Analysis of bone structure in PEROMYSCUS: Effects of burrowing behavior.

We compare the effects of burrowing behavior on appendicular bone structure in two Peromyscus (deer mouse) species. P. polionotus creates complex burrows in their territories, while P. eremicus is a non-burrowing nesting mouse. We examined museum specimens' bones of wild-caught mice of the two species and lab-reared P. polionotus not given the opportunity to burrow. Bones were scanned using micro-computed tomography, and cortical and trabecular bone structural properties were quantified. Wild P. polionotus mice had a larger moment of area in the ulnar and tibial cortical bone compared with their lab-reared counterparts, suggesting developmental adaptation to bending resistance. Wild P. polionotus had a larger normalized second moment of area and cross-sectional area in the tibia compared with P. eremicus. Tibial trabecular analysis showed lower trabecular thickness and spacing in wild P. polionotus than in P. eremicus and femoral analysis showed wild P. polionotus had lower thickness than P. eremicus and lower spacing than lab-reared P. polionotus, suggesting adaptation to high loads from digging. Results lay the groundwork for future exploration of the ontogenetic and evolutionary basis of mechanoadaptation in Peromyscus.

Lost in research translation: Female athletes are not male athletes, especially at the hip.

Altered shape of the proximal femur (cam morphology) or acetabulum (pincer morphology) is indicative of femoroacetabular impingement, which can result in hip pain and osteoarthritis of the hip. As mechanical load during growth affects the resulting bone shape, there is strong evidence in males that cam morphology develops during skeletal growth while physes are open, rather than as an adaptation after growth plates are closed (skeletal maturity). This adaptation is particularly evident in athletes who participate at elite levels prior to skeletal maturity. The research providing this evidence, however, has primarily focused on male athletes. Despite the lack of inclusion in the research, females consistently comprise two thirds of the clinical and surgical populations with structural hip pain or pathology. Knowledge gained from male-dominated cohorts may not appropriately transfer to female athletes, especially at the hip. This perspectives article briefly reviews differences between females and males in femoral and acetabular structure, hormones, timing of puberty/maturation, hypermobility, activity level and movement control-factors which affect hip structure development and loading. Without female-focused research, the application of research findings from male athletes to female athletes may lead to ineffective or even inappropriate recommendations and treatments. Thus, there is a critical need for investment in research to promote life-long hip health for females.

E-cigarette aerosol exposure effect on bone biomechanical properties in murine models.

Numerous studies have shown the detrimental health effects of tobacco smoking on bone volume and strength in human and animal models. Little is known regarding the impacts of e-cigarettes, a form of smoke-less nicotine intake, despite their growing population of users. This study uses murine models to evaluate the effects of exposure to e-cigarette aerosols (JUUL) on bone structure and strength through micro-CT imaging and mechanical testing. JUUL mice had more trabecular bone in thickness and volume, yet lower ultimate stress and modulus values in the cortical bone than the control mice. These outcomes suggest that, although vaping can result in a higher bone volume, this bone is weaker than average. E-cigarettes should be examined more closely regarding adolescence and long-term consequences on skeletal health.

Determining the effects of AR/VR HMD design parameters (mass and inertia) on cervical spine joint torques.

This study aimed to determine gravitational and dynamic torques and muscle activity of the neck across a series of design parameters of head mounted displays (mass, center of mass, and counterweights) associated with virtual and augmented reality (VR/AR). Twenty young adult participants completed five movement types (Slow and Fast Flexion/Extension and Rotation, and Search) while wearing a custom-designed prototype headset that varied the three design parameters: display mass (0, 200, 500, and 750 g), distance of the display's center of mass in front of the eyes (approximately 1, 3, and 5 cm anteriorly), and counterweights of 0, 166, 332, and 500 g to balance the display mass of 500 g at 7 cm. Inverse dynamics of a link segment model of the head and headset provided estimates of the torques about the joint between the skull and the occiput-first cervical vertebrae (OC1) and joint between the C7 and T1 vertebrae (C7). Surface electromyography (EMG) measured bilateral muscle activity of the splenius and upper trapezius muscles. Adding 750 g of display mass nearly doubled root mean square joint torques across all movement types. Increasing the distance of the display mass in front of the eyes by 4 cm increased torques about OC1 for the Slow and Fast Rotation and Search movements by approximately 20%. Adding a counterweight decreased torques about OC1 during the rotation and search tasks but did not decrease the torques experienced in the lower cervical spine (C7). For the flexion/extension axis, the magnitude of the dynamic torque component was 20% or less of the total torque experienced whereas for the rotation axis the magnitude of the dynamic torque component was greater than 50% of the total torque. Surface EMG root mean square values significantly varied across movement types with the fast rotation having the largest values; however, they did not vary significantly across the headset configurations.

Material properties in regenerating axolotl limbs using inverse finite element analysis.

BACKGROUND: The extracellular mechanical environment plays an important role in the skeletal development process. Characterization of the material properties of regenerating tissues that recapitulate development, provides insights into the mechanical environment experienced by the cells and the maturation of the matrix. In this study, we estimated the viscoelastic material properties of regenerating forelimbs in the axolotl (Ambystoma mexicanum) at three different regeneration stages: 27 days post-amputation (mid-late bud) and 41 days post-amputation (palette stage), and fully-grown time points. A stress-relaxation indentation test followed by two-term Prony series viscoelastic inverse finite element analysis was used to obtain material parameters. Glycosaminoglycan (GAG) content was estimated using a 1,9- dimethyl methylene blue assay. RESULTS: The instantaneous and equilibrium shear moduli significantly increased with regeneration while the short-term stress relaxation time significantly decreased with limb regeneration. The long-term stress relaxation time in the fully-grown time point was significantly lower than 27 and 41 DPA groups. The GAG content was not significantly different between 27 and 41 DPA but the GAG content of cartilage in the fully-grown group was significantly greater than in 27 and 41 DPA. CONCLUSIONS: The mechanical environment of the proliferating cells changes drastically during limb regeneration. Understanding how the tissue's mechanical properties change during limb regeneration is critical for linking molecular-level matrix production of the cells to tissue-level behavior and mechanical signals.

Automated measurement of lamellar thickness in human bone using polarized light microscopy.

Lamellar bone formed in individuals with moderate and severe osteogenesis imperfecta (OI) is often composed of lamellae that are structurally abnormal. Measuring the thickness of these lamellae can be helpful in assessing the effect of specific collagen and collagen-related mutations on OI bone synthesis. Manual measurement of lamellar thicknesses in large quantities is very time consuming. The method for automated measurement described in this article utilizes an image processing script to identify the average thickness of multiple lamellae automatically from histologic images of bone. This allows for faster measurements that are less prone to human error and can account for variability in the thickness of a lamella along its length.•OI and control bone samples are prepared per the glycol methacrylate resin (JB-4 plastic) technique and viewed using polarized light microscopy.•Ideal bone regions for measurement are identified using specific qualitative criteria designed to ensure uniform and accurate thickness measurements.•The method was validated with dataset containing 211 lamellae from control bone and 212 lamellae from OI bone.

Arthritis Foundation/HSS Workshop on Hip Osteoarthritis, Part 1: Epidemiology, Early Development, and Cohorts From Around the World.

Far more publications are available for osteoarthritis of the knee than of the hip. Recognizing this research gap, the Arthritis Foundation, in partnership with the Hospital for Special Surgery, convened an in-person meeting of thought leaders to review the state of the science of and clinical approaches to hip osteoarthritis. This article summarizes the recommendations and clinical research gaps gleaned from 5 presentations given in the "how hip osteoarthritis begins" session of the 2023 Hip Osteoarthritis Clinical Studies Conference, which took place on February 17 and 18, 2023, in New York City.

Neonatal Hip Loading in Developmental Dysplasia: Finite Element Simulation of Proximal Femur Growth and Treatment.

Background: Abnormal prenatal hip joint loading can lead to compromised hip joint function. Early intervention is crucial for favorable outcomes. Purpose: This study investigates the impact of treatment timing (initiation and duration) on cartilage growth and ossification in the proximal femur of infants with developmental dysplasia of the hip, a condition affecting newborns. Methods: We used a mechanobiological model to simulate proximal femur growth during treatment durations of 3 months, 6 months, and a late-start treatment. Results: The findings indicate that the timing of treatment initiation is crucial, while a longer treatment duration does not contribute to improved morphological development of the hip joint. Conclusions: Mechanobiological models of growth can be used to develop treatments and therapies that correct loading conditions. Growing bone is particularly sensitive to loading conditions, and altered loading during growth can affect bone shape and functionality.

Frontal plane pelvic kinematics during high velocity running: Association with hamstring injury history.

INTRODUCTION: Hamstring injuries are the most prevalent non-contact soft tissue injury in sports, with a larger portion of injuries being recurrent. The sagittal plane running kinematics correlated to hamstring injury history has been well documented. However, analysis of frontal plane kinematics allows for observation of stability and symmetry. This study aimed to examine the frontal plane running kinematics of elite collegiate level sprinters, with and without previous hamstring injury, compared to healthy counterparts. METHODS: Thirty-nine participants performed three 50-m sprints, with three inertial measurement unit sensors placed on the pelvis: one on each iliac crest and one on the sacrum. Participants were classified based on sex, competitive status, and injury history. To investigate differences based on group classification, the data were used to analyze mediolateral motion (relative magnitude of mediolateral acceleration) and asymmetry (difference in acceleration between right and left iliac crests) during each stance phase throughout the run. RESULTS: Injured sprinters displayed significantly greater mediolateral motion and asymmetry during stances than healthy counterparts. CONCLUSIONS: This study demonstrates that frontal plane running stance dynamics are different in athletes with previous hamstring injury than healthy athletes. These athletes may benefit from rehabilitation strategies targeting postural control and stability during dynamic tasks.

Can pelvic tilt cause cam morphology? A computational model of proximal femur development mechanobiology.

Cam deformity of the proximal femur is a risk factor for early osteoarthritis. While cam morphology is related to mechanical force at a formative time in skeletal growth, the specific problematic forces contributing to the development of cam morphology remain unknown. Individuals with femoroacetabular impingement syndrome exhibit an increased anterior pelvic tilt during walking, which alters their hip joint forces. This study aims to investigate the influence of altered joint force caused by anterior pelvic tilt on proximal femur epiphyseal growth and the potential association between increased anterior pelvic tilt and the development of cam morphology. A computational model is utilized to simulate the endochondral ossification in the proximal femur and predict cam formation. Cartilage growth and ossification patterns for a gait cycle with and without anterior pelvic tilt were modeled. The simulated growth results indicated an increased alpha angle (53° for typically developing to 68° for anterior pelvic tilt) and aspherical femoral head in the model with anterior pelvic tilt. We conclude that anterior pelvic tilt may be sufficient to cause the formation of the cam morphology. Identifying the critical mechanical conditions that increase the risk of cam deformity could help prevent this condition by adjusting the physical activities before skeletal maturity.

Using Finite Element Modeling in Bone Mechanoadaptation.

PURPOSE OF THE REVIEW: Bone adapts structure and material properties in response to its mechanical environment, a process called mechanoadpatation. For the past 50 years, finite element modeling has been used to investigate the relationships between bone geometry, material properties, and mechanical loading conditions. This review examines how we use finite element modeling in the context of bone mechanoadpatation. RECENT FINDINGS: Finite element models estimate complex mechanical stimuli at the tissue and cellular levels, help explain experimental results, and inform the design of loading protocols and prosthetics. FE modeling is a powerful tool to study bone adaptation as it complements experimental approaches. Before using FE models, researchers should determine whether simulation results will provide complementary information to experimental or clinical observations and should establish the level of complexity required. As imaging technics and computational capacity continue increasing, we expect FE models to help in designing treatments of bone pathologies that take advantage of mechanoadaptation of bone.

Lamellar thickness measurements in control and osteogenesis imperfecta human bone, with development of a method of automated thickness averaging to simplify quantitation.

Lamellar bone that forms in moderate and severe osteogenesis imperfecta (OI) is composed of structurally irregular lamellae compared to those in control bone. OI and control cortical bone fragments were prepared for light microscopy in standardized fashion: decalcified, embedded in plastic, sectioned and stained with toluidine blue. Polarization light microscopy (PLM) was used to demonstrate and quantify bright and dark lamellar thicknesses in cortical bone fragments from 5 patients with moderate to severe OI in whom type I collagen structural/molecular defects were detected and in control bone from 5 patients. Rigid selection criteria identified lamellar regions for quantification. Thicknesses of bright and dark lamellae were measured manually at 20X magnification using a histomorphometric image analysis system. A method of automated thickness averaging was developed to determine lamellar thicknesses from PLM images to make measurement faster. Our study demonstrates, for the first time, that in OI bone from patients with type I collagen structural/molecular defects mean lamellar thickness measurements (along with the bright and dark lamellar thicknesses) were less than those in control bone by statistically highly significant differences. The mean value for bright lamellae was less than that for dark lamellae in both control and OI bone. The ratio of mean values for bright/dark lamellar thicknesses was the same in control and OI bone. The automated method obtained similar results to the manual method. Lamellar bone in moderate and severe OI with type I collagen defects is composed of thinner and less structurally regular lamellae than those in control bone. This finding indicates that lamellar thickness measurements can be helpful in assessing the effect of specific collagen and collagen-related mutations on OI bone synthesis and warrant inclusion in research and clinical histomorphometric assessments.

Adolescent obesity incurs adult skeletal deficits in murine induced obesity model.

Adolescent obesity has risen dramatically in the last few decades. While adult obesity may be osteoprotective, the effects of obesity during adolescence, which is a period of massive bone accrual, are not clear. We used a murine model of induced adolescent obesity to examine the structural, mechanical, and compositional differences between obese and healthy weight bone in 16-week-old female C57Bl6 mice. We also examined the effects of a return to normal weight after skeletal maturity (24 weeks old). We found obese adolescent bone exhibited decreased trabecular bone volume, increased cortical diameter, increased ultimate stress, and increased brittleness (decreased plastic energy to fracture), similar to an aging phenotype. The trabecular bone deficits remained after return to normal weight after skeletal maturity. However, after returning to normal diet, there was no difference in ultimate stress nor plastic energy to fracture between groups as the normal diet group increased ultimate stress and brittleness. Interestingly, compositional changes appeared in the former high-fat diet mice after skeletal maturity with a lower mineral to matrix ratio compared to normal diet mice. In addition there was a trend toward increased fluorescent advanced glycation endproducts in the former high-fat diet mice compared to normal diet mice but this did not reach significance (p < 0.05) due to the large variability. The skeletal consequences of adolescent obesity may have lasting implications for the adult skeleton even after return to normal weight. Given the rates of adolescent obesity, skeletal health should be a concern.

The role of computational models in mechanobiology of growing bone.

Endochondral ossification, the process by which long bones grow in length, is regulated by mechanical forces. Computational models, specifically finite element models, have been used for decades to understand the role of mechanical loading on endochondral ossification. This perspective outlines the stages of model development in which models are used to: 1) explore phenomena, 2) explain pathologies, 3) predict clinical outcomes, and 4) design therapies. As the models progress through the stages, they increase in specificity and biofidelity. We give specific examples of models of endochondral ossification and expect models of other mechanobiological systems to follow similar development stages.

Manipulating load-induced fluid flow in vivo to promote bone adaptation.

Mechanical stimulation is critical to maintaining bone mass and strength. Strain has been commonly thought of as the mechanical stimulus driving bone adaptation. However, numerous studies have hypothesized that fluid flow in the lacunar-canalicular system plays a role in mechanoadaptation. The role of fluid flow compared to strain magnitude on bone remodeling has yet to be characterized. This study aimed to determine the contribution of fluid flow velocity compared to strain on bone adaptation. We used finite element modeling to design in vivo experiments, manipulating strain and fluid flow contributions. Using a uniaxial compression tibia model in mice, we demonstrated that high fluid flow velocity results in significant bone adaptation even under low strain magnitude. In contrast, high strain magnitude paired with low fluid velocity does not trigger a bone response. These findings support previous hypotheses stating that fluid flow is the principal mechanical stimulus driving bone adaptation. Moreover, they give new insights regarding bone adaptative response and provide new pathways toward treatment against age-related mechanosensitivity loss in bone.

Usability of deep learning pipelines for 3D nuclei identification with Stardist and Cellpose.

Segmentation of 3D images to identify cells and their molecular outputs can be difficult and tedious. Machine learning algorithms provide a promising alternative to manual analysis as emerging 3D image processing technology can save considerable time. For those unfamiliar with machine learning or 3D image analysis, the rapid advancement of the field can make navigating the newest software options confusing. In this paper, two open-source machine learning algorithms, Cellpose and Stardist, are compared in their application on a 3D light sheet dataset counting fluorescently stained proliferative cell nuclei. The effects of image tiling and background subtraction are shown through image analysis pipelines for both algorithms. Based on our analysis, the relative ease of use of Cellpose and the absence of need to train a model leaves it a strong option for 3D cell segmentation despite relatively longer processing times. When Cellpose's pretrained model yields results that are not of sufficient quality, or the analysis of a large dataset is required, Stardist may be more appropriate. Despite the time it takes to train the model, Stardist can create a model specialized to the users' dataset that can be iteratively improved until predictions are satisfactory with far lower processing time relative to other methods.

Local mechanical stimuli correlate with tissue growth in axolotl salamander joint morphogenesis.

Movement-induced forces are critical to correct joint formation, but it is unclear how cells sense and respond to these mechanical cues. To study the role of mechanical stimuli in the shaping of the joint, we combined experiments on regenerating axolotl (Ambystoma mexicanum) forelimbs with a poroelastic model of bone rudiment growth. Animals either regrew forelimbs normally (control) or were injected with a transient receptor potential vanilloid 4 (TRPV4) agonist during joint morphogenesis. We quantified growth and shape in regrown humeri from whole-mount light sheet fluorescence images of the regenerated limbs. Results revealed significant differences in morphology and cell proliferation between groups, indicating that TRPV4 desensitization has an effect on joint shape. To link TRPV4 desensitization with impaired mechanosensitivity, we developed a finite element model of a regenerating humerus. Local tissue growth was the sum of a biological contribution proportional to chondrocyte density, which was constant, and a mechanical contribution proportional to fluid pressure. Computational predictions of growth agreed with experimental outcomes of joint shape, suggesting that interstitial pressure driven from cyclic mechanical stimuli promotes local tissue growth. Predictive computational models informed by experimental findings allow us to explore potential physical mechanisms involved in tissue growth to advance our understanding of the mechanobiology of joint morphogenesis.

Marrow aspiration in aged mice: intramedullary osteogenesis, reduced mechano-adaptation, increased marrow fat.

Introduction: With age, the number of adipocytes and osteoclasts increases, the number of osteoblasts decreases, and mechano-adaptation is impaired.Objectives: Using marrow aspiration, which has a known osteogenic effect in young mice, we sought to recruit osteoblast progenitors to mediate the mechano-adaptive response to in vivo tibial loading.Methods: First, we assessed bone formation and marrow adiposity in the tibiae of old mice (>20 months) sacrificed 1, 2, and 4 weeks after unilateral marrow aspiration. Then, we examined the effects of marrow aspiration on mechano-adaptation in aged mice using tibial loading.Results: Two weeks after aspiration, aspirated tibiae had more bone than contralateral tibiae due to the formation of bone in the medullary canal. Two weeks and four weeks after marrow aspiration, the volume of marrow adipose tissue was higher in the aspirated tibiae, compared to contralateral tibiae. Histomorphometry indicated that aspiration increased non-periosteal (endosteal, intracortical, intramedullary) bone formation, compared to the contralateral tibia.  Mice with marrow aspiration had reduced periosteal bone formation in the contralateral tibia, compared to mice that had loading alone. Loading-induced periosteal bone formation was higher in mice that had loading alone, compared to mice that had aspiration + loading, indicating that aspiration further reduced the mechano-adaptive response.Conclusion: These data demonstrate that, in old mice, bone forms in the medullary canal following aspiration. Adiposity is increased following marrow aspiration, and periosteal mechano-adaptation is reduced.

Computational model of endochondral ossification: Simulating growth of a long bone.

Mechanical loading is a crucial factor in joint and bone development. Using a computational model, we investigated the role of mechanics on cartilage growth rate, ossification of the secondary center, formation of the growth plate, and overall bone shape. A computational algorithm was developed and implemented into finite element models to simulate the endochondral ossification for symmetric and asymmetric motion in a generic diarthrodial joint. Under asymmetric loading condition the secondary center ossifies asymmetrically leaning toward the external load and results in tilted growth plate. Also the mechanics seems to have greater influence in the early onset of the ossification of the secondary center rather than later progression of the center. While previous models have simulated select stages of skeletal development, our model can simulate growth and ossification during the entirety of post-natal development. Such computational models of skeletal development may provide insight into specific loading conditions that cause bone and joint deformities, and the required timing for rehabilitative repair.

Mechanoadaptation of the bones of mice with high fat diet induced obesity in response to cyclical loading.

An upward trend in childhood obesity implies a great need to determine its effects, both immediate and long-term. Obesity is osteoprotective in adults, but we know very little about the effects of obesity on the growing skeleton, particularly its ability to adapt to load. The objective of this research is to assess bone mechanoadaptation in adolescent obese mice. Ten mice were fed a high-fat diet (HFD) from 4 to 16 weeks of age, while a control group of the same size received a normal diet (ND). At 14 weeks of age, right tibiae were cyclically loaded with a 12 N peak load for HFD mice and a 9 N peak load for ND mice three times a week for two weeks, resulting in equal peak strains of about 2500 microstrain. At 16 weeks of age, mice were sacrificed, and tibiae and gonadal fat pads were dissected. Fat pads were weighed as an obesity indicator, and tibiae were imaged with microCT to measure bone structure. The left tibiae (nonloaded) were subsequently decalcified, stained with osmium, and scanned to quantify marrow fat. Results showed that HFD mice had larger tibial cross-sectional areas compared to ND mice, as well as greater marrow adiposity. However, there was no significant difference in the amount of bone adaptation in the cortical or trabecular bone between the two groups. This indicates that the bones of HFD and ND mice adapt equally well to loading.