Serum Levels Of Receptor Activator Of Nuclear factor-kappaB Ligand Research Articles

Evaluation of bone formation using recombinant human bone morphogenetic proteins-7 in small maxillofacial bony defects.

Context:Bone morphogenetic proteins (BMP) are multifunctional molecules of transforming growth factor-β superfamily that induces the differentiation of fibroblasts into osteoblasts to form bone.Aims:This study was undertaken to evaluate the effects of recombinant human BMP-7 (rhBMP-7) in bone healing of small maxillofacial bone defects and assess the serum levels of osteopontin (OPN) and receptor activator of nuclear factor kappa-B ligand (RANKL) biomarkers for bone remodeling.Materials and Methods:Twenty patients with small maxillofacial bony defects were enrolled in this study and randomly allocated to two groups; wherein after apicoectomy of the involved teeth, the control group had defect filled with collagen sponge only while the experimental group had rhBMP-7 impregnated collagen sponge placed in the defect.Results:The clinical parameters showed no significant difference between the two groups (P > 0.05). The radiographic parameters showed a significantly slower rate of reduction in bone defect volume (P < 0.01) in control group than the experimental group when followed at 2, 4 and 24 postoperative weeks. RANKL and OPN serum levels showed no significant changes in pre- and post-operative stage.Conclusion:This study confirms that rhBMP-7 in collagen definitely accelerates bone healing in maxillofacial bone defects and minimizes postoperative complications. RANKL and OPN biomarkers in serum may not show bone remodeling, hence tissue samples may be used to assess their levels.

A STUDY OF SERUM LEVELS OF RANKL AND OPG INPATIENTS WITH ACUTE CORONARY SYNDROME WITH AND WITHOUT PSORIATIC ARTHRITIS

Receptor Activator of Nuclear Factor Kappa B Ligand (RANKL) is a member of the tumor necrosis factor alpha(TNFa) family.It signals by its RANK receptor on monocyte/macrophage progenitors and stimulates the formation of bone-resorbing osteoclasts [1]. In psoriatic arthritis (PsA) RANKL is produced by activated Th17 cells, synovial fibroblasts, macrophages, dendritic cells and activated lymphocytes [2,3]. These are cells involved in systemic and local inflammation and atherogenesis.Osteoprotegerin (OPG) is a glycoprotein first described in 1997 by Simonet et al. [1]who refer to it as a bone-protecting protein. OPG is a new member of theTNFa superfamily,a soluble receptor which acts as a decoy for RANKL and other ligands. It is also synthesized in many other tissues such as kidney, placenta, bones, vessels, lungs, and vascular structures. It circulates in serum and in plasma, although the concentration is much lower than locally in the bone and arterial tissue[4,5,6]. It has been found that vascular smooth muscle cellssynthesize it as a result of various stimuli. OPG also protects large blood vessels from medialcalcification[7].The biological effect of OPG is to oppose RANKL and thus prevent subsequent stimulation of the receptor, therefore the level of OPG increases and acts as a mechanism of self-defense.The role of RANKL and OPG in cardiovascular disease is still insufficiently defined and remains contradictory[8]. Also, the relationship between high serum levels of OPG in acute coronary syndrome( ACS) and their relation to inflammatory joint disease (IJD) is still discussed [9]. Evidence has been collected demonstrating the active involvement of RANKL and OPG in vascular pathology, including atherogenesis, arterial calcification and plaque instability.Epidemiological studies show that elevated OPG levels in the circulation have a prognostic function in cardiovascular mortality and morbidity and may be a serum prognostic marker for future vascular events. On the other hand, the involvement ofRANKL and OPG in the pathogenesis of local and systemic inflammation in ACS and IJD could link them to the amplification of cardiovascular risk and the severity of acute coronary syndromes, especially in patients with PsA. It is possible that the serum OPG level is increased in response to vascular wall damage and ongoing inflammation process within the atherosclerotic plaque lesion as a component of a complex compensatory mechanism in which RANKL plays a central role [11]. Serum OPG levels may be indicative of persistent damage of endothelial cells as well as the activation of VSMCs in advanced atherosclerotic plaque lesion.Together with other inflammatory mediators, RANKL and OPG can play a crucial role in the formation of the atherosclerotic plaque, its maturation and calcification[12, 13]. The present study aimed to examine and compare levels of sRANKL and sOPG in patients with ACS, divided into two groups, with and without PsA.Associations with cardiovascular mortality risk measured by the Global Registry of Acute Coronary Events (GRACE) were sought.ACS patients with and without PsA were compared in relation to sRANKL (24 and 48hrs) and sOPG (24 and 48hrs) to explore the effect of ACS.

Effect of esculetin on bone metabolism in ovariectomized rats

OBJECTIVETo determine the effect of an esculetin formulation (at 97.4% purity) on osteoporosis, and to investigate the potential underlying molecular mechanism(s). METHODSSixty specific pathogen free-grade female Wistar rats were randomly assigned to three groups: blank control (n = 12), sham (n = 12), and model (n = 36). The model group were bilaterally ovariectomized. The sham group had the tissue surrounding the ovaries removed, while the ovaries were retained. After 3 months, the model group was randomly divided into three subgroups: OVX (n = 12), positive control (n = 12), and esculetin (n = 12). The positive control group and the esculetin group were intragastrically administered diethylstilbestrol (0.046 mg • kg−1 • d−1) or esculetin (384 mg • kg−1 • d−1), respectively, once per day for 6 consecutive days; medication administration was then stopped for 1 d, before being administered for another 6 consecutive days. All rats were treated for 3 months. Samples were collected at the end of the treatment period. An Osteocore3 Digital 2D bone densitometer was used to test the bone mineral density, and histomorphometric analysis was performed to measure bone mass, bone formation, and bone resorption. Enzyme-linked immunosorbent assay analysis was used to measure the serum concentrations of interleukin-6 (IL-6), osteoprotegerin (OPG), and receptor activator of nuclear factor-kappa B ligand (RANKL). Immunohistochemistry and in situ hybridization were performed to detect the protein and mRNA expressions of OPG and RANKL in osteoblasts and bone marrow stromal cells. RESULTSCompared with the OVX group, the esculetin group had significantly greater femoral bone mineral density and tibial trabecular bone volume, and significantly smaller trabecular resorption surface. The percentage of trabecular formation surface, average osteoid width, trabecular bone mineralization rate, and cortical bone mineralization rate did not significantly differ between groups. Compared with the sham group, the esculetin group had significantly decreased serum levels of IL-6 and RANKL, and significant downregulation of RANKL protein and mRNA expression levels in osteoblasts and bone marrow stromal cells; however, there was no significant difference between groups in OPG. CONCLUSIONEsculetin can increase bone mass by upregulating RANKL expression in osteoblasts and bone marrow stromal cells, and decreasing serum IL-6 concentration. This indicates that the therapeutic effect of esculetin on osteoporosis occurs via decreased bone resorption.

Influences of donor and host age on human muscle-derived stem cell-mediated bone regeneration

BackgroundHuman muscle-derived stem cells (hMDSCs) have been shown to regenerate bone efficiently when they were transduced with Lenti-viral bone morphogenetic protein 2 (LBMP2). However, whether the age of hMDSCs and the animal host affect the bone regeneration capacity of hMDSCs and mechanism are unknown which prompted the current study.MethodsWe isolated three gender-matched young and old populations of skeletal muscle stem cells, and tested the influence of cells’ age on in vitro osteogenic differentiation using pellet culture before and after Lenti-BMP2/green fluorescent protein (GFP) transduction. We further investigated effects of the age of hMDSCs and animal host on hMDSC-mediated bone regeneration in a critical-size calvarial bone defect model in vivo. Micro-computer tomography (CT), histology, and immunohistochemistry were used to evaluate osteogenic differentiation and mineralization in vitro and bone regeneration in vivo. Western blot, quantitative polymerase chain reaction (PCR), and oxidative stress assay were performed to detect the effects of age of hMDSCs on cell survival and osteogenic-related genes. Serum insulin-like growth factor 1 (IGF1) and receptor activator of nuclear factor-kappa B ligand (RANKL) were measured with an enzyme-linked immunosorbent assay (ELISA).ResultsWe found LBMP2/GFP transduction significantly enhanced osteogenic differentiation of hMDSCs in vitro, regardless of donor age. We also found old were as efficient as young LBMP2/GFP-transduced hMDSCs for regenerating functional bone in young and old mice. These findings correlated with lower phosphorylated p38MAPK expression and similar expression levels of cell survival genes and osteogenic-related genes in old hMDSCs relative to young hMDSCs. Old cells exhibited equivalent resistance to oxidative stress. However, both young and old donor cells regenerated less bone in old than young hosts. Impaired bone regeneration in older hosts was associated with high bone remodeling due to higher serum levels of RANKL and lower level of IGF-1.ConclusionhMDSC-mediated bone regeneration was not impaired by donor age when hMDSCs were transduced with LBMP2/GFP, but the age of the host adversely affected hMDSC-mediated bone regeneration. Regardless of donor and host age, hMDSCs formed functional bone, suggesting a promising cell resource for bone regeneration.

Berberine alleviates oxidative stress in rats with osteoporosis through receptor activator of NF-kB/receptor activator of NF-kB ligand/osteoprotegerin (RANK/RANKL/OPG) pathway.

Previous studies suggested that oxidative stress is related to the onset and development of osteoporosis. Moreover, it was demonstrated that berberine has a protective effect against oxidative stress-induced injuries. In this study, we aimed to investigate the effect and mechanism of action of berberine on rats with induced osteoporosis. Sixty 8-week-old female Wistar rats were randomly divided into the following 6 groups: control saline-treated, osteoporosis saline-treated, 3 osteoporosis berberine-treated groups (Ber 5, 10, and 20 mg/kg/body weight, respectively), and osteoporosis alendronate-treated (ALD) group. Osteoporosis was induced by bilateral ovariectomy. All treatments were performed for 8 weeks. The bone mineral density (BMD), serum alkaline phosphatase (ALP), osteocalcin, calcium, phosphorus, superoxide dismutase (SOD), methylenedioxyamphetamine (MDA), and glutathione peroxidase (GSH-Px) level was determined in the rat femur tissue. The gene and protein expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) was analyzed by quantitative reverse transcription PCR and Western blot, respectively. The BMD, SOD and GSH⁃Px levels, and the expression of OPG were significantly lower in osteoporosis compared to control group (all p < 0.05). The serum levels of osteocalcin, ALP, and MDA, and the expression of RANKL were significantly higher in osteoporosis compared to control group (all p < 0.05). Berberine, especially the high doses of berberine, effectively increased SOD, GSH⁃Px, and OPG levels as well as decreased serum osteocalcin, ALP, MDA and RANKL levels in berberine-treated osteoporosis groups (all p < 0.05). To conclude, oxidative stress may promote the development of osteoporosis in rats through the RANK/RANKL/OPG pathway. The antioxidative effect of berberine reduces the development of osteoporosis in rats to some extent.

Abstract P6-07-11: Serum levels of RANKL are increased in primary breast cancer patients in the presence of disseminated tumor cells in the bone marrow

Abstract Background: Receptor activator of nuclear factor kappa-B ligand (RANKL) is an essential protein for osteoclast regulation that is associated with benign and malignant bone disease. The activity of RANKL is controlled by its soluble decoy receptor osteoprotegerin (OPG). There is increasing evidence that RANKL may also directly affect breast cancer progression and metastasis to the bone. This study was aimed to assess the levels of RANKL and OPG in 509 newly diagnosed breast cancer patients with regard to the presence of disseminated tumor cells (DTCs) in the bone marrow (BM), circulating tumor cells (CTCs) in blood and clinical parameters. Patients and Methods: 509 patients with first diagnosis of breast cancer between Aug 2006 and Dec 2009 were included in our study. Blood and BM sampling was performed before surgery in an adjuvant setting. Blood was collected from each patient and sRANKL as well as OPG in the serum were measured by ELISA (Immunodiagnostic, Vienna, Austria). Two BM aspirates were analyzed for DTCs by immunocytochemistry using the pan-cytokeratin antibody A45-B/B3. In a subgroup of 364 patients, 2 x 5 ml blood was studied for CTCs using the AdnaTest BreastCancer (QIAGEN, Hannover GmbH, Germany) for the detection of EpCAM, MUC-1, HER-2, and beta-Actin transcripts. Results: Mean serum values for RANKL and OPG were 0.23 ± 0.20 pmol/l and 4.24 ± 1.70 pmol/l, respectively. RANKL levels were significantly lower in women above 60 years of age (0.19 pmol/l vs 0.26 pmol/l; p &amp;lt; 0.0001). This finding was reflected by higher RANKL serum levels and RANKL/OPG ratios in premenopausal patients compared to peri- (p&amp;lt;0.05) and postmenopausal patients (p&amp;lt;0.001), respectively. RANKL/OPG ratios were also higher in patients with lymph node involvement (N1-N3, p=0.03). All other clinical parameters did not influence RANKL or OPG levels. DTCs were detected in 213/509 (42%) patients and CTCs in 81/364 (22%) patients, respectively. However, while RANKL levels were unchanged in patients with detectable CTCs, they significantly increased by 33% (p&amp;lt;0.0001) in patients with DTCs. There was no difference in OPG levels, resulting in an increased RANKL to OPG ratio in patients with DTCs in the BM (0.087 vs. 0.060; p &amp;lt; 0.0001). Conclusion: In conclusion, we show that RANKL serum levels and RANKL/OPG ratios are increased in patients with detectable DTCs in the BM, prior to the establishment of detectable bone metastases. This finding warrants further investigation as it may provide a rational for novel diagnostic or therapeutic approaches. Citation Format: Kasimir-Bauer S, Bittner A-K, Goebel A, Hoffmann O, Browne AJ, Rauner M, Hofbauer LC, Wimberger P, Kimmig R, Rachner TD. Serum levels of RANKL are increased in primary breast cancer patients in the presence of disseminated tumor cells in the bone marrow [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P6-07-11.

Chemotherapy effectively suppresses interleukin-20, receptor activator of nuclear factor kappa-B ligand, and osteoprotegerin levels in patients with lung adenocarcinoma and bone metastasis.

Bone metastasis (BM) is common in patients with lung cancer. Osteolysis is caused by increased osteoclast activity. Interleukin-20 (IL-20) and receptor activator of nuclear factor kappa-B ligand (RANKL) are crucial for osteoclast formation. Osteoprotegerin (OPG) inhibits a receptor activator of RANKL/RANK signaling. The aims of this study were to analyze the serum levels of IL-20, OPG, and RANKL in patients with and without BM and to observe the effect of chemotherapy on these cytokines. A total 54 cases of pathologically confirmed lung adenocarcinoma (ADC) and 18 healthy individuals (Control) were enrolled in this study. Eligible patients were divided into three groups (18 patients per group): ADC without BM (ADC), ADC with BM (ADC + BM), and ADC with BM treated with chemotherapy (ADC + BM + Chemo). Serum IL-20, RANKL, and OPG levels were analyzed by enzyme-linked immunosorbent assay. Serum IL-20, RANKL, and OPG levels in ADC + BM patients were significantly elevated compared with that in the Control or ADC groups (both P < 0.001). The serum cytokine levels were significantly lower following chemotherapy compared with that in patients who did not receive chemotherapy (P < 0.001). Serum IL-20, RANKL, and OPG levels increase in patients with lung cancer and BMs. Chemotherapy suppresses the elevation of these cytokines.

The Effects of Dietary Supplements of Calcium, Vitamin D and Estrogen Hormone on Serum Levels of OPG and RANKL Cytokines and their Relationship with Increased Bone Density in Rats.

Osteoprotegerin (OPG)-Receptor activator of nuclear factor kappa-B ligand (RANKL) pathway is one of the contributing factors in the regulation of osteogenesis and bone resorption routes. The purpose of this study was to evaluate the effects of various dietary supplements on this pathway. The samples for this study (24 newborn rats) were divided in three groups according to the experiment applied for each group. Rats were given special diet according to their group plan for six weeks. Blood samples were collected to measure their serum levels of OPG and RANKL and all organs of rats were used to measure their bone density too. The results were analysed using appropriate statistical analysing tests. Levels of whole-body bone mineral density in calcium plus vitamin D plus Estrogen (Ca + D + E) group and calcium plus vitamin D (Ca + D) group were significantly increased compared to control group. Mineral density was highest in calcium plus vitamin D plus Estrogen group and was about 0.1357 g/cm2. RANKL had a significant decrease in calcium plus vitamin D plus Estrogen group compared to control and calcium plus vitamin D groups. There was a significant increase in the mean calcium and OPG in both experimental groups rather than control. Also, significant increase in estrogen was observed in Ca + D group than the control group. The results showed that intake of calcium and vitamin D and estrogen at determined dose led to an increase in OPG and RANKL cytokines reduction which ultimately led to an increase in bone mineral density. But Ca, D and E synergies were more effective in increasing bone mineral density compared to only the use of Ca and D.

Effect of Astragalus polysaccharides on ovariectomy-induced osteoporosis in mice.

Postmenopausal osteoporosis, a common type of osteoporosis in women, has become a serious public health issue. Astragalus polysaccharides (APS), possessing various pharmacological activities, are the active ingredients of Radix Astragali. It can be advantageous in the treatment of postmenopausal osteoporosis. In the present study, we evaluated the potential therapeutic effects of APS on postmenopausal osteoporosis by using a mice model induced by ovariectomy (OVX). Forty-eight female 6-week-old outbred ICR mice were randomly divided into six groups (N = 8): Sham group, OVX group, 17 β-estradiol (E2, 0.1 mg/kg)-treated OVX group, and APS (at three doses: 100, 200, and 400 mg/kg)-treated OVX groups. The effect of APS on the bone mineral density (BMD) was determined using dual-energy X-ray absorptiometry. The serum levels of receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), osteocalcin, and tumor necrosis factor (TNF)-α were measured using ELISA kits. The results revealed that APS exerted significant anti-osteoporotic activity by increasing the BMD considerably in a dose-dependent manner. APS treatment reduced the serum RANKL levels considerably and increased the serum OPG levels, thereby lowering the ratio of RANKL/OPG. Furthermore, APS also markedly reduced osteocalcin and TNF-α concentration in OVX-induced postmenopausal osteoporosis mice model. These results showed that APS exerts a protective effect on bone loss in OVX mice. The molecular mechanism underlying this effect be the reduction of bone resorption and inhibition of osteoclastogenesis. Our findings suggest that APS may be a potential strategy for the prevention and treatment of postmenopausal osteoporosis.

Serum level of receptor activator of nuclear factor kappa-B ligand in patients with psoriasis.

Psoriasis usually precedes the onset of psoriatic arthritis by an average of 10 years in about 70% of patients. Receptor activator of nuclear factor kappa-B ligand (RANKL) is a natural and necessary surface-bound molecule, which is a ligand for osteoprotegrin and functions as a key factor for osteoclast differentiation and activation. Evaluation of the serum level of RANKL in psoriasis and psoriatic arthritis and its correlation with severity of the disease as a trial to predict the occurrence of psoriatic arthritis in such patients. This study included 80 subjects; 40 patients had chronic plaque psoriasis, 20 patients had chronic plaque psoriasis with psoriatic arthritis, and 20 were healthy controls. Patients with psoriasis were divided into three subgroups according to psoriasis area severity index score: mild, moderate, and severe. Serum RANKL levels were estimated for all subjects using enzyme-linked immunosorbent assay. Serum RANKL level in psoriatic arthritis was significantly higher compared to both patients with plaque psoriasis and control groups. Serum RANKL level significantly correlated to the severity of psoriasis, with a very high correlation coefficient in groups I and II. The diagnostic performance of serum RANKL level for the early prediction of psoriatic arthritis in patients with psoriasis was >170 pg/ml. Serum RANKL level could be considered as a useful diagnostic marker for the early prediction of psoriatic arthritis in patients with moderate and severe psoriasis. Assessment of soluble RANKL in psoriasis could identify those at increased risk for psoriatic arthritis, and anti-RANKL agents may be effective in decreasing incidence of psoriatic arthritis.

Serum osteopontin levels are upregulated and predict disability after an ischaemic stroke.

After an acute ischaemic stroke (AIS), several inflammatory biomarkers have been investigated, but their predictive role on functional recovery remains to be validated. Here, we investigated the prognostic relevance of biomarkers related to atherosclerotic plaque calcification, such as osteopontin (OPN), osteoprotegerin (OPG) and the receptor activator of nuclear factor kappa-B ligand (RANKL) in a cohort of patients with AIS (n = 90) during 90-day follow-up. Radiological and clinical examinations as well as blood sampling were performed at admission and at days 1, 7 and 90 from the event. Validated scores [such as modified Rankin scale (mRS) and the National Institutes of Health Stroke Scale (NIHSS)] were used to assess poststroke outcome. Serum levels of OPN, OPG and RANKL were measured by colorimetric enzyme-linked immunosorbent assay (ELISA). When compared to the admission, OPN serum levels increased at day 7. Serum OPN levels at this time point were positively correlated with both ischaemic lesion volume and NIHSS at days 7 and 90. A cut-off of 30.53 ng/mL was identified for serum OPN by receiver operating characteristic (ROC) curve analysis. Adjusted logistic regression showed that serum OPN levels at day 7 predicted worse mRS at day 90 [OR 4.13 (95% CI 1.64-10.36); P = 0.002] and NIHSS [1.49 (95% CI 1.16-1.99); P = 0.007], independently of age, gender, hypertension and thrombolysis. Serum levels of OPN, but not OPG and RANKL, peaked at day 7 after AIS and predicted worse neurological scores. Therefore, OPN might have a pathophysiological and clinical relevance after AIS.

Osteoprotegerin and RANKL in the pathogenesis of rheumatoid arthritis-induced osteoporosis

Osteoporosis represents an important cause of morbidity in adult rheumatoid arthritis (RA) patients who exhibit increased fracture risk. It is thought that osteoclast and its dysfunction which mediated by many cytokines are the principal pathogenesis of this bone disease, although the mechanisms are still not fully understood. Osteoprotegerin (OPG) and receptor activator of nuclear factor-kappa B ligand (RANKL) have been revealed in the pathogenesis of primary osteoporosis and other metabolic bone diseases. Thus, the aim of this study was to investigate the possible role of the OPG/RANKL system in RA-related bone loss. A total of 64 Chinese patients with RA and 60 healthy control subjects were involved. Serum levels of OPG and RANKL were measured by ELISA. BMD of nondominant forearm, lumbar spine(L(1-4)) and proximal femur, including femoral neck, Wards triangle, greater trochanter were assessed using dual-energy X-ray absorptiometry. RA patients had a higher incidence of osteoporosis (23/64, 35.9%) than that in healthy controls (9/60, 15.0%) (P < 0.0001). They displayed lower BMD values than controls at positions of all detected region. Compared with healthy controls, RA group showed significantly higher serum levels of RANKL (48.4 ± 12.5 vs. 23.0 ± 11.2 pmol/l, P < 0.0001), lower serum levels of OPG (106.2 ± 40.6 vs. 231.6 ± 65.6 pg/ml, P < 0.0001), and OPG/RANKL ratio (2.4 ± 0.7 vs. 7.0 ± 1.1, P < 0.0001). Multiple linear regression analysis revealed that in RA group, plasma rheumatoid factor concentration (β = -0.187, P = 0.031), swollen joint count (β = 0.567, P = 0.029), BMD at forearm (β = 0.324, P = 0.002), femoral Wards triangle (β = 0.370, P < 0.0001), and lumbar spine (β = 0.313, P = 0.003) were the contributors for serum OPG (R ( 2 ) = 0.718, P < 0.0001). Age (β = 0.241, P = 0.042) and BMD at femoral Wards triangle (β = -0.441, P < 0.0001) and lumbar spine (β = -0.320, P = 0.013) were the determinants for serum RANKL (R ( 2 ) = 0.616, P < 0.0001), while swollen joint count (β = 1.029, P = 0.019) and BMD at femoral neck (β = 0.285, P = 0.042) for serum OPG/RANKL ratio (R ( 2 ) = 0.279, P < 0.011). Analysis of logistic regression showed age (P = 0.004, OR = 1.156, 95% CI: 1.047-1.276) and the level of C-reactive protein (P = 0.028, OR = 1.019, CI 95%: 1.002-1.036) in peripheral blood of RA were the risk factors for the occurrence of osteoporosis in RA, while OPG/RANKL ratio (P = 0.007, OR = 0.035, CI 95%: 0.003-0.400) was the unique protective factor. These data suggest that, in Chinese RA patients, an altered modulation of the OPG/RANKL system resulting in increased RANKL and decreased OPG in peripheral blood, could contribute to the bone loss characteristic and the generation of osteoporosis in these patients. Changes of ratio of OPG/RANKL might be a protective mechanism against the accelerated bone loss in RA.

Soluble receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin in ankylosing spondylitis: OPG is associated with poor physical mobility and reflects systemic inflammation

The objective of the study was to investigate the role of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in ankylosing spondylitis (AS). Serum levels of soluble RANKL (sRANKL) and OPG were measured in 42 AS patients and 26 healthy controls. We evaluated the AS patient's disease activity, functional ability, global assessment, and physical mobility and tested markers of systemic inflammation, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) levels. Serum levels of sRANKL [mean (SD), 4.75 (1.88) vs. 3.70 (1.14) pmol/l, p = 0.015] and OPG [mean (SD), 5.18 (1.19) vs. 4.52 (0.85) pmol/l, p = 0.026] were significantly higher in the 42 AS patients than the 26 healthy controls. Interestingly, serum OPG levels correlated significantly with ESR (r = 0.417, p = 0.007), CRP (r = 0.524, p < 0.001), tragus-to-wall distance (r = 0.556, p < 0.001), fingertip-to-floor distance (r = 0.423, p = 0.007), and occiput-to-wall distance (r = 0.465, p = 0.002) and correlated inversely with modified Schober index (r = -0.525, p = 0.001), cervical rotation (r = -0.403, p = 0.022), lateral lumbar flexion (r = -0.587, p < 0.001), and chest expansion (r = -0.553, p < 0.001). Moreover, in the AS patients with higher (> or =4.925 pmol/l, n = 21) serum OPG levels, there were significant increases in the tragus-to-wall distance (p = 0.007), fingertip-to-floor distance (p = 0.023), and CRP levels (p = 0.014) and decreased in the modified Schober index (p = 0.012), lateral lumbar flexion (p = 0.019), and chest expansion (p = 0.005). Serum levels of sRANKL and OPG are increased in the AS patients and may participate in the disease process of AS. Production of OPG has association with poor physical mobility and may reflect systemic inflammation in AS.

Osteoclastogenesis in Children with 21-Hydroxylase Deficiency on Long-Term Glucocorticoid Therapy: The Role of Receptor Activator of Nuclear Factor-κB Ligand/Osteoprotegerin Imbalance

Children with 21-hydroxylase deficiency (21-OHD) need chronic glucocorticoid (cGC) therapy to replace congenital deficit of cortisol synthesis. cGC therapy is the most frequent and severe form of drug-induced osteoporosis, and different mechanisms have been proposed to explain its pathogenesis. We investigated the osteoclastogenic potential of peripheral blood mononuclear cells (PBMCs) from 18 children with 21-OHD on cGC therapy and 25 controls who never received GCs. We also evaluated the presence of circulating osteoclast precursors (OCPs) and the role of T cells in osteoclast formation. Spontaneous osteoclastogenesis, without adding macrophage-colony stimulating factor and receptor activator of nuclear factor-kappaB ligand (RANKL), and significantly higher osteoclasts resorption activity occurred in 21-OHD patients. Conversely, macrophage-colony stimulating factor and RANKL were essential to trigger and sustain osteoclastogenesis in controls. Furthermore, in 21-OHD patients, we identified a significant percentage of CD11b-CD51/CD61 and CD51/61-RANK-positive cells, which are OCPs strongly committed. Additionally, we demonstrated a T cell-dependent osteoclastogenesis from 21-OHD patients' PBMCs. T cells from patients expressed high levels of RANKL and low levels of osteoprotegerin (OPG) with respect to controls. Moreover, 21-OHD patients had higher soluble RANKL and lower OPG serum levels compared with controls; thus, soluble RANKL to OPG ratio was significantly higher in patients than controls. The present study showed for the first time a high osteoclastogenic potential of PBMCs from 21-OHD patients on cGC therapy. This spontaneous osteoclastogenesis seems to be supported by both the presence of circulating OCPs and factors released by T cells.

The effect of risedronate treatment on serum cytokines in postmenopausal osteoporosis: a 6-month randomized and controlled study

There is much evidence suggesting that the decline in ovarian function after menopause is associated with spontaneous increases in proinflammatory cytokines. Treatment with risedronate is accompanied by significant changes in bone turnover and bone mineral density. The objective of this study was to determine the effects of risedronate treatment on the level of serum cytokines including receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin among postmenopausal women with osteoporosis. The study group consisted of 61 postmenopausal women with osteoporosis. Patients were randomly divided in two groups: In group 1 (n = 41) postmenopausal women received oral risedronate (35 mg/week), calcium (1,000 mg/day), and vitamin D (400 IU/day) for 12 months. In group 2 (control group; n = 20) patients received only oral calcium (1,000 mg/day) and vitamin D (400 IU/day). Bone mineral density (BMD) of lumbar spine (L1-L4) and proximal femur were determined using dual X-ray absorptiometry at baseline and after one year. Venous blood samples were obtained for determination of serum cytokines including interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), RANKL, osteoprotegerin, and markers of bone formation and resorption. Levels of serum cytokines were measured before therapy and after three and 6 months. Markers of bone metabolism were studied before therapy and after 6 months. In group 1 (risedronate plus calcium/vitamin D-treated patients), serum levels of RANKL and IL-1beta significantly decreased and the level of osteoprotegerin significantly increased after three and 6 months, but no significant difference was found in TNF-alpha level. In group 2, however, the level of serum cytokines did not change after three and 6 months. In cases of bone turnover, both markers of bone resorption and formation significantly decreased after 6 months in group 1. In conclusion risedronate could improve osteoporosis by increasing osteoprotegerin and reducing RANKL and IL-1beta.

Relationships between OPG, RANKL, bone metabolism, and bone mineral density in biliary atresia

Osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL) have been implicated in osteoclastogenesis. However, the relationship between the OPG-RANKL system and bone status in biliary atresia (BA) has not, as yet, been clarified. Thus, the aim of this study has been to evaluate the relationship between the OPG-RANKL system and bone mineral metabolism in patients with BA. Fifty patients with BA and 13 healthy controls were investigated. The mean age of BA patients and controls was 7.3 +/- 0.6 and 8.0 +/- 1.1 years, respectively. Serum levels of OPG, RANKL, osteocalcin, and C-terminal telopeptide of type I collagen (CTX) were measured by sandwich enzyme-linked immunosorbent assay. Bone mineral density (BMD) of the lumbar spine was determined by dual energy X-ray absorptiometry. Biliary atresia patients had significantly elevated serum OPG levels compared with controls (4.0 +/- 0.3 vs. 3.0 +/- 0.3 pmol/L, P = 0.02) and serum OPG levels in BA patients with jaundice were higher than in those without jaundice (4.6 +/- 0.4 vs. 3.6 +/- 0.4 pmol/L, P = 0.04). Likewise, serum RANKL levels were significantly higher in BA patients than in controls (2.9 +/- 0.2 vs. 1.2 +/- 0.7 pmol/L, P = 0.001). In addition, serum RANKL levels were increased in BA patients with jaundice compared to those without jaundice, but this difference was not statistically significant (3.2 +/- 0.3 vs. 2.7 +/- 0.2 pmol/L, P = 0.2). The serum osteocalcin levels in BA patients were not significantly different from those in the healthy controls, whereas the serum CTX levels were elevated in BA patients compared with the controls (0.4 +/- 0.1 vs. 0.2 +/- 0.1 ng/mL, P = 0.02). Furthermore, BMD of BA children with jaundice was significantly lower than that of BA children without jaundice (P = 0.0005). BMD of BA patients was inversely correlated with serum levels of OPG (r = -0.452, P < 0.001). Elevated serum OPG levels are associated with reduced BMD and the outcome of BA. The increase of serum OPG in BA patients with severe disease could reflect a compensatory response to bone loss.

Changes in serum levels of receptor activator of nuclear factor-κB ligand, osteoprotegerin, IL-6 and TNF-α in patients with a concomitant head injury and fracture

Several reports indicated that interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF- alpha) play important regulatory roles in bone remodeling and homeostasis. In addition, receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) have been shown to be important regulators of osteoclastogenesis during bone remodeling, and their expressions were examined during fracture healing in a mouse model of tibial fracture. However, studies linking RANKL, OPG, IL-6 and TNF-alpha in patients with head injury and fracture are lacking. Within the first few hours of admission to hospital and at 4, 8, and 12 weeks after the injury, we evaluated changes in serum levels of RANKL, OPG, IL-6 and TNF-alpha in 24 male patients with a concomitant head injury and fracture and in 26 male patients with fracture only. These levels were compared with those found in 36 healthy controls. The RANKL/OPG ratios were found to significantly lower in patients with a concomitant head injury and fracture than in the controls immediately after admission and at 4, 8, and 12 weeks after the injury. In addition, RANKL/OPG ratios were significantly lower in patients with a concomitant head injury and fracture than in those with fracture at 8 and 12 weeks after the injury. The serum IL-6 levels were significantly higher in patients with a concomitant head injury and fracture than in the controls upon admission, and at 4, 8, and 12 weeks after the injury. Moreover, the serum IL-6 levels were significantly higher in patients with a head injury and fracture than in those with just a fracture at 4, 8, and 12 weeks after the injury. Based on these changes in the profiles of RANKL, OPG, and IL-6 and the RANKL/OPG ratio, altered repair of a fracture can occur in patients with a concomitant head injury and fracture.

An acute injection of Porphyromonas gingivalis lipopolysaccharide modulates the OPG/RANKL system and interleukin‐6 in an ovariectomized mouse model

In the present study, we attempted to develop a simulated model to explore the causal effects of periodontal pathogens on skeletal homeostasis in postmenopausal osteoporosis. Fifty-three female adult ICR mice were randomly assigned to an experimental group (ovariectomized) or a control group. A single injection of Porphyromonas gingivalis lipopolysaccharide (P. gingivalis-LPS, ATCC 33277) or Escherichia coli lipopolysaccharide (E. coli-LPS) was administered intraperitoneally 4 weeks after an ovariectomy. Concentrations of interleukin-6 (IL-6), osteoprotegerin (OPG), and the receptor activator of nuclear factor-kappaB ligand (RANKL) in serum were subsequently analyzed using an enzyme-linked immunosorbent assay (ELISA). Under stimulation with P. gingivalis-LPS or E. coli-LPS, the concentration of OPG rose in both groups. The serum level of RANKL showed a decreasing trend 24 h after the injection in both groups. After injection of P. gingivalis-LPS in both the experimental and control animals, the OPG : RANKL ratio increased 24 h after the booster (22.26-620.99, P < 0.05). The serum level of IL-6 in the experimental group significantly increased 1-6 h after administration of E. coli-LPS and 1-3 h after administration of P. gingivalis-LPS (P < 0.05). A single booster injection of P. gingivalis-LPS induced short-term changes in OPG, RANKL, and IL-6 serum levels in this ovariectomized mouse model.

Relationships between insulin-like growth factor-I (IGF-I) and OPG, RANKL, bone mineral density in healthy Chinese women

Serum IGF-I level was negatively correlated with OPG and OPG/RANKL ratio, but positively correlated with RANKL. Serum OPG level in the highest quintile of IGF-I was significantly lower than that in the lowest. We conclude that the effect of IGF-I on bone remodeling may be mediated by the OPG/RANKL system. Insulin-like growth factor I (IGF-I) is an important factor in coupling bone remodeling, activating both formation and resorption. Compared with the many studies on the role of IGF-I in bone formation, the information regarding its effects on bone resorption is limited and conflicting. The balance of the two peptides produced by osteoblasts, osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL), is critical for the bone resorption process. Our study was designed to analyze the relationships of serum concentrations of IGF-I with OPG, RANKL, OPG/RANKL ratio as well as BMDs in healthy Chinese women. BMDs at lumbar spine and proximal femur in 504 pre- and postmenopausal women were measured by DXA. Serum levels of IGF-I, OPG and RANKL were also measured. Pearson's correlation and partial correlation analysis, ANOVA, covariance analysis and stepwise multiple regression analysis were used as appropriate. Age was negatively correlated with serum levels of IGF-I (r = -0.702, p < 0.001). IGF-I was negatively correlated with OPG and OPG/RANKL ratio, but positively correlated with RANKL. The relationship between IGF-I and BMDs disappeared after adjustment for age. In postmenopausal women, IGF-I was lower in women with osteoporosis than in those with normal BMD (p = 0.056), but no differences were found among OPG, RANKL and OPG/RANKL ratio. Serum levels of OPG in the highest quintile of IGF-I were significantly lower than those in the lowest quintile of IGF-I, while no difference was found in RANKL. In the multiple regression analysis model, serum levels of IGF-I were the main determinants of the bone mass in Chinese women. In conclusion, the relationship between decreasing IGF-I and BMDs in healthy Chinese women influenced by age, whereas the effect of IGF-I on bone remodeling (bone resorption) may be mediated by the OPG/RANKL system.

Increased augmentation index and central aortic blood pressure in osteoporotic postmenopausal women

Osteoporosis has been associated with cardiovascular disease. We found increased augmentation index, a measure of wave reflections and arterial stiffness, and central pressures in osteoporotic postmenopausal women. They also showed a higher estimated aortic pulse wave velocity, indicating a stiffer aorta. These changes may increase cardiovascular risk in postmenopausal osteoporosis. Evidence suggests a link between osteoporosis and cardiovascular disease. We investigated whether augmentation index (AIx), a measure of pulse wave reflections and arterial stiffness, is increased and related to the osteoprotegerin (OPG)/receptor activator of nuclear factor-kappa B ligand (RANKL) system in postmenopausal osteoporosis. AIx and central aortic haemodynamics were assessed using pulse wave analysis in 182 cardiovascular disease-free osteoporotic postmenopausal women and in 160 controls. Statistical analysis was performed by unpaired t test, Mann-Whitney test, Spearman's correlation coefficient, and multivariate linear regression analysis. AIx (37.2 +/- 7.0 vs. 29.6 +/- 9.2 %, P < 0.0001) and central aortic systolic (117.5 +/- 12.1 vs. 111.4 +/- 12.2 mmHg, P < 0.0001) and pulse (40.5 +/- 10.3 vs. 36.4 +/- 8.1 mmHg, P = 0.0007) pressures were significantly higher in osteoporotic patients than in controls. The estimated aortic pulse wave velocity (PWV) was also significantly higher in the osteoporotic group. In multivariate analysis for osteoporotic patients, femoral neck and lumbar spine bone mineral density T scores were independent negative predictors of AIx (P < 0.0001). AIx was not correlated with serum levels of OPG and RANKL. Osteoporotic postmenopausal women show increased AIx and central aortic pressures, and a higher estimated aortic PWV, indicating a stiffer aorta. Such alterations may increase cardiovascular risk in postmenopausal osteoporosis.