JOURNAL OF AMERICAN GERIATRICS SOCIETY, Issue 11 2008
Andre B. Araujo PhD
OBJECTIVES: To examine the association between aging and physical function in men by testing a theoretically based model of aging, hormones, body composition, strength, and physical function with data obtained from men enrolled in the Boston Area Community Health/Bone (BACH/Bone) Survey. DESIGN: Cross-sectional, observational survey. SETTING: Population-based. PARTICIPANTS: Eight hundred ten black, Hispanic, and white randomly selected men from the Boston area aged 30 to 79. MEASUREMENTS: Testosterone, estradiol, sex hormone,binding globulin, lean and fat mass, grip strength, and summated index of physical function (derived from walk and chair stand tests). RESULTS: Measures of grip strength and physical function declined strongly with age. For instance, 10 years of aging was associated with a 0.49-point difference (scale 0,7) in physical function. Age differences in total testosterone and estradiol concentrations were smaller than age differences in their free fractions. Weak or nonsignificant age-adjusted correlations were observed between hormones and measures of physical function, although path analysis revealed a positive association between testosterone and appendicular lean mass and a strong negative association between testosterone and total fat mass. Lean and fat mass, in turn, were strongly associated with grip strength and physical function, indicating the possibility that testosterone influences physical function via indirect associations with body composition. CONCLUSION: The age-related decline in serum testosterone concentration in men has a weak association with physical strength and functional outcomes through its associations with lean and fat mass. [source]

FLUCLOXACILLIN ASSOCIATED NEUTROPENIA IN CHILDREN TREATED FOR BONE AND JOINT INFECTIONS (J. Paediatr.

JOURNAL OF PAEDIATRICS AND CHILD HEALTH, Issue 9-10 2005
Child Health 2005; 41: 4
No abstract is available for this article. [source]

HN09P ASSESSMENT OF FREE FIBULAR BONE IN THE RECONSTRUCTED MANDIBLE USING THREE-DIMENSIONAL COMPUTER GENERATED IMAGES

ANZ JOURNAL OF SURGERY, Issue 2007
H. Nabi
We report the preliminary results of the Royal Adelaide Hospital experience with multidimensional simulated views of the fibula-flap reconstructed mandible. The free fibular flap is a well recognised option for mandibular reconstruction. What is not well understood however is how the fibula behaves in comparison to the dentate mandible. To date, skeletal remodelling and bone atrophy has only been assessed using standard orthopantogram films. For many years three-dimensional (3D) computer generated models using data from CT scans have been utilised for craniofacial reconstruction. We proposed that these images will enable us to more accurately visualise the integration of the transplanted graft within the mandible. We recalled and CT scanned patients from 2004 to 2006 that underwent free fibular flaps for reconstruction of mandibular malignancy and performed 3D reconstruction of these images. This is the first reported series of multidimensional computer generated images to assess bone in the reconstructed mandible. [source]

TOCOTRIENOL OFFERS BETTER PROTECTION THAN TOCOPHEROL FROM FREE RADICAL-INDUCED DAMAGE OF RAT BONE

CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 9 2005
NS Ahmad
SUMMARY 1.,Free radicals generated by ferric nitrilotriacetate (FeNTA) can activate osteoclastic activity and this is associated with elevation of the bone resorbing cytokines interleukin (IL)-1 and IL-6. In the present study, we investigated the effects of 2 mg/kg FeNTA (2 mg iron/kg) on the levels of serum IL-1 and IL-6 with or without supplementation with a palm oil tocotrienol mixture or ,-tocopherol acetate in Wistar rats. 2.,The FeNTA was found to elevate levels of IL-1 and IL-6. Only the palm oil tocotrienol mixture at doses of 60 and 100 mg/kg was able to prevent FeNTA-induced increases in IL-1 (P < 0.01). Both the palm oil tocotrienol mixture and ,-tocopherol acetate, at doses of 30, 60 and 100 mg/kg, were able to reduce FeNTA-induced increases in IL-6 (P < 0.05). Therefore, the palm oil tocotrienol mixture was better than pure ,-tocopherol acetate in protecting bone against FeNTA (free radical)-induced elevation of bone-resorbing cytokines. 3.,Supplementation with the palm oil tocotrienol mixture or ,-tocopherol acetate at 100 mg/kg restored the reduction in serum osteocalcin levels due to ageing, as seen in the saline (control) group (P < 0.05). All doses of the palm oil tocotrienol mixture decreased urine deoxypyridinoline cross-link (DPD) significantly compared with the control group, whereas a trend for decreased urine DPD was only seen for doses of 60 mg/kg onwards of ,-tocopherol acetate (P < 0.05). 4.,Bone histomorphometric analyses have shown that FeNTA injections significantly lowered mean osteoblast number (P < 0.001) and the bone formation rate (P < 0.001), but raised osteoclast number (P < 0.05) and the ratio of eroded surface/bone surface (P < 0.001) compared with the saline (control) group. Supplementation with 100 mg/kg palm oil tocotrienol mixture was able to prevent all these FeNTA-induced changes, but a similar dose of ,-tocopherol acetate was found to be effective only for mean osteoclast number. Injections of FeNTA were also shown to reduce trabecular bone volume (P < 0.001) and trabecular thickness (P < 0.05), whereas only supplementation with 100 mg/kg palm oil tocotrienol mixture was able to prevent these FeNTA-induced changes. [source]

ON ISOTOPES AND OLD BONES*

ARCHAEOMETRY, Issue 6 2008
J. A. LEE-THORP
This review charts the developments and progress made in the application of stable light isotope tools to palaeodietary adaptations from the 1970s onwards. It begins with an outline of the main principles governing the distribution of stable light isotopes in foodwebs and the quality control issues specific to the calcified tissues used in these analyses, and then proceeds to describe the historical landmark studies that have marked major progress, either in their archaeological applications or in enhancing our understanding of the tools. They include the adoption of maize agriculture, marine-focused diets amongst coastal hunter,gatherers, trophic level amongst Glacial-period modern humans and Neanderthals, and the use of savannah resources by early hominins in Africa. Particular attention is given to the progress made in addressing the challenges that have arisen out of these studies, including issues related to the routing of dietary nutrients. I conclude with some firm, and some more speculative, pointers about where the field may be heading in the next decade or so. [source]

Performance evaluation of LIBTA/hybrid time-slot selection algorithm for cellular systems,

INTERNATIONAL JOURNAL OF COMMUNICATION SYSTEMS, Issue 6 2001
Jyh-Horng Wen
Abstract This paper studies the performance of radio assignment algorithms for portable access in cellular systems. Several channel access procedures are proposed and simulated using block oriented network simulator (BONeS) simulation of a model 36-port system. Simulation results exhibit that load-sharing system with LIBTA algorithm is better than directed retry system with the same algorithm by around 0.9 erlangs while better than quasi-fixed channel assignment (QFCA) system by around 2 erlangs if the grade of service (GOS) is constrained to less than 10 per cent. Plus, a hybrid time-slot selection procedure is proposed to enhance the system performance. It is observed that systems with hybrid time-slot selection perform better than those with LIBTA algorithm in GOS under heavy load. It is also observed that load sharing system with hybrid time-slot selection algorithm is better than directed retry system with the same algorithm by around 0.7 erlangs and better than QFCA system by around 2 erlangs. Copyright © 2001 John Wiley & Sons, Ltd. [source]

Bone marrow-derived cells expand memory CD8+ T,cells in response to viral infections of the lung and skin

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 2 2006
Gabrielle
Abstract While naive CD8+ T,cells have been shown to require bone marrow-derived dendritic cells (DC) to initiate immunity, such a requirement for memory CD8+ T,cells has had limited assessment. By generating bone marrow chimeras that express the appropriate antigen-presenting molecules on either radiation-sensitive bone marrow-derived or radiation-resistant non-bone marrow-derived compartments, we showed that both primary and secondary immune responses to influenza virus infection of the lung were initiated in the draining LN. This required cells of bone marrow origin, most likely DC, for optimal expansion within the secondary lymphoid compartment. This was similarly the case with HSV-1 infection of the skin. As Langerhans cells are radioresistant, unlike other DC populations, these studies also demonstrate that the radiosensitive DC responsible for secondary expansion of HSV-specific memory are not Langerhans cells. [source]

Expression of milk fat globule epidermal growth factor,8 in immature dendritic cells for engulfment of apoptotic cells

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 5 2004
Kay Miyasaka
Abstract Milk fat globule epidermal growth factor,8 (MFG-E8) is a protein that stimulates the engulfment of apoptotic cells by phagocytes. Here, we show that mouse immature dendritic cells (DC) generated in vitro by culturing bone marrow progenitors in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF), and Langerhans cells present in the skins, expressed MFG-E8. Bone marrow-derived macrophages generated by M-CSF did not express MFG-E8. MFG-E8 expressed in immature DC was found to be secreted as exosomes. The expression of MFG-E8 was significantly suppressed when the immature DC were induced to mature by treating them with lipopolysaccharides. This expression of MFG-E8 was well correlated with the ability of the cells to engulf apoptotic cells. That is,immature DC phagocytosed apoptotic cells more efficiently than did mature DC or bone marrow-derived macrophages. The ability of immature DC to engulf apoptotic cells was severely reduced when the immature DC were prepared from MFG-E8-deficient mice. These results indicated that MFG-E8 plays an essential role in the engulfment of apoptotic cells by bone marrow-derived immature DC. [source]

Bonding of a Silorane-Based Composite System to Bone

ADVANCED ENGINEERING MATERIALS, Issue 11 2009
Xiaohong Wu
This work was to analyze by Weibull statistics the shear bond strength of a low-shrink Silorane-based composite system to bone. The etching abilities of the adhesives were investigated by scanning electron microscopy. Results suggest that an effective and reliable bond to bone could be achieved by the Silorane-based composite system, showing the potential of this system to be used as a bone cement. [source]

Biocompatibility of Lotus-type Stainless Steel and Titanium in Alveolar Bone

ADVANCED ENGINEERING MATERIALS, Issue 9 2006
Y. Higuchi
Abstract Lotus-type porous stainless steel (SUS304L) and porous titanium were fabricated by unidirectional solidification in a mixture gas of hydrogen and argon. The porous metals which were cut into 5,mm cubes (non-dehydrogenated) and 3.4,mm,,×,5,mm cylinders (dehydrogenated) were implanted into the canine mandible alveolar bone for two, four and eight weeks for animal experiments. The changes in the tissues were observed using SEM. For porous stainless steel (cylindrical; dehydrogenated) new formation of bones was observed around the sample in two weeks without any sign of bony ingrowth into the pores. The osteogenesis was found in shallow areas in the pores in four weeks and deep in the pores in eight weeks. Porous titanium, on the other hand, showed deep ingrowth of new bones in four weeks. Our observations allowed us to expect application of the porous metals as biomaterials. They maintain mechanical strength and are lighter in weight so that it is expected to be applied for dental implants and core materials of artificial bones. [source]

Osteocytes in the pathogenesis of osteoporosis

GERIATRICS & GERONTOLOGY INTERNATIONAL, Issue 4 2008
Kyoji Ikeda
Bone is continuously renewed by bone resorption and subsequent bone formation, a coupling process that maintains the quality as well as the quantity of bone. It is widely accepted that osteoporosis develops when bone resorption exceeds bone formation, and the treatment as well as diagnosis has been targeted to two major cell types, osteoclasts and osteoblasts. Inside bone is a network of the third cell type, osteocytes, the physiological function of which has long remained an enigma. We have developed a transgenic mouse model in which inducible and specific ablation of osteocytes can be achieved in vivo, and here use it to demonstrate that osteocytes serve an important function in regulating the activities of osteoblasts and osteoclasts, while sensing and transducing the mechanical forces exerted on bone. Thus, osteocytes should provide an attractive target for the development of new types of mechanotransduction-based therapeutics and diagnostics for osteoporosis. [source]

Inside Front Cover: A Unique Microcracking Process Associated with the Inelastic Deformation of Haversian Bone (Adv. Funct.

ADVANCED FUNCTIONAL MATERIALS, Issue 1 2009
Mater.
Human cortical bone is capable of adapting to the mechanical environment through dynamic remodeling of the Haversian systems. The presence of Haversian canals, however, also introduces stress concentration and could have detrimental effects on the fracture resistance of bone. How is the hierarchical structure in bone designed to alleviate such stress concentrations? On page 57, Vincent Ebacher and Rizhi Wang report a unique and stable microcracking process accompanying the inelastic deformation of Haversian bone. The results lead to the critical role of the well-organized bone lamellae surrounding each Haversian canal. [source]

A Unique Microcracking Process Associated with the Inelastic Deformation of Haversian Bone

ADVANCED FUNCTIONAL MATERIALS, Issue 1 2009
Vincent Ebacher
Abstract Since the discovery of the Haversian system in human bone over three hundred years ago, researchers have been wondering about its mechanical advantages. Despite positive experimental evidences on the intervention of Haversian systems in the fracture process, the contributions of Haversian systems to bone fracture have been obscure. Here a unique microcracking process accompanying the inelastic deformation of Haversian bone is reported that may shine light on its structural advantages over other bones. When compressed transversely, the concentric bone lamellae surrounding each Haversian canal allow multiple radial microcracks and arc-shaped cracks to develop intralamellarly. Groups of circumferential arc-shaped microcracks develop in high shear zones and radiate out in oblique directions from each Haversian canal. At the cortical bone level, where the Haversian systems are randomly distributed within the interstitial matrix, multiple nucleations and stable development of such arc-shaped cracks happen to most Haversian systems progressively. As a result, Haversian bone is not sensitive to the presence of Haversian canals and demonstrates high inelastic strains at macroscopic level. [source]

Hepatic differentiation of human bone marrow-derived UE7T-13 cells: Effects of cytokines and CCN family gene expression

HEPATOLOGY RESEARCH, Issue 12 2007
Takashi Shimomura
Aim:, Bone marrow-derived mesenchymal stem cells (MSC) are expected to be an excellent source of cells for transplantation. We aimed to study the culture conditions and involved genes to differentiate MSC into hepatocytes. Methods:, The culture conditions to induce the efficient differentiation of human bone marrow-derived UE7T-13 cells were examined using cytokines, hormones, 5-azacytidine and type IV collagen. Results:, We found that combination of acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) with type IV collagen coating induced hepatic differentiation of UE7T-13 cells at over 30% frequency, where expression of albumin mRNA was increased over 20-fold. The differentiated cells had functions of albumin production, glycogen synthesis and urea secretion as well as expressing hepatocyte-specific genes. In addition, these cellshave binuclear and cuboidal morphology, which is a characteristic feature of hepatocytes. During hepatic differentiation, UE7T-13 cells showed depressed expression of WISP1 and WISP2 genes, members of the CCN family. Conversely, knockdown of WISP1 or WISP2 gene by siRNA stimulated hepatic differentiation. The effect of aFGF/bFGF/HGF/type IV collagen coating and WISP1-siRNA on hepatic differentiation was additive. Conclusion:, The present study suggests that aFGF/bFGF/HGF/type IV collagen coating is the efficient condition for hepatic differentiation of UE7T-13 cells, and that WISP1 and WISP2 play an important role in hepatic transdifferentiation of these cells. [source]

Murine TNF,ARE Crohn's disease model displays diminished expression of intestinal Ca2+ transporters

INFLAMMATORY BOWEL DISEASES, Issue 6 2008
Sylvie Huybers MSc
Abstract Background: Patients suffering from Crohn's disease (CD) show increased incidence of low bone mineral density. Investigating this complication is difficult because the exact etiology of CD remains elusive. Mice carrying a deletion in the tumor necrosis factor (TNF) AU-rich elements (ARE) are reported as a model for human CD and are characterized by elevated TNF-, levels and inflammations in the terminal ileum. To evaluate whether these mice have a Ca2+ handling problem, this study analyzed the Ca2+ homeostasis in heterozygous TNF,ARE mice (TNF,ARE/+) in comparison to wildtype littermates. Methods: Beside serum Ca2+ and vitamin D levels, the expression of Ca2+ transporters was analyzed in intestine, kidney and bone using quantitative real-time PCR, Western blot and immunohistochemistry. Bone scans were performed to measure bone parameters. Results: Ca2+ transporters in duodenum (TRPV6, calbindin-D9K, PMCA1b) and kidney (TRPV5, calbindin-D28K, NCX1) showed significantly reduced mRNA expression levels in TNP,ARE/+ mice, except for renal TRPV5. In bone, only calbindin-D9K mRNA displayed a significant down-regulation. These findings were supported by declined duodenal calbindin-D9K and renal calbindin-D28K protein values. Likely, this down-regulation of Ca2+ transporters in TNP,ARE/+ mice is mediated by the 58 ± 9% reduction in serum 1,25(OH)2D3 levels. Diminished expression of Ca2+ transporters combined with unchanged serum Ca2+ levels assumes Ca2+ loss from bone to compensate for the body's overall Ca2+ shortage. Indeed, microcomputed tomography scanning demonstrated reduced trabecular and corticol bone thickness and volume in TNF,ARE/+ mice. This finding is further supported by increased total deoxypyridinoline in serum. Conclusions: Our results imply that TNF,ARE/+ mice have a disturbed Ca2+ homeostasis characterized by reduced duodenal and renal Ca2+ transporters, diminished 1,25(OH)2D3 levels, and increased bone resorption associated with profound bone abnormalities. (Inflamm Bowel Dis 2008) [source]

The Effect of NaF In Vitro on the Mechanical and Material Properties of Trabecular and Cortical Bone

ADVANCED MATERIALS, Issue 4 2009
Philipp J. Thurner
High doses of sodium fluoride in bones lead to severe softening, by weakening interfacial properties between the inorganic minerals and the organic components, while leaving mineralization unchanged. This leads to reduction of microdamage and associated stress-whitening pointing to a change in failure mode. Accordingly, elastic modulus, failure stress, and indentation-distance increase are decreased, whereas failure strain is increased. [source]

The FRAX tool in French women: How well does it describe the real incidence of fracture in the OFELY cohort

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2010
Elisabeth Sornay-Rendu
Abstract The FRAX tool estimates an individual's fracture probability over 10 years from clinical risk factors with or without bone mineral density (BMD) measurement. The aim of our study was to compare the predicted fracture probabilities and the observed incidence of fracture in French women during a 10-year follow-up. The probabilities of fracture at four major sites (hip, clinical spine, shoulder, or wrist) and at the hip were calculated with the FRAX tool in 867 women aged 40 years and over from the Os des Femmes de Lyon (OFELY) cohort. The incidence of fracture was observed over 10 years. Thus 82 women sustained 95 incident major osteoporotic (OP) fractures including 17 fractures at the hip. In women aged at least 65 years (n,=,229), the 10-year predicted probabilities of fracture with BMD were 13% for major OP fractures and 5% for hip fractures, contrasting with 3.6% and 0.5% in women younger than 65 years (p,<,.0001). The predicted probabilities of both major OP and hip fractures were significantly higher in women with osteoporosis (n,=,77, 18% and 10%) and osteopenia (n= 390, 6% and 2%) compared with women with normal BMD (n,=,208, 3% and <1%; p,<,.0001. The predicted probabilities of fracture were two and five times higher in women who sustained an incident major OP fracture and a hip fracture compared with women who did not (p,<,.0001). Nevertheless, among women aged at least 65 years with low BMD values (T -score , ,1; n,=,199), the 10-year predicted probability of major OP fracture with BMD was 48% lower than the observed incidence of fractures (p,<,.01). A 10-year probability of major OP fracture higher than 12% identified more women with incident fractures than did BMD in the osteoporotic range (p,<,.05). In French women from the OFELY cohort, the observed incidence of fragility fractures over 10 years increased with age following a pattern similar to the predicted probabilities given by the FRAX tool. However, in women aged at least 65 years with low BMD, the observed incidence of fractures was substantially higher than the predicted probability. © 2010 American Society for Bone and Mineral Research. [source]

Glucocorticoids maintain human osteoclasts in the active mode of their resorption cycle

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2010
Kent Søe
Abstract Osteoclasts are known to exert their resorptive activity through a so-called resorption cycle consisting of alternating resorption and migration episodes and resulting typically in the formation of increasing numbers of discrete round excavations on bone slices. This study shows that glucocorticoids deeply modify this resorptive behavior. First, glucocorticoids gradually induce excavations with a trenchlike morphology while reducing the time-dependent increase in excavation numbers. This indicates that glucocorticoids make osteoclasts elongate the excavations they initiated rather than migrating to a new resorption site, as in control conditions. Second, the round excavations in control conditions contain undegraded demineralized collagen as repeatedly reported earlier, whereas the excavations with a trenchlike morphology generated under glucocorticoid exposure appear devoid of leftovers of demineralized collagen. This indicates that collagenolysis proceeds generally at a lower rate than demineralization under control conditions, whereas collagenolysis rates are increased up to the level of demineralization rates in the presence of glucocorticoids. Taking these observations together leads to a model where glucocorticoid-induced increased collagenolysis allows continued contact of osteoclasts with mineral, thereby maintaining resorption uninterrupted by migration episodes and generating resorption trenches. In contrast, accumulation of demineralized collagen, as prevails in controls, acts as a negative-feedback loop, switching resorptive activity off and promoting migration to a new resorption site, thereby generating an additional resorption pit. We conclude that glucocorticoids change the osteoclastic resorption mode from intermittent to continuous and speculate that this change may contribute to the early bone fragilization of glucocorticoid-treated patients. © 2010 American Society for Bone and Mineral Research. [source]

GATA-3 transduces survival signals in osteoblasts through upregulation of bcl-xL gene expression,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2010
Ruei-Ming Chen
Abstract GATA-3, a transcription factor, participates in regulating cell development, proliferation, and death. This study was aimed at evaluating the roles of GATA-3 in protecting osteoblasts against oxidative stress,induced apoptotic insults and their possible mechanisms. Pretreatment with nitric oxide (NO) for 24 hours protected osteoblasts, prepared from neonatal rat calvaria, against oxidative stress,induced apoptotic insults. Such protection involved enhancement of Bcl-XL messenger (m)RNA and protein syntheses and the translocation of this antiapoptotic protein from the cytoplasm to mitochondria. GATA-3 was detected in rat osteoblasts, and GATA-3-specific DNA-binding elements exist in the promoter region of the bcl-xL gene. NO preconditioning attenuated oxidative stress,caused suppression of GATA-3 mRNA and protein synthesis and the translocation of this transcription factor from the cytoplasm to nuclei. Application of GATA-3 small interfering (si)RNA into osteoblasts decreased the levels of this transcription factor and simultaneously inhibited Bcl-XL mRNA synthesis. Pretreatment with NO lowered the oxidative stress,caused alteration in the binding of GATA-3 to its specific DNA motifs. Oxidative stress,inhibited Runx2 mRNA expression, but NO preconditioning decreased such inhibition. NO pretreatment time-dependently enhanced the association of GATA-3 with Runx2. Knocking down the translation of GATA-3 using RNA interference significantly decreased the protection of NO preconditioning against oxidative stress,induced alterations of cell morphologies, DNA fragmentation, and cell apoptosis. In comparison, overexpression of GATA-3 could promote NO preconditioning,involved Bcl-XL expression and cell survival. Therefore, this study shows that GATA-3 plays critical roles in mediating survival signals in osteoblasts, possibly through upregulating bcl-xL gene expression. © 2010 American Society for Bone and Mineral Research. [source]

High-phosphate-induced calcification is related to SM22, promoter methylation in vascular smooth muscle cells

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2010
Addy Montes de Oca
Abstract Hyperphosphatemia is closely related to vascular calcification in patients with chronic kidney disease. Vascular smooth muscle cells (VSMCs) exposed to high phosphate concentrations in vitro undergo phenotypic transition to osteoblast-like cells. Mechanisms underlying this transdifferentiation are not clear. In this study we used two in vitro models, human aortic smooth muscle cells and rat aortic rings, to investigate the phenotypic transition of VSMCs induced by high phosphate. We found that high phosphate concentration (3.3,mmol/L) in the medium was associated with increased DNA methyltransferase activity and methylation of the promoter region of SM22,. This was accompanied by loss of the smooth muscle cell,specific protein SM22,, gain of the osteoblast transcription factor Cbfa1, and increased alkaline phosphatase activity with the subsequent in vitro calcification. The addition of a demethylating agent (procaine) to the high-phosphate medium reduced DNA methyltransferase activity and prevented methylation of the SM22, promoter, which was accompanied by an increase in SM22, expression and less calcification. Additionally, downregulation of SM22,, either by siRNA or by a methyl group donor (S -adenosyl methionine), resulted in overexpression of Cbfa1. In conclusion, we demonstrate that methylation of SM22, promoter is an important event in vascular smooth muscle cell calcification and that high phosphate induces this epigenetic modification. These findings uncover a new insight into mechanisms by which high phosphate concentration promotes vascular calcification. © 2010 American Society for Bone and Mineral Research [source]

Aged mice have enhanced endocortical response and normal periosteal response compared with young-adult mice following 1 week of axial tibial compression

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2010
Michael D Brodt
Abstract With aging, the skeleton may lose its ability to respond to positive mechanical stimuli. We hypothesized that aged mice are less responsive to loading than young-adult mice. We subjected aged (22 months) and young-adult (7 months) BALB/c male mice to daily bouts of axial tibial compression for 1 week and evaluated cortical and trabecular responses using micro,computed tomography (µCT) and dynamic histomorphometry. The right legs of 95 mice were loaded for 60 rest-inserted cycles per day to 8, 10, or 12,N peak force (generating mid-diaphyseal strains of 900 to 1900 µ, endocortically and 1400 to 3100 µ, periosteally). At the mid-diaphysis, mice from both age groups showed a strong anabolic response on the endocortex (Ec) and periosteum (Ps) [Ec.MS/BS and Ps. MS/BS: loaded (right) versus control (left), p,<,.05]. Generally, bone formation increased with increasing peak force. At the endocortical surface, contrary to our hypothesis, aged mice had a significantly greater response to loading than young-adult mice (Ec.MS/BS and Ec.BFR/BS: 22 months versus 7 months, p,<,.001). Responses at the periosteal surface did not differ between age groups (p,>,.05). The loading-induced increase in bone formation resulted in increased cortical area in both age groups (loaded versus control, p,<,.05). In contrast to the strong cortical response, loading only weakly stimulated trabecular bone formation. Serial (in vivo) µCT examinations at the proximal metaphysis revealed that loading caused a loss of trabecular bone in 7-month-old mice, whereas it appeared to prevent bone loss in 22-month-old mice. In summary, 1 week of daily tibial compression stimulated a robust endocortical and periosteal bone-formation response at the mid-diaphysis in both young-adult and aged male BALB/c mice. We conclude that aging does not limit the short-term anabolic response of cortical bone to mechanical stimulation in our animal model. © 2010 American Society for Bone and Mineral Research [source]

Replication of previous genome-wide association studies of bone mineral density in premenopausal American women

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 8 2010
Shoji Ichikawa
Abstract Bone mineral density (BMD) achieved during young adulthood (peak BMD) is one of the major determinants of osteoporotic fracture in later life. Genetic variants associated with BMD have been identified by three recent genome-wide association studies. The most significant single-nucleotide polymorphisms (SNPs) from these studies were genotyped to test whether they were associated with peak BMD in premenopausal American women. Femoral neck and lumbar spine BMD were determined by dual-energy X-ray absorptiometry in two groups of premenopausal women: 1524 white women and 512 black women. In premenopausal white women, two SNPs in the C6orf97/ESR1 region were significantly associated with BMD (p,<,4.8,×,10,4), with suggestive evidence for CTNNBL1 and LRP5 (p,<,.01). Evidence of association with one of the two SNPs in the C6orf97/ESR1 region also was observed in premenopausal black women. Furthermore, SNPs in SP7 and a chromosome 4 intergenic region showed suggestive association with BMD in black women. Detailed analyses of additional SNPs in the C6orf97/ESR1 region revealed multiple genomic blocks independently associated with femoral neck and lumbar spine BMD. Findings in the three published genome-wide association studies were replicated in independent samples of premenopausal American women, suggesting that genetic variants in these genes or regions contribute to peak BMD in healthy women in various populations. © 2010 American Society for Bone and Mineral Research [source]

Genetic variation in the RANKL/RANK/OPG signaling pathway is associated with bone turnover and bone mineral density in men

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 8 2010
Delnaz Roshandel
Abstract The aim of this study was to determine if single-nucleotide polymorphisms (SNPs) in RANKL, RANK, and OPG influence bone turnover and bone mineral density (BMD) in men. Pairwise tag SNPs (r2,,,0.8) were selected for RANKL, RANK, and OPG and their 10-kb flanking regions. Selected tag SNPs plus five SNPs near RANKL and OPG, associated with BMD in published genome-wide association studies (GWAS), were genotyped in 2653 men aged 40 to 79 years of age recruited for participation in a population-based study of male aging, the European Male Ageing Study (EMAS). N-terminal propeptide of type I procollagen (PINP) and C-terminal cross-linked telopeptide of type I collagen (CTX-I) serum levels were measured in all men. BMD at the calcaneus was estimated by quantitative ultrasound (QUS) in all men. Lumbar spine and total-hip areal BMD (BMDa) was measured by dual-energy X-ray absorptiometry (DXA) in a subsample of 620 men. Multiple OPG, RANK, and RANKL SNPs were associated with bone turnover markers. We also identified a number of SNPs associated with BMD, including rs2073618 in OPG and rs9594759 near RANKL. The minor allele of rs2073618 (C) was associated with higher levels of both PINP (,,=,1.83, p,=,.004) and CTX-I (,,=,17.59, p,=,4.74,×,10,4), and lower lumbar spine BMDa (,,=,,0.02, p,=,.026). The minor allele of rs9594759 (C) was associated with lower PINP (,,=,,1.84, p,=,.003) and CTX-I (,,=,,27.02, p,=,6.06,×,10,8) and higher ultrasound BMD at the calcaneus (,,=,0.01, p,=,.037). Our findings suggest that genetic variation in the RANKL/RANK/OPG signaling pathway influences bone turnover and BMD in European men. © 2010 American Society for Bone and Mineral Research [source]

Guidelines for assessment of bone microstructure in rodents using micro,computed tomography

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2010
Mary L Bouxsein
Abstract Use of high-resolution micro,computed tomography (µCT) imaging to assess trabecular and cortical bone morphology has grown immensely. There are several commercially available µCT systems, each with different approaches to image acquisition, evaluation, and reporting of outcomes. This lack of consistency makes it difficult to interpret reported results and to compare findings across different studies. This article addresses this critical need for standardized terminology and consistent reporting of parameters related to image acquisition and analysis, and key outcome assessments, particularly with respect to ex vivo analysis of rodent specimens. Thus the guidelines herein provide recommendations regarding (1) standardized terminology and units, (2) information to be included in describing the methods for a given experiment, and (3) a minimal set of outcome variables that should be reported. Whereas the specific research objective will determine the experimental design, these guidelines are intended to ensure accurate and consistent reporting of µCT-derived bone morphometry and density measurements. In particular, the methods section for papers that present µCT-based outcomes must include details of the following scan aspects: (1) image acquisition, including the scanning medium, X-ray tube potential, and voxel size, as well as clear descriptions of the size and location of the volume of interest and the method used to delineate trabecular and cortical bone regions, and (2) image processing, including the algorithms used for image filtration and the approach used for image segmentation. Morphometric analyses should be based on 3D algorithms that do not rely on assumptions about the underlying structure whenever possible. When reporting µCT results, the minimal set of variables that should be used to describe trabecular bone morphometry includes bone volume fraction and trabecular number, thickness, and separation. The minimal set of variables that should be used to describe cortical bone morphometry includes total cross-sectional area, cortical bone area, cortical bone area fraction, and cortical thickness. Other variables also may be appropriate depending on the research question and technical quality of the scan. Standard nomenclature, outlined in this article, should be followed for reporting of results. © 2010 American Society for Bone and Mineral Research [source]

Genome-wide pleiotropy of osteoporosis-related phenotypes: The framingham study

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2010
David Karasik
Abstract Genome-wide association studies offer an unbiased approach to identify new candidate genes for osteoporosis. We examined the Affymetrix 500K,+,50K SNP GeneChip marker sets for associations with multiple osteoporosis-related traits at various skeletal sites, including bone mineral density (BMD, hip and spine), heel ultrasound, and hip geometric indices in the Framingham Osteoporosis Study. We evaluated 433,510 single-nucleotide polymorphisms (SNPs) in 2073 women (mean age 65 years), members of two-generational families. Variance components analysis was performed to estimate phenotypic, genetic, and environmental correlations (,P, ,G, and ,E) among bone traits. Linear mixed-effects models were used to test associations between SNPs and multivariable-adjusted trait values. We evaluated the proportion of SNPs associated with pairs of the traits at a nominal significance threshold ,,=,0.01. We found substantial correlation between the proportion of associated SNPs and the ,P and ,G (r,=,0.91 and 0.84, respectively) but much lower with ,E (r,=,0.38). Thus, for example, hip and spine BMD had 6.8% associated SNPs in common, corresponding to ,P,=,0.55 and ,G,=,0.66 between them. Fewer SNPs were associated with both BMD and any of the hip geometric traits (eg, femoral neck and shaft width, section moduli, neck shaft angle, and neck length); ,G between BMD and geometric traits ranged from ,0.24 to +0.40. In conclusion, we examined relationships between osteoporosis-related traits based on genome-wide associations. Most of the similarity between the quantitative bone phenotypes may be attributed to pleiotropic effects of genes. This knowledge may prove helpful in defining the best phenotypes to be used in genetic studies of osteoporosis. © 2010 American Society for Bone and Mineral Research [source]

Hip geometry variation is associated with bone mineralization pathway gene variants: The framingham study

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2010
Ching-Lung Cheung
Abstract Mineralization of bone matrix is an important process in bone formation; thus defects in mineralization have been implicated in bone mineral density (BMD) and bone structure alterations. Three central regulators of phosphate balance, ALPL, ANKH, and ENPP1, are central in the matrix mineralization process; therefore, the genes encoding them are considered important candidates genes for BMD and bone geometry. To test for an association between these three candidate genes and BMD and bone geometry traits, 124 informative single-nucleotide polymorphisms (SNPs) were selected and genotyped in 1513 unrelated subjects from the Framingham offspring cohort. Initial results showed that SNP rs1974201 in the gene ENPP1 was a susceptibility variant associated with several hip geometric indices, with the strongest p value of 3.8,×,10,7 being observed for femoral neck width. A few modest associations were observed between SNPs in or near ALPL and several bone traits, but no association was observed with ANKH. The association signals observed for SNPs around rs1974201 were attenuated after conditional analysis on rs1974201. Transcription factor binding-site prediction revealed that the HOXA7 binding site was present in the reference sequence with the major allele, whereas this potential binding site is lost in the sequence with the minor allele of rs1974201. In conclusion, we found evidence for association of bone geometry variation with an SNP in ENPP1, a gene in the mineralization pathway. The alteration of a binding site of the deregulator of extracellular matrix HOXA7 warrants further investigation. © 2010 American Society for Bone and Mineral Research [source]

Cell-based immunotherapy with mesenchymal stem cells cures bisphosphonate-related osteonecrosis of the jaw,like disease in mice

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2010
Takashi Kikuiri
Abstract Patients on high-dose bisphosphonate and immunosuppressive therapy have an increased risk of bisphosphonate-related osteonecrosis of the jaw (BRONJ); despite the disease severity, its pathophysiology remains unknown, and appropriate therapy is not established. Here we have developed a mouse model of BRONJ-like disease that recapitulates major clinical and radiographic manifestations of the human disease, including characteristic features of an open alveolar socket, exposed necrotic bone or sequestra, increased inflammatory infiltrates, osseous sclerosis, and radiopaque alveolar bone. We show that administration of zoledronate, a potent aminobisphosphonate, and dexamethasone, an immunosuppressant drug, causes BRONJ-like disease in mice in part by suppressing the adaptive regulatory T cells, Tregs, and activating the inflammatory T-helper-producing interleukin 17 cells, Th17. Most interestingly, we demonstrate that systemic infusion with mesenchymal stem cells (MSCs) prevents and cures BRONJ-like disease possibly via induction of peripheral tolerance, shown as an inhibition of Th17 and increase in Treg cells. The suppressed Tregs/Th17 ratio in zoledronate- and dexamethasone-treated mice is restored in mice undergoing salvage therapy with Tregs. These findings provide evidence of an immunity-based mechanism of BRONJ-like disease and support the rationale for in vivo immunomodulatory therapy using Tregs or MSCs to treat BRONJ. © 2010 American Society for Bone and Mineral Research [source]

Origin matters: Differences in embryonic tissue origin and Wnt signaling determine the osteogenic potential and healing capacity of frontal and parietal calvarial bones

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2010
Natalina Quarto
Abstract Calvarial bones arise from two embryonic tissues, namely, the neural crest and the mesoderm. In this study we have addressed the important question of whether disparate embryonic tissue origins impart variable osteogenic potential and regenerative capacity to calvarial bones, as well as what the underlying molecular mechanism(s). Thus, by performing in vitro and in vivo studies, we have investigated whether differences exist between neural crest,derived frontal and paraxial mesodermal,derived parietal bone. Of interest, our data indicate that calvarial bone osteoblasts of neural crest origin have superior potential for osteogenic differentiation. Furthermore, neural crest,derived frontal bone displays a superior capacity to undergo osseous healing compared with calvarial bone of paraxial mesoderm origin. Our study identified both in vitro and in vivo enhanced endogenous canonical Wnt signaling in frontal bone compared with parietal bone. In addition, we demonstrate that constitutive activation of canonical Wnt signaling in paraxial mesodermal,derived parietal osteoblasts mimics the osteogenic potential of frontal osteoblasts, whereas knockdown of canonical Wnt signaling dramatically impairs the greater osteogenic potential of neural crest,derived frontal osteoblasts. Moreover, fibroblast growth factor 2 (FGF-2) treatment induces phosphorylation of GSK-3, and increases the nuclear levels of ,-catenin in osteoblasts, suggesting that enhanced activation of Wnt signaling might be mediated by FGF. Taken together, our data provide compelling evidence that indeed embryonic tissue origin makes a difference and that active canonical Wnt signaling plays a major role in contributing to the superior intrinsic osteogenic potential and tissue regeneration observed in neural crest,derived frontal bone. © 2010 American Society for Bone and Mineral Research [source]

rBMP represses Wnt signaling and influences skeletal progenitor cell fate specification during bone repair

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 6 2010
Steve Minear
Abstract Bone morphogenetic proteins (BMPs) participate in multiple stages of the fetal skeletogenic program from promoting cell condensation to regulating chondrogenesis and bone formation through endochondral ossification. Here, we show that these pleiotropic functions are recapitulated when recombinant BMPs are used to augment skeletal tissue repair. In addition to their well-documented ability to stimulate chondrogenesis in a skeletal injury, we show that recombinant BMPs (rBMPs) simultaneously suppress the differentiation of skeletal progenitor cells in the endosteum and bone marrow cavity to an osteoblast lineage. Both the prochondrogenic and antiosteogenic effects are achieved because rBMP inhibits endogenous ,-catenin-dependent Wnt signaling. In the injured periosteum, this repression of Wnt activity results in sox9 upregulation; consequently, cells in the injured periosteum adopt a chondrogenic fate. In the injured endosteum, rBMP also inhibits Wnt signaling, which results in the runx2 and collagen type I downregulation; consequently, cells in this region fail to differentiate into osteoblasts. In muscle surrounding the skeletal injury site, rBMP treatment induces Smad phosphorylation followed by exuberant cell proliferation, an increase in alkaline phosphatase activity, and chondrogenic differentiation. Thus different populations of adult skeletal progenitor cells interpret the same rBMP stimulus in unique ways, and these responses mirror the pleiotropic effects of BMPs during fetal skeletogenesis. These mechanistic insights may be particularly useful for optimizing the reparative potential of rBMPs while simultaneously minimizing their adverse outcomes. © 2010 American Society for Bone and Mineral Research [source]

Identification of genes influencing skeletal phenotypes in congenic P/NP rats

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 6 2010
Imranul Alam
Abstract We previously showed that alcohol-preferring (P) rats have higher bone density than alcohol-nonpreferring (NP) rats. Genetic mapping in P and NP rats identified a major quantitative trait locus (QTL) between 4q22 and 4q34 for alcohol preference. At the same location, several QTLs linked to bone density and structure were detected in Fischer 344 (F344) and Lewis (LEW) rats, suggesting that bone mass and strength genes might cosegregate with genes that regulate alcohol preference. The aim of this study was to identify the genes segregating for skeletal phenotypes in congenic P and NP rats. Transfer of the NP chromosome 4 QTL into the P background (P.NP) significantly decreased areal bone mineral density (aBMD) and volumetric bone mineral density (vBMD) at several skeletal sites, whereas transfer of the P chromosome 4 QTL into the NP background (NP.P) significantly increased bone mineral content (BMC) and aBMD in the same skeletal sites. Microarray analysis from the femurs using Affymetrix Rat Genome arrays revealed 53 genes that were differentially expressed among the rat strains with a false discovery rate (FDR) of less than 10%. Nine candidate genes were found to be strongly correlated (r2,>,0.50) with bone mass at multiple skeletal sites. The top three candidate genes, neuropeptide Y (Npy), , synuclein (Snca), and sepiapterin reductase (Spr), were confirmed using real-time quantitative PCR (qPCR). Ingenuity pathway analysis revealed relationships among the candidate genes related to bone metabolism involving ,-estradiol, interferon-,, and a voltage-gated calcium channel. We identified several candidate genes, including some novel genes on chromosome 4 segregating for skeletal phenotypes in reciprocal congenic P and NP rats. © 2010 American Society for Bone and Mineral Research [source]