets from d 0 to 42. The inclusion of SDP increased the BWG (P < 0.001), FI (P < 0.001), and FCR (P < 0.05) during d 0 to 10 of broiler life but not effect of SDP was observed for the whole 0-42 d period. Carcass yield increased with HND (P < 0.001) and dietary SDP (P = 0.002). However, HND feeding significantly decreased liver (P < 0.001), bursa of Fabricius (P = 0.002), abdominal fat (P < 0.001), proventriculus (P < 0.001) and gizzard weight (P < 0.001), but increased heart weight (P = 0.013), although spleen weight remained unaffected (P > 0.05) on d 42. Tibial bone morphometric and mechanical properties improved (P < 0.05) with SDP supplementation, and bone ash, Ca, and P remained unaffected (P > 0.05) on d 14. With the exception at d 28 (P = 0.037), the antibody titer to ND virus was similar among all treatment groups (P > 0.05) at d 0, 14, and 42. In conclusion, HND diets improve performance of broilers during the whole period and SDP supplementation during starter phase improve performance at this period, but also increased carcass yield, and tibial quality. Therefore, inclusion of SDP in the starter diet could be a beneficial nutritional strategy to improve the health and production of broilers provided feeding strategies using various nutrient densities.

A graph model to describe the network connectivity of trabecular plates and rods.

Introduction: The trabecular network is perceived as a collection of interconnected plate- (P) and rod-like (R) elements. Previous research has highlighted how these elements and their connectivity influence the mechanical properties of bone, yet further work is required to elucidate better the deeply interconnected nature of the trabecular network with distinct element formations conducting forces per their mechanical boundary conditions. Within this network, forces act through elements: a rod or plate with force applied to one end will transmit this force to a component connected to the other end, defining the boundary conditions for the loading of each element. To that end, this study has two aims: First, to investigate the connectivity of individually segmented elements of trabecular bone with respect to their local boundary conditions as defined by the surrounding trabecular network and linking them directly to the bone's overall mechanical response during loading using a mathematical graph model of the plate and rod (PR) Network. Second, we use this model to quantify side artifacts, a known artifact when testing an excised specimen of trabecular bone, where vertical trabeculae lose their load-bearing capacity due to a loss of connectivity, ultimately resulting in a change of the trabecular network topology. Resuts: Connected elements derived from our model predicted apparent elastic modulus by fitting a linear regression (R 2 = 0.81). In comparison, prediction using conventional bone volume fraction results in a lower accuracy (R 2 = 0.72), demonstrating the ability of the PR Network to estimate compressive elastic modulus independent of specimen size or loading boundary condition. Discussion: PR Network models are a novel approach to describing connectivity within the trabecular network and incorporating mechanical boundary conditions within the morphological analysis, thus enabling the study of intrinsic material properties of trabecular bone. Ultimately, PR Network models may be an early predictor or provide further insights into osteo-degenerative diseases.

Zoledronate reduces loading-induced microdamage in cortical ulna of ovariectomized rats

As a daily physiological mechanism in bone, microdamage accumulation dissipates energy and helps to prevent fractures. However, excessive damage accumulation might bring adverse effects to bone mechanical properties, which is especially problematic among the osteoporotic and osteopenic patients treated by bisphosphonates. Some pre-clinical studies in the literature applied forelimb loading models to produce well-controlled microdamage in cortical bone. Ovariectomized animals were also extensively studied to assimilate human conditions of estrogen-related bone loss. In the present study, we combined both experimental models to investigate microdamage accumulation in the context of osteopenia and zoledronate treatment. Three-month-old normal and ovariectomized rats treated by saline or zoledronate underwent controlled compressive loading on their right forelimb to create in vivo microdamage, which was then quantified by barium sulfate contrast-enhanced micro-CT imaging. Weekly in vivo micro-CT scans were taken to evaluate bone (re)modeling and to capture microstructural changes over time. After sacrifice, three-point-bending tests were performed to assess bone mechanical properties. Results show that the zoledronate treatment can reduce cortical microdamage accumulation in ovariectomized rats, which might be explained by the enhancement of several bone structural properties such as ultimate force, yield force, cortical bone area and volume. The rats showed increased bone formation volume and surface after the generation of microdamage, especially for the normal and the ovariectomized groups. Woven bone formation was also observed in loaded ulnae, which was most significant in ovariectomized rats. Although all the rats showed strong correlations between periosteal bone formation and microdamage accumulation, the correlation levels were lower for the zoledronate-treated groups, potentially because of their lower levels of microdamage. The present study provides insights to further investigations of pharmaceutical treatments for osteoporosis and osteopenia. The same experimental concept can be applied in future studies on microdamage and drug testing.

Rat bone responses to hindlimb unloading-reloading: Composition, BMD and mechanical properties

IntroductionBone loss in lower limbs under space microgravity condition can persist even after the astronauts return to Earth's gravitational environment. It is crucial to determine the extent and timing of bone recovery after spaceflight, especially for long-term flights. However, there is a lack of comprehensive descriptions of the recovery of various aspects of bone quality, particularly in terms of compositional characteristics that reflect changes in bone quality. Therefore, the objective of this study is to investigate the alterations in bone mineral density, composition characteristics, and mechanical properties of rat lower limbs during unloading (tail suspension) and the subsequent recovery loading period. MethodsRats were subjected to tail suspension for four weeks (TS4) followed by reloading for an additional four weeks. Bone mineral density (BMD) was measured by μ-CT and analyzed. The mechanical properties of the tibia were assessed by a three-point bending test, and the composition characteristics of bone tissue were evaluated using Raman spectroscopy. Regression analysis was performed to identify main component related to mechanical property. ResultsIn the metaphysis, BMD of the tibia significant decreased, particularly in trabecular bone. Upon reloading, only the cortical bone showed recovery. In the diaphysis, cortical bone remained unchanged during unloading but demonstrated delayed bone loss during the reloading period. Simultaneously, the mechanical properties of tibia experienced a substantial reduction after four weeks of unloading, The stiffness and maximum strength did not fully recover, though the elastic modulus and the ultimate strength were restored following the reloading process. Raman spectroscopy characterization found an increase in the carbonate substitution and crystal crystallinity of the diaphysis after 4 weeks of unloading. There was an intriguing alteration in the mineralized collagen ratio, that an increase was observed in the diaphysis while a decrease was observed in the metaphysis. During 4 weeks of reloading, the compositional characteristics of the diaphysis recovered earlier than those of the metaphysis, with the exception of crystallinity which remained unrecovered. Regression analysis indicated that carbonate replacement has a strong correlation to bone mechanical properties during the 4 weeks of unloading and subsequent 4 weeks of reloading. ConclusionsThe data obtained in this study highlight the incomplete recovery of bone mineral density, mechanical properties, and compositional characteristics during reloading in tail-suspended rats. These findings deepen our understanding of microgravity-induced bone loss and provide valuable information to develop interventions not only for astronauts but also for persons on earth during and after prolonged periods of unloading.

Determining the Mechanical Properties of Super-Elastic Nitinol Bone Staples Through an Integrated Experimental and Computational Calibration Approach.

Super-elastic bone staples have emerged as a safe and effective alternative for internal fixation. Nevertheless, several biomechanical aspects of super-elastic staples are still unclear and require further exploration. Within this context, this study presents a combined experimental and computational approach to investigate the mechanical characteristics of super-elastic staples. Two commercially available staples with distinct geometry, characterized by two and four legs, respectively, were examined. Experimental four-point bending tests were conducted to evaluate staple performance in terms of generated forces. Subsequently, a finite element-based calibration procedure was developed to capture the unique super-elastic behavior of the staple materials. Finally, a virtual bench testing framework was implemented to separate the effect of geometry from that of the material characteristics on the mechanical properties of the devices, including generated force, strain distribution, and fatigue behavior. The experimental tests indicated differences in the force vs. displacement curves between staples. The material calibration procedure revealed marked differences in the super-elastic properties of the materials employed in staple 1 and staple 2. The results obtained from the virtual bench testing framework have showed that both geometric features and material characteristics had a substantial impact on the mechanical properties of the device, especially on the generated force, whereas their effect on strain distribution and fatigue behavior was comparatively less pronounced. To conclude, this study advances the biomechanical understanding of Nitinol super-elastic staples by separately investigating the impact of geometry and material characteristics on the mechanical properties of two commercially available devices.

Titanium Alloy Implants with Lattice Structures for Mandibular Reconstruction.

In recent years, the field of mandibular reconstruction has made great strides in terms of hardware innovations and their clinical applications. There has been considerable interest in using computer-aided design, finite element modelling, and additive manufacturing techniques to build patient-specific surgical implants. Moreover, lattice implants can mimic mandibular bone's mechanical and structural properties. This article reviews current approaches for mandibular reconstruction, their applications, and their drawbacks. Then, we discuss the potential of mandibular devices with lattice structures, their development and applications, and the challenges for their use in clinical settings.

Differential expression of the inflammatory ciita gene may be accompanied by altered bone properties in intact sex steroid-deficient female rats

ObjectiveThe class II transactivator (CIITA), encoded by the CIITA gene, controls expression of immune response regulators, which affect bone homeostasis. Previously, we investigated a functional CIITA polymorphism in elderly women. Women carrying the allele associated with lower CIITA levels displayed higher bone mineral density (BMD), but also higher bone loss. The present exploratory study in a rat model sought to investigate effects of differential expression of Ciita on bone structural integrity and strength. Two strains DA (normal-to-high expression) and DA.VRA4 (lower expression) underwent ovariectomy (OVX) or sham-surgery at ~ 14-weeks of age (DA OVX n = 8, sham n = 4; DA.VRA4 OVX n = 10, sham n = 2). After 16-weeks, femoral BMD and bone mineral content (BMC) were measured and morphometry and biomechanical testing performed.ResultsIn DA.VRA4 rats, BMD/BMC, cross-sectional area and biomechanical properties were lower. Ciita expression was accompanied by OVX-induced changes to cross-sectional area and femoral shaft strength; DA rats had lower maximum load-to-fracture. Thus, while lower Ciita expression associated with lower bone mass, OVX induced changes to structural and mechanical bone properties were less pronounced.ConclusionThe data tentatively suggests association between Ciita expression and structural and mechanical bone properties, and a possible role in bone changes resulting from estrogen deficiency.

Activin A marks a novel progenitor cell population during fracture healing and reveals a therapeutic strategy.

Insufficient bone fracture repair represents a major clinical and societal burden and novel strategies are needed to address it. Our data reveal that the transforming growth factor-β superfamily member Activin A became very abundant during mouse and human bone fracture healing but was minimally detectable in intact bones. Single-cell RNA-sequencing revealed that the Activin A-encoding gene Inhba was highly expressed in a unique, highly proliferative progenitor cell (PPC) population with a myofibroblast character that quickly emerged after fracture and represented the center of a developmental trajectory bifurcation producing cartilage and bone cells within callus. Systemic administration of neutralizing Activin A antibody inhibited bone healing. In contrast, a single recombinant Activin A implantation at fracture site in young and aged mice boosted: PPC numbers; phosphorylated SMAD2 signaling levels; and bone repair and mechanical properties in endochondral and intramembranous healing models. Activin A directly stimulated myofibroblastic differentiation, chondrogenesis and osteogenesis in periosteal mesenchymal progenitor culture. Our data identify a distinct population of Activin A-expressing PPCs central to fracture healing and establish Activin A as a potential new therapeutic tool.

Does tissue fixation change the mechanical properties of dry ovine bone extracellular matrix?

Tissue fixation is a prevalent method for bone conservation. Bone biopsies are typically fixed in formalin, dehydrated in ethanol, and infiltrated with polymethyl methacrylate (PMMA) Since some experiments can only be performed on fixed bone samples, it is essential to understand how fixation affects the measured material properties. The aim of this study was to quantify the influence of tissue fixation on the mechanical properties of cortical ovine bone at the extracellular matrix (ECM) level with state-of-the-art micromechanical techniques.A small section from the middle of the diaphysis of two ovine tibias (3.5 and 5.5 years old) was cut in the middle and polished on each side, resulting in a pair of mirrored surfaces. For each pair, one specimen underwent a fixation protocol involving immersion in formalin, dehydration with ethanol, and infiltration with PMMA. The other specimen (mirrored) was air-dried. Six osteons were selected in both pairs, which could be identified in both specimens. The influence of fixation on the mechanical properties was first analyzed using micropillar compression tests and nanoindentation in dry condition. Additionally, changes in the degree of mineralization were evaluated with Raman spectroscopy in both fixed and native bone ECM. Finally, micro tensile experiments were conducted in the 3.5-year fixed ovine bone ECM and compared to reported properties of unfixed dry ovine bone ECM.Interestingly, we found that tissue fixation does not alter the mechanical properties of ovine cortical bone ECM compared to experiments in dry state. However, animal age increases the degree of mineralization (p = 0.0159) and compressive yield stress (p = 0.041). Tissue fixation appears therefore as a valid conservation technique for investigating the mechanical properties of dehydrated bone ECM.

The Application of Advanced Bone Imaging Technologies in Sports Medicine

Until recently, the evaluation of bone health and fracture risk through imaging has been limited to dual-energy X-ray absorptiometry (DXA) and plain radiographs, with a limited application in the athletic population. Several novel imaging technologies are now available for the clinical assessment of bone health, including bone injury risk and healing progression, with a potential for use in sports medicine. Among these imaging modalities is high-resolution peripheral quantitative computed tomography (HR-pQCT) which is a promising technology that has been developed to examine the bone microarchitecture in both cortical and trabecular bone at peripheral anatomical sites. Technologies that do not expose patients to ionizing radiation are optimal, particularly for athletes who may require frequent imaging. One such alternative is diagnostic ultrasound, which is preferable due to its low cost and lack of radiation exposure. Furthermore, ultrasound, which has not been a common imaging modality for monitoring fracture healing, has been shown to potentially demonstrate earlier signs of union compared to conventional radiographs, including callus mineralization and density at the healing site. Through the use of conventional magnetic resonance imaging (MRI), finite element analysis (FEA) can be used to simulate the structural and mechanical properties of bone. On the other hand, the ultrashort echo time (UTE) MRI can evaluate cortical bone quality by detecting water bound to the organic bone matrix and free water, providing important information about bone porosity. Several novel bone imaging techniques originally developed for osteoporosis assessment have great potential to be utilized to improve the standard of care in bone fracture risk assessment and healing in sports medicine with much greater precision and less adverse radiation exposure.

Expanding the mechanical and mass-transport combination for bone scaffolds: Through stretched structure

Optimizing the mechanical and mass-transport properties of porous biomaterials simultaneously can be challenging, as they often correspond one-to-one through structure. For instance, increasing permeability by reducing relative density frequently leads to lower mechanical properties. In this study, we introduced a stretched structure to optimize the two properties simultaneously. Initially, the stretched scaffolds were evaluated at the same porosity, revealing that the elastic modulus and permeability of the stretched scaffold are approximately twice that of the original type. Subsequently, the orientation dependence of these properties was investigated. The results suggest that the stiffness of the stretched structure in the width direction is compromised, aligning with the mechanical properties of cortical bone in shaft bones. The permeability of the stretched structure exhibits significant anisotropy, with values much higher than those of original structure. Moreover, our biological experiments demonstrate that extremely stretched structures exhibit low and uneven curvature, potentially impeding cell growth. Hence, striking a balance between curvature and the stretching method employed is crucial. The stretching method can also be applied to other strut-based structures, enabling greater design flexibility in achieving a desirable mechanical and mass-transport combination, providing a foundation for high-performance artificial bone prostheses.

Dietary supplementation with nacre reduces cortical bone loss in aged female mice

Aging is associated with detrimental bone loss leading to fragility fractures in both men and women. Notably, a majority of bone loss with aging is cortical, as well as a large number of fractures are non-vertebral and at the non-hip sites. Nacre is a product of mollusks composed of calcium carbonate embedded in organic components. As our previous study demonstrated the protective effect of nacre supplementation on trabecular bone loss in ovariectomized rats, we sought to evaluate the effect of dietary nacre on bone loss related to aging in female mice which do not suffer true menopause as observed in women. The current study compared the effect of a 90-day long nacre-supplemented diet to that of Standard or CaCO3 diets on both bone mass and strength in 16-month-old C57BL/6 female mice. Multiple approaches were performed to assess the microarchitecture and mechanical properties of long bones, analyze trabecular histomorphometry, and measure bone cell-related gene expressions, and bone turnover markers. In the cortex, dietary nacre improved cortical bone strength in line with lower expression levels of genes reflecting osteoclasts activity compared to Standard or CaCO3 diets (p < 0.05). In the trabeculae, nacre-fed mice were characterized by a bone remodeling process more active than the other groups as shown by greater histomorphometric parameters and osteoblast-related gene expressions (p < 0.05). But these differences were not exhibited at the level of the trabecular microarchitecture at this age. Collectively, these data suggest that dietary nacre should be a potential candidate for reducing aging-associated cortical bone loss in the elderly.