Cortical Bone Microarchitecture Research Articles

Novel roles of Atrogin‐1 in cardiac disease, lipid metabolism and bone microstructure

Ubiquitination plays a critical role in controlling protein turnover via proteasomal degradation, altering subcellular localization, and regulating protein activity. Functional role(s) for the Atrogin‐1 (MAFbx) ubiquitin ligase in regulating transcription factors have been found in skeletal muscle and the heart, but not in other organs. Within the myocyte, Atrogin‐1 localizes to the sarcomere and nucleus. We have previously reported that Atrogin‐1 is a critical regulator of pathologic and physiologic cardiac hypertrophy in vivo, involving specific interaction and ubiquitination of transcription factors central the signaling pathways driving these hypertrophic responses, including FOXO1/3. Recent studies have linked cardiomyocyte Atrogin‐1 to the regulation of the extracellular matrix. With evidence that cardiomyocyte Atrogin‐1 can affect cells other than myocytes, we analyzed Atrogin‐1−/− mice for their effects on metabolism and bone. DEXA whole body analysis of Atrogin‐1−/− mice revealed significantly decreased fat mass (15.6% vs. 35.8% fat, p<0.05) with a corresponding increase in lean mass (84.4% vs. 64.2%, p<0.05) at the age of 11–15 months, but not at 5 months, when compared to strain‐ and age‐matched wildtype (WT) control mice. Since the Atrogin‐1−/− mice weighed significantly less than WT mice (29±3 vs. 40±2 g, p<0.05), the net fat loss was 4.6 g (vs. 14.3 g in age‐matched WT) and the lean body mass identical between groups (25.7 vs. 24.6 g). Together these findings suggest Atrogin‐1’s role in regulating fat metabolism. Analysis of bone microarchitecture and mechanical properties of adult Atrogin‐1−/− mice was undertaken at 17–21 weeks of age, a time point where no phenotype has been previously observed in skeletal muscle or heart. Femurs were dissected and subsequently assessed by μCT (SkyScan 1172) for bone microarchitecture in the distal femur metaphysis (1 mm) and mid‐diaphysis region and challenged by a three‐point bending test to assess their material and structural properties. No differences in cortical or trabecular bone microarchitecture were identified and nor were any changes in mechanical properties. Together, these findings illustrate novel biological roles of Atrogin‐1 in regulating systemic fat metabolism, possibly involving crosstalk between skeletal muscle/heart and systemic fat metabolism. Our analysis provides an essential framework for the potential therapeutic targeting of ubiquitin ligases like Atrogin‐1 in the context of striated muscle and metabolic disease.Support or Funding InformationSupport: Lilly Foundation/Indiana Center for Musculoskeletal Health/ IU School of Medicine, Physician Scientist Initiative, Scientific Research Initiative

Freeze-Dried Watermelon Supplementation Has Modest Effects on Bone and Lipid Parameters of Ovariectomized Mice.

This study investigated the effects of two doses of freeze-dried watermelon (WM) on bone and lipid parameters in ovariectomized (OVX) mice, a model of post-menopausal osteoporosis. Three-month-old C57BL/6 female mice (n=46) were sham-operated (SHAM) or OVX and randomly assigned to the control or WM diets for 12 weeks: SHAM-control, OVX-control, OVX+1%, or 10% (wt/wt) freeze-dried WM. All diets were isocaloric and isonitrogenous, and had the same calcium and phosphorus concentrations. Freeze-dried WM supplementation was not able to prevent the decrease in whole body, tibial, and lumbar bone mineral density due to estrogen deficiency. Micro-computed tomography analyses showed that WM was also not able to modulate changes in tibial trabecular and cortical bone microarchitecture due to ovariectomy. However, the lumbar trabecular micro-architecture analyses revealed that the WM-10% group had a similar connectivity density, trabecular number, trabecular separation, and structure model index as the SHAM group. Supplementation with 10% WM reduced plasma cholesterol and total liver lipids to the level of the SHAM group but was still similar to that of the OVX-control group. Supplementation with 10% WM increased liver catalase (CAT) mRNA levels but had no effects on mRNA levels of glutathione peroxidase (GPX) and the pro-inflammatory cytokine interleukin-6. There were no differences in plasma activity of the antioxidant enzymes GPX and CAT between all treatment groups. Our findings demonstrate some positive effects of watermelon for modulating lipids and attenuating lumbar vertebral bone loss arising from ovarian hormone deficiency.

Prospective Study Evaluating Changes in Bone Quality in Premenopausal Women With Breast Cancer Undergoing Adjuvant Chemotherapy

BackgroundOvarian suppression from chemotherapy results in bone loss in premenopausal women with breast cancer (BC). Less is known about bone microarchitecture changes. We used high-resolution peripheral quantitative computed tomography (HR-pQCT) to measure volumetric bone density and trabecular and cortical microarchitecture in this population. Materials and MethodsThe primary endpoint was to assess changes in cortical thickness and trabecular bone density by HR-pQCT. Premenopausal women with stage I to III BC undergoing adjuvant chemotherapy underwent a bone mineral density (BMD) dual energy x-ray absorptiometry scan and HR-pQCT (voxel size, 82 microns) at baseline and 12 months. Paired t tests were used to observe the change over time in bone microarchitecture and areal and volumetric density. ResultsEighteen patients were evaluated, of which 12 patients had baseline and matched 12-month imaging. The mean age was 45.2 years (range, 35-51 years), 17 (94%) patients had hormone receptor-positive BC, and 16 (89%) initiated tamoxifen. At 12 months, there was a significant decrease in femoral neck (P < .05) and lumbar spine and total hip (P < .01) BMD. Changes detected by HR-pQCT at 12 months included significant decreases in cortical thickness and area at the tibia (P < .05), and total and cortical volumetric BMD at the radius and tibia (P < .01), as well as an increase in tibial trabecular area (P < .05). ConclusionPremenopausal women undergoing chemotherapy experience BMD decline and trabecular and cortical bone microarchitecture deterioration. In this population, future efforts should focus on therapy-induced bone loss and optimizing bone density-related management.

Global and site-specific analysis of bone in a rat model of spinal cord injury-induced osteoporosis

Micro-Computed Tomography bone analysis is the gold standard method for assessing trabecular and cortical bone microarchitecture in small animal bones. This technique reports morphometric parameters as averages over selected volumes of interest (VOIs). This study proposes the introduction of an additional global 2D morphometric step into the analysis process, that provides a survey of the underlying morphometric variation present throughout both trabecular and cortical bone. The visualisation of these morphometric distributions provides a systematic approach to VOI selection that provides rationale and adds confidence to subsequent 3D morphometric analysis. To test the applicability and value of this methodological addition it was applied to the distal femur of a rat model of spinal cord injury (SCI)-induced osteoporosis. The 2D morphometric variation of both trabecular and cortical bone was quantified as a function of bone length. SCI-induced osteoporosis was localised in i) trabecular bone, where metaphyseal bone was more severely affected than epiphyseal bone, and there was a significant reduction in Distal Femoral Trabecular Extent, a new parameter defined here that quantifies how far trabecular bone penetrates in to the marrow cavity, ii) cortical bone, where diaphyseal bone underwent significant lowering of both cortical area and thickness, while distal-metaphyseal bone did not. Theses site-specific changes were validated, further elucidated and compared with follow-up conventional 3D analysis. The techniques applied here are equally applicable to other long bones (tibia, humerus, radius, ulna), other types of imaging modality and other types of experimental design including the effects of rehabilitation, aging, loading, gene knockout and pharmacological intervention.

GIP analogues augment bone strength by modulating bone composition in diet-induced obesity in mice

Receptors to glucose-dependent insulinotropic polypeptide (GIP), have been identified on bone and GIP receptor (GIPr) knockout mice exhibit reduced bone strength and quality. Despite this, little is known on the potential beneficial bone effects of exogenous GIP on bone physiology. The aim of the present study was to assess whether stable GIP analogues were capable of ameliorating bone strength in mice with diet-induced obesity. The stable GIP analogue (D-Ala²)-GIP, and (D-Ala²)-GIP-Tag, a specific GIP analogue homing exclusively to bone, were employed. In vitro studies were used to assess effects of (D-Ala²)-GIP and (D-Ala²)-GIP-Tag on bone mineralization, lysyl oxidase activity, collagen maturity as well as osteoclast formation and activity. Subsequent in vivo studies employed obese-prediabetic Swiss NIH mice subjected to a 42-day period of daily administration of saline, (D-Ala²)-GIP or (D-Ala²)-GIP-Tag. In vitro studies confirmed that (D-Ala²)-GIP and (D-Ala²)-GIP-Tag had similar beneficial biological effects on bone cells. Administration of (D-Ala²)-GIP and (D-Ala²)-GIP-Tag resulted in lower blood glucose levels without any effects on body weight. Both GIP analogues augmented bone strength to a similar extent. Trabecular or cortical bone microarchitecture were not changed over the time course of the study. However, (D-Ala²)-GIP and (D-Ala²)-GIP-Tag augmented enzymatic collagen crosslinking as well as the heterogeneity of enzymatic collagen crosslinking, mineral-to-matrix ratio and significantly reduced the heterogeneity in mineral bone crystallite size. This study demonstrates that activation of skeletal GIPr by stable GIP analogues enhance bone strength in prediabetes and suggest that these analogues may be beneficial in the treatment of bone disease.

HR-pQCT Measures of Bone Microarchitecture Predict Fracture: Systematic Review and Meta-Analysis.

High-resolution peripheral quantitative computed tomography (HR-pQCT) is a noninvasive imaging modality for assessing volumetric bone mineral density (vBMD) and microarchitecture of cancellous and cortical bone. The objective was to (1) assess fracture-associated differences in HR-pQCT bone parameters; and (2) to determine if HR-pQCT is sufficiently precise to reliably detect these differences in individuals. We systematically identified 40 studies that used HR-pQCT (39/40 used XtremeCT scanners) to assess 1291 to 3253 and 3389 to 10,687 individuals with and without fractures, respectively, ranging in age from 10.9 to 84.7 years with no comorbid conditions. Parameters describing radial and tibial bone density, microarchitecture, and strength were extracted and percentage differences between fracture and control subjects were estimated using a random effects meta-analysis. An additional meta-analysis of short-term in vivo reproducibility of bone parameters assessed by XtremeCT was conducted to determine whether fracture-associated differences exceeded the least significant change (LSC) required to discern measured differences from precision error. Radial and tibial HR-pQCT parameters, including failure load, were significantly altered in fracture subjects, with differences ranging from -2.6% (95% confidence interval [CI] -3.4 to -1.9) in radial cortical vBMD to -12.6% (95% CI -15.0 to -10.3) in radial trabecular vBMD. Fracture-associated differences reported by prospective studies were consistent with those from retrospective studies, indicating that HR-pQCT can predict incident fracture. Assessment of study quality, heterogeneity, and publication biases verified the validity of these findings. Finally, we demonstrated that fracture-associated deficits in total and trabecular vBMD and certain tibial cortical parameters can be reliably discerned from HR-pQCT-related precision error and can be used to detect fracture-associated differences in individual patients. Although differences in other HR-pQCT measures, including failure load, were significantly associated with fracture, improved reproducibility is needed to ensure reliable individual cross-sectional screening and longitudinal monitoring. In conclusion, our study supports the use of HR-pQCT in clinical fracture prediction. © 2019 American Society for Bone and Mineral Research.

Inferring bone attribution to species through micro-Computed Tomography: A comparison of third metapodials from Homo sapiens and Ursus americanus

Abstract Gross similarities between human hand/foot bones and bear paws have been well-documented. Macroscopic skeletal analyses provide insight into species origin when whole bones are recovered but are frequently rendered inapplicable when bones are fragmented. In these scenarios, histological techniques are often applied; though specific research focusing on the quantification of bear bone microstructure remains scarce. We hypothesized that 3D analysis of bear cortical bone microarchitecture provides a more representative and accurate means to infer bone attribution to species from fragmented metapodials. Methods included visualizing and quantifying bone microstructural parameters using micro-Computed Tomography (µCT). Third metacarpals and metatarsals from mature black bears and humans were assessed using 3D analyses. Micro-CT experiments were carried out using a laboratory X-ray system at The University of Akron. Projections were reconstructed and cylindrical Volumes of Interest (VOIs) were identified within each bone sample. Variables measured within the VOIs included: total volume (TV), total canal volume (Ca.V), canal number (Ca.N), average canal diameter (Ca.Dm), and cortical porosity (Ca.V/TV). Between-species t-tests revealed that both Ca.N and Ca.Dm significantly differed between human and bear metapodials. Qualitative features including osteon banding and resorption bays were more prevalent in bear metapodials. The 3D data for this study were obtained non-destructively and reveal the usefulness of laboratory µCT as a diverse and novel tool for the anthropologist. Results demonstrated differences between the human and black bear third metapodials, supporting the hypothesis that a microstructural comparison is necessary for fragmentary bone identification of human and bear metapodials.

Biomechanical and tomographic differences in the microarchitecture and strength of trabecular and cortical bone in the early stage of male osteoporosis.

Osteoporosis is a continuous process of loss of bone tissue. Compared to women, osteoporosis in men is associated with greater morbidity and mortality. In this study, we conducted tomographic and biomechanical evaluations of trabecular and cortical bone in the early stage of male osteoporosis. Male Wistar rats were subjected to orchiectomy and sham operation. Four weeks after being castrated, decreased levels of testosterone in plasma were found and resulted in concurrent bone loss. Separately, the orchiectomy led to significant tomographic alterations in the trabecular bone number, trabecular separation, and trabecular pattern factor. Data of a mechanistic compression test further showed that the orchiectomy diminished the maximum loading force, displacement at maximum load, energy at maximum load, and ultimate stress. Interestingly, orchiectomy-triggered changes in the maximum loading force and tomographic parameters were highly correlated. In contrast, tomographic and biomechanical analyses showed that 4 weeks after rats were orchiectomized, the thickness, area, maximum loading force, bone stiffness, energy at maximum load, and ultimate stress of the cortical bone were not changed. Taken together, this study showed specific differences in the microarchitecture and strength of trabecular bone in the early stage of male osteoporosis.

Reproducibility and Radiation Effect of High-Resolution In Vivo Micro Computed Tomography Imaging of the Mouse Lumbar Vertebra and Long Bone.

A moderate radiation dose, in vivo µCT scanning protocol was developed and validated for long-term monitoring of multiple skeletal sites (femur, tibia, vertebra) in mice. A customized, 3D printed mouse holder was designed and utilized to minimize error associated with animal repositioning, resulting in good to excellent reproducibility in most cortical and trabecular bone microarchitecture and density parameters except for connectivity density. Repeated in vivo µCT scans of mice were performed at the right distal femur and the 4th lumbar vertebra every 3weeks until euthanized at 9weeks after the baseline scan. Comparing to the non-radiated counterparts, no radiation effect was found on trabecular bone volume fraction, osteoblast and osteoblast number/surface, or bone formation rate at any skeletal site. However, trabecular number, thickness, and separation, and structure model index were sensitive to ionizing radiation associated with the µCT scans, resulting in subtle but significant changes over multiple scans. Although the extent of radiation damage on most trabecular bone microarchitecture measures are comparable or far less than the age-related changes during the monitoring period, additional considerations need to be taken to minimize the confounding radiation factors when designing experiments using in vivo µCT imaging for long-term monitoring of mouse bone.

Consumption of Rabbiteye Blueberry Results in Accumulation of Hippuric Acid in the Bone Marrow and Increased Bone Deposition in Ovariectomized Rats but Few Other Bone Benefits (P06-064-19)

ObjectivesDetermine the effects of an 8-wk chronic consumption of two different blueberry varieties at varying doses on calcium balance, calcium kinetics, bone microarchitecture and polyphenol metabolism and distribution in ovariectomized rats. MethodsEighty 5-mo old ovariectomized rats were sorted by weight and block-randomized to an 8-wk chronic feeding treatment of a polyphenol-free (control), 2.5% or 5% `Montgomery ́ (Mont) rabbiteye blueberry (V. ashei) or 5% wild lowbush blueberry (Wild BB) (V. angustifolium) (% w/w). During week 0 and week 8, rats went through a calcium balance study to determine calcium absorption and retention. At the end of week 8, rats were dosed with 45Ca and serial blood draws were done from baseline up to 48 h post-dose to map Ca kinetics. At sacrifice, the right femur was collected to determine differences in bone strength and microarchitecture of trabecular and cortical bone through MicroCT and a three-point bending test. A 24 h urine collection was done at baseline and during week 8 of treatment to determine urinary phenolic acid excretion and the left femur bone marrow phenolic acid accumulation through UPLC-MS/MS. ResultsA blueberry enriched diet had no effect on cortical bone microarchitecture, with a trend towards increased trabecular bone protection (p = 0.08). No differences were observed in bone strength. During baseline, a 2.5% Mont diet significantly increased Ca retention, but the effect was lost after the 8 weeks of treatment when rats had stabilized to treatment. Calcium kinetics showed that Mont 5% significantly increased Ca absorption and bone turnover (p < 0.05), while Mont 2.5% and Wild BB 5% did not. An accumulation of hippuric acid in the bone marrow was detected with the Mont blueberry treatments and it was significantly and positively correlated with bone deposition. The metabolism of phenolic acids was significantly affected by a chronic consumption of the treatment diets, resulting in significant phenolic acid profiles and excretion shifts. ConclusionsA blueberry-enriched diet had minimal effects on bone after stabilized to ovariectomy and the effects differ depending on blueberry variety and dose. Funding SourcesNational Institute of Health (NIH) and National Center for Complementary and Integrative Health (NCCIH).

High FGF23 levels are associated with impaired trabecular bone microarchitecture in patients with osteoporosis.

Fibroblast growth factor 23 (FGF23) is an endocrine hormone-regulating phosphate and vitamin D metabolism. While its mode of action is well understood in diseases such as hereditary forms of rickets or tumor-induced osteomalacia, the interpretation of FGF23 levels in patients with osteoporosis with regard to bone microarchitecture is less clear. C-terminal FGF23 levels and bone turnover markers were assessed in 82 patients with osteoporosis (i.e., DXA T-score ≤ - 2.5 at the lumbar spine or total hip). Bone microarchitecture was measured by high-resolution peripheral quantitative computed tomography (HR-pQCT) at the distal radius and tibia. Data were analyzed in a cross-sectional design using correlation and regression models. We found a significant negative logarithmic correlation between FGF23 levels and trabecular but not cortical bone microarchitecture at both skeletal sites. Furthermore, using a multiple linear regression model, we confirmed FGF23 as a predictor for reduced trabecular parameters even when adjusting for confounding factors such as age, BMI, phosphate, bone-specific alkaline phosphatase, vitamin D3, and PTH. Taken together, high FGF23 levels are associated with impaired trabecular bone microarchitecture in osteoporosis patients, and this association seems to occur after adjustment of confounding variables including phosphate and vitamin D. Future longitudinal studies are now needed to validate our findings and investigate FGF23 in relation to fracture risk.

Dietary docosahexaenoic acid contributes to increased bone mineral accretion and strength in young female Sprague-Dawley rats

Growing evidence suggests that docosahexaenoic acid (DHA: 22:6n-3) enhances bone mineral content (BMC) and bone mineral density (BMD) in adulthood and during aging, however the effects during and after sexual maturation are unclear. The aim of this study was to examine the dose-response of BMC, BMD and microarchitectural properties of bone to dietary DHA in healthy growing female rats during acquisition of peak bone mass (PBM). Female Sprague-Dawley rats (n = 12/diet) were randomized to receive a control diet (AIN-93 M, 60 g soybean oil/kg diet) or an experimental diet containing 0.1, 0.4, 0.8 and 1.2% DHA (w/w of total diet) for 10 weeks. Dietary DHA increased the whole body, lumbar spine and long bone BMC compared to the control, in addition to higher aBMD and also BMD. Additionally, an increase in cortical bone microarchitecture parameters of lumbar spine as well as peak force were observed in dietary DHA diet groups. Dietary DHA contributes to PBM when consumed during and after sexual maturation, however higher doses of DHA do not provide further benefits.

Microcomputed tomography of the femur of diabetic rats: alterations of trabecular and cortical bone microarchitecture and vasculature\u2014a feasibility study

BackgroundTo better understand bone fragility in type 2 diabetes mellitus and define the contribution of microcomputed tomography (micro-CT) to the evaluation of bone microarchitecture and vascularisation, we conducted an in vitro preliminary study on the femur of Zucker diabetic fatty (ZDF) rats and Zucker lean (ZL) rats. We first analysed bone microarchitecture, then determined whether micro-CT allowed to explore bone vascularisation, and finally looked for a link between these parameters.MethodsEight ZDF and six ZL rats were examined for bone microarchitecture (group 1), and six ZDF and six ZL rats were studied for bone vascularisation after Microfil® perfusion which is a radiopaque casting agent (group 2). In group 1, we used micro-CT to examine the trabecular and cortical bone microarchitecture of the femoral head, neck, shaft, and distal metaphysis. In group 2, micro-CT was used to study the blood vessels in the head, neck, and distal metaphysis.ResultsCompared to ZL rats, the ZDF rats exhibited significantly lower trabecular bone volume and number and higher trabecular separation in the three locations (p = 0.02, p = 0.02, p = 0.003). Cortical porosity was significantly higher in the ZDF rats at the neck and shaft (p = 0.001 and p = 0.005). We observed a dramatically poorer bone vascularisation in the femur of ZDF rats, especially in distal metaphysis (p < 0.047).ConclusionsMicro-CT demonstrated not only significant alterations in the bone microarchitecture of the femurs of ZDF rats, but also significant alterations in bone vascularisation. Further studies are required to demonstrate the causal link between poor vascularisation and impaired bone architecture.

Paget\u2019s Disease of Long Bones: Microstructural Analyses of Historical Bone Samples

Although Paget’s disease of bone (PDB) is the second most common metabolic bone disease, there is only limited information about the microarchitecture of affected bones. Therefore, the aim of this study was to determine cortical and trabecular bone properties in clinically relevant locations by microcomputed tomography (µCT). Ten femora and ten tibiae affected by Paget’s disease taken from the Natural History Museum Vienna were compared to 13 femora and 10 tibiae of non-affected body donors. Digitization of the cortical and trabecular bone microarchitecture was performed with an X-ray-based µCT scanner. Additionally, semi-quantitative gradings of trabecular and cortical architectural parameters of the femora and the tibiae were generated. Microcomputed tomography images showed changes in the thickness of cortices, cortical porosity, and trabecularization of cortical structures. Moreover, severe disorganization of trabecular structures, trabecular defects, and thickening of (remaining) trabeculae were detected. Numerical cortical analyses showed lower total bone volume (BV) and lower BV in the outer region (66–100%) (− 36%, p = 0.004, and − 50%, p < 0.001, respectively), lower total volume (TV) in the outer region (66–100%) (− 42%, p < 0.001), lower total bone volume fraction (BV/TV) and BV/TV in the outer region (66–100%) (− 23%, and − 12%, p < 0.001, respectively), higher BV and TV in the middle region (33–66%) and higher BV/TV in the inner region (0–33%) (123%, p = 0.011, 147%, p = 0.010, and 33%, p = 0.025, respectively) in Pagetic compared to non-affected bones. Trabecular analyses showed higher BV/TV (96%, p = 0.008) and Tb.Th (43%, p = 0.004) in Pagetic compared to non-affected bones. There is a major and consistent structural alteration of PDB at cortical and trabecular sites in weight-bearing long bones. Our findings are relevant for the differential diagnosis of PDB and for the pathogenesis of associated complications, since the disorder produces abnormalities in the structure that might lead to bone fragility.

Maternal Transient Receptor Potential Vanilloid 6 (Trpv6) Is Involved In Offspring Bone Development.

Embryonic growth and bone development depend on placental Ca2+ transport across the feto-maternal barrier to supply minerals to the fetus. The individual factors and cellular mechanisms that regulate placental Ca2+ transfer, however, are only beginning to emerge. We find that the Ca2+ -selective transient receptor potential vanilloid 6 (TRPV6) channel is expressed in trophoblasts of the fetal labyrinth, in the yolk sac, and in the maternal part of the placenta. Lack of functional TRPV6 channels in the mother leads to a reduced Ca2+ content in both placenta and embryo. Ca2+ uptake in trophoblasts is impaired in the absence of Trpv6. Trpv6-deficient embryos are smaller, have a lower body weight, and shorter and less calcified femurs. The altered cortical bone microarchitecture persists in adulthood. We show that TRPV6's Ca2+ -conducting property causes this embryonic and bone phenotype. Our results show that TRPV6 is necessary for the Ca2+ uptake in trophoblasts and that TRPV6 deficiency in the placenta leads to reduced embryo growth, minor bone calcification, and impaired bone development. © 2019 American Society for Bone and Mineral Research.

The Association of Vitamin D in Youth and Early Adulthood with Bone Mineral Density and Microarchitecture in Early Adulthood.

This study aimed to describe the association of vitamin D status at different stages of growth with bone measures in adolescence and early adulthood. There were 415 participants followed from age 8 to 16, and 201 further followed to age 25. Areal bone mineral density (BMD) at the spine, hip and total body was measured by dual-energy X-ray absorptiometry at ages 16 and 25, and tibial and radial trabecular and cortical bone microarchitecture by high resolution peripheral quantitative computerised tomography at age 25. Serum 25-hydroxyvitamin D (25OHD) concentrations were measured at ages 8, 16 and 25. Multivariable linear regression was used to analyse the association of 25OHD concentrations at three timepoints with bone measures at ages 16 and 25. The proportion of participants with vitamin D deficiency (< 50nmol/L) was 11%, 43% and 41% at three timepoints, respectively. Serum 25OHD concentrations at age 8 were not significantly associated with any bone measures at age 16 or 25. Serum 25OHD concentrations at age 16 had a significant association with higher BMD at nearly all sites at ages 16 and 25 as well as lower radial porosity and more compact trabecular microarchitecture (higher density, increased number and reduced separation) at both the radius and tibia at age 25. Serum 25OHD concentrations at age 25 were only associated with hip BMD. Higher vitamin D concentrations in adolescence, to a lesser extent at age 25, have beneficial associations with BMD and bone microarchitecture in early adulthood. Optimising vitamin D status particularly during adolescence should be a priority.

Cortical and trabecular bone microarchitecture as an independent predictor of incident fracture risk in older women and men in the Bone Microarchitecture International Consortium (BoMIC): a prospective study

Although areal bone mineral density (aBMD) assessed by dual-energy x-ray absorptiometry (DXA) is the clinical standard for determining fracture risk, most older adults who sustain a fracture have T scores greater than -2·5 and thus do not meet the clinical criteria for osteoporosis. Importantly, bone fragility is due to low BMD and deterioration in bone structure. We assessed whether indices of high-resolution peripheral quantitative CT (HR-pQCT) were associated with fracture risk independently of femoral neck aBMD and the Fracture Risk Assessment Tool (FRAX) score. We assessed participants in eight cohorts from the USA (Framingham, Mayo Clinic), France (QUALYOR, STRAMBO, OFELY), Switzerland (GERICO), Canada (CaMos), and Sweden (MrOS). We used Cox proportional hazard ratios (HRs) to estimate the association between HR-pQCT bone indices (per 1 SD of deficit) and incident fracture, adjusting for age, sex, height, weight, and cohort, and then additionally for femoral neck DXA aBMD or FRAX. 7254 individuals (66% women and 34% men) were assessed. Mean baseline age was 69 years (SD 9, range 40-96). Over a mean follow-up of 4·63 years (SD 2·41) years, 765 (11%) participants had incident fractures, of whom 633 (86%) had femoral neck T scores greater than -2·5. After adjustment for age, sex, cohort, height, and weight, peripheral skeleton failure load had the greatest association with risk of fracture: tibia HR 2·40 (95% CI 1·98-2·91) and radius 2·13 (1·77-2·56) per 1 SD decrease. HRs for other bone indices ranged from 1·12 (95% CI 1·03-1·23) per 1 SD increase in tibia cortical porosity to 1·58 (1·45-1·72) per 1 SD decrease in radius trabecular volumetric bone density. After further adjustment for femoral neck aBMD or FRAX score, the associations were reduced but remained significant for most bone parameters. A model including cortical volumetric bone density, trabecular number, and trabecular thickness at the distal radius and a model including these indices plus cortical area at the tibia were the best predictors of fracture. HR-pQCT indices and failure load improved prediction of fracture beyond femoral neck aBMD or FRAX scores alone. Our findings from a large international cohort of men and women support previous reports that deficits in trabecular and cortical bone density and structure independently contribute to fracture risk. These measurements and morphological assessment of the peripheral skeleton might improve identification of people at the highest risk of fracture. National Institutes of Health National Institute of Arthritis Musculoskeletal and Skin Diseases.

Montmorency tart cherry protects against age-related bone loss in female C57BL/6 mice and demonstrates some anabolic effects

Age-related bone loss is a consequence of endocrine and immune changes that disrupt bone remodeling. Functional foods (e.g., tart cherries) with antioxidant, anti-inflammatory and prebiotic activity can potentially counter this age-related phenomenon. The aim of this study was to determine if Montmorency tart cherry protects against early age-related bone loss and the culpable alterations in bone metabolism. Female, 5-month-old, C57BL/6 mice were assigned to baseline or treatment groups: AIN-93M diet supplemented with 0, 1, 5, or 10% tart cherry for 90 days. Bone mineral density (BMD) and trabecular and cortical bone microarchitecture were assessed. Treatment effects on bone metabolism and regulators of bone formation, resorption and mineralization were determined. Mice consuming the 5% and 10% doses had higher vertebral and tibial BMD (p < 0.05) compared to controls. The age-related decrease in trabecular bone volume (BV/TV) of the distal femur was prevented with these doses. Vertebral trabecular BV/TV and cortical bone thickness of the femur mid-diaphysis were greater (p < 0.05) in the groups receiving the 5% and 10% cherry than the control diet. Notably, these improvements were significantly greater than the baseline controls, consistent with an anabolic response. Although no differences in systemic biomarkers of bone formation or resorption were detected at 90days, local increases in Phex and decreases in Ppar-γ suggest a bone environment that supports increased mineralization. These findings demonstrate that cherry supplementation (5% and 10%) improves BMD and some indices of trabecular and cortical bone microarchitecture; these effects are likely attributed to increased bone mineralization.

Effect of ionizing radiation after-therapy interval on bone: histomorphometric and biomechanical characteristics.

This study aimed to evaluate the effects of radiotherapy on biomechanical, histomorphometric, and microstructural characteristics of bone, in diverse periods, compared with intact bone tissue. Eighteen adult male New Zealand rabbits were treated with a single radiation dose of 30Gy. The animals were randomly divided into six groups: NoIr, control group, no radiation, and five irradiated groups sacrificed after 24h (Ir24h), 7 (Ir7d), 14 (Ir14d), 21 (Ir21d), and 28 (Ir28d) days. After these periods, the animals were sacrificed and their tibias (n = 6) evaluated using three-point bending test to calculate the ultimate force, work to failure, and bone stiffness. Dynamic indentation test was used to quantify Vickers hardness and elasticity modulus of bone tissue. Micro-CT was used to analyze the cortical volume (CtV), cortical thickness (CtTh), and porosity (Ct.Po). Histomorphometric assessment was based on the lacunarity of bone tissue. Data were analyzed using one-way ANOVA and Kruskal-Wallis tests followed by Tukey, Dunnet, and Dunn's post-tests (P < 0.05). The ultimate force, work to failure, stiffness, elastic modulus, and Vickers hardness values of irradiated bone were significantly lower that non-irradiated bone. Irradiated bone showed significantly lower CtTh and CtV values and higher CtPo than non-irradiated bone. No significant difference was found for lacunarity between non-irradiated bone and irradiated bone. Ionizing radiation decreases normal anisotropy on microarchitecture of cortical bone, and increases bone fragility compared with non-irradiated bone. Further, these changes were seen after longer periods (e.g., 14 and 21days), and not immediately after radiation therapy. The radiotherapy reduces bone mechanical properties and the normal structure of organic and inorganic bone matrix. For studying the protocols to protect the radiotherapy effect using rabbit model, the use of the sacrificing period between 14 and 21days is recommended.

Micro-CT evaluation of the cortical bone micro-architecture in the anterior and posterior maxilla and the maxillary sinus floor

ObjectivesTo perform a within-subject comparison of the cortical bone micro-architecture of the maxillary sinus floor (MSF) to that of the buccal aspect of the anterior and posterior maxilla.MethodsMicro-CT scans of the buccal aspect of the anterior and posterior maxilla and of the MSF in 14 human anatomical specimens were recorded. Within-subject comparisons were performed for cortical thickness (Ct.Th) and porosity (Ct.Po), average pore volume (AvgPo.V), and pore density (Po.Dn).ResultsThe MSF presented the lowest and the anterior maxilla the highest Ct.Th, while Ct.Po was significantly higher at the MSF compared to the posterior maxilla (p = 0.021). No relevant differences were recorded for AvgPo.V and Po.Dn among regions (p > 0.067). Further, an increased Ct.Th at the MSF was significantly associated with a lower Po.Dn, while a higher Ct.Th and an increased AvgPo.V in the anterior maxilla were associated with a higher Ct.Th and an increased AvgPo.V, respectively, in the posterior maxilla and MSF. Finally, within each region, the AvgPo.V was associated positively with Ct.Po and negatively with Po.Dn.ConclusionsThe cortical bone of the MSF is slightly less thick and slightly more porous compared to the cortical bone at the buccal aspect of the anterior and posterior maxilla.Clinical relevanceDuring lateral and vertical bone augmentation procedures, the cortical recipient bone is perforated several times to open the bone marrow compartment to facilitate provision of osteoinductive cells and molecules in the augmented space. Whether it is meaningful to approach the MSF in a similar way during MSF augmentation procedures or whether the slightly more porous structure of the MSF observed herein reduces the cortical barrier function already sufficiently has to be assessed in future clinical trials.