Cortical Bone Volume Research Articles

Failure Analysis Following Osteochondroplasty for Hip Impingement in Osteoporotic and Non-Osteoporotic Bones

Femoral osteochondroplasty is the most common treatment for femoroacetabular impingement (FAI). The risk of femoral neck fracture is increased following surgery and increases further when the bone is osteoporotic. The current requirement to undertake osteochondroplasty on patients with osteoporosis is forecast to increase; however, the effect of osteoporosis on the risk of post operative fracture is currently unknown. We developed three three-dimensional (3D) finite element models using computerised tomography (CT) scan data for a hip with cam-type impingement and used them to investigate the association between osteoporosis and the increased possibility of femoral neck fracture after femoral osteochondroplasty. Femoral osteochondroplasty was performed “virtually” on the intact hip model to two different resection depths, a ‘standard’ (6 mm) and a ‘critical’ resection (12 mm) depth, corresponding to 18% and 36% of the overall femoral neck diameter, respectively. Cortical and trabecular bone were included in the intact and resection hip models, and material properties representing both non-osteoporotic and osteoporotic cases employed, overall, 18 scenarios were analysed. Loading corresponding to “descending stairs” and “stumbling” activities were applied in the models enabling fracture propensity to be estimated. Our model predicted that fracture propagation can occur in the bone of osteoporotic patients following osteochondroplasty during typical daily activities, such as descending stairs. The level of damage increases significantly when patients are subjected to high load conditions and activities, even in non-osteoporotic patients, indicating an increased likelihood of fracture occurring. In the “stumbling activity” simulation, osteoporotic trabecular bone damage volume approached 50% for the 6 mm resection, rising to 70% at a resection depth of 12 mm. The corresponding rise in osteoporotic cortical bone volume damage was from 6% to 10%. Our findings support the recommendation for protected weight-bearing in patients in the postoperative phase and suggest an extended period of protected weight-bearing in osteoporotic patients could be considered.

Manipulation of Ovarian Function Significantly Influenced Trabecular and Cortical Bone Volume, Architecture and Density in Mice at Death

Previously, transplantation of ovaries from young, cycling mice into old, postreproductive-age mice increased life span and decreased cardiomyopathy at death. We anticipated that the same factors that increased life span and decreased cardiomyopathy could also influence the progression of orthopedic disease. At 11 months of age, prepubertally ovariectomized and ovary-intact mice (including reproductively cycling and acyclic mice) received new 60-day-old ovaries. At death, epiphyseal bone in the proximal tibia and the distal femur and mid-shaft tibial and femoral diaphyseal bone was analyzed with micro-computed tomography. For qualitative analysis of osteophytosis, we also included mineralized connective tissue within the stifle joint. Prepubertal ovariectomy had the greatest influence on bone volume, ovarian transplantation had the greatest influence on bone architecture and both treatments influenced bone density. Ovarian transplantation increased cortical, but not trabecular bone density and tended to increase osteophytosis and heterotopic mineralization, except in acyclic recipients. These effects may have been dictated by the timing of the treatments, with ovariectomy appearing to influence early development and ovarian transplantation limited to influencing only the postreproductive period. However, major differences observed between cycling, acyclic and ovariectomized recipients of new ovaries may have been, in part due to differences in the levels of hormone receptors present and the responsiveness of specific bone processes to hormone signaling. Changes that resulted from these treatments may represent a compensatory response to normal age-associated, negative, orthopedic changes. Alternatively, differences between treatments may simply be the 'preservation' of unblemished orthopedic conditions, prior to the influence of negative, age-associated effects. These findings may suggest that in women, tailoring hormone replacement therapy to the patient's current reproductive status may improve therapy effectiveness and that beginning therapy earlier may help preserve trabecular bone mineral density that would otherwise be lost during perimenopause.

Deletion of the membrane complement inhibitor CD59a drives age and gender-dependent alterations to bone phenotype in mice

Degenerative joint diseases such as osteoarthritis are characterised by aberrant region-specific bone formation and abnormal bone mineral content. A recent study suggested a role for the complement membrane attack complex in experimental models of osteoarthritis. Since CD59a is the principal regulator of the membrane attack complex in mice, we evaluated the impact of CD59a gene deletion upon maintenance of bone architecture.In vivo bone morphology analysis revealed that male CD59a-deficient mice have increased femur length and cortical bone volume, albeit with reduced bone mineral density. However, this phenomenon was not observed in female mice. Histomorphometric analysis of the trabecular bone showed increased rates of bone homeostasis, with both increased bone resorption and mineral apposition rate in CD59a-deficient male mice. When bone cells were studied in isolation, in vitro osteoclastogenesis was significantly increased in male CD59a-deficient mice, although osteoblast formation was not altered.Our data reveal, for the first time, that CD59a is a regulator of bone growth and homeostasis. CD59a ablation in male mice results in longer and wider bones, but with less density, which is likely a major contributing factor for their susceptibility to osteoarthritis. These findings increase our understanding of the role of complement regulation in degenerative arthritis.

The y6 receptor suppresses bone resorption and stimulates bone formation in mice via a suprachiasmatic nucleus relay

The neuropeptide Y system is known to play an important role in the regulation of bone homeostasis and while the functions of its major receptors, Y1R and Y2R, in this process have become clearer, the contributions of other Y-receptors, like the y6 receptor (y6R), are unknown. Y6R expression is restricted to the suprachiasmatic nucleus (SCN) of the hypothalamus, an area known to regulate circadian rhythms, and the testis. Here we show that lack of y6R signalling, results in significant reduction in bone mass, but no changes in bone length. Male and female y6R knockout (KO) mice display reduced cortical and cancellous bone volume in axial and appendicular bones. Mechanistically, the reduction in cancellous bone is the result of an uncoupling of bone remodelling, leading to an increase in osteoclast surface and number, and a reduction in osteoblast number, osteoid surface, mineralizing surface and bone formation rate. y6R KO mice displayed increased numbers of osteoclast precursors and produced greater numbers of osteoclasts in RANKL-treated cultures. They also produced fewer CFU-ALP osteoblast precursors in the marrow and showed reduced mineralization in primary osteoblastic cultures, as well as reduced expression for the osteoblast lineage marker, alkaline phosphatase, in bone isolates. The almost exclusive location of y6Rs in the hypothalamus suggests a critical role of central neuronal pathways controlling this uncoupling of bone remodelling which is in line with known actions or other Y-receptors in the brain. In conclusion, y6R signalling is required for maintenance of bone mass, with loss of y6R uncoupling bone remodelling and resulting in a negative bone balance. This study expands the scope of hypothalamic regulation of bone, highlighting the importance for neural/endocrine coordination and their marked effect upon skeletal homeostasis.

Transgenic expression of Dspp partially rescued the long bone defects of Dmp1-null mice

Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) belong to the Small Integrin-Binding Ligand N-linked Glycoprotein (SIBLING) family. In addition to the features common to all SIBLING members, DMP1 and DSPP share several unique similarities in chemical structure, proteolytic activation and tissue localization. Mutations in, or deletion of DMP1, cause autosomal recessive hypophosphatemic rickets along with dental defects; DSPP mutations or its ablation are associated with dentinogenesis imperfecta. While the roles and functional mechanisms of DMP1 in osteogenesis have been extensively studied, those of DSPP in long bones have been studied only to a limited extent. Previous studies by our group revealed that transgenic expression of Dspp completely rescued the dentin defects of Dmp1-null (Dmp1−/−) mice. In this investigation, we assessed the effects of transgenic Dspp on osteogenesis by analyzing the formation and mineralization of the long bones in Dmp1−/− mice that expresses a transgene encoding full-length DSPP driven by a 3.6-kb rat Col1a1 promoter (referred as “Dmp1−/−;Dspp-Tg mice”). We characterized the long bones of the Dmp1−/−;Dspp-Tg mice at different ages and compared them with those from Dmp1−/− and Dmp1+/− (normal control) mice. Our analyses showed that the long bones of Dmp1−/−;Dspp-Tg mice had a significant increase in cortical bone thickness, bone volume and mineral density along with a remarkable restoration of trabecular thickness compared to those of the Dmp1−/− mice. The long bones of Dmp1−/−;Dspp-Tg mice underwent a dramatic reduction in the amount of osteoid, significant improvement of the collagen fibrillar network, and better organization of the lacunocanalicular system, compared to the Dmp1−/− mice. The elevated levels of biglycan, bone sialoprotein and osteopontin in Dmp1−/− mice were also noticeably corrected by the transgenic expression of Dspp. These findings suggest that DSPP and DMP1 may function synergistically within the complex milieus of bone matrices.

Polycythemia is associated with bone loss and reduced osteoblast activity in mice.

Increased fragility has been described in humans with polycythemia vera (PV). Herein, we describe an osteoporotic phenotype associated with decreased osteoblast activity in a mouse model of PV and another mouse of polycythemia and elevated circulating erythropoietin (EPO). Our results are important for patients with PV or those treated with recombinant EPO (rEPO). PV and other myeloproliferative syndromes have been recently associated with an increased risk for fractures. However, the presence of osteoporosis in these patients has not been well documented. EPO, a hormone primarily known to stimulate erythropoiesis, has been shown recently to regulate bone homeostasis in mice. The aim of this study was to examine the bone phenotype of a mouse model of PV and compare it to that of animals with polycythemia caused by elevated circulating EPO. Bone mass and remodeling were evaluated by micro-computed tomography and histomorphometry. The JAK2(V617F) knock-in mouse, a model of human PV, manifests polycythemia and low circulating EPO levels. Results from this mouse were compared to wild type (wt) controls and the tg6 transgenic mouse that shows polycythemia caused by increased constitutive expression of EPO. Compared to wt, both JAK2(V617F) and tg6 mice had a decrease in trabecular bone mass. Tg6 mice showed an additional modest decrease in cortical thickness and cortical bone volume per tissue volume (P < 0.01) suggesting a more severe bone phenotype than JAK2(V617F). Decreased osteoblast numbers and bone formation along with normal osteoclast numbers and activity were found in both mice. This study indicates that PV is associated with low bone mass and decreased osteoblast activity in mice. Our results support future studies of osteoporosis in affected humans. Polycythemia caused by chronically elevated circulating EPO also results in bone loss, and implications on patients treated with rEPO should be evaluated.

Anti-Sclerostin Treatment Prevents Multiple Myeloma Induced Bone Loss and Reduces Tumor Burden

Multiple myeloma (MM) is a malignancy of plasma cells and is characterized by unrestricted tumor cell growth in bone marrow (BM). MM causes destructive osteolytic lesions causing bone fracture, bone pain, hypercalcaemia, and nerve-compression, resulting from increased bone resorption and suppressed bone formation. Despite the introduction of agents to inhibit bone resorption, such as bisphosphonates, which prevent further bone loss, approaches to preventing osteoblast suppression and repair bone lesions are limited and there are no agents available clinically.The wnt/β-catenin pathway plays a critical role in the regulation of bone formation. Production of the soluble wnt antagonist dickkopf1 (Dkk1) by MM cells has been implicated in MM inhibition of bone formation. As such, Anti-Dkk1 treatment prevents bone disease in pre-clinical models of MM and is in early clinical development. However, Dkk1 is not expressed by all myeloma cells; hence only a proportion of patients may respond to anti-Dkk1 therapy. Sclerostin (Scl) is a soluble wnt antagonist whose expression, unlike Dkk1, is restricted to osteocytes; therefore Scl targeted agents may have less off target effects. Anti-Sclerostin (Anti-Scl) treatment increased bone formation and bone volume in experimental models of osteoporosis, and increased bone mineral density in phase II osteoporosis clinical trials. However, Anti-Scl treatment effects on myeloma bone disease are unknown. Further, cells of the BM such as osteoblasts have been implicated in the regulation of MM cell survival and growth. Thus, in the present study we explored the potential for Anti-Scl therapy to prevent MM induced bone loss and inhibit MM growth in both murine and human xenograft MM models.Female C57BLKalwRij mice (n=8) were injected i.v, with 5TGM1/eGFP murine MM cells (1×106) and female SCID/beige mice (n=10) were injected i.v. with MM1S/Luc/eGFP human MM cells (4 × 106). 24 hours later, naïve mice (without tumor cells) or mice bearing MM cells were treated with anti-sclerostin antibody (Anti-Scl) (100mg/kg i.v) or control antibody. Mice were sacrificed at day 21 (MM1S) or day 28 (5TGM1) and the effect of Anti-Scl on bone structure in the femora and vertebrae were determined by microCT analysis. The effect of Anti-Scl on MM burden was determined by bioluminescent imaging (BLI) performed twice weekly from week 1 using a Xenogen IVIS system, whereas MM burden in 5TGM1/eGFP bearing mice was examined by FACS analysis.Anti-Scl treatment in naïve C57BLKalwRij mice increased trabecular bone volume fraction (BV/TV, 39%, p<0.01, Fig. 1B) in the femur, which was mediated by increases in trabecular thickness (Tb.Th, 42%, p<0.01). Treatment also increased cortical bone volume (22%, p<0.01) in the femur and increased trabecular BV/TV in the vertebra (32%, p<0.01). This demonstrated the potent bone anabolic effect of Anti-Scl independent of myeloma cells. Injection of 5TGM1 cells resulted in a decrease in femoral trabecular BV/TV (30%, p<0.01) through a 30% reduction in trabecular number (TbN) (p<0.01), but no effect on Tb.Th (Figs. 1A and B), whilst also reducing cortical bone volume (BV) by 6% (p<0.05). Vertebrae were also impacted by 5TGM1 tumor growth with a 29% reduction in Tb BV/TV through a 23% reduction in Tb.Th (p<0.01) and also a 15% reduction in cortical BV (p<0.01). Treatment of 5TGM1-bearing mice with Anti-Scl increased trabecular BV/TV (46%, p<0.01) and Tb.Th (30%, p<0.01) to values equivalent to femora of naïve, non-tumor bearing, control mice. Treatment with Anti-Scl also increased cortical BV by 16% (p<0.01), vertebral Tb BV/TV by 29% and cortical BV by 36% in 5TGM1 burdened mice (p<0.01).Treatment of 5TGM1-bearing mice with Anti-Scl had no effect on the proportion 5TGM1/eGFP cells in the BM or spleen. However Anti-Scl treatment significantly suppressed tumor progression in the MM1S model at 3, 3.5 and 4 weeks post cell injection, as determined by BLI imaging (p=0.02, wk 3; p=0.0019, wk 3.5, and p=0.0068, wk 4, 2-tailed t-tests, Fig. 1C).These data demonstrate that Anti-Scl antibody treatment can prevent development of myeloma bone disease. Furthermore, Anti-Scl treatment also suppressed tumor growth, supporting the possibility that targeting the BM microenvironment with this agent may slow disease progression. Our findings highlight the potential clinical application of Anti-Scl antibody treatment in patients with MM and other bone destructive cancers. [Display omitted] DisclosuresKneissel:Novartis Institutes for Biomedical Research, Novartis Pharma AG: Employment. Kramer:Novartis Pharma AG: Employment. Brooks:Spouse works for Boston Biomedical Inc: Employment.

Computer modelling integrated with micro-CT and material testing provides additional insight to evaluate bone treatments: Application to a beta-glycan derived whey protein mice model

The primary aim of this study was to evaluate the influence of a whey protein diet on computationally predicted mechanical strength of murine bones in both trabecular and cortical regions of the femur. There was no significant influence on mechanical strength in cortical bone observed with increasing whey protein treatment, consistent with cortical tissue mineral density (TMD) and bone volume changes observed. Trabecular bone showed a significant decline in strength with increasing whey protein treatment when nanoindentation derived Young׳s moduli were used in the model. When microindentation, micro-CT phantom density or normalised Young׳s moduli were included in the model a non-significant decline in strength was exhibited. These results for trabecular bone were consistent with both trabecular bone mineral density (BMD) and micro-CT indices obtained independently. The secondary aim of this study was to characterise the influence of different sources of Young׳s moduli on computational prediction. This study aimed to quantify the predicted mechanical strength in 3D from these sources and evaluate if trends and conclusions remained consistent. For cortical bone, predicted mechanical strength behaviour was consistent across all sources of Young׳s moduli. There was no difference in treatment trend observed when Young׳s moduli were normalised. In contrast, trabecular strength due to whey protein treatment significantly reduced when material properties from nanoindentation were introduced. Other material property sources were not significant but emphasised the strength trend over normalised material properties. This shows strength at the trabecular level was attributed to both changes in bone architecture and material properties.

Reproducibility of compartmental subchondral bone morphometry in the mouse tibiofemoral joint

AimEvidence suggests that subchondral bone can be used as a predictor for the onset of osteoarthritis. As such, there is a need to accurately and reproducibly quantify subchondral bone in areas where osteoarthritis develops. In this paper, we present a novel technique for the segmentation of subchondral bone in the tibiofemoral joint and assess the reproducibility of this method with multiple measures and users. MethodsThe right hind leg of seven C57BL/6 mice were excised and imaged in μCT. The menisci and patella were manually segmented and the image data was Gaussian filtered and binarized. An in-house algorithm was used to generate cortical and epiphyseal volumes of interest and standard morphometric indices for bone were computed. The intraclass correlation coefficient (ICC), absolute precision error (PE(SD)), and precision error as a percentage of the coefficient of variation of the repeated measurements (PE(%CV)) were calculated for each index. Additionally, an inter-user study was performed using the same indices and statistics. ResultsFor repeated measures, ICC ranged from 0.869 (cortical bone volume fraction, femur) to 0.994 (degree of anisotropy, femur). Similarly, PE(%CV) ranged from 0.84% (cortical bone volume fraction, femur) to 5.11% (connectivity density, tibia). For repeated users, no effect was seen in the femur with a slight effect in the tibia. ConclusionsA novel method for the automatic segmentation of cortical and epiphyseal bone is presented and is shown to be reproducible in C57BL/6 mice. This tool will allow for high-throughput studies of osteoarthritis in animal models.

Low-carbohydrate, high-fat diets have sex-specific effects on bone health in rats.

Studies in humans suggest that consumption of low-carbohydrate, high-fat diets (LC-HF) could be detrimental for growth and bone health. In young male rats, LC-HF diets negatively affect bone health by impairing the growth hormone/insulin-like growth factor axis (GH/IGF axis), while the effects in female rats remain unknown. Therefore, we investigated whether sex-specific effects of LC-HF diets on bone health exist. Twelve-week-old male and female Wistar rats were isoenergetically pair-fed either a control diet (CD), "Atkins-style" protein-matched diet (LC-HF-1), or ketogenic low-protein diet (LC-HF-2) for 4weeks. In females, microcomputed tomography and histomorphometry analyses were performed on the distal femur. Sex hormones were analysed with liquid chromatography-tandem mass spectrometry, and endocrine parameters including GH and IGF-I were measured by immunoassay. Trabecular bone volume, serum IGF-I and the bone formation marker P1NP were lower in male rats fed both LC-HF diets versus CD. LC-HF diets did not impair bone health in female rats, with no change in trabecular or cortical bone volume nor in serum markers of bone turnover between CD versus both LC-HF diet groups. Pituitary GH secretion was lower in female rats fed LC-HF diet, with no difference in circulating IGF-I. Circulating sex hormone concentrations remained unchanged in male and female rats fed LC-HF diets. A 4-week consumption of LC-HF diets has sex-specific effects on bone health-with no effects in adult female rats yet negative effects in adult male rats. This response seems to be driven by a sex-specific effect of LC-HF diets on the GH/IGF system.

Flapless cortical bone damage has no effect on medullary bone mesial to teeth being moved

In this study, we evaluated the effects of bone awl-induced damage to bone surrounding a tooth that was moved. A randomized split-mouth design with 7 foxhounds was performed to evaluate protraction of the mandibular third premolars for 56 days with 200 g of orthodontic force. Before initiating tooth movements, a bone awl was used on the experimental side to create 60 buccal and lingual microfracture injuries to the cortical bone without a periosteal flap. Tooth movements were performed on the control and experimental sides. Microcomputed tomography and histology were used to assess bone morphology and modeling. Radiographic and caliper measures were used to assess tooth movements. The awl-induced injuries produced significant damage and microfractures (95 mm(3)). Buccal and lingual cortical bone volume fractions and densities were significantly less and cortical modeling was significantly greater on the experimental sides than on the control sides. Bone volume fractions and densities mesial to the third premolars were the same on the experimental and control sides. Experimental side tooth movements (1.40 ± 0.25 mm) were statistically the same as the control side tooth movements (1.57 ± 0.45 mm). The effects of flapless, bone awl-induced damage were limited to the cortical bone. Because there was no effect on the medullary bone mesial to the tooth being moved, no differences in tooth movements were produced.

Adipose Tissue Infiltration of Bone Marrow and Skeletal Muscle in Children with Mild Spastic Cerebral Palsy

Non-ambulatory children with severe cerebral palsy (CP) have a high degree of adipose tissue (AT) infiltration which could play a role in the development of insulin resistance and fractures. The objective of this study was to determine the degree of AT infiltration within the bone marrow cavity of the middle tibia and within the surrounding muscle in ambulatory children with mild spastic CP (5-11 y) compared to typically developing controls matched for age and sex (n = 8/group). The concentration of AT in the bone marrow and muscle, and the intermuscular, subfascial and subcutaneous AT volume, muscle volume, cortical bone volume and bone strength (i.e., section modulus, Z and polar moment of inertia, J) were estimated using magnetic resonance imaging. Children with CP vs. controls had a higher (P < 0.05) concentration of AT in the bone marrow (83 ± 2 % vs. 81 ± 2 %) and muscle (27 ± 7 % vs. 18 ± 5 %). Children with CP vs. controls also had higher (P < 0.05) subfascial AT (357 %), and lower muscle volume (40%), cortical bone volume (26 %), Z (31 %) and J (38 %). Although not statistically significant (P = 0.07), children with CP vs. controls also had higher intermuscular AT (391 %). Whether the high concentration of AT in bone marrow and muscle during childhood are early indicators of insulin resistance and future fracture requires further investigation. Supported by the NIH(Grant HD071397-01).

Effect of daily lithium chloride administration on bone mass and strength in growing broiler chickens,

The objective was to determine the effects of oral lithium chloride supplementation on bone strength and mass in broiler chickens. Ninety-six broilers were assigned to 1 of 2 treatment groups (lithium chloride or control; n = 48/treatment). Beginning at 1 or 3 wk of age, chickens were administered lithium chloride (20 mg/kg body weight) or water daily by oral gavage. At 6 wk of age, chickens were euthanized and bone and muscle samples were collected. A 24 h lithium chloride (20 mg/kg body weight) challenge determined that serum lithium chloride increased within 2 h and cleared the system within 24 h, demonstrating the effective delivery of lithium chloride. Treatment did not influence body weight (P ≥ 0.20) or feed intake (P ≥ 0.81), demonstrating that lithium chloride did not negatively affect broiler growth. To determine bone strength, 3-point bending was performed on the femora and tibiae obtained from control and lithium chloride-treated birds in the 1 wk group. Lithium chloride-treated birds had a 22% reduction in stiffness compared with control in the femora (P = 0.02) without a corresponding reduction in elastic modulus. No differences were observed in yield or ultimate load and in the corresponding calculations of stresses (P ≥ 0.26). The toughness of tibiae was not altered in lithium chloride compared with control (P = 0.11). Bone length and micro-CT imaging were performed on the tibiae of control and lithium chloride groups. No differences (P ≥ 0.52) in bone length, cortical or trabecular bone volume, trabecular thickness, number, or spacing were observed. Lithium chloride treatment did not affect pectoralis muscle color or lipid oxidation (P > 0.05). In conclusion, lithium chloride treatment in broilers did not negatively affect growth or meat quality. A reduction in bone stiffness of the femur with lithium chloride treatment was observed, however unlike the mouse model, the dosages of lithium chloride used in the current study did not result in anabolic effects on broiler long bones.

Individual and combining effects of anti-RANKL monoclonal antibody and teriparatide in ovariectomized mice

We examined the individual and combined effects of teriparatide and anti-RANKL (receptor activator of nuclear factor κB ligand) monoclonal antibody in ovariectomized mice. Three-month-old female C57BL/6 mice were ovariectomized (OVX) or sham operated. Four weeks after OVX, they were assigned to 3 different groups to receive anti-RANKL monoclonal antibody (Ab) alone (5mg/kgsingle injection at 4weeks after OVX, Ab group), teriparatide alone (80μg/kg daily injection for 4weeks from 4weeks after OVX, PTH group), or mAb plus teriparatide (Ab+PTH group). Mice were sacrificed 8weeks after OVX. Bone mineral density (BMD) was measured at the femur and lumbar spine. Hind limbs were subjected to histological and histomorphometric analysis. Serum osteocalcin and CTX-I levels were measured to investigate the bone turnover. Compared with Ab group, Ab+PTH group showed a significant increase in BMD at distal femur and femoral shaft. Cortical bone volume was significantly increased in PTH and Ab+PTH groups compared with Ab group. Bone turnover in Ab+PTH group was suppressed to the same degree as in Ab group. The number of TRAP-positive multinucleated cells was markedly reduced in Ab and Ab+PTH groups. These results suggest that combined treatment of teriparatide with anti-RANKL antibody has additive effects on BMD in OVX mice compared with individual treatment.

High fructose and high fat diet increased bone volume of trabecular and cortical bone in growing female rats

Purpose: The objective of this study was to investigate the effects of a high fructose and fat diet on bone growth and maturation in growing female rats. Methods: Three-week-old female SD rats were randomly assigned to four experimental groups; the control group (CON: fed control diet based on AIN-93G, n = 8); the high-fructose diet group (HFrc: fed control diet with 30% fructose, n = 8); the high-fat diet group (Hfat: fed control diet with 45 kcal% fat, n = 8); and the high-fat diet plus high fructose group (HFrc + HFat: fed diets 45 kcal% fat with 30% fructose, n = 8). Each group was assigned their respective diets for the remaining eight weeks. Bone-related parameters (bone mineral density (BMD) and structural parameters, osteocalcin (OC), deoxypyridinoline (DPD)) and morphologic changes of kidney were analyzed at the end of the experiment. Results: Final body weights and weight gain were higher in the HFat and HFrc + HFat groups and showed higher tendency in the HFrc group compared with those of the CON group (p < 0.05); however, no significant difference in caloric intake was observed among the four experimental groups. The serum OC levels of the HFrc and HFrc + HFat groups were lower than those of the CON and HFat groups (p < 0.05). Urinary levels of DPD did not differ among the experimental groups. BV/TV and Tb.N of trabecular bone were higher in the HFrc + HFat group and showed a higher tendency in the HFrc group than those of the CON and HFat groups (p < 0.05). Tb.Pf of trabecular bone were lower in the HFrc + HFat group than those in the CON and HFat groups (p < 0.05). However, no difference in trabecular BMD was observed among the experimental groups. Cortical bone volume was higher in the HFat and HFrc + HFat groups than in the CON and HFrc groups (p < 0.05). No morphology change in kidney was observed among the experimental groups. Conclusion: Our study suggests that 8 weeks of high-fructose and high fat intake could improve the bone quality (Structural parameters) of trabecular and cortical bone of tibia in growing female rats.

Bone cement grafting increases implant primary stability in circumferential cortical bone defects.

Implant beds with an insufficient amount of cortical bone or a loss of cortical bone can result in the initial instability of a dental implant. Thus, the objective of this study was to evaluate the effect of bone cement grafting on implant initial stability in areas with insufficient cortical bone. Two different circumferential defect depths (2.5 mm and 5 mm) and a control (no defect) were prepared in six bovine rib bones. Fourteen implants of the same type and size (4 mm×10 mm) were placed in each group. The thickness of the cortical bone was measured for each defect. After the implant stability quotient (ISQ) values were measured three times in four different directions, bone cement was grafted to increase the primary stability of the otherwise unstable implant. After grafting, the ISQ values were measured again. As defect depth increased, the ISQ value decreased. In the controls, the ISQ value was 85.45±3.36 (mean±standard deviation). In circumferential 2.5-mm and 5-mm defect groups, the ISQ values were 69.42±7.06 and 57.43±6.87, respectively, before grafting. These three values were significantly different (P<0.001). After grafting the bone cement, the ISQ values significantly increased to 73.72±8.00 and 67.88±10.09 in the 2.5-mm and 5.0-mm defect groups, respectively (P<0.05 and P<0.001). The ISQ value increased to more than double that before grafting in the circumferential 5-mm defect group. The ISQ values did not significantly differ when measured in any of the four directions. The use of bone cement remarkably increased the stability of the implant that otherwise had an insufficient level of stability at placement, which was caused by insufficient cortical bone volume.

Effect of Low-intensity Pulsed Ultrasound (LIPUS) with Different Frequency on Bone Defect Healing

Low-intensity pulsed ultrasound (LIPUS) is known to promote bone defect healing. The frequency of the LIPUS is known to influence directivity and depth of penetration, but the differences on bone healing remains unknown. This study was to investigate the effect of LIPUS with different frequencies on bone defect healing. Bone defects of 1.6 mm in diameter were created in femurs of ten-week-old male Long-Evans rats (n=36). Experimetal groups were exposed LIPUS (intensity: 30 mW/cm2, burst width: 200 μs, time: 15 min/day) and divided into a low frequency (LF, 1.5 MHz) group and a high frequency (HF, 3.0 MHz) group. Non-exposed LIPUS group were used as control. After 3, 5, 7, and 10 days, femurs were removed and radiological, histomorphological and molecular biological evaluations were conducted. Micro-CT images showed that the depression in cortical bone was reduced in LF and HF groups. 3D bone morphological analysis at 10 days revealed that LIPUS increased cortical bone volume / tissue volume (BV/TV) and decreased BV/TV in the lower layer of cancellous bone (P<0.05). Histomorphologically, clot retraction was seen in the both LIPUS groups but not in the control group at 3 days. These effects were observed at a deeper layer in the LF group than in the HF group. No significant difference in osteopontin (OPN) gene expression was observed. However, osteocalcin (OCN) gene expression was significantly elevated in the HF group relative to the control group at 10 days (P<0.05). Immunohistochemical staining revealed that newly formed bone exhibited a positive reaction to OPN and OCN in both LIPUS groups. Healing of the bone defect area was noted in both LIPUS groups, but there was no clear difference in histomorphology between the LF and HF groups. LIPUS frequencies of 1.5 MHz and 3.0 MHz promote increased cortical bone mass and remodeling of cancellous bone in rat femurs with bone defects.

PTH Signaling During Exercise Contributes to Bone Adaptation.

Improving the structural integrity of bone reduces fracture risk and development of osteoporosis later in life. Exercise can increase the mechanical properties of bone, and this increase is often attributed to the dynamic loading created during exercise. However, the increase in systemic parathyroid hormone (PTH) levels during exercise gives reason to hypothesize that PTH signaling also regulates bone adaptation in response to exercise. Therefore, the first aim of this study was to establish the impact PTH signaling has on bone adaptation during exercise by inhibiting PTH signaling with PTH(7-34); the second aim was to determine whether increasing PTH levels during exercise with PTH(1-34) can augment bone adaptation. Thirty minutes after a single bout of running on a treadmill, mice exhibited a twofold increase in systemic PTH levels. Under the same exercise regimen, the influence of PTH signaling on bone adaptation during exercise was then evaluated in mice after 21 consecutive days of exercise and treatment with PTH(7-34), PTH(1-34), or vehicle. Exercise alone caused a significant increase in trabecular bone volume with adaptation to a more platelike structure, which was inhibited with PTH(7-34) during exercise. Changes in structural-level and tissue-level mechanical properties during exercise occurred in the absence of significant changes to cortical bone geometry. Inhibition of PTH signaling during exercise attenuated the changes in structural-level mechanical properties, but not tissue-level properties. Enhanced PTH signaling during exercise with PTH(1-34) increased trabecular and cortical bone volume, but had little effect on the structural-level and tissue-level mechanical properties compared to exercise alone. Our study is the first to demonstrate that bone adaptation during exercise is not only a function of dynamic loading, but also PTH release, and that PTH signaling contributes differently at the structural and tissue levels.

Netrin-1 is a critical autocrine/paracrine factor for osteoclast differentiation.

Bone metabolism is a vital process that involves resorption by osteoclasts and formation by osteoblasts, which is closely regulated by immune cells. The neuronal guidance protein Netrin-1 regulates immune cell migration and inflammatory reactions, but its role in bone metabolism is unknown. During osteoclast differentiation, osteoclast precursors increase expression of Netrin-1 and its receptor Unc5b. Netrin-1 binds, in an autocrine and paracrine manner, to Unc5b to promote osteoclast differentiation in vitro, and absence of Netrin-1 or antibody-mediated blockade of Netrin-1 or Unc5b prevents osteoclast differentiation of both murine and human precursors. We confirmed the functional relationship of Netrin-1 in osteoclast differentiation in vivo using Netrin-1-deficient (Ntn1(-/-) ) or wild-type (WT) bone marrow transplanted mice. Notably, Ntn1(-/-) chimeras have markedly diminished osteoclasts, as well as increased cortical and trabecular bone density and volume compared with WT mice. Mechanistic studies revealed that Netrin-1 regulates osteoclast differentiation by altering cytoskeletal assembly. Netrin-1 increases regulator of Rho-GEF subfamily (LARG) and repulsive guidance molecule (RGMa) association with Unc5b, which increases expression and activation of cytoskeletal regulators RhoA and focal adhesion kinase (FAK). Netrin-1 and its receptor Unc5b likely play a role in fusion of osteoclast precursors because Netrin-1 and DC-STAMP are tightly linked. These results identify Netrin-1 as a key regulator of osteoclast differentiation that may be a new target for bone therapies.

Where does hip fracture initiate?

Where does hip fracture initiate?