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Bone microarchitecture assessed by high-resolution peripheral quantitative computed tomography varies across populations of different origin. The study presents a reference dataset of microarchitectural parameters in a homogeneous group of participants aged within 22-27 range determined by a discriminant analysis of a larger cross-sectional cohort of 339 women. INTRODUCTION: High-resolution peripheral quantitative computed tomography (HR-pQCT) non-invasively measures three-dimensional bone microarchitectural parameters and volumetric bone mineral density. Previous studies established normative reference HR-pQCT datasets for several populations, but there were few data assessed in a reference group of young women with Caucasian ethnicity living in Western Europe. It is important to obtain different specific reference dataset for a valid interpretation of cortical and trabecular microarchitecture data. The aim of our study was to find the population with the most optimal bone status in order to establish a descriptive reference HR-pQCT dataset in a young and healthy normal-weight female cohort living in a European area including Geneva, Switzerland, Lyon and Saint-Etienne, France. METHODS: We constituted a cross-sectional cohort of 339 women aged 19-41 years with a BMI > 18 and < 30 kg/m2. All participants had HR-pQCT measurements at both non-dominant distal radius and tibia sites. RESULTS: We observed that microarchitectural parameters begin to decline before the age of 30 years. Based on a discriminant analysis, the optimal bone profile in this population was observed between the age range of 22 to 27 years. Consequently, we considered 43 participants aged 22-27 years to establish a reference dataset with median values and percentiles. CONCLUSION: This is the first study providing reference values of HR-pQCT measurements considering specific age bounds in a Franco-Swiss female cohort at the distal radius and tibia sites.
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Densidad Ósea , Etnicidad , Adulto , Anciano , Estudios Transversales , Femenino , Humanos , Radio (Anatomía)/diagnóstico por imagen , Suiza , Tibia , Adulto JovenRESUMEN
We reviewed the experimental and clinical evidence that hip bone strength estimated by BMD and/or finite element analysis (FEA) reflects the actual strength of the proximal femur and is associated with hip fracture risk and its changes upon treatment. INTRODUCTION: The risk of hip fractures increases exponentially with age due to a progressive loss of bone mass, deterioration of bone structure, and increased incidence of falls. Areal bone mineral density (aBMD), measured by dual-energy X-ray absorptiometry (DXA), is the most used surrogate marker of bone strength. However, age-related declines in bone strength exceed those of aBMD, and the majority of fractures occur in those who are not identified as osteoporotic by BMD testing. With hip fracture incidence increasing worldwide, the development of accurate methods to estimate bone strength in vivo would be very useful to predict the risk of hip fracture and to monitor the effects of osteoporosis therapies. METHODS: We reviewed experimental and clinical evidence regarding the association between aBMD and/orCT-finite element analysis (FEA) estimated femoral strength and hip fracture risk as well as their changes with treatment. RESULTS: Femoral aBMD and bone strength estimates by CT-FEA explain a large proportion of femoral strength ex vivo and predict hip fracture risk in vivo. Changes in femoral aBMD are strongly associated with anti-fracture efficacy of osteoporosis treatments, though comparable data for FEA are currently not available. CONCLUSIONS: Hip aBMD and estimated femoral strength are good predictors of fracture risk and could potentially be used as surrogate endpoints for fracture in clinical trials. Further improvements of FEA may be achieved by incorporating trabecular orientations, enhanced cortical modeling, effects of aging on bone tissue ductility, and multiple sideway fall loading conditions.
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Fracturas de Cadera , Huesos Pélvicos , Absorciometría de Fotón , Anciano , Densidad Ósea , Estudios de Casos y Controles , Femenino , Fémur , Análisis de Elementos Finitos , Fracturas de Cadera/epidemiología , Fracturas de Cadera/etiología , Humanos , MasculinoRESUMEN
We investigated the interaction between periostin SNPs and the SNPs of the genes assumed to modulate serum periostin levels and bone microstructure in a cohort of postmenopausal women. We identified an interaction between LRP5 SNP rs648438 and periostin SNP rs9547970 on serum periostin levels and on radial cortical porosity. PURPOSE: The purpose of this study is to investigate the interaction between periostin gene polymorphisms (SNPs) and other genes potentially responsible for modulating serum periostin levels and bone microstructure in a cohort of postmenopausal women. METHODS: In 648 postmenopausal women from the Geneva Retirees Cohort, we analyzed 6 periostin SNPs and another 149 SNPs in 14 genes, namely BMP2, CTNNB1, ESR1, ESR2, LRP5, LRP6, PTH, SPTBN1, SOST, TGFb1, TNFRSF11A, TNFSF11, TNFRSF11B and WNT16. Volumetric BMD and bone microstructure were measured by high-resolution peripheral quantitative computed tomography at the distal radius and tibia. RESULTS: Serum periostin levels were associated with radial cortical porosity, including after adjustment for age, BMI, and years since menopause (p = 0.036). Sixteen SNPs in the ESR1, LRP5, TNFRSF11A, SOST, SPTBN1, TNFRSF11B and TNFSF11 genes were associated with serum periostin levels (p range 0.03-0.001) whereas 26 SNPs in 9 genes were associated with cortical porosity at the radius and/or at the tibia. WNT 16 was the gene with the highest number of SNPs associated with both trabecular and cortical microstructure. The periostin SNP rs9547970 was also associated with cortical porosity (p = 0.04). In particular, SNPs in LRP5, ESR1 and near the TNFRSF11A gene were associated with both cortical porosity and serum periostin levels. Eventually, we identified an interaction between LRP5 SNP rs648438 and periostin SNP rs9547970 on serum periostin levels (interaction p = 0.01) and on radial cortical porosity (interaction p = 0.005). CONCLUSION: These results suggest that periostin expression is genetically modulated, particularly by polymorphisms in the Wnt pathway, and is thereby implicated in the genetic variation of bone microstructure.
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Densidad Ósea/genética , Moléculas de Adhesión Celular/genética , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad/genética , Polimorfismo de Nucleótido Simple , Anciano , Densidad Ósea/fisiología , Moléculas de Adhesión Celular/sangre , Estudios de Cohortes , Femenino , Humanos , Porosidad , Posmenopausia/sangre , Posmenopausia/genética , Radio (Anatomía)/anatomía & histología , Radio (Anatomía)/diagnóstico por imagen , Radio (Anatomía)/fisiología , Tibia/anatomía & histología , Tibia/diagnóstico por imagen , Tibia/fisiología , Tomografía Computarizada por Rayos X , Vía de Señalización Wnt/genética , Vía de Señalización Wnt/fisiologíaRESUMEN
A longitudinal analysis of bone microstructure in postmenopausal women of the Geneva Retirees Cohort indicates that age-related cortical bone loss is attenuated at non-bearing bone sites in fermented dairy products consumers, not in milk or ripened cheese consumers, independently of total energy, calcium, or protein intakes. INTRODUCTION: Fermented dairy products (FDP), including yogurts, provide calcium, phosphorus, and proteins together with prebiotics and probiotics, all being potentially beneficial for bone. In this prospective cohort study, we investigated whether FDP, milk, or ripened cheese consumptions influence age-related changes of bone mineral density (BMD) and microstructure. METHODS: Dietary intakes were assessed at baseline and after 3.0 ± 0.5 years with a food frequency questionnaire in 482 postmenopausal women enrolled in the Geneva Retirees Cohort. Cortical (Ct) and trabecular (Tb) volumetric (v) BMD and microstructure at the distal radius and tibia were assessed by high-resolution peripheral quantitative computerized tomography, in addition to areal (a) BMD and body composition by dual-energy X-ray absorptiometry, at the same time points. RESULTS: At baseline, FDP consumers had lower abdominal fat mass and larger bone size at the radius and tibia. Parathyroid hormone and ß-carboxyterminal cross-linked telopeptide of type I collagen levels were inversely correlated with FDP consumption. In the longitudinal analysis, FDP consumption (mean of the two assessments) was associated with attenuated loss of radius total vBMD and of Ct vBMD, area, and thickness. There was no difference in aBMD and at the tibia. These associations were independent of total energy, calcium, or protein intakes. For other dairy products categories, only milk consumption was associated with lower decrease of aBMD and of failure load at the radius. CONCLUSION: In this prospective cohort of healthy postmenopausal women, age-related Ct bone loss was attenuated at non-bearing bone sites in FDP consumers, not in milk or ripened cheese consumers, independently of total energy, calcium, or protein intakes. STUDY REGISTRATION: ISRCTN11865958 ( http://www.isrctn.com ).
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Calcio de la Dieta/administración & dosificación , Productos Lácteos Cultivados/estadística & datos numéricos , Proteínas en la Dieta/administración & dosificación , Conducta Alimentaria/fisiología , Osteoporosis/prevención & control , Absorciometría de Fotón/métodos , Anciano , Biomarcadores/sangre , Composición Corporal/efectos de los fármacos , Composición Corporal/fisiología , Densidad Ósea/efectos de los fármacos , Densidad Ósea/fisiología , Remodelación Ósea/efectos de los fármacos , Remodelación Ósea/fisiología , Calcio de la Dieta/farmacología , Dieta/estadística & datos numéricos , Proteínas en la Dieta/farmacología , Ingestión de Energía/fisiología , Femenino , Humanos , Osteoporosis/epidemiología , Osteoporosis/fisiopatología , Posmenopausia/fisiología , Suiza/epidemiologíaRESUMEN
Fragility fractures are increasingly recognized as a complication of both type 1 and type 2 diabetes, with fracture risk that increases with disease duration and poor glycemic control. Yet the identification and management of fracture risk in these patients remains challenging. This review explores the clinical characteristics of bone fragility in adults with diabetes and highlights recent studies that have evaluated bone mineral density (BMD), bone microstructure and material properties, biochemical markers, and fracture prediction algorithms (i.e., FRAX) in these patients. It further reviews the impact of diabetes drugs on bone as well as the efficacy of osteoporosis treatments in this population. We finally propose an algorithm for the identification and management of diabetic patients at increased fracture risk.
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Diabetes Mellitus Tipo 1/complicaciones , Diabetes Mellitus Tipo 2/complicaciones , Fracturas Osteoporóticas/etiología , Conservadores de la Densidad Ósea/uso terapéutico , Remodelación Ósea , Diabetes Mellitus Tipo 1/epidemiología , Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/fisiopatología , Humanos , Osteoporosis/tratamiento farmacológico , Osteoporosis/epidemiología , Osteoporosis/etiología , Osteoporosis/fisiopatología , Fracturas Osteoporóticas/epidemiología , Fracturas Osteoporóticas/fisiopatología , Fracturas Osteoporóticas/prevención & control , Factores de RiesgoRESUMEN
Discontinuation of denosumab (Dmab) therapy is associated with lower serum CTX levels in osteoporotic patients previously exposed to bisphosphonates compared to those who were not. INTRODUCTION: Discontinuation of Dmab therapy is followed by a transient increase of bone turnover markers (BTMs) above pretreatment values, together with accelerated bone loss, and potentially an increased risk of multiple vertebral fractures. Since a substantial proportion of patients discontinuing Dmab have previously been exposed to bisphosphonates (BPs), we hypothesized that previous BP therapy could attenuate this increase in bone turnover because of the prolonged biological effects of BPs on bone. METHODS: In a retrospective observation, we assessed serum CTX levels between 7 and 24 months after the last Dmab injection in 37 patients (33 women and 4 men, aged 50 to 84 years). CTX levels were analyzed according to the number of Dmab injections (1 or multiple) and previous exposure to BPs. RESULTS: In 8 patients who had received only 1 Dmab injection, 7 out of 8 were previously on BPs and none of them showed CTX values above the premenopausal range after Dmab discontinuation. CTX also remained in the premenopausal range in 14 out of 17 patients who discontinued Dmab after multiple (4.1 ± 1.4, range 2-7) injections but were previously exposed to BPs (mean exposure 6.9 ± 5.8 years, range 11 months-15 years; mean time interval between BP exposure and Dmab initiation 25 ± 10 months, range 0-48). In contrast, in 12 patients who discontinued Dmab after multiple (5, range 3-9) injections without prior exposure to BPs, mean CTX levels as measured on average 11.3 months (range 6-23) after the last Dmab injection were above the upper limit of premenopausal range (mean +114%, range 28-320%, p = 0.003-0.005 vs previous BPs). CONCLUSION: The higher CTX levels occurring after Dmab discontinuation in patients who have received multiple injections may be prevented by prior exposure to BPs. This observation may be related to the persistent effects of BPs on bone that prevent the resorbing activity of newly formed osteoclasts when RANK Ligand is no more antagonized.
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Conservadores de la Densidad Ósea/uso terapéutico , Colágeno Tipo I/sangre , Denosumab/uso terapéutico , Difosfonatos/uso terapéutico , Osteoporosis/tratamiento farmacológico , Péptidos/sangre , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Conservadores de la Densidad Ósea/administración & dosificación , Conservadores de la Densidad Ósea/farmacología , Remodelación Ósea/efectos de los fármacos , Denosumab/administración & dosificación , Denosumab/farmacología , Deprescripciones , Difosfonatos/administración & dosificación , Difosfonatos/farmacología , Esquema de Medicación , Quimioterapia Combinada , Femenino , Humanos , Masculino , Persona de Mediana Edad , Osteoporosis/sangre , Osteoporosis/fisiopatología , Estudios Retrospectivos , Privación de TratamientoRESUMEN
The exact role of biochemical markers of bone turnover in the management of metabolic bone diseases remains a topic of controversy. In this consensus paper, the Belgian Bone Club aimed to provide a state of the art on the use of these biomarkers in different clinical or physiological situations like in postmenopausal women, osteoporosis in men, in elderly patients, in patients suffering from bone metastasis, in patients with chronic renal failure, in pregnant or lactating women, in intensive care patients, and in diabetics. We also gave our considerations on the analytical issues linked to the use of these biomarkers, on potential new emerging biomarkers, and on the use of bone turnover biomarkers in the follow-up of patients treated with new drugs for osteoporosis.
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Biomarcadores/análisis , Densidad Ósea , Enfermedades Óseas Metabólicas/diagnóstico , Remodelación Ósea , Osteoporosis/diagnóstico , Bélgica , Neoplasias Óseas , Consenso , Femenino , Humanos , Lactancia , Masculino , Osteoporosis Posmenopáusica/diagnóstico , Embarazo , Insuficiencia Renal CrónicaRESUMEN
Activation of beta2-adrenergic receptors inhibits osteoblastic bone formation and enhances osteoclastic bone resorption. Whether beta-blockers inhibit ovariectomy-induced bone loss and decrease fracture risk remains controversial. To further explore the role of beta-adrenergic signaling in skeletal acquisition and response to estrogen deficiency, we evaluated mice lacking the three known beta-adrenergic receptors (beta-less). Body weight, percent fat, and bone mineral density were significantly higher in male beta-less than wild-type (WT) mice, more so with increasing age. Consistent with their greater fat mass, serum leptin was significantly higher in beta-less than WT mice. Mid-femoral cross-sectional area and cortical thickness were significantly higher in adult beta-less than WT mice, as were femoral biomechanical properties (+28 to +49%, P < 0.01). Young male beta-less had higher vertebral (1.3-fold) and distal femoral (3.5-fold) trabecular bone volume than WT (P < 0.001 for both) and lower osteoclast surface. With aging, these differences lessened, with histological evidence of increased osteoclast surface and decreased bone formation rate at the distal femur in beta-less vs. WT mice. Serum tartrate-resistance alkaline phosphatase-5B was elevated in beta-less compared with WT mice from 8-16 wk of age (P < 0.01). Ovariectomy inhibited bone mass gain and decreased trabecular bone volume/total volume similarly in beta-less and WT mice. Altogether, these data indicate that absence of beta-adrenergic signaling results in obesity and increased cortical bone mass in males but does not prevent deleterious effects of estrogen deficiency on trabecular bone microarchitecture. Our findings also suggest direct positive effects of weight and/or leptin on bone turnover and cortical bone structure, independent of adrenergic signaling.
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Densidad Ósea/genética , Ovariectomía/efectos adversos , Receptores Adrenérgicos beta/deficiencia , Envejecimiento/fisiología , Animales , Fenómenos Biomecánicos , Composición Corporal/fisiología , Femenino , Fémur/anatomía & histología , Fémur/diagnóstico por imagen , Fémur/fisiología , Vértebras Lumbares/diagnóstico por imagen , Masculino , Ratones , Ratones Noqueados , Columna Vertebral/anatomía & histología , Columna Vertebral/fisiología , Tomografía Computarizada por Rayos XRESUMEN
Evidence that leptin regulates bone turnover in part through a central nervous system (CNS)/beta-adrenergic system relay has driven attention towards the potential therapeutic benefits of beta-adrenergic blockade to improve bone mass and strength. beta2- adrenergic receptor-mediated signaling in osteoblasts inhibits bone formation and triggers RANKL-mediated osteoclastogenesis and bone resorption. Mouse models of adrenergic-deficiency, particularly the mouse lacking the beta2-adrenergic receptor, have increased bone mass, more specifically increased trabecular bone volume. In turn, beta-blockers, such as propranolol, were reported to inhibit ovariectomy-induced bone loss. In contrast, a number of experiments in mice and rats suggest that inhibition of beta-adrenergic receptor-mediated signaling does not improve, and could actually be detrimental, for bone mass and microstructure. In humans, epidemiological observations suggested that users of beta-blockers have higher bone mineral density (BMD) and/or a reduced risk of fractures, yet not all studies were concordant. Here we review the evidence for a role of the adrenergic system in the regulation of bone metabolism in vitro and in vivo and provide some new evidence for a dual role of beta-adrenergic receptors 1 and 2 on bone turnover. Furthermore, we will examine the similarities and disparities that may exist in the effects of beta-adrenergic and PTH stimulation on bone metabolism.
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Adrenérgicos/uso terapéutico , Remodelación Ósea/fisiología , Osteoporosis/tratamiento farmacológico , Receptores Adrenérgicos/metabolismo , Transducción de Señal/fisiología , Agonistas Adrenérgicos/farmacología , Antagonistas Adrenérgicos/farmacología , Animales , Resorción Ósea/fisiopatología , Huesos/anatomía & histología , Huesos/efectos de los fármacos , Huesos/metabolismo , Huesos/fisiología , Humanos , Osteoclastos/efectos de los fármacos , Hormona Paratiroidea/metabolismo , Receptores Adrenérgicos beta 1/metabolismo , Receptores Adrenérgicos beta 2/metabolismoRESUMEN
Increased bone formation by PTH mainly results from activation of osteoblasts, an effect largely mediated by the cAMP-PKA pathway. Other pathways, however, are likely to be involved in this process. In this study we investigated whether PTH can activate p38 MAPK and the role of this kinase in osteoblastic cells. Bovine PTH(1-34) and forskolin markedly increased alkaline phosphatase (ALP) activity and doubled osteocalcin (Oc) expression in early differentiating MC3T3-E1 cells. These effects were associated with increase in cellular cAMP and activation of the MAP kinases ERK and p38. Activation of these MAP kinases was detectable after 1 h incubation with 10(-7) M PTH and lasted 1-2 h. Activation of p38 was mimicked by 10 microM forskolin and prevented by H89 suggesting a cAMP-PKA-dependent mechanism of p38 activation. Interestingly, PTH-induced ALP stimulation was dose-dependently inhibited by a specific p38 inhibitor with no change in the generation of cAMP and the production of osteocalcin. Similar inhibitory effect was obtained in cells stably expressing a dominant-negative p38 molecule. Finally, treatment of MC3T3-E1 cells with PTH for 3 weeks significantly enhanced matrix mineralization and this effect was markedly reduced by a selective p38 but not a specific MEK inhibitor. In conclusion, data presented in this study indicate that PTH can activate p38 in early differentiating osteoblastic cells. Activation of p38 is cAMP-PKA-dependent and mediates PTH-induced stimulation of ALP which plays a critical role for the calcification of the bone matrix.
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Fosfatasa Alcalina/metabolismo , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Hormona Paratiroidea/farmacología , Fragmentos de Péptidos/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Células 3T3 , Animales , Matriz Ósea/efectos de los fármacos , Matriz Ósea/metabolismo , Bovinos , Diferenciación Celular , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Imidazoles/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Osteoblastos/citología , Osteocalcina/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Piridinas/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidoresRESUMEN
Subjects with type 1 diabetes mellitus (T1DM) have decreased bone mineral density and an up to sixfold increase in fracture risk. Yet bone fragility is not commonly regarded as another unique complication of diabetes. Both animals with experimentally induced insulin deficiency syndromes and patients with T1DM have impaired osteoblastic bone formation, with or without increased bone resorption. Insulin/IGF1 deficiency appears to be a major pathogenetic mechanism involved, along with glucose toxicity, marrow adiposity, inflammation, adipokine and other metabolic alterations that may all play a role on altering bone turnover. In turn, increasing physical activity in children with diabetes as well as good glycaemic control appears to provide some improvement of bone parameters, although robust clinical studies are still lacking. In this context, the role of osteoporosis drugs remains unknown.
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Enfermedades Óseas Metabólicas/etiología , Diabetes Mellitus Tipo 1/complicaciones , Fracturas Óseas/etiología , Adiposidad/fisiología , Densidad Ósea , Enfermedades Óseas Metabólicas/metabolismo , Enfermedades Óseas Metabólicas/terapia , Médula Ósea/metabolismo , Médula Ósea/patología , Remodelación Ósea/fisiología , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 1/terapia , Fracturas Óseas/metabolismo , Fracturas Óseas/prevención & control , Humanos , Factores de RiesgoRESUMEN
Intermittent PTH administration increases bone turnover, resulting in net anabolic effects on bone. These effects are primarily mediated by intracellular cAMP signaling. However, the molecular mechanisms that regulate PTH activity in bone remain incompletely understood. beta-Arrestin2, a G protein-coupled receptor regulatory protein, inhibits PTH-stimulated cAMP accumulation in vitro. Using beta-arrestin2(-/-) (KO) and wild-type (WT) mice, we investigated the response to PTH in primary osteoblasts (POB) and the effects of intermittent PTH administration on bone mass and microarchitecture in vivo. Compared with that in WT mice, PTH-stimulated intracellular cAMP was increased and sustained in KO POB. Intermittent exposure of POB to PTH significantly decreased the ratio of osteoprotegerin (OPG) receptor activator of nuclear factor-kappaB ligand (RANKL) mRNA expression in KO POB, whereas it increased this ratio in WT POB. Total body bone mass and cortical and trabecular bone parameters were 5-10% lower in male KO mice compared with WT, and these differences were magnified upon in vivo administration of intermittent PTH (80 mug/kg.d) for 1 month. Thus, PTH significantly increased total body bone mineral content as well as vertebral trabecular bone volume and thickness in WT, but not KO mice. The anabolic response to PTH in cortical bone was also slightly more pronounced in WT than KO mice. Histomorphometry indicated that PTH prominently stimulated indexes of bone formation in both WT and KO mice, whereas it significantly increased indexes of bone resorption (i.e. osteoclast number and surface) in KO mice only. In conclusion, these results suggest that beta-arrestins may specify the activity of intermittent PTH on the skeleton by limiting PTH-induced osteoclastogenesis.
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Arrestinas/fisiología , Huesos/efectos de los fármacos , Hormona Paratiroidea/farmacología , Animales , Densidad Ósea/efectos de los fármacos , Calcio/sangre , Proteínas Portadoras/genética , Glicoproteínas/genética , Masculino , Glicoproteínas de Membrana/genética , Ratones , Ratones Endogámicos C57BL , Osteoblastos/efectos de los fármacos , Osteoprotegerina , Fosfatos/sangre , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Receptores Citoplasmáticos y Nucleares/genética , Receptores del Factor de Necrosis Tumoral , beta-ArrestinasRESUMEN
Mutations in the low-density lipoprotein receptor-related protein 5 gene (LRP5) have demonstrated the role of LRP5 in bone mass acquisition. LRP5 variants were recently reported to contribute to the population-based variance in vertebral bone mass and size in males. To investigate whether LRP5 variants are implicated in idiopathic male osteoporosis, we studied 78 men with low BMD (<2.5 T score or < -2 Z score) aged less than 70 years (mean +/- SD: 50 +/- 16 years) in whom secondary causes of osteoporosis had been excluded and 86 controls (51 +/- 10 years). Genotypes and haplotypes were based on LRP5 missense substitutions in exons 9 (c.2047G > A, p.V667M) and 18 (c.4037C > T, p.A1330V), and their association with osteoporosis evaluated after adjustment for multiple clinical and environmental variables using logistic regression. The presence of osteoporosis was significantly associated with LRP5 haplotypes (P = 0.0036) independent of age (P = 0.006), weight (P = 0.004), calcium intake (P = 0.002), alcohol (P = 0.005) and tobacco (P = 0.004) consumption. Accordingly, the odds ratio for osteoporosis was 3.78 (95% CI 1.27-11.26, P < 0.001) in male carriers of haplotype 3 (c.2047A-4037T, n = 20 cases and 12 controls) versus homozygous carriers of haplotype 1 (c.2047G-4037C, n = 42 cases and 61 controls). In conclusion, these data indicate beyond a significant role for environmental factors, an association between LRP5 variants and idiopathic osteoporosis in males, pointing to a role of LRP5 in this disease.
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Proteínas Relacionadas con Receptor de LDL/genética , Osteoporosis/genética , Polimorfismo Genético , Adulto , Anciano , Sustitución de Aminoácidos , Estudios de Casos y Controles , Predisposición Genética a la Enfermedad , Haplotipos , Humanos , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad , Masculino , Persona de Mediana Edad , Mutación MissenseRESUMEN
PTH promotes endocytosis of human PTH receptor 1 (PTH1Rc) by activating protein kinase C and recruiting beta-arrestin2. We examined the role of beta-arrestin2 in regulating the cellular distribution and cAMP signaling of two constitutively active PTH1Rc mutants, H223R and T410P. Overexpression of a beta-arrestin2-green fluorescent protein (GFP) conjugate in COS-7 cells inhibited constitutive cAMP accumulation by H223R and T410P in a dose-dependent manner, as well as the response to PTH of both mutant and wild-type PTH1Rcs. The cellular distribution of PTH1Rc-GFP conjugates, fluorescent ligands, and ssarrestin2-GFP was analyzed by fluorescence microscopy in HEK-293T cells. In cells expressing either receptor mutant, a ligand-independent mobilization of beta-arrestin2 to the cell membrane was observed. In the absence of ligand, H223R and wild-type PTH1Rcs were mainly localized on the cell membrane, whereas intracellular trafficking of T410P was also observed. While agonists promoted beta-arrestin2-mediated endocytosis of bot PTH1Rc mutants, antagonists were rapidly internalized only with T410P. The protein kinases inhibitor, staurosporine, significantly decreased internalization of ligand-PTH1Rc mutant complexes, although the recruitment of beta-arrestin2 to the cell membrane was unaffected. Moreover, in cells expressing a truncated wild-type PTH1Rc lacking the C-terminal cytoplasmic domain, agonists stimulated translocation of beta-arrestin2 to the cell membrane followed by ligand-receptor complex internalization without associated beta-arrestin2. In conclusion, cAMP signaling by constitutively active mutant and wild-type PTH1Rcs is inhibited by a receptor interaction with beta-arrestin2 on the cell membrane, possibly leading to uncoupling from G(s)alpha. This phenomenon is independent from protein kinases activity and the receptor C-terminal cytoplasmic domain. In addition, there are differences in the cellular localization and internalization features of constitutively active PTH1Rc mutants H223R and T410P.
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Arrestinas/farmacología , AMP Cíclico/metabolismo , Mutación , Receptores de Hormona Paratiroidea/análisis , Transducción de Señal/efectos de los fármacos , Animales , Arrestinas/genética , Arrestinas/metabolismo , Células COS , Membrana Celular/química , Membrana Celular/metabolismo , Endocitosis/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Proteínas Fluorescentes Verdes , Humanos , Proteínas Luminiscentes/genética , Microscopía Fluorescente , Inhibidores de Proteínas Quinasas , Receptor de Hormona Paratiroídea Tipo 1 , Receptores de Hormona Paratiroidea/genética , Receptores de Hormona Paratiroidea/metabolismo , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/metabolismo , Estaurosporina/farmacología , Transfección , beta-ArrestinasRESUMEN
Parathyroid hormone-related protein (PTHrP) plays a prominent role in the pathogenesis of humoral hypercalcemia of malignancy. However, it is also expressed in various nonmalignant tissues, particularly during fetal organogenesis and tissue differentiation. Thus, PTHrP is synthesized in skin, placenta, and mammary gland during lactation. Little is known, however, about the regulation of PTHrP synthesis and release in nontumoral cells. We investigated the regulation of PTHrP production by epidermal growth factor (EGF), a factor of major importance in the development of lactating breast, in primary cultures of rat mammary epithelial cells. EGF stimulated the production of immunoreactive and bioactive PTHrP in a time- and concentration-dependent manner. A 12 h incubation with 10 ng/ml of EGF increased PTHrP production by 36.0 +/- 7.1% (n = 7 experiments, p < 0.01). This was accompanied by an increase in PTHrP mRNA steady-state levels. The production of PTHrP was stimulated by the protein kinase C (PKC) activator phorbol-12-myristate-13-acetate (PMA) by 82.9 +/- 9.7% (n = 4 experiments, p < 0.01). The effects of PMA and EGF were additive. The EGF-induced stimulation appeared to be independent of extracellular calcium concentration, prostaglandin, or cAMP synthesis, but may have involved tyrosine kinase-mediated mechanisms. These results indicate that EGF was capable of increasing the production of PTHrP by cultured mammary epithelial cells. They also suggest that factors activating the PKC pathway are involved in the upregulation of PTHrP expression in mammary epithelial cells.
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Factor de Crecimiento Epidérmico/farmacología , Glándulas Mamarias Animales/efectos de los fármacos , Biosíntesis de Proteínas , Animales , Calcio/metabolismo , Células Cultivadas , Medios de Cultivo Condicionados , AMP Cíclico/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Indometacina/farmacología , Lactancia , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/metabolismo , Proteína Relacionada con la Hormona Paratiroidea , Prostaglandinas/metabolismo , Proteínas/efectos de los fármacos , Proteínas/genética , Ratas , Ratas Wistar , Acetato de Tetradecanoilforbol/farmacología , Regulación hacia ArribaRESUMEN
Whether vitamin D receptor (VDR) gene polymorphisms are associated with osteoporosis is highly controversial. The relationship between VDR gene polymorphisms and bone mineral density (BMD) might, however, be modified by age-related and/or environmental factors. We studied the potential association between BMD and VDR genotypes in females from prepuberty to premenopause and prospectively investigated the interaction of VDR genotypes with dietary calcium and BMD changes during childhood. Bsm I VDR gene polymorphisms and BMD at the lumbar spine (LS) and femur (neck [FN] and midshaft [FS]) were assessed in 369 healthy Caucasian females, aged 7-56 years (143 prepubertal girls, 54 peri- and postpubertal adolescents, and 172 premenopausal adults). Femoral trochanter (FT) and distal radius BMD (metaphysis and diaphysis) were also measured in 101 of the prepubertal girls who participated in a 1-year, double-blind, randomized study of calcium supplementation (850 mg/day) versus placebo on bone mineral mass accrual. Among all females, 150 (40.7%) had bb, 167 (45.3%) Bb, and 52 (14%) BB VDR genotypes. In prepubertal and adolescent girls altogether, LS BMD (Z scores) was associated with VDR genotypes and was significantly lower in BB than in Bb or bb subjects. Trends for a similar difference were also detected at the FN level as well as on the mean BMD (Z scores) of the three sites measured (LS, FN, and FS). By contrast, no BMD differences were detectable among VDR genotypes in the adults. In 101 prospectively studied prepubertal girls, calcium supplementation significantly increased BMD at most skeletal sites, except LS. After segregation for VDR genotypes (40 bb, 47 Bb, and 14 BB), a significant calcium effect was present in Bb but not bb girls, whereas in BB girls there was a positive but nonsignificant trend for a calcium effect. Moreover, dietary calcium intake was significantly correlated with BMD changes at various independent bone sites in Bb girls but not in bb girls. In contrast, BMD gain in bb girls appeared to be higher than among the other genotypes when the dietary calcium intake was low, i.e., in the absence of calcium supplements. BMD was significantly associated with VDR gene polymorphisms only before puberty, BB girls having significantly lower BMD (Z scores) than the other genotypes. By increasing dietary calcium intake, BMD accrual was increased in Bb and possibly BB prepubertal girls, whereas bb subjects had the highest spontaneous BMD accrual and remained unaffected by calcium supplements. Taking into account complex interactions between VDR gene polymorphisms and environmental factors, including calcium intake, may thus help to understand the discordant relationships between BMD and VDR gene polymorphisms.
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Envejecimiento/fisiología , Densidad Ósea/fisiología , Calcio de la Dieta/administración & dosificación , Suplementos Dietéticos , Receptores de Calcitriol/genética , Adolescente , Adulto , Envejecimiento/genética , Estatura , Peso Corporal , Niño , Estudios de Cohortes , Estudios Transversales , Femenino , Fémur , Cuello Femoral , Genotipo , Humanos , Vértebras Lumbares , Persona de Mediana Edad , Polimorfismo Genético , Premenopausia/genética , Estudios Prospectivos , Pubertad/genética , Radio (Anatomía) , SuizaRESUMEN
Cadherins are a family of cell surface adhesion molecules that play an important role in tissue differentiation. A limited repertoire of cadherins has been identified in osteoblasts, and the role of these molecules in osteoblast function remains to be elucidated. We recently cloned an osteoblast-derived N-cadherin gene from a rat osteoblast complementary DNA library. After in situ hybridization of rat bone and immunohistochemistry of human osteophytes, N-cadherin expression was localized prominently in well-differentiated (lining) osteoblasts. Northern blot hybridization in primary cultures of fetal rat calvaria and in human SaOS-2 and rat ROS osteoblast-like cells showed a relationship between N-cadherin messenger RNA expression and cell-to-cell adhesion, morphological differentiation, and alkaline phosphatase and osteocalcin gene expression. Treatment with a synthetic peptide containing the His-Ala-Val (HAV) adhesion motif of N-cadherin significantly decreased bone nodule formation in primary cultures of fetal rat calvaria and inhibited cell-to-cell contact in rat osteoblastic TRAB-11 cells. HAV peptide also regulated the expression of specific genes such as alkaline phosphatase and the immediate early gene zif268 in SaOS-2 cells. Transient transfection of SaOS-2 cells with a dominant-negative N-cadherin mutant (NCADdeltaC) significantly inhibited their morphological differentiation. In addition, aggregation of NCTC cells derived from mouse connective tissue stably transfected with osteoblast-derived N-cadherin was inhibited by either treatment with HAV or transfection with NCADdeltaC. Together, these results strongly support a role for N-cadherin, in concert with other previously identified osteoblast cadherins, in the late stages of osteoblast differentiation.
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Cadherinas/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , Osteoblastos/citología , Osteoblastos/fisiología , Animales , Diferenciación Celular/fisiología , Línea Celular , Humanos , Inmunohistoquímica , Hibridación in Situ , Ratones , ARN Mensajero/análisis , Ratas , TransfecciónRESUMEN
IL-6 is a pleiotropic cytokine that plays a critical role in bone resorption. We describe two allelic variants in the IL-6 promoter, -572 and -174 G-->C, that alone and in combination influence IL-6 activity in vitro and in vivo. The association of IL-6 -572 genotypes and -572/-174 G-->C haplotypes with serum C-reactive protein (CRP), serum and urinary C-terminal cross-linking of type I collagen (a marker of bone resorption), and osteocalcin (a marker of bone formation) was investigated in a cohort of healthy postmenopausal women (n = 495; mean age +/- SD, 72 +/- 5.7 yr). Among IL-6 -572 genotypes, CRP was 37% higher (P = 0.02) and urinary C-terminal cross-linking of type I collagen was 20% higher (P = 0.01) in the presence of the C allele, whereas serum osteocalcin was not different. IL-6 -572/-174 haplotypes (G/C, G/G, and C/G) were significantly associated with all biochemical markers, and additive effects of the two polymorphic loci were found. Thus, there was a significant increase in the level of CRP (up to +79%; P = 0.007) and bone resorption markers (up to +32%; P = 0.017) with a decreasing number (from four to one) of IL-6 protective alleles -572G and -174C. In addition, there was a trend for lower age-adjusted bone mineral density at the lumbar spine in subjects with less IL-6 protective alleles (up to -9.6%; P = 0.037; P = 0.08 after further adjustment for weight). In conclusion, we describe two functional polymorphisms in the IL-6 gene regulatory region associated with IL-6 activity in postmenopausal women, both systemically (CRP) and locally in bone. As such, IL-6 polymorphisms are able to influence the risk of osteoporosis as well as other chronic disorders involving IL-6 activity.
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Proteína C-Reactiva/análisis , Regulación de la Expresión Génica/genética , Interleucina-6/genética , Osteoporosis Posmenopáusica/sangre , Osteoporosis Posmenopáusica/genética , Polimorfismo Genético/fisiología , Regiones Promotoras Genéticas/genética , Anciano , Alelos , Sustitución de Aminoácidos , Biomarcadores/sangre , Densidad Ósea/fisiología , Remodelación Ósea/genética , Remodelación Ósea/fisiología , Cisteína , Femenino , Genotipo , Glicina , Haplotipos , Humanos , Técnicas In VitroRESUMEN
Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue. At a given age, bone mass results from the amount of bone acquired during growth, i.e. the peak bone mass (Bonjour et al., 1991, Theintz et al. 1992) minus the age-related bone loss which particularly accelerates after menopause. The rate and magnitude of bone mass gain during the pubertal years and of bone loss in later life may markedly differ from one skeletal site to another, as well as from one individual to another. Bone mass gain is mainly related to increases in bone size, that is in bone external dimensions, with minimal changes in bone microarchitecture. In contrast, postmenopausal and age-related decreases in bone mass result from thinning of both cortices and trabeculae, from perforation and eventually disappearance of the latter, leading to significant alterations of the bone microarchitecture (Fig. 1).