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Elevated follicle-stimulating hormone (FSH) activity is proposed to directly cause bone loss independent of estradiol deficiency in aging women. Using transgenic female mice expressing human FSH (TgFSH), we now reveal that TgFSH dose-dependently increased bone mass, markedly elevating tibial and vertebral trabecular bone volume. Furthermore, TgFSH stimulated a striking accrual of bone mass in hypogonadal mice lacking endogenous FSH and luteinizing hormone (LH) function, showing that FSH-induced bone mass occurred independently of background LH or estradiol levels. Higher TgFSH levels increased osteoblast surfaces in trabecular bone and stimulated de novo bone formation, filling marrow spaces with woven rather than lamellar bone, reflective of a strong anabolic stimulus. Trabecular bone volume correlated positively with ovarian-derived serum inhibin A or testosterone levels in TgFSH mice, and ovariectomy abolished TgFSH-induced bone formation, proving that FSH effects on bone require an ovary-dependent pathway. No detectable FSH receptor mRNA in mouse bone or cultured osteoblasts or osteoclasts indicated that FSH did not directly stimulate bone. Therefore, contrary to proposed FSH-induced bone loss, our findings demonstrate that FSH has dose-dependent anabolic effects on bone via an ovary-dependent mechanism, which is independent of LH activity, and does not involve direct FSH actions on bone cells. animal model | gonadotropin | osteosclerosis | microcomputed tomography doi/ 10.1073/pnas.1012141108

Nanocrystals of apatitic calcium phosphate impart the organic-inorganic nanocomposite in bone with favorable mechanical properties. So far, the factors preventing crystal growth beyond the favorable thickness of ca. 3 nm have not been identified. Here we show that the apatite surfaces are studded with strongly bound citrate molecules, whose signals have been identified unambiguously by multinuclear magnetic resonance (NMR) analysis. NMR reveals that bound citrate accounts for 5.5 wt% of the organic matter in bone and covers apatite at a density of about I molecule per [(2 nm).sup.2], with its three carboxylate groups at distances of 0.3 to 0.45 nm from the apatite surface. Bound citrate is highly conserved, being found in fish, avian, and mammalian bone, which indicates its critical role in interfering with crystal thickening and stabilizing the apatite nanocrystals in bone. nanocrystal stabilization | nanocomposite | organic-inorganic interface | bone composition | solid-state NMR doi/ 10.1073/pnas.1009219107

Runx2 is indispensable for osteoblast lineage commitment and early differentiation but also blocks osteoblast maturation, thereby causing bone loss in Runx2 transgenic mice. Zinc finger protein 521 (Zfp521) antagonizes Runx2 in vivo. Eliminating one Zfp521 allele mitigates the cleidocranial dysplasia--like phenotype of newborn [Runx.sup.2+/-] mice, whereas overexpressing Zfp521 exacerbates it. Overexpressing Zfp521 also reverses the severe osteopenia of adult Runx2 transgenic mice. Zfp521 binds to both Runx2 and histone deacetylase 3 (HDAC3), promotes their association, and antagonizes Runx2 transcriptional activity in an HDAC3-dependent manner. Mutating the Zfp521 zinc finger domains 6 and 26 reduces the binding of Zfp521 to Runx2 and inhibition of Runx2 activity. These data provide evidence that Zfp521 antagonizes Runx2 in vivo and thereby regulates two stages of osteoblast development, early during mesenchymal cell lineage commitment and later during osteoblast maturation. Thus, the balance and molecular interplay between Zfp521 and Runx2 contribute to the control of osteoblast differentiation, skeletal development, and bone homeostasis. doi/ 10.1083/jcb.201009107

Soybeans are a good source of bone-healthy nutrients. Epidemiological studies in Asia evaluating diets containing traditional whole soy foods show a positive association with bone mineral density and fracture protection. Smaller scale intervention studies in Western nations mainly feature isolated soy protein (SP) and purified or concentrated soy isoflavones (SI) rather than whole soy foods and they have produced inconsistent results. Consumption of SP does not alter calcium (Ca) retention even though urinary Ca excretion is less in diets with SP compared with proteins higher in sulfur-containing amino acids. SI, often consumed at higher concentrations than would be available in traditional Asian diets, are not yielding the type of incontrovertible evidence that might be expected in support of their benefit to bone health. This forces one to ask whether whole soy might provide a superior effect on bone. J. Nutr. 140:2312S-2317S, 2010. doi: 10.3945/jn.110.124008.

In cross-sectional geometric (CSG) studies, both the subperiosteal and endosteal contours are considered important factors in determining bone bending rigidity. Recently, regression equations predicting CSG properties from a section's external dimensions were developed in a world-wide sample of human long bones. The results showed high correlations between some subperiosteally derived and actual CSG parameters. We present a theoretical model that further explores the influence of endosteal dimensions on CSG properties. We compare two hypothetical femoral midshaft samples with the same total subperiosteal area but with percentages of cortical bone at the opposite ends of published human variation for population sample means. Even in this relatively uncommon scenario, the difference between the samples in the resultant means for predicted femoral polar second moment of area (J) appears to be modest: power analysis indicates that a minimum sample size of 61 is needed to detect the difference 90% of the time via a t-test. Moreover, endosteal area can be predicted--although with substantial error--from periosteal area. Despite this error, including this relationship in subperiosteally derived estimates of J produces sample mean estimates close to true mean values. Power analyses reveal that when similar samples are used to develop prediction equations, a minimum sample of hundreds or more may be needed to distinguish a predicted mean J from the true mean J. These results further justify the use of regression equations estimating J from periosteal contours when analyzing behaviorally induced changes in bone rigidity in ancient populations, when it is not possible to measure endosteal dimensions. However, in other situations involving comparisons of individual values, growth trends, and senescence, where relative cortical thickness may vary greatly, inclusion of endosteal dimensions is still important. Am J Phys Anthropol 143:612-624, 2010. [c] 2010 Wiley-Liss, Inc. KEY WORDS bone mechanical properties; periosteal CSG; external dimensions DOI 10.1002/ajpa.21361

Induced osteogenesis includes a program of microRNAs (miRs) to repress the translation of genes that act as inhibitors of bone formation. How expression of bone-related miRs is regulated remains a compelling question. Here we report that Runx2, a transcription factor essential for osteoblastogenesis, negatively regulates expression of the miR cluster 23a~27a~24-2. Overexpression, reporter, and chromatin immunoprecipitation assays established the presence of a functional Runx binding element that represses expression of these miRs. Consistent with this finding, exogenous expression of each of the miRs suppressed osteoblast differentiation, whereas antagomirs increased bone marker expression. The biological significance of Runx2 repression of this miR cluster is that each miR directly targets the 3' UTR of SATB2, which is known to synergize with Runx2 to facilitate bone formation. The findings suggest Runx2-negative regulation of multiple miRs by a feed-forward mechanism to cause derepression of SATB2 to promote differentiation. We find also that miR-23a represses Runx2 in the terminally differentiated osteocyte, representing a feedback mechanism to attenuate osteoblast maturation. We provide direct evidence for an interdependent relationship among transcriptional inhibition of the miR cluster by Runx2, translational repression of Runx2 and of SATB2 by the cluster miRs during progression of osteoblast differentiation. Furthermore, miR cluster gain of function (i.e., inhibition of osteogenesis) is rescued by the exogenous expression of SATB2. Taken together, we have established a regulatory network with a central role for the miR cluster 23a~27a~24-2 in both progression and maintenance of the osteocyte phenotype. bone formation by miRs | Runx2-regulation of microRNA cluster | phenotype attenuation by miRs | feed forward-feedback miR control | SATB2 targeted by cluster miRs doi/ 10.1073/pnas.1007698107

Bone may contain an intraskeletal reservoir of sex steroids that is capable of producing biological effects. The purposes of these experiments were to 1) establish and validate methods to extract and measure intraskeletal sex hormones, 2) compare serum and intraskeletal sex hormone abundance, and 3) determine the impact of testosterone-enanthate administration and orchiectomy on intraskeletal sex hormone concentrations. Tibiae from male F344 rats were crushed, suspended in an aqueous buffer, disrupted mechanically and sonically, extracted with organic solvents, dried, and reconstituted in assay buffer appropriate for measurement of testosterone, dihydrotestosterone, and estradiol by immunoassay. Prior to extraction, bone homogenate was spiked with [[sup.3]H]testosterone, [[sup.3]H]dihydrotestosterone, or [[sup.3]H]estradiol, and >80% of each [sup.3]H-labeled sex hormone was recovered. Extracted bone samples were also assayed with and without known amounts of unlabeled sex hormones, and >97% of the expected hormone concentrations were measured. Administration of testosterone-enanthate increased intraskeletal testosterone 11-fold and intraskeletal dihydrotestosterone by 82% without altering intraskeletal estradiol (P < 0.01). Conversely, orchiectomy did not alter intraskeletal testosterone or estradiol but increased intraskeletal dihydrotestosterone by 39% (P < 0.05). In intact rats, intraskeletal testosterone and dihydrotestosterone were directionally higher than in serum, whereas intraskeletal estradiol was directionally lower than serum. Serum androgens were positively correlated with intraskeletal androgens (r = 0.74-0.96, P < 0.001); however, neither serum nor intraskeletal androgens nor serum estradiol were correlated with intraskeletal estradiol. We report the validation of a novel method for measuring intraskeletal sex hormones. Our findings demonstrate that the intraskeletal sex steroid reservoirs are modifiable and only partially influenced by circulating sex hormones. testosterone; estrogen; intracrine; dihydrotestosterone; estradiol doi: 10.1152/ajpendo.00384.2010.

Bone mass is determined by a continuous remodeling process, whereby the mineralized matrix is being removed by osteoclasts and subsequently replaced with newly formed bone tissue produced by osteoblasts. Here we report the presence of endogenous amides of long-chain fatty acids with amino acids or with ethanolamine (N-acyl amides) in mouse bone. Of these compounds, N-oleoyl-L-serine (OS) had the highest activity in an osteoblast proliferation assay. In these cells, OS triggers a Gi-protein-coupled receptor and Erk1/2. It also mitigates osteoclast number by promoting osteoclast apoptosis through the inhibition of Erk1/2 phosphorylation and receptor activator of nuclear-[kappa]B ligand (RANKL) expression in bone marrow stromal cells and osteoblasts. In intact mice, OS moderately increases bone volume density mainly by inhibiting bone resorption. However, in a mouse ovariectomy (OVX) model for osteoporosis, OS effectively rescues bone loss by increasing bone formation and markedly restraining bone resorption. The differential effect of exogenous OS in the OVX vs. intact animals is apparently a result of an OVX-induced decrease in skeletal OS levels. These data show that OS is a previously unexplored lipid regulator of bone remodeling. It represents a lead to antiosteoporotic drug discovery, advantageous to currently available therapies, which are essentially either proformative or antiresorptive. fatty acyl amides doi/ 10.1073/pnas.0912479107

Background. Higher protein diets are promoted for effective weight loss. Striated tissues in omnivorous diets contain high-quality protein, but limited data exist regarding their effects on bone. Methods. To examine the effects of energy restriction--induced weight loss with higher protein omnivorous diets versus lower protein vegetarian diets on bone mineral density in overweight postmenopausal women, two randomized controlled feeding studies were conducted. In Study 1.28 women consumed 750 kcal/day energy deficit diets with 18% energy from protein via lacto-ovo vegetarian sources (normal protein, n = 15) or 30% energy from protein with 40% of protein from lean pork (higher protein, n = 13, omnivorous) for 12 weeks. In Study 2, 54 women consumed their habitual diet (control, n = 11) or 1,250 kcal/day diets with 16% energy from nonmeat protein sources (n = 14) or 26% energy from protein, including chicken (n = 15) or beef (n = 14) for 9 weeks. Results. Study I: With weight loss (normal protein -11.2%, higher protein -10.1%), bone mineral density was not significantly changed in normal protein (-0.003 [+ or -] 0.003 g/[cm.sup.2], -0.3 %) but decreased in higher protein (- 0.0167 [+ or -] 0.004 g/[cm.sup.2], -1.4%, group-by-time p < .05). Study 2: The control, nonmeat, chicken, and beef groups lost 1.5%, 7.7%, 10.4%, and 8.1% weight and 0.0%, 0.4%, 1.1%, and 1.4% bone mineral density, respectively. The change of bone mineral density was significant for chicken and beef compared with the control (group-by-time, p < .05). Markers of calcium metabolism and bone homeostasis in blood and urine were not changed over time or differentially affected by diet. Conclusion. Consumption of higher protein omnivorous diets promoted decreased bone mineral density after weight loss in overweight postmenopausal women. Key Words: Protein--Weight loss--Bone mineral density. doi: 10.1093/geron/glq083

Osteogenesis imperfecta (OI) is a clinically and genetically heterogeneous disease due primarily to mutations in the type I procollagen genes, COL1A1 and COL1A2, causing bone deformity and numerous lifetime fractures. OI murine (oim) model mice carry a mutation in the col1a2 gene causing aberrant production of homotrimeric type I collagen [[alpha]1[(I).sub.3] ], leading to bone fragility and glomerular accumulation of type I collagen. Previous studies demonstrated that heterozygous (+/oim) and homozygous (oim/oim) mice have elevated tibiae fluoride concentrations but reduced femoral biomechanics. However, it is unclear whether these 2 variables are causally related, because impaired renal function could reduce urinary fluoride excretion, thus elevating bone fluoride concentrations regardless of disease status. Our goal in this study was to determine whether dietary fluoride restriction would improve femoral biomechanics in oim mice. Wild-type, +/oim, and oim/oim mice were fed a control (5 mg/kg fluoride) or fluoride-restricted diet (0 mg/kg fluoride) for ~ 13 wk, at which time plasma and femora were analyzed for fluoride concentrations and bone biomechanical properties. In wild-type, +/oim, and oim/oim mice, dietary fluoride restriction reduced femoral fluoride burden by 54-74%, respectively (P < 0.05), without affecting glomerular collagen deposition. Oim/oim mice fed the fluoride-restricted diet had reduced material tensile strength (P < 0.05) compared with oim/oim mice fed the control diet. However, dietary fluoride restriction did not affect stiffness or whole bone femoral breaking strength, regardless of genotype. These data suggest that oim mice have reduced bone strength due to homotrimeric type I collagen, independent of bone fluoride content. J. Nutr. 140:1752-1756, 2010. doi: 10.3945/jn.109.120261.

Extracellular regulation of signaling by transforming growth factor (TGF)--[beta] family members is emerging as a key aspect of organ formation and tissue remodeling. In this study, we demonstrate that fibrillin-1 and -2, the structural components of extracellular microfibrils, differentially regulate TGF-[beta] and bone morphogenetic protein (BMP) bioavailability in bone. Fibrillin-2--null ([Fbn2.sup.-/-]) mice display a low bone mass phenotype that is associated with reduced bone formation in vivo and impaired osteoblast maturation in vitro. This [Fbn2.sup.-/-] phenotype is accounted for by improper activation of latent TGF-[beta] that selectively blunts expression of osterix, the transcriptional regulator of osteoblast maturation, and collagen I, the structural template for bone mineralization. Cultured osteoblasts from Fbn1-/- mice exhibit improper latent TGF-[beta] activation as well, but mature faster because of increased availability of otherwise matrix-bound BMPs. Additional in vitro evidence excludes a direct role of microfibrils in supporting mineral deposition. Together, these findings identify the extracellular microfibrils as critical regulators of bone formation through the modulation of endogenous TGF-[beta] and BMP signaling. doi/ 10.1083/jcb.201003089

The activator protein-1 (AP-1) transcription factor complex, in particular the Fos proteins, is an important regulator of bone homeostasis. Fra-2 (Fosl2), a Fos-related protein of the AP-1 family, is expressed in bone cells, and newborn mice lacking Fra-2 exhibit defects in chondrocytes and osteoclasts. Here we show that Fra-2-deficient osteoblasts display a differentiation defect both in vivo and in vitro. Moreover, Fra-2--overexpressing mice are osteosclerotic because of increased differentiation of osteoblasts, which appears to be cell autonomous. Importantly, the osteoblast-specific osteocalcin (Oc) gene and collagen 1[alpha]2 (col1[alpha]2) are transcriptional targets of Fra-2 in both murine and human bone cells. In addition, Fra-2, Oc, and col1 are expressed in stromal cells of human chondroblastic and osteoblastic osteosarcomas (Os's) as well as during osteoblast differentiation of human Os cell lines. These findings reveal a novel function of Fra-2/AP-1 as a positive regulator of bone and matrix formation in mice and humans. doi/ 10.1083/jcb.201002111

Sherk, VD, Bemben, MG, and Bemben, DA. Comparisons of bone mineral density and bone quality in adult rock climbers, resistance trained men, and untrained men. J Strength Cond Res 24(9): 2468-2474, 2010--The nature of muscular contractions and episodes of impact loading during technical rock climbing are often varied and complex, and the resulting effects on bone health are unclear. The purpose of this study was to compare total body, lumbar spine, proximal femur, and forearm areal bone mineral density (aBMD) and tibia and forearm bone quality in male rock climbers (RC) (n = 15), resistance trained men (RT) (n = 16), and untrained male controls (CTR) (n = 16). Total body, anteroposterior (AP) lumbar spine, proximal femur, and forearm aBMD and body composition were measured using dual-energy X-ray absorptiometry (DXA) (Lunar Prodigy, v. 10.50.086; GE Healthcare, Waukesha, Wisconsin, U.S.A.). Volumetric BMD (vBMD), bone content, bone area, and muscle cross-sectional area (MCSA) of the tibia and forearm were measured using pQCT (peripheral quantitative computed tomography; Stratec XCT 3000, Pforzheim, Germany). No significant group differences were seen in bone-free lean body mass. CTR had significantly (p < 0.05) greater body fat % than RC and RT and significantly (p < 0.05) greater fat mass than RC. Lumbar spine and femoral neck aBMD were significantly (p < 0.05) greater in RT compared to both RC and CTR. RC had significantly (p < 0.05) lower aBMD at the 33% radius site than CTR. Forearm MCSA was significantly (p < 0.05) lower in CTR than in the other groups. No significant differences were seen between groups for vBMD or bone area of the tibia and forearm. In conclusion, resistance-trained men had higher bone density at the central skeletal sites than rock climbers; however, bone quality variables of the peripheral limbs were similar in rock climber and resistance-trained groups. KEY WORDS bone mass, mechanical loading, pQCT, fat mass, bone-free lean body mass

Glutamatergic intercellular communication is involved in many aspects of metabolic homeostasis in normal bone. In bone metastasis, the balance between bone formation and degradation is disrupted. Although the responsible mechanisms are not clear, we have previously identified that cancer cell lines used in bone tumour models secrete glutamate, suggesting that tumour-derived glutamate may disrupt sensitive signalling systems in bone. This study examines the role of glutamate in mature osteoclastic bone resorption, osteoblast differentiation, and bone nodule formation. Glutamate was found to have no effect on the survival or activity of mature osteoclasts, although glutamate transporter inhibition and receptor blockade increased the number of bone resorption pits. Furthermore, transporter inhibition increased the area of resorbed bone while significantly decreasing the number of osteoclasts. Alkaline phosphatase activity and extracellular matrix mineralization were used as measurements of osteoblast differentiation. Glutamate significantly increased osteoblast differentiation and mineralization, but transport inhibitors had no effect. These studies support earlier findings suggesting that glutamate may be more important for osteoclastogenesis than for osteoclast proliferation or functions. Since glutamate is capable of changing the differentiation and activities of both osteoclast and osteoblast cell types in bone, it is reasonable to postulate that tumour-derived glutamate may impact bone homeostasis in bone metastasis. Key words: bone, cancer, glutamate, metastasis, osteoblast, osteoclast, transporter. La communication intercellulaire glutamatergique participe a de nombreux aspects de l'homeoslasie metabolique dans l'os normal. Dans la melastase osseuse, l'equilibre entre la formation et la degradation de l'os est rompu. Bien que les mecanismes a la base de ce desequilibre ne soient pas clairs, nous avons indique dans des travaux anterieurs que les lignees de cellules cancereuses utilisees dans les modeles de tumeurs osseuses secretent du glutamate, ce autorise a penser que le glutamate tumoral pourrait perturber les systemes de signalisation sensibles dans l'os. La presente etude examine le role du glutamate dans la resorption de l'osteoclaste mature, la differenciation osteoblastique et la formation de nodules osseux. Le glutamate n'a pas eu d'effet sur la survie ou l'activite des osteoclastes matures, quoique l'inhibition du transporteur de glutamate et le blocage du recepteur aient augmente les puits de resorption osseuse. De plus, l'inhibition du transporteur a augmente la surface de l'os resorbe tout en diminuant significativement le nombre d'osteoclastes. L'activite de la phosphatase alcaline et la mineralisation de la matrice extracellulaire ont servi de mesures de la differenciation osteoblastique. Le glutamate a augmente de maniere significative la differenciation et la mineralisation des osteoclastes, mais les inhibiteurs du transport n'ont eu aueira effet. Ces observations conforteni les resultats anterieurs suggerant que le glutamate pourrait etre plus important pour l'osteoclastogenese que les fonctions ou la proliferation des osteoclastes. Comme le glutamate est capable de modifier la differenciation et les activites des deux types de cellules dans l'os, il est raisonnable de postuler que le glutamate provenant des tumeurs pourrait avoir un impact sur l'homeostasie du tissu osseux dans la melastase osseuse. Mots-cles : os, cancer, glutamate, melastase, osteoblast, osteoclast, transporter. [Traduit par la Redaction], Introduction A variety of factors participate in the highly coordinated balancing of osteoclast (Oc) and osteoblast (Ob) cell functions in bone remodelling. Recent evidence suggests that the neurotransmitter L-glutamate is [...]

The study identifies the gene defect causing autosomal-recessive Setleis syndrome (type III focal facial dermal dysplasias or FFDDs). Findings reveal that homozygous TWIST2 nonsense mutations cause an FFDD, demonstrating the conserved role of the gene in mammalian skin and bone development.

The ATDC5 cell line exhibits a multistep process of chondrogenic differentiation analogous to that observed during endochondral bone formation. Previous investigators have induced ATDC5 cells to differentiate by exposing them to insulin at high concentrations. We have observed spontaneous differentiation of ATDC5 cells maintained in ascorbic acid-containing [alpha]-MEM. A comparison of the differentiation events in response to high-dose insulin vs. ascorbic acid showed similar expression patterns of key genes, including collagen II, Runx2, Sox9, Indian hedgehog, and collagen X. We took advantage of the action of ascorbic acid to examine signaling events associated with differentiation. In contrast to high-dose insulin, which downregulates both IGF-I and insulin receptors, there were only minimal changes in the abundance of these receptors during ascorbic acid-induced differentiation. Furthermore, ascorbic acid exposure was associated with ERK activation, and ERK inhibition attenuated ascorbic acid-induced differentiation. This was in contrast to the inhibitory effect of ERK activation during IGF-I-induced differentiation. Inhibition of collagen formation with a proline analog markedly attenuated the differentiating effect of ascorbic acid on ATDC5 cells. When plates were conditioned with ATDC5 cells exposed to ascorbic acid, ATDC5 cells were able to differentiate in the absence of ascorbic acid. Our results indicate that matrix formation early in the differentiation process is essential for ascorbic acid-induced ATDC5 differentiation. We conclude that ascorbic acid can promote the differentiation of ATDC5 cells by promoting the formation of collagenous matrix and that matrix formation mediates activation of the ERK signaling pathway, which promotes the differentiation program. ATDC5 cells; chondrocyte differentiation; collagen; extracellular matrix doi: 10.1152/ajpendo.00145.2010.

Carboxypeptidase E (CPE) is a prohormone/proneuropeptide processing enzyme, and mice bearing CPE mutations exhibit an obese and diabetic phenotype. Studies on CPE knockout (KO) mice revealed poor prohormone processing, resulting in deficiencies in peptide hormones/neuropeptides such as insulin, gonadotropin-releasing hormone, and cocaine-and amphetamine-regulated transcript (CART). Here, we show that CPE KO mice, an obese animal model, have low bone mineral density (BMD) accompanied by elevated plasma CTX-1 (carboxy-terminal collagen crosslinks), and osteocalcin, indicators of increased bone turnover. Receptor activator for NF-[kappa]B ligand (RANKL) expression was elevated ~2-fold relative to osteoprotegerin in the femur of KO animals, suggesting increased osteoclastic activity in the KO mice. In the hypothalamus, mature CART, a peptide involved in eating behavior and implicated in bone metabolism, was undetectable. The melanocortin and neuropeptide Y (NPY) systems in the hypothalamus have also been implicated in bone remodeling, since MC4R KO and NPY KO mice have increased BMD. However, reduction of [alpha]-MSH, the primary ligand of MC4R by up to 94% and the lack of detectable NPY in the hypotbalamus of CPE KO do not recapitulate the single-gene KO phenotypes. This study highlights the complex physiological interplay between peptides involved in energy metabolism and bone formation and furthermore suggests the possibility that patients, bearing CPE and CART mutations leading to inactive forms of these molecules, may be at a higher risk of developing osteoporosis. doi: 10.1152/ajpendo.00516.2009.

A combination of homozygosity mapping and candidate gene approach was used to identify a homozygous single base pair deletion in SP7/Osterix (OSX) in an Egyptian child with recessive osteogenesis imperfecta. The findings provide insight into the spectrum of genes associated with osteogenesis imperfecta and the significant role of SP7/OSX towards the human bone development.

This is a novel descriptive study to characterize oft-season, preseason, and postseason bone and body composition measures in women collegiate athletes. From 2006 through 2008, 67 women collegiate athletes from 5 sports, softball (n = 17), basketball (n = 10), volleyball (n = 7), swimming (n = 16), and track jumpers and sprinters (n = 17) were scanned using dual energy X-ray absorptiometry (DXA) at 3 seasonal periods: (a) oft-season = before preseason training, (b) preseason = after preseason training, and (c) postseason = after competitive season. Dual energy X-ray absorptiometry scans were analyzed for total body mass, lean mass (LM), fat mass (FM), percent body fat (%BF), bone mineral content, bone mineral density (BMD), arm BMD, leg BMD, pelvis BMD, and spine BMD. Data were analyzed between sports using analysis of variance (ANOVA) with Tukey post hoc follow-ups, and within each sport using repeated-measures ANOVA and LSD; [alpha] < 0.05. Significant oft-season to preseason or postseason changes in %BF, LM, and BMD within each sport were as follows, respectively: softball, - 7, +4, +1%; basketball, -11, +4, +1%; volleyball, unchanged, unchanged, +2%; swimming, unchanged, +2.5%, unchanged; track jumpers and sprinters, -7, +3.5, +1%. Comparisons among athletes in each sport showed bone measurements of swimmers averaged 4-19% lower than that of athletes in any other sport, whereas for track jumpers and sprinters, %BF and FM averaged 36 and 43% lower compared with other sports at all seasonal periods. Values for athletes playing basketball and volleyball were most similar, whereas softball athletes' values fell between all other athletes. These data serve as sport-specific reference values for comparisons at in-season and oft-season training periods among women collegiate athletes in various sports. KEY WORDS DXA, division I, season, female, morphology

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.jada.2010.03.013 Byline: Shreela V. Sharma, Deanna M. Hoelscher, Steven H. Kelder, Pamela Diamond, R. Sue Day, Albert Hergenroeder Abstract: Calcium intake has been associated with promoting bone health in children and adolescents, thus preventing osteoporosis later in life. Behavior change such as increased calcium intake, as well as physiological factors such as bone quality, may be facilitated by psychosocial and environmental factors. The purpose of this study was to identify pathways by which psychosocial factors influence calcium intake and bone quality in middle school girls. The study design was cross-sectional. Baseline data from the Incorporating More Physical Activity and Calcium in Teens (IMPACT) study, collected in 2001-2003, were used. IMPACT was a 1.5-year nutrition and physical activity intervention study, designed to improve bone density in 717 middle school girls in Texas. Main outcome measures were calcium intake determined using mean milligrams of calcium consumed per day and number of glasses of milk consumed per day, and bone quality determined using a calcaneal stiffness index. Confirmatory factor analysis and path analysis were performed to identify the direct and indirect pathways used by various psychosocial factors such as knowledge, self-efficacy, outcome expectations, and milk availability at home, to influence calcium intake and bone quality. Results showed that knowledge of osteoporosis and calcium-rich foods had an indirect effect on calcium intake, with outcome expectations as the mediating variable ([beta]=.035 and [beta]=.03, respectively; P<0.05). Calcium self-efficacy had a significant indirect effect on calcium intake, with outcome expectations as the mediator ([beta]=.085, P<0.05). None of the variables significantly influenced bone quality. Thus, several direct and indirect pathways used to influence calcium intake among adolescent girls were identified. These findings are critical for the development of effective interventions to promote calcium intake in this population. Article History: Accepted 30 September 2009

Evolution of flight in maniraptoran dinosaurs is marked by the acquisition of distinct avian characters, such as feathers, as seen in Archaeopteryx from the Solnhofen limestone. These rare fossils were pivotal in confirming the dinosauria-avian lineage. One of the key derived avian characters is the possession of feathers, details of which were remarkably preserved in the Lagerstatte environment. These structures were previously simply assumed to be impressions; however, a detailed chemical analysis has, until now, never been completed on any Archaeopteryx specimen. Here we present chemical imaging via synchrotron rapid scanning X-ray fluorescence (SRS-XRF) of the Thermopolis Archaeopteryx, which shows that portions of the feathers are not impressions but are in fact remnant body fossil structures, maintaining elemental compositions that are completely different from the embedding geological matrix. Our results indicate phosphorous and sulfur retention in soft tissue as well as trace metal (Zn and Cu) retention in bone. Other previously unknown chemical details of Archaeopteryx are also revealed in this study including: bone chemistry, taphonomy (fossilization process), and curation artifacts. SRS-XRF represents a major advancement in the study of the life chemistry and fossilization processes of Archaeopteryx and other extinct organisms because it is now practical to image the chemistry of large specimens rapidly at concentration levels of parts per million. This technique has wider application to the archaeological, forensic, and biological sciences, enabling the mapping of 'unseen' compounds critical to understanding biological structures, modes of preservation, and environmental context. trace elements | X-ray absorption spectroscopy doi/ 10.1073/pnas.1001569107

The continuously forming fin bony rays of zebrafish represent a simple bone model system in which mineralization is temporally and spatially resolved. The mineralized collagen fibrils of the fin bones are identical in structure to those found in all known bone materials. We study the continuous mineralization process within the tissue by using synchrotron microbeam x-ray diffraction and small-angle scattering, combined with cryo-scanning electron microscopy. The former provides information on the mineral phase and the mineral particles size and shape, whereas the latter allows high-resolution imaging of native hydrated tissues. The integration of the two techniques demonstrates that new mineral is delivered and deposited as packages of amorphous calcium phosphate nanospheres, which transform into platelets of crystalline apatite within the collagen matrix. biomineralization | carbonated hydroxyapatite | cryo-SEM | microbeam x-ray diffraction | small-angle x-ray scattering doi/10.1073/pnas.0914218107

Neonatal exposure to soy isoflavones at levels similar to that of infants fed soy protein formula resulted in higher bone mineral density (BMD), improved bone structure, and greater bone strength at young adulthood in female CD-1 mice (1,2). Our objective in this study was to determine whether these improvements in bone quantity and quality at 4 mo of age provide protection against the deterioration of bone tissue that occurs after a decline in endogenous sex steroid production. Male and female CD-1 mice (n = 8-18 pups per group per gender) were randomized to subcutaneous injections of corn oil [negative control (CON)], daidzein + genistein (DG; 7 mg. kg body [weight.sup.-1] x [d.sup.-1]), or diethylstilbestrol [(DES); positive control, 2 mg x kg body [weight.sup.-1] x [d.sup.-1]) from postnatal d 1 to 5. At 4 mo of age, mice were ovariectomized (females) or orchidectomized (males) and studied to 8 mo of age. Females treated with DG had higher (P < 0.05) femur and vertebral bone mineral content (BMC) and BMD compared with the CON group. Microstructural analysis revealed that improvements in BMD induced by DG and DES were coupled with greater trabecular thickness at the lumbar spine. Importantly, structural improvements resulted in bones that were more resistant to fracture, as the peak load of the femoral midpoint and lumbar vertebra 2 were higher (P < 0.05) with DG compared with CON. Effects in males were not significant. In conclusion, short-term neonatal exposure to isoflavones provides protection against the deterioration of bone tissue in females but not males after a decline of endogenous sex steroid production. doi: 10.3945/jn.109.116343

Recent studies have indicated that follicle-stimulating hormone (FSH) promotes bone loss. The present study tested the hypothesis that FSH enhances the activity of bone-resorbing cytokines [interleukin (IL)- 1[beta], tumor necrosis factor (TNF)-[alpha], and IL-6], either by inducing their secretion or by altering their receptor expression. Thirty-six women between the ages of 20 and 50 were assessed for bone mineral density (BMD), reproductive hormone, cytokine ligand and soluble receptor concentrations, and surface expression of cytokine receptors on monocytes. In addition, isolated mononuclear cells were incubated in vitro with exogenous FSH. Univariate regression analyses indicated that BMD was inversely related to serum FSH (r = -0.29 to -0.51, P = 0.030.001, depending upon the skeletal site). Physical activity and body composition were also identified as significant factors by multiple regressions. Exogenous FSH induced isolated cells to secrete IL-1[beta], TNF-[alpha], and IL-6 in proportion to the surface expression of FSH receptors on the monocytes. Endogenous (serum) FSH concentrations correlated with the circulating concentrations of these cytokines. None of these individual cytokines was related to BMD, but the IL-1[beta] to IL-1 receptor antagonist (IL-lRa) ratio was inversely related to BMD (r = -0.53, P = 0.002) in all but the most physically active women, who had significantly lower expression of IL-1 type I receptors relative to type II (decoy receptors, P = 0.01). Physical activity also correlated positively with secretion of inhibitory soluble IL-I receptors (r = 0.53, P = 0.003). Moreover, IL-1Ra correlated strongly with percent body fat (r = 0.66, P < 0.0001). These results indicate that BMD is related to FSH concentration, physical activity, and body composition. Although each of these factors likely has direct effects on bone, the present study suggests that each may also influence BMD by modulating the activity of the osteoresorptive cytokine IL-1[beta]. leptin: inhibin; physical activity doi:10.1152/ajpregu.00728.2009

Parathyroid hormone (PTH) restores bone mass to the osteopenic skeleton, but significant questions remain as to the underlying mechanisms. The receptor for advanced glycation end products (RAGE) is a multiligand receptor of the immunoglobulin superfamily; however, recent studies indicate a role in bone physiology. We investigated the significance of RAGE to hormone-induced increases in bone by treating 10-wk-old female Rage-knockout (KO) and wild-type (WT) mice with human PTH-(1-34) at 30 [micro]g x [kg.sup.-1] x [day.sup.-1] or vehicle control, 7 days/wk, for 7 wk. PTH produced equivalent relative gains in bone mineral density (BMD) and bone mineral content (BMC) throughout the skeleton in both genotypes. PTH-mediated relative increases in cortical area of the midshaft femur were not compromised in the null mice. However, the hormone-induced gain in femoral cancellous bone was significantly attenuated in Rage-KO mice. The loss of RAGE impaired PTH-mediated increases in femoral cancellous bone volume, connectivity density, and trabecular number but did not impact increases in trabecular thickness or decreases in trabecular spacing. Disabling RAGE reduced femoral expression of bone formation genes, but their relative PTH-responsiveness was not impaired. Neutralizing RAGE did not attenuate vertebral cancellous bone response to hormone. Rage-null mice exhibited an attenuated accrual rate of bone mass, with the exception of the spine, and an enhanced accrual rate of fat mass. We conclude that RAGE is necessary for key aspects of the skeleton's response to anabolic PTH. Specifically, RAGE is required for hormone-mediated improvement of femoral trabecular architecture but not intrinsically necessary for increasing cortical thickness. receptor for advanced glycation end products; anabolic; high-mobility group box 1 protein; osteoblast; osteoporosis; osteoimmunology doi:10.1152/ajpendo.00564.2009.

Interest in the development of new sources of transplantable materials for the treatment of injury or disease has led to the convergence of tissue engineering with stem cell technology. Bone and joint disorders are expected to benefit from this new technology because of the low self-regenerating capacity of bone matrix secreting cells. Herein, the differentiation of stem cells to bone cells using active multilayered capsules is presented. The capsules are composed of poly-L-glutamic acid and poly-L-lysine with active growth factors embedded into the multilayered film. The bone induction from these active capsules incubated with embryonic stem cells was demonstrated in vitro. Herein, we report the unique demonstration of a multilayered capsule-based delivery system for inducing bone formation in vivo. This strategy is an alternative approach for in vivo bone formation. Strategies using simple chemistry to control complex biological processes would be particularly powerful, as they make production of therapeutic materials simpler and more easily controlled. active biomaterials | layer-by-layer | nanostructured capsules | stem cells | tissue engineering doi/10.1073/pnas.0908531107

A genome-wide association study and follow-up replication studies to identify genetic variants that influence bone mineral density (BMD) in different ethnic groups reveal that rs2273061 of the Jagged1 (JAG1) gene is associated with high BMD and osteoporotic fractures. The results reveal that the JAG1 gene is involved in BMD regulation in different ethnic groups, and is also a possible important factor in fracture pathogenesis.

Energy restriction decreases bone mineral density (BMD), and pidemiological studies suggest that the risk of weight loss-induced bone loss is greater in lean than in heavier individuals. Our goal in this study was to determine how bone density and geometry respond to energy restriction in mature obese rats compared with lean rats. At 6 mo of age, 36 diet- induced obese and lean female Sprague-Dawley rats were allocated to control (CTL; ad libitum; n = 18) and energy-restricted (EnR; 40% restriction; n = 18) diets. After 10 wk of dietary intervention, obese EnR rats lost more weight (-61 [+ or -] 14 g) than lean EnR rats (-91 [+ or -] 34 g) (P < 0.02), whereas body weight did not change significantly in the 2 CTL groups (14 [+ or -] 23 g). Only the lean EnR (and not obese EnR) rats showed lower BMD compared with CTL rats at the tibia, distal, and proximal femur and femoral neck, and trabecular bone volume (P < 0.05). Serum estradiol declined in lean EnR rats compared with baseline (P< 0.05) but not in the obese EnR rats. In addition, the final serum 25-hydroxyvitamin D (250HD) concentration was higher (P < 0.05) in obese than in lean EnR rats. Serum parathyroid hormone decreased (P < 0.05) from baseline to final in lean and obese CTL, but not EnR rats. These data support the hypothesis that energy restriction in lean rats compared with obese rats is more detrimental to bone, and it is possible that the greater decline in estrogen and lower levels of 250HD contribute to this effect. J. Nutr. 140: 31-37, 2010. doi: 10.3945/jn.109.111450.

J Strength Cond Res 24(1): 213-21 7,2010--This study evaluated impulse (I), peak ground reaction forces (GRF), and the rate of force development (RFD) of a variety of exercise modes for the purpose of estimating the magnitude and rate of mechanical loading as a measure of osteogenic potential. Twenty-three subjects participated in this study (mean [+ or -] SD, age 21.2 [+ or -] 1.4 years; body mass 77.8 [+ or -] 16.2 kg). Kinetic data were obtained via a force platform for the test exercises modes, which included walking, jogging, depth jumps, loaded jump squats, and the back squat. Repeated measures analysis of variance revealed significant main effects for I, GRF, and RFD (p [less than or equal to] 0.001). Bonferroni-adjusted post hoc analyses demonstrated that I and GRF were different between each exercise mode and that RFD was different between all exercise modes except for jogging and the back squat. The depth jump demonstrated the highest GRF and RFD, while the back squat produced the highest I. The jump squat produced the second highest value for all the variables assessed. Thus, the depth jump, jump squat, and back squat appear to offer the greatest potential as osteogenic stimuli and a mixed mode training strategy including exercises such as these is recommended. These results suggest that walking and jogging may have less osteogenic potential. KEY WORDS ground reaction forces, resistance training, plyometrics, maximal power training, osteogenesis, bone

Jan Zalasiewicz enjoys Brian Switek's exploration of how the human scaffold -- and our ideas about it -- evolved. Jan Zalasiewicz enjoys Brian Switek's exploration of how the human scaffold -- and our ideas about it -- evolved., Author(s): Jan Zalasiewicz Author Affiliations: The skeleton articulated Our bones have evolved over millions of years. [see PDF for image] Credit: Nick Veasey X-ray sequence of a human skeleton running [...]

Adult human mesenchymal stromal cells (hMSCs) have the potential to differentiate into chondrogenic, adipogenic, or osteogenic lineages, providing a potential source for tissue regeneration. An important issue for efficient bone regeneration is to identify factors that can be targeted to promote the osteogenic potential of hMSCs. Using transcriptome analysis, we found that integrin [alpha]5 (ITGA5) expression is up-regulated during dexamethasone-induced osteoblast differentiation of hMSCs. Gain-of-function studies showed that ITGA5 promotes the expression of osteoblast phenotypic markers and in vitro osteogenesis of hMSCs. Down-regulation of endogenous ITGA5 using specific shRNAs blunted osteoblast marker gene expression and osteogenic differentiation. Molecular analyses showed that the enhanced osteoblast differentiation induced by ITGA5 was mediated by activation of focal adhesion kinase/ERK1/2-MAPKs and PI3K signaling pathways. Remarkably, activation of endogenous ITGA5 using agonists such as a specific antibody that primes the integrin or a peptide that specifically activates ITGA5 was sufficient to enhance ERK1/2-MAPKs and PI3K signaling and to promote osteoblast differentiation and osteogenic capacity of hMSCs. Importantly, we demonstrated that hMSCs engineered to overexpress ITGA5 exhibited a marked increase in their osteogenic potential in vivo. Taken together, these findings not only reveal that ITGA5 is required for osteoblast differentiation of adult hMSCs but also provide a targeted strategy using ITGA5 agonists to promote the osteogenic capacity of hMSCs. This may be used for tissue regeneration in bone disorders where the recruitment or capacity of hMSCs is compromised. mesenchymal stem cells | bone formation | agonist www.pnas.org/cgi/doi/10.1073/pnas.0812334106

The growth, development, and maintenance of bone are influenced by genetic and environmental variables. Understanding variability in bone microstructure among primates may help illuminate the factors influencing the number and size of secondary osteons. The purpose of this study is to assess the bone microstructure in 8 humeral and 12 femoral sections of 12 juvenile chimpanzees, aged 2-15.3 years, and one adult chimp. Secondary osteons were counted and measured for 16 fields per section. Results indicate that the femur exhibits a mean osteon population density (OPD) of 4.46 [+ or -] 2.34/[mm.sup.2], mean Haversian canal area of 0.0016 [+ or -] 0.0007 [mm.sup.2], and mean osteon area of 0.033 [+ or -] 0.006 [mm.sup.2]. The humerus has a mean OPD of 4.72 [+ or -] 1.57/[mm.sup.2], mean Haversian canal area of 0.0013 [+ or -] 0.0003 [mm.sup.2], and mean osteon area of 0.033 [+ or -] 0.005 [mm.sup.2]. Differences are not significant between the humerus and femur, possibly indicating similar mechanical demands during locomotion. Osteon population density exhibits a moderate correlation with age (r = 0.498) in the femur of the juvenile chimps, but the adult chimp has an OPD of 10.28/[mm.sup.2], suggesting that osteons likely accumulate with age. Females exhibit higher osteon densities in the periosteal envelope compared to males in the humerus, indicating more remodeling during periosteal expansion. Overall similarities between chimpanzees and humans as well as previously published data on Late Pleistocene hominids (Abbott et al.: Am J Phys Anthropol 99 [1996] 585-601) suggest that bone microstructure has been stable throughout human evolution. KEY WORDS osteon; Hominoidea; histomorphometry; primate

This study attempts to fill a persistent gap in the literature by documenting the timing of epiphyseal union at the vertebral end of the ribs in a sample of modern Portuguese skeletons. The skeletal remains of 53 females and 45 males, between the ages of 11 and 30, were taken from the Lisbon documented skeletal collection. Individuals in the sample have been previously described as being representative of a middle-to-low socioeconomic segment of the early 20th century Lisbon population. Three anatomical locations were examined for epiphyseal union: the head, the articular tubercle and the nonarticular tubercle. The first epiphysis to show partial union is that of the nonarticular tubercle (females, 11-19 years; males, 11-19 years), followed by the epiphysis of the articular tubercle (females, 11-20 years; males, 16-20 years), and finally by the head epiphysis (females, 15-24 years; males, 16-22 years), which can still show incomplete epiphyseal closure at 25 and 24 years for females and males, respectively. A trend for earlier female maturation was observed, but the statistical tests only confirmed this result for some ribs and age groups. No directional asymmetry was found, but a significant fluctuating asymmetry was observed in all three epiphyses. A preliminary analysis showed that the asymmetric group of individuals in the study sample includes all the rural-to-urban migrants, relative to the symmetric group. KEY WORDS skeletal age; rib maturation; Lisbon collection; fluctuating asymmetry

Consumption of green tea may reduce body weight gain. Although many disorders are related to obesity, bone mass is positively correlated with body mass. Therefore, our purpose in this study was to determine the effects of green tea extract (GTE) on bone mass and architecture in rapidly growing lean [C57BL/6 wild type (WT)] and genetically obese, leptin-deficient (ob/ob) male mice. Five-week-old lean and ob/ob mice were assigned to diets containing GTE at 0, 1, or 2% for 6 wk. Femoral and lumbar vertebral bone volume and architecture were evaluated by micro-computed tomography ([mu]CT). Following [mu]CT analysis, femora were ashed to determine bone mineral content and density. Compared with WT mice, ob/ ob mice had shorter femora (P < 0.001), lower femoral bone volume (P < 0.001), and lower femoral bone mineral content (P < 0.001), but higher cancellous bone volume in lumbar vertebrae (P < 001). Neither genotype nor treatment affected femoral bone mineral density, indicating normal mineralization. GTE consumption resulted in lower femur length, volume, mineral content, cortical volume, and cortical thickness (P < 0.001), as well as lower cancellous bone volume/tissue volume (P < 0.008) and trabecular thickness (P < 0.004) in lumbar vertebrae. The results indicate that leptin is not essential for the reduced gains in body weight and bone mass due to GTE in growing mice and suggest that consumption of large quantities of green tea may reduce the rate of bone accumulation during growth.

Soy isoflavones and their metabolites, with estrogenic activity, have been considered candidates for reducing postmenopausal bone loss. In this study, we examined the effect of dietary equol, a bioactive metabolite of the soy isoflavone daidzein, on equol tissue distribution, bone parameters, and reproductive tissue activity using an adult ovariectomized (OVX) rat model. An 8-wk feeding study was conducted to compare 4 dietary treatments of equol (0, 50, 100, 200 mg/kg diet) in 6-mo-old OVX female Sprague-Dawley rats. A dose response increase in tissue equol concentrations was observed for serum, liver, kidney, and heart, and a plateau occurred at 100 mg equol/kg diet for intestine. In OVX rats receiving 200 mg equol/kg diet, femoral calcium concentration was greater than those receiving lower doses but was still less than SHAM (P < 0.05), and other bone measures were not improved. Tibia calcium concentrations were lower in OVX rats receiving 100 and 200 mg equol/kg diet compared with the OVX control rats. Trabecular bone mineral density of tibia was also lower in equol-fed OVX rats. At this dietary equol intake, uterine weight was higher (P < 0.05) than in other OVX groups but lower than the SHAM-operated intact rats. The 200 mg/kg diet dose of dietary equol significantly increased proliferative index in the uterine epithelium. Dietary equol had no stimulatory effect on mammary gland epithelium. We conclude that in OVX rats, a dietary equol dose that had modest effect on bone also exerts mild uterotropic effects.

Relatively few studies have examined the effects of formula feeding relative to breast-feeding on bone in the neonate. Using peripheral quantitative CT scan and histomorphometric analysis, we demonstrated that neonatal piglets fed with soy-based formula (SF) and cow milk-based formula (MF) for 21 or 35 d had greater bone mineral density and content than breast-fed piglets (BF) (P < 0.05). Osteoblast numbers and bone formation rate at postnatal d 35 were greater in SF compared with other groups (P < 0.05), whereas osteoclast numbers were lower in both MF and SF groups than in the BF group (P < 0.05). Osteoblastogenesis was greater in ex vivo bone marrow cell cultures from SF than in MF or BF piglets (P < 0.05). Bone formation markers in serum were greater, whereas bone resorption markers were lower in the MF- and SF-fed groups than in the BF group (P < 0.05). Bone morphogenic protein (BMP) 2 and alkaline phosphatase mRNAs were upregulated in the MF and SF groups compared with the BF group (P < 0.05), whereas receptor activator of NF-[kappa]B ligand was downregulated (P < 0.05). Extracellular signal-regulated kinase, p38, Smad1/5/8 phosphorylation, and runt-related transcription factor 2 expression were greater in bone from the MF and SF groups compared with the BF group (P < 0.05). In vitro studies showed that 2.5% serum from SF- or MF-fed piglets was able to stimulate osteoblast differentiation but not in the presence of the BMP blocker noggin. Therefore, formula feeding promoted bone growth compared with BF. SF piglets had the highest bone volume over tissue volume. This suggests that SF-fed piglets may have the best quality bone. The anabolic effects of SF on bone appear to be mediated through enhanced B MP signaling.

Calcium (Ca) supplements, especially Ca carbonate (CaC[O.sub.3]), are the main alternative sources of dietary Ca and an important part of a treatment regimen for osteoporosis, the most common metabolic bone disorder of aging and menopause. In a female ovariectomized (OVX) rat model for studying postmenopausal osteoporosis, we tested the hypothesis that a small compared with a large particle size of GAG[O.sub.3] (13.0- vs. 18.5-[micro]m geometric diameter) would result in increased Ca balance and subsequently bone mass and that this would be affected by dietary Ca level. We used 6-moold rats that were OVX either at 6 or 3 mo of age as models of early or stable menopausal status, respectively. The rats received semipurified diets that contained either 0.4 or 0.2% dietary Ca provided from CaC[O.sub.3] of 2 particle sizes. A group of Sham-operated rats with intact ovaries served as control and were fed 0.4% dietary Ca from large particles. Estrogen deficiency as a result of ovariectomy had an adverse effect on bone density, mineral content, and bone mechanical properties (P < 0.001 ). Reducing dietary Ca from 0.4 to 0.2% resulted in significant adverse effects on bone density and mechanical properties (P < 0.001). The particle size of CaC[O.sub.3] did not affect total Ca balance, bone dual energy X-ray absorptiometry and peripheral quantitative computed tomography indices, bone ash and Ca content, or the mechanical determinants of bone strength. We conclude that a decrease in particle size of CaC[O.sub.3] to 70% of that typically found in Ca supplements does not provide a benefit to overall Ca metabolism or bone characteristics and that the amount of Ca consumed is of greater influence in enhancing Ca nutrition and skeletal strength.

The aim of the present study was to evaluate the effect of dietary lactoferrin on bone metabolism in vivo using a postmenopausal animal model. We investigated whether bovine lactoferrin (bLF) ingestion could prevent bone loss in ovariectomized mice. Twelve-week-old female C3H mice either ovariectomized or sham operated were fed for 27 wk with the control diet (AIN-93M with 140 g of total milk protein as a protein source per kg of diet). Four groups of ovariectomized mice received diets including different concentrations of bLF (1, 5, 10, or 20 g of total milk protein were replaced by bLF). Ovariectomy induced a decreased uterine weight and a smaller gain of bone mineral density. Immunoreactive bLF was detected in the peripheral blood, and its concentration was related to the amount of bLF ingestion, bLF supplementation to the diet improved bone mineral density (BMD) and femoral failure load in a dose-dependent manner. We confirmed the direct effects of bLF in vitro using established and primary cultures of murine bone cells. Addition of bLF to the culture medium at a concentration of between 1 and 1,000 [micro]g/ml stimulated both cell growth and differentiation of osteoblastic MC3T3 cells while inhibiting the growth of preosteoclastic RAW 267.4 cells. In primary culture of mixed bone cells, an enhanced osteoblast differentiation was associated with an inhibition of osteoclast differentiation at lower bLF concentrations (1-10 [micro]g/ml). In conclusion, these findings suggest that dietary lactoferrin supplementation can have a beneficial effect on postmenopausal bone loss by modulating bone formation and resorption. osteoblast; osteoclast; bone mineral density

Our goal in this cross-sectional study was to investigate the influence of low-vitamin D status on bone mass, bone turnover, and muscle strength in 301 healthy Chinese adolescent girls. Blood plasma 25-hydroxyvitamin D [25(OH)D] was measured by RIA and plasma and urine biomarkers of bone turnover were measured. Bone mineral content (BMC) and density and bone area for the whole body and the distal and proximal forearm were measured by dual energy X-ray absorptiometry. When vitamin D deficiency was defined as a serum 25(OH)D concentration of [less than or equal to] 50 nmol/L and severe deficiency as

Lactoferrin (LF) is reported to stimulate osteoblast proliferation and inhibit osteoclast activity in bone cell culture. However, the effect of oral LF on bone in osteoporosis needs to be explored. Three-month-old female Sprague-Dawley rats (n = 70) were assigned to the following groups: sham-operated, ovariectomized (OVX) untreated, OVX + bovine serum albumin (BSA; 85 mg/kg body weight), OVX + LF (0.85 mg/kg, 8.5 mg/kg, and 85 mg/kg body weight), and OVX + 17[beta]-estradiol (E2; 10 [micro]g/kg body weight). After 3 mo of treatment, [E.sub.2] completely prevented the OVX-induced bone loss. OVX rats treated with LF were protected against the OVX-induced reduction of bone volume, trabecular number, and thickness, and the elevation of trabecular separation was prevented. LF also increased bone mineral density and increased the parameters of mechanical strength at 8.5- and 85-mg/kg doses. Greater bone formation and reduced bone resorption, as assessed by biochemical markers of bone remodeling, occurred in rats administered LF. LF at 8.5- and 85-mg/kg concentrations caused a significant decrease in serum calcium, but this reduction did not occur in rats fed 0.85 mg/kg LF. In addition, serum tumor necrosis factor-[alpha] and interleukin-6 production were suppressed and serum calcitonin was elevated significantly in LF-fed rats at all 3 doses. These findings indicated that oral LF not only preserved bone mass but also improved bone microarchitecture. The absorption of LF peptides and their effects on bone ceils could to some extent account for the osteogenic function of oral LF.

We report that oxytocin (OT), a primitive neurohypophyseal hormone, hitherto thought solely to modulate lactation and social bonding, is a direct regulator of bone mass. Deletion of OT or the OT receptor (Oxtr) in male or female mice causes osteoporosis resulting from reduced bone formation. Consistent with low bone formation, OT stimulates the differentiation of osteoblasts to a mineralizing phenotype by causing the up-regulation of BMP-2, which in turn controls Schnurri-2 and 3, Osterix, and ATF-4 expression. In contrast, OT has dual effects on the osteoclast. It stimulates osteoclast formation both directly, by activating NF-[kappa]B and MAP kinase signaling, and indirectly through the up-regulation of RANK-L. On the other hand, OT inhibits bone resorption by mature osteodasts by triggering cytosolic [Ca.sup.2+] release and NO synthesis. Together, the complementary genetic and pharmacologic approaches reveal OT asa novel anabolic regulator of bone mass, with potential implications for osteoporosis therapy. osteoblast | osteoclast | osteoporosis | pituitary hormones | bone density

Genetic evidence indicates that Wnt signaling is critically involved in bone homeostasis. In this study, we investigated the functions of canonical Wnts on differentiation of adult multipotent human mesenchymal stem cells (hMSCs) in vitro and in vivo. We observe differential sensitivities of hMSCs to Wnt inhibition of osteogenesis versus adipogenesis, which favors osteoblastic commitment under binary in vitro differentiation conditions. Wnt inhibition of osteogenesis is associated with decreased expression of osteoblastic transcription factors and inhibition of c-Jun N-terminal kinase and p38 mitogen-activated protein kinase activation, which are involved in osteogenic differentiation. An hMSC subpopulation exhibits high endogenous Wnt signaling, the inhibition of which enhances osteogenic and adipogenic differentiation in vitro. In an in vivo bone formation model, high levels of Wnt signaling inhibit de novo bone formation by hMSCs. However, hMSCs with exogenous expression of Wntl but not stabilized [beta]-catenin markedly stimulate bone formation by naive hMSCs, arguing for an important role of a canonical Wnt gradient in hMSC osteogenesis in vivo.