Bone Stiffness Research Articles

Markers of bone turnover and biomechanical properties of the third metacarpal bone of growing pigs subjected to the different dietary phosphorus and calcium content

Abstract Eighteen weaned piglets were subjected to deficiency, adequate or excess of dietary phosphorus and calcium (group C−50, C and C+50, respectively) during growth from 33 to 110 day of age. However, the ratio of calcium/digestible phosphorus in all feeds was the same and consistent with the nutritional recommendation. Performance, bone growth, concentration of osteocalcin (OC) and carboxyterminal telopeptide of type I collagen (CTX-I), densitometry, geometry and mechanical properties of the 3rd metacarpal bone were measured. Growth rate and final body weight did not differ between C and C+50 animals, however, they grew approximately 5.9% faster and were heavier by about 4.0% than C−50 pigs. In turn, mass and length of bones were similar in C−50 and C+50 groups, however, they were approximately 11% lighter and 4% shorter compared to C pigs. Serum content of OC took the following order: C−50 < C < C+50 group (P<0.01). Content of CTX-I did not differ between C and C+50 animals, however they were lower by 36% (P<0.01) than in the C−50 pigs. All measured geometry parameters (cortical wall thickness, cross section area and cortical index) did not differ between C and C+50 pigs and were higher than in C−50 pigs (P<0.01). Bones of C−50 pigs contained less minerals than bones of the C and C+50 pigs (5.24 g vs mean 6.14 g, P<0.01). The 3rd metacarpal bone mineral density (g/cm2) was the lowest in pigs in group C−50, higher in pigs in group C, and the highest in animals in group C+50 (0.398, 0.431 and 0.475, respectively, P<0.01). Maximum strength of bones was similar in the C and C+50 pigs, however it was 13% greater (P<0.01) compared to C−50 animals. In turn, elastic strength and elastic deflection did not differ between the C and C−50 animals and were over 20% lower (P<0.01) than in the C+50 pigs. Whereas, there was no significant difference between groups in bone stiffness (mean 27.13 mm). The results of the present study proved that even a strong excess of phosphorus and calcium content in the diet for growing pigs is inadvisable and does not improve performance and bones parameters. While excessively lowering the dietary phosphorus and calcium concentration (e.g. to reduce environmental pollution) may adversely affect growth rate and bone characteristics.

Anti-osteoporotic effects of Yi Mai Jian on bone metabolism of ovariectomized rats.

Yi Mai Jian herbal formula (YMJ) is formulated with Eucommiae Folium, Astragali Radix, Ligustri Lucidi Fructus, and Elaeagnus Fructus to improve bone function in traditional Chinese medicine. The anti-osteoporotic effects of YMJ in bone metabolism were evaluated in ovariectomized (OVX) rats. The skeletal structure of the femur and vertebrae was analyzed after treating OVX rats with YMJ for 114days. The results showed that YMJ significantly increased the bone mineral density (BMD) and trabecular number (Tb. N) of the femur and 5th lumbar vertebrae and reduced trabecular separation (Tb. Sp). Moreover, trabecular bone volume/total tissue volume (BV/TV), bone stiffness, and maximum femur load were significantly increased. The serum concentrations of NTX1 and PYD were significantly decreased. According to these results, YMJ could ameliorate osteoporosis in ovariectomized rats. Eucommiae Folium and Elaeagnus Fructus inhibited osteoclast differentiation, Ligustri Lucidi Fructus inhibited calcium reabsorption, Astragali Radix stimulated osteoblast proliferation, and Astragali Radix and Eucommiae Folium stimulated mineralization. Therefore, the combination of the four herbs into one formula, YMJ, could alleviate bone remodeling caused by low estrogen levels. We suggest that YMJ could be a healthy food candidate for preventing post-menopausal osteoporosis.

The primary stability of a cementless PEEK femoral component is sensitive to BMI: A population-based FE study

The use of polyetheretherketone (PEEK) for cementless femoral total knee arthroplasty (TKA) components is of interest due to several potential advantages, e.g. the use in patients with metal hypersensitivity. Additionally, the stiffness of PEEK closer resembles the stiffness of bone, and therefore, peri-prosthetic stress-shielding may be avoided. When introducing a new implant material for cementless TKA designs, it is important to study its effect on the primary fixation, which is required for the long-term fixation. Finite element (FE) studies can be used to study the effect of PEEK as implant material on the primary fixation, which may be dependent on patient factors such as age, gender and body weight index (BMI). Therefore, the research objectives of this study were to investigate the effect of PEEK vs cobalt-chrome (CoCr) and patient characteristics on the primary fixation of a cementless femoral component. 280 FE models of 70 femora were created with varying implant material and gait and squat activity. Overall, the PEEK models generated larger peak micromotions than the CoCr models. Distinct differences were seen in the micromotion distributions between the PEEK and CoCr models for both the gait and squat models. The micromotions of all femoral models significantly increased with BMI. Neither gender nor age of the patients had a significant effect on the micromotions. This population study gives insights into the primary fixation of a cementless femoral component in a cohort of FE models with varying implant material and patient characteristics.

Interleukin-19 in Bone Marrow Contributes to Bone Loss Via Suppressing Osteogenic Differentiation Potential of BMSCs in Old Mice.

Cellular senescence is an important process related to the pathogenic mechanism of different disorders, especially bone loss. During senescence, bone marrow stromal cells (BMSCs) lose their self-renewal and functional differentiation abilities. Therefore, finding signals opposing the osteogenic differentiation of BMSCs within bone marrow microenvironment is the important for elucidating these above-mentioned mechanisms. Inflammatory cytokines affect bone physiology and remodeling. However, the function of interleukin-19 (IL-19) in skeletal system remains unclear. The mouse model of IL-19 knockout was established through embryonic stem cell injection for analyzing how IL-19 affected bone formation. Micro-CT examinations were performed to evaluate bone microstructures. We performed a three-point bending test to measure bone stiffness and the ultimate force. Antibody arrays were performed to detect interleukin family members in bone marrow aspirates. BMSCs were cultured and induced for osteogenic differentiation. According to our findings, there was increased IL-19 accumulation within bone marrow in old mice relative to that in their young counterparts, resulting in bone loss via the inhibition of BMSCs osteogenic differentiation. Among Wnt/β-catenin pathway members, IL-19 strongly upregulated sFRP1 via STAT3 phosphorylation. The inhibition of STAT3 and sFRP1 abolished IL-19's inhibition against the BMSCs osteogenic differentiation. To sum up, IL-19 inhibited BMSCs osteogenic differentiation in old mice. Our findings shed novel lights on pathogenic mechanism underlying age-related bone loss and laid a foundation for further research on identifying novel targets to treat senile osteoporosis.

The loading direction dramatically affects the mechanical properties of the mouse tibia.

Introduction: The in vivo tibial loading mouse model has been extensively used to evaluate bone adaptation in the tibia after mechanical loading treatment. However, there is a prevailing assumption that the load is applied axially to the tibia. The aim of this in silico study was to evaluate how much the apparent mechanical properties of the mouse tibia are affected by the loading direction, by using a validated micro-finite element (micro-FE) model of mice which have been ovariectomized and exposed to external mechanical loading over a two-week period. Methods: Longitudinal micro-computed tomography (micro-CT) images were taken of the tibiae of eleven ovariectomized mice at ages 18 and 20weeks. Six of the mice underwent a mechanical loading treatment at age 19weeks. Micro-FE models were generated, based on the segmented micro-CT images. Three models using unitary loads were linearly combined to simulate a range of loading directions, generated as a function of the angle from the inferior-superior axis (θ, 0°-30° range, 5° steps) and the angle from the anterior-posterior axis (ϕ, 0°: anterior axis, positive anticlockwise, 0°-355° range, 5° steps). The minimum principal strain was calculated and used to estimate the failure load, by linearly scaling the strain until 10% of the nodes reached the critical strain level of -14,420με. The apparent bone stiffness was calculated as the ratio between the axial applied force and the average displacement along the longitudinal direction, for the loaded nodes. Results: The results demonstrated a high sensitivity of the mouse tibia to the loading direction across all groups and time points. Higher failure loads were found for several loading directions (θ = 10°, ϕ 205°-210°) than for the nominal axial case (θ = 0°, ϕ = 0°), highlighting adaptation of the bone for loading directions far from the nominal axial one. Conclusion: These results suggest that in studies which use mouse tibia, the loading direction can significantly impact the failure load. Thus, the magnitude and direction of the applied load should be well controlled during the experiments.

The Effect of Patient-Related Factors on the Primary Fixation of PEEK and Titanium Tibial Components: A Population-Based FE Study.

Polyetheretherketone (PEEK) is of interest as implant material for cementless tibial total knee arthroplasty (TKA) components due to its potential advantages. One main advantage is that the stiffness of PEEK closely resembles the stiffness of bone, potentially avoiding peri-prosthetic stress-shielding. When introducing a new implant material for cementless TKA designs, it is essential to study its effect on the primary fixation. The primary fixation may be influenced by patient factors such as age, gender, and body mass index (BMI). Therefore, the research objectives of this finite element (FE) study were to investigate the effect of material (PEEK vs. titanium) and patient characteristics on the primary fixation (i.e., micromotions) of a cementless tibial tray component. A total of 296 FE models of 74 tibiae were created with either PEEK or titanium material properties, under gait and squat loading conditions. Overall, the PEEK models generated larger peak micromotions than the titanium models. Differences were seen in the micromotion distributions between the PEEK and titanium models for both the gait and squat models. The micromotions of all tibial models significantly increased with BMI, while gender and age did not influence micromotions.

Combining PTH(1-34) and mechanical loading has increased benefit to tibia bone mechanics in ovariectomised mice.

Combined treatment with PTH(1-34) and mechanical loading confers increased structural benefits to bone than monotherapies. However, it remains unclear how this longitudinal adaptation affects the bone mechanics. This study quantified the individual and combined longitudinal effects of PTH(1-34) and mechanical loading on the bone stiffness and strength evaluated in vivo with validated micro-finite element (microFE) models. C57BL/6 mice were ovariectomised at 14-week-old and treated either with injections of PTH(1-34), compressive tibia loading or both interventions concurrently. Right tibiae were in vivo microCT-scanned every 2 weeks from 14 until 24-week-old. MicroCT images were rigidly registered to reference tibia and the cortical organ level (whole bone) and tissue level (midshaft) morphometric properties and bone mineral content were quantified. MicroCT images were converted into voxel-based homogeneous, linear elastic microFE models to estimate the bone stiffness and strength. This approach allowed us for the first time to quantify the longitudinal changes in mechanical properties induced by combined treatments in a model of accelerated bone resorption. Both changes of stiffness and strength were higher with co-treatment than with individual therapies, consistent with increased benefits with the tibia bone mineral content and cortical area, properties strongly associated with the tibia mechanics. The longitudinal data shows that the two bone anabolics, both individually and combined, had persistent benefit on estimated mechanical properties, and that benefits (increased stiffness and strength) remained after treatment was withdrawn.

Relationship between DXA measured systemic bone mineral density and subchondral bone cysts in postmenopausal female patients with knee osteoarthritis: a cross-sectional study

BackgroundIndividuals with high systemic bone mineral density (BMD) may have an increased risk of incident knee osteoarthritis (OA). Besides that, radiographic osteophytes are strongly associated with BMD. Because of these reasons, the aim of the study was to investigate the possible association between radiological subchondral bone cyst (SBC) grade and systemic BMD and vitamin D status in the postmenopausal female patients with knee OA in a crosss-sectional study.MethodsThis study included of 48 osteoporosis treatment-free postmenopausal patients diagnosed with symptomatic medial compartment knee OA. BMD analysis was performed using dual-energy X-ray absorptiometry (DXA) and serum vitamin D levels were measured after recording patients’ findings. Each knee was scanned using computed tomography (CT), and categorical SBC scores were graded for the medial and lateral tibiofemoral (TF) and patellofemoral (PF) compartments and further calculated as compartmental total, total TF and grand total of both TF compartments. SBC scores were analysed with correlation analysis.ResultsThe patient population was characterized by radiographic joint space narrowing, obesity and low vitamin D status. Median medial total and grand total TF SBC scores were significantly different between the patient groups according to the Kellgren-Lawrence (KL) radiographic grading (p = 0.006 and p = 0.007, respectively). There were no correlations between femoral BMD values and SBC scores. However, positive correlations were detected significantly between L1 − 4 DXA values and TF SBC scores, but not with PF SBC scores (p = 0.005 for the correlation between L1 − 4 BMD and medial compartments total TF SBC score, p = 0.021 for the correlation between L1 − 4 BMD and grand total TF SBC score). No significant correlations were found with Vitamin D levels.ConclusionsDevelopment of TF OA high-grade SBCs may be linked to systemic bone mass as represented by trabecular bone-rich lumbar vertebrae. The relationship might point to the importance of bone stiffness as an acting factor in knee OA possibly with mechanical energy transfer to the joint.

The impact of complexation or complex coacervation of lactoferrin and osteopontin on simulated infant gastrointestinal digestion, intestinal inflammation, and in vivo bone development.

Lactoferrin (LF) and osteopontin (OPN) are bioactive milk proteins which can form heteroprotein complexes and complex coacervates. This research studied the effect of LF-OPN complexation and complex coacervation on the simulated infant gastrointestinal digestion of LF with subsequent examination of gut and bone health bioactivities in preclinical models. In an infant digestion model, the proteolytic profile of LF was unaltered by the pre-association of LF and OPN. Gastric proteolysis of LF was increased when the model gastric pH was reduced from 5.3 to 4.0, but less so when complexed with OPN. In a model of intestinal inflammation, undigested (79% inhibition) and gastric digestates (26% inhibition) of LF, but not gastrointestinal digestates, inhibited lipopolysaccharide (LPS)-induced NF-κB activation in intestinal epithelial cells. LF-OPN complexation sustained the inhibitory effect (21-43% of the undigested effect, depending on the type of complex) of LF after gastrointestinal digestion, suggesting that the peptides produced were different. In a neonatal rodent model used to study bone development, coacervating LF and OPN improved bone structures with a significant increase of trabecular proportion (BV/TV increase by 21.7%). This resulted in an 11.3% increase in stiffness of bones. Feeding the LF and OPN proteins in coacervate format also increased the levels of OPN, P1NP and M-CSF in blood, signifying a more pronounced impact on bone development. This research demonstrated that LF-OPN complexation and complex coacervation can delay simulated infant gastrointestinal digestion of LF and protect or improve the bioactivity of the proteins.

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.

Comparing the biomechanical properties of conventional suture and all-suture anchors using patient-specific and realistic osteoporotic and non-osteoporotic phantom using 3D printing

Conventional suture anchors (CAs) and all-suture anchors (ASAs) are used for rotator cuff repair. Pull-out strength (POS) is an important factor that affects surgical outcomes. While the fixation mechanism differs between the anchor types and relies on the quality, few studies have compared biomechanical properties of anchors based on bone quality. This study aimed to compare the biomechanical properties of anchors using osteoporotic bone (OB) and non-osteoporotic bone (NOB) simulators. Humerus simulators were fabricated using fused deposition modeling of 3D printing and acrylonitrile butadiene styrene adjusting the thickness of cortical bone and density of cancellous bone based on CT images. Cyclic loading from 10 to 50 N, 10 to 100 N, and 10 to 150 N for 10 cycles was clinically determined at each anchor because the supraspinatus generates a force of 67–125 N in daily activities of normal control. After cyclic loading, the anchor was extruded at a load of 5 mm/min. Displacement, POS, and stiffness were measured. In OB simulators, CAs revealed bigger gap displacement than ASAs with cyclic loading of 10–150 N. ASA showed higher values for POS and stiffness. In NOB simulators, ASAs revealed bigger gap displacement than CAs with cyclic loading of 10–150 N. ASA showed higher values for POS and CA showed higher values for stiffness. POS of anchors depends on anchors ‘displacement and bone stiffness. In conclusion, ASA demonstrated better biomechanical performance than CA in terms of stability under cyclic loading and stiffness with similar POS in OB.

Quantitative osteoporosis diagnosis of porous cancellous bone using poroelastodynamic modal analysis.

Osteoporosis is a skeletal disease characterized by reduced bone mass and microarchitectural deterioration, leading to increased fragility. This study presents a novel three-dimensional poroelastodynamic model for analyzing cancellous bone free vibration responses. The model incorporates the Navier-Stokes equations of linear elasticity and the Biot theory of porous media, allowing the investigation of osteoporosis-related changes. The analysis considers parameters like porosity, density, elasticity, Poisson ratio, and viscosity of bone marrow within the porous medium. Our findings indicate that natural frequencies of cancellous bone play a crucial role in osteoporosis prediction. By incorporating experimental data from 12 mouse femurs, we unveil insights into osteoporosis prediction. Increased porosity reduces bone stiffness, lowering natural frequencies. However, it also increases bone mass loss relative to stiffness, leading to higher frequencies. Therefore, the natural frequencies of osteoporotic bone are always higher than the natural frequencies of normal bone. Additionally, an increase in bone marrow within the pores, while increasing damping effects, also increases natural frequencies, which is another indication of osteoporosis growth in bone. The presence of bone marrow within the pores further influences natural frequencies, providing additional insights into osteoporosis growth. Thinner and smaller bones are found to be more susceptible to osteoporosis compared to larger and bigger bones due to their higher natural frequencies at equivalent porosity levels.

Repair of Defects of the Nasal Tip After Mohs Surgery.

Mohs Micrographic Surgery (MMS) is a treatment option for high-risk facial nonmelanoma skin cancer with high cure rates. Especially on the nasal tip, the tissue sparing properties of MMS are appealing. The nasal tip is a common location of nonmelanoma skin cancer and can be a challenging anatomical structure for reconstructive surgery due to its prominent location in the face, the shortage of spare tissue, as well as the stiffness and composition of different skin types, cartilage and bone. The aim of the present paper is to review and demonstrate how reconstruction of the nasal tip can be done successfully to improve the care for patients undergoing MMS in this area. Using selected literature on the area and the surgeons experience, each method of repair are described including their individual advantages and challenges. Pictures and consent were selected from one patient who underwent each repair method and three photos are presented in this paper: one after tumor resection, one immediately after repair, and one minimum 6 months post-surgery. We present eight surgical methods as well as pictures from previous surgeries. The results demonstrate obtainable results using very different surgical methods and the importance of an individualized approach to repairing cutaneous defects of the nasal tip.

Hand grip strength, vitamin D status, and diets as predictors of bone health in 6–12 years old school children

BackgroundVitamin D and calcium-rich foods, exposure to sunlight, and physical activities (PA) play a pivotal role in promoting the production of sufficient vitamin D and improving grip strength needed for better bone health among school children.PurposeThis study aimed to determine the effects of hand grip muscle strength (HGS), vitamin D in addition to diets, and PA on bone health status among 6–12 years old schoolchildren.MethodsThis study was based on a cross-sectional observational design, which was descriptive in nature. A diverse sample of 560 elementary school children aged 6–12 years old were invited to participate in this descriptive cross-sectional study. The Dual-Energy X-Ray Absorptiometry (DXA), QUS technique, and ACTi graph GT1M accelerometer were used respectively as a valid tools to identify BMD, BMC, and other parameters of bone health like c-BUA values and bone stiffness (SI), and physical activity (PA) of all individuals participated in this study. In addition, a hydraulic dynamometer was used to measure hand grip strength among the participants. Moreover, an immunoassay technique was used to measure the serum levels of vitamin 25(OH)D level, and bone metabolism markers; NTX, DPD, Ca, and sBAP in all participants. Bone loss (osteoporosis) was cross-sectionally predicted in 19.64% of the total population, most of whom were girls (14.3% vs. 5.4% for boys; P = 0.01). Compared to boys, the incidence of osteoporosis was higher and significantly correlated in girls with lower HGS, deficient vitamin D, inadequate vitamin D and Ca intake, greater adiposity, poor PA, and lower sun exposure. Also, in girls, lower vitamin 25(OH)D levels, and poor HGS were shown to be significantly associated with lower values of BMD, BMC, SI, and higher values of bone resorption markers; NTX, DPD, and sBAP and lower serum Ca than do in boys. The findings suggested that deficient vitamin D, lower HGS, adiposity, PA, and sun exposure as related risk factors to the pravelence of bone loss among school children, particularly in girls. In addition, these parameters might be considered diagnostic non-invasive predictors of bone health for clinical use in epidemiological contexts; however, more studies are required.

Femurs segmentation by machine learning from CT scans combined with autonomous finite elements in orthopedic and endocrinology applications

Efficient, accurate and reliable segmentation of femurs from CT-scans is of major importance for patient-specific autonomous finite element analysis (AFE) to determine bone's stiffness and strength. We present a fully automated segmentation algorithm for whole and partial femurs with or without tumors, and clinical applications of the AFE [1] in clinical practice.The segmentation is based on an U-Net convolutional neural network, resulting a 3D mask representing the desired femur in a CT scan. It is robust, independent of the scanning parameters such as slice spacing, pixel size, scanner manufacturer or the femoral length available in the scan. The U-Net was trained on 178 manually segmented femurs (23,721 images) and tested on 43. The performance evaluation resulted in a Dice similarity score (DSC) of 0.9924, intersection over union (IoU) of 0.9849, Hausdorff distance of 4.3315 mm and symmetric average surface distance (ASD) of 0.1326 mm. The algorithm is competitive with the best state-of-the-art femoral segmentation methodologies available.Based on the segmentation an automatic p-FE mesh is generated and physiological boundary conditions representing sidewise falls or stance are being applied automatically to improve the performance of the AFE described in [1]. New examples of the usage of the AFE in endocrinology and orthopedic oncology demonstrate this disruptive technology in actual clinical practice. We present the use of AFE for predicting hip fracture risk in the elderly population due to a sidewise fall and the identification of patients who require a prophylactic surgery due to metastatic tumors in their femurs.

Genetic Variants in WNT16 and PKD2L1 Locus Affect Heel Ultrasound Bone Stiffness: Analyses from the General Population and Patients Evaluated for Osteoporosis.

Osteoporosis, a complex chronic disease with increasing prevalence, is characterised by reduced bone mineral density (BMD) and increased fracture risk. The high heritability of BMD suggests substantial impact of the individual genetic disposition on bone phenotypes and the development of osteoporosis. In the past years, genome-wide association studies (GWAS) identified hundreds of genetic variants associated with BMD or osteoporosis. Here, we analysed 1103 single nucleotide polymorphisms (SNPs), previously identified as associated with estimated BMD (eBMD) in the UK Biobank. We assessed whether these SNPs are related to heel stiffness index obtained by quantitative ultrasound in 5665 adult participants of the Study of Health in Pomerania (SHIP). We confirmed 45 significant associations after correction for multiple testing. Next, we analysed six selected SNPs in 631 patients evaluated for osteoporosis [rs2707518 (CPED1/WNT16), rs3779381 (WNT16), rs115242848 (LOC101927709/EN1), rs10239787 (JAZF1), rs603424 (PKD2L1) and rs6968704 (JAZF1)]. Differences in minor allele frequencies (MAF) of rs2707518 and rs3779381 between SHIP participants (higher MAF) and patients evaluated for osteoporosis (lower MAF) indicated a protective effect of the minor allele on bone integrity. In contrast, differences in MAF of rs603424 indicated a harmful effect. Co-localisation analyses indicated that the rs603424 effect may be mediated via stearoyl-CoA desaturase (SCD) expression, an enzyme highly expressed in adipose tissue with a crucial role in lipogenesis. Taken together, our results support the role of the WNT16 pathway in the regulation of bone properties and indicate a novel causal role of SCD expression in adipose tissue on bone integrity.

THU346 Repairing Skeletal Deterioration In Diabetes With Bone Anabolic Therapies

Abstract Disclosure: S. Marino: None. N. Akel: None. S. Li: None. M. Cregor: None. B. Perez: None. G. Troncoso: None. J. Meeks: None. S. Stuart: None. A. Sato: None. I. Nookaew: None. T. Bellido: None. Treatments for skeletal fragility in diabetes are sorely needed as antiresorptive agents, the current standard of care, stop bone loss but do not rebuild bone nor restore the deteriorated bone microarchitecture. Herein, we demonstrate bone restoration in a preclinical model of established diabetes by two bone anabolic pathways: activation of the PTH1 receptor, with equimolar doses of either PTH (1-34) or Abaloparatide/ABL (100/95 µg/kg/day), and neutralization of the osteocyte-derived inhibitor of bone formation sclerostin with romosozumab/Scl-Ab (100mg/kg/w). Type 2 diabetes (T2D) was induced in skeletally mature male C57BL/6J mice by high fat diet combined with streptozotocin; and, after established diabetes, mice were treated for 4 weeks with the bone anabolic agents. At the end of the study, T2D mice exhibited decreased total (−5%), femoral (−7%) and spinal (−11%) BMD vs nondiabetic mice. PTH/ABL restored the bone lost and increased BMD to a similar extent: total 8%, femoral 11% and spinal 9% vs vehicle-treated T2D mice. Scl-Ab also increased BMD at all sites, but the effects were superior: total 24%, femoral 26%, and spinal 40%. T2D mice exhibited low bone formation (BF) and high bone resorption (BR) circulating markers throughout the disease progression. However, at the tissue level BF and BR were suppressed as evidenced by histomorphometric indexes (BFR −70%; Oc.S/BS -34%) and bone transcriptome GO profiles. PTH/ABL increased BF (BFR ∼700% and P1NP 130/244%) as well as BR (Oc.S/BS unchanged/34%; TRAP5b ∼200%) and corrected the diabetic low bone remodeling signature towards high bone remodeling with bone gain. In contrast, Scl-Ab increased formation (BFR 500% and P1NP 50%) but drastically decreased resorption (TRAP5b −190% and Oc.S/BS −500%). T2D mice exhibited damaged bone microarchitecture. The reduced femoral and spinal cancellous bone BV/TV (−22 and −10%) was corrected by ABL (40 and 42%), Scl-Ab (130 and 83%), and PTH only in the spine (43%). Further, femoral and spinal trabecular thickness were increased equally by PTH/ABL (35 and 24%) and also by Scl-Ab (60 and 66%). Moreover, the decreased cortical thickness (−7%) and BA/TA (−5%) at the femoral mid-diaphysis of T2D mice were corrected and further increased by PTH/ABL (10 and 11%) and Scl-Ab (23 and 19%). Yet, whereas PTH reduced marrow area, ABL increased tissue area TA, and Scl-Ab exerted both effects, resulting in advantageous bone distribution by ABL and Scl-Ab leading to increased energy to ultimate load (60 and 90%) and toughness (50 and 82%). Nevertheless, all agents corrected the low stiffness of T2D bone (-19%) and also increased ultimate load. These findings demonstrate the therapeutic value of bone anabolic agents to treat diabetes-induced bone disease by increasing bone formation and improving bone microarchitecture and show the benefit of simultaneously inhibiting resorption to achieve superior bone gain and strength. Presentation: Thursday, June 15, 2023

The microarchitecture and chemical composition of the femur neck of senescent female rats after different physical training protocols.

A sedentary lifestyle, coupled with a decrease in estrogen, impairs bone homeostasis, favoring to the development of osteopenia and osteoporosis, both recognized as risk factors for fractures. Here, we investigated the quality of the femur, particularly the femur neck region, and the ambulation performance of senescent rats subjected to three different physical training protocols during the periestropause period. Forty-eight female rats, 18 months of age, were subjected to a 120-day training period, three times a week. The rats were distributed into four groups: aerobic training (AT), strength training (ST), concurrent training (CT), or no training (NT). After the experimental period, at 21 months of age, ambulation performance and femur were analyzed using microtomography, Raman stereology, densitometry, and mechanical strength tests. The results demonstrated greater remodeling activity and improvement in resistance and bone microarchitecture in the femur neck of senescent female rats after undergoing physical training. Our verified higher intensities of bands related to collagen, phosphate, amide III, and amide I. Furthermore, the analysis of the secondary collagen structures indicated alterations in the collagen network due to the exercise, resulting in increased bone strength. Both AT and strength-based training proved beneficial, with AT showing greater adaptations in bone density and stiffness in the femur, while strength-based training greater adaptations in trabecular and cortical structure. These insights contribute to the understanding of the potential interventions for preventing osteopenia and osteoporosis, which are critical risk factors for fractures.

Application of Optical Photothermal Infrared (O-PTIR) Spectroscopy for Assessment of Bone Composition at the Submicron Scale.

The molecular basis of bone structure and strength is mineralized collagen fibrils at the submicron scale (∼500 nm). Recent advances in optical photothermal infrared (O-PTIR) spectroscopy allow the investigation of bone composition with unprecedented submicron spatial resolution, which may provide new insights into factors contributing to underlying bone function. Here, we investigated (i) whether O-PTIR-derived spectral parameters correlated to standard attenuated total reflection (ATR) Fourier transform infrared spectroscopy spectral data and (ii) whether O-PTIR-derived spectral parameters, including heterogeneity of tissue, contribute to the prediction of proximal femoral bone stiffness. Analysis of serially demineralized bone powders showed a significant correlation (r = 0.96) between mineral content quantified using ATR and O-PTIR spectroscopy, indicating the validity of this technique in assessing bone mineralization. Using femoral neck sections, the principal component analysis showed that differences between O-PTIR and ATR spectra were primarily attributable to the phosphate ion (PO4) absorbance band, which was typically shifter toward higher wavenumbers in O-PTIR spectra. Additionally, significant correlations were found between hydrogen phosphate (HPO4) content (r = 0.75) and carbonate (CO3) content (r = 0.66) quantified using ATR and O-PTIR spectroscopy, strengthening the validity of this method to assess bone mineral composition. O-PTIR imaging of individual trabeculae at 500 nm pixel resolution illustrated differences in submicron composition in the femoral neck from bones with different stiffness. O-PTIR analysis showed a significant negative correlation (r = -0.71) between bone stiffness and mineral maturity, reflective of newly formed bone being an important contributor to bone function. Finally, partial least squares regression analysis showed that combining multiple O-PTIR parameters (HPO4 content and heterogeneity, collagen integrity, and CO3 content) could significantly predict proximal femoral stiffness (R2 = 0.74, error = 9.7%) more accurately than using ATR parameters. Additionally, we describe new findings in the effects of bone tissue orientation in the O-PTIR spectra. Overall, this study highlights a new application of O-PTIR spectroscopy that may provide new insights into molecular-level factors underlying bone mechanical competence.

Anti-Siglec-15 Antibody Prevents Marked Bone Loss after Acute Spinal Cord Injury-Induced Immobilization in Rats.

Rapid and extensive sublesional bone loss after spinal cord injury (SCI) is a difficult medical problem that has been refractory to available interventions except the antiresorptive agent denosumab (DMAB). While DMAB has shown some efficacy in inhibiting bone loss, its concurrent inhibition of bone formation limits its use. Sialic acid-binding immunoglobulin-like lectin (Siglec)-15 is expressed on the cell surface of mature osteoclasts. Anti-Siglec-15 antibody (Ab) has been shown to inhibit osteoclast maturation and bone resorption while maintaining osteoblast activity, which is distinct from current antiresorptive agents that inhibit the activity of both osteoclasts and osteoblasts. The goal of the present study is to test a Siglec-15 Ab (NP159) as a new treatment option to prevent bone loss in an acute SCI model. To this end, 4-month-old male Wistar rats underwent complete spinal cord transection and were treated with either vehicle or NP159 at 20 mg/kg once every 2 weeks for 8 weeks. SCI results in significant decreases in bone mineral density (BMD, -18.7%), trabecular bone volume (-43.1%), trabecular connectivity (-59.7%), and bone stiffness (-76.3%) at the distal femur. Treatment with NP159 almost completely prevents the aforementioned deterioration of bone after SCI. Blood and histomorphometric analyses revealed that NP159 is able to greatly inhibit bone resorption while maintaining bone formation after acute SCI. In ex vivo cultures of bone marrow cells, NP159 reduces osteoclastogenesis while increasing osteoblastogenesis. In summary, treatment with NP159 almost fully prevents sublesional loss of BMD and metaphysis trabecular bone volume and preserves bone strength in a rat model of acute SCI. Because of its unique ability to reduce osteoclastogenesis and bone resorption while promoting osteoblastogenesis to maintain bone formation, Siglec-15 Ab may hold greater promise as a therapeutic agent, compared with the exclusively antiresorptive or anabolic agents that are currently used, in mitigating the striking bone loss that occurs after SCI or other conditions associated with severe immobilization. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.