Monosodium Urate in the Presence of RANKL Promotes Osteoclast Formation through Activation of c-Jun N-Terminal Kinase

The aim of this study was to clarify the role of monosodium urate (MSU) crystals in receptor activator of nuclear factor kB ligand- (RANKL-) RANK-induced osteoclast formation. RAW 264.7 murine macrophage cells were incubated with MSU crystals or RANKL and differentiated into osteoclast-like cells as confirmed by staining for tartrate-resistant acid phosphatase (TRAP) and actin ring, pit formation assay, and TRAP activity assay. MSU crystals in the presence of RANKL augmented osteoclast differentiation, with enhanced mRNA expression of NFATc1, cathepsin K, carbonic anhydrase II, and matrix metalloproteinase-9 (MMP-9), in comparison to RAW 264.7 macrophages incubated in the presence of RANKL alone. Treatment with both MSU crystals and RANKL induced osteoclast differentiation by activating downstream molecules in the RANKL-RANK pathway including tumor necrosis factor receptor-associated factor 6 (TRAF-6), JNK, c-Jun, and NFATc1. IL-1b produced in response to treatment with both MSU and RANKL is involved in osteoclast differentiation in part through the induction of TRAF-6 downstream of the IL-1b pathway. This study revealed that MSU crystals contribute to enhanced osteoclast formation through activation of RANKL-mediated pathways and recruitment of IL-1b. These findings suggest that MSU crystals might be a pathologic causative agent of bone destruction in gout.

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Optimized Extract from Corylopsis coreana Uyeki (Hamamelidaceae) Flos Inhibits Osteoclast Differentiation

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/6302748 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9

Author(s):

Yongjin Lee ◽

Jung-Eun Kim ◽

Kwang-Jin Kim ◽

Seung-Sik Cho ◽

Young-Jin Son

Keyword(s):

Osteoclast Differentiation ◽

Cathepsin K ◽

Inhibitory Effect ◽

Osteoclast Formation ◽

Tartrate Resistant Acid Phosphatase ◽

Alcoholic Extract ◽

Nuclear Factor ◽

Activated T Cells ◽

Fractures Of The Spine ◽

The Stability

Osteoporosis is a metabolic disorder that decreases the stability against fractures of the spine, femur, and radius by weakening the strength and integrity of bones. Receptor activator of nuclear factor-kappa B ligand signaling ultimately activated nuclear factor-activated T cells c1, a major transcription factor for osteoclast formation. This study researched the effects of Corylopsis coreana (C. coreana) Uyeki flos extracts on the antiosteoclastic potential of macrophages and the phytochemicals contained therein. The alcoholic extract of C. coreana Uyeki flos inhibited the differentiation of osteoclast. We carried out the experiments of the pattern of differentiation of osteoclasts based on the alcoholic percentage of extracts. Among them, 80% alcoholic extract showed the highest inhibitory effect. The alcoholic extract was composed of phytochemicals such as bergenin, quercetin, and quercitrin. This extract inhibited not only mRNA expression levels of NFATc1, osteoclast-associated receptor (OSCAR), cathepsin K, and tartrate-resistant acid phosphatase (TRAP), but also the translational expression of NFATc1. The inhibitory effect for osteoclast differentiation of the alcoholic extract was confirmed using the resorption pit assay. This is the first scientific report of the antiosteoclastic effects of C. coreana Uyeki flos extract, which can be applied therapeutically for the treatment of osteoporosis.

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The Prenyl Group Contributes to Activities of Phytoestrogen 8-Prenynaringenin in Enhancing Bone Formation and Inhibiting Bone Resorption In Vitro

Endocrinology ◽

10.1210/en.2012-2086 ◽

2013 ◽

Vol 154 (3) ◽

pp. 1202-1214 ◽

Cited By ~ 28

Author(s):

Lei-Guo Ming ◽

Xiang Lv ◽

Xiao-Ni Ma ◽

Bao-Feng Ge ◽

Ping Zhen ◽

...

Keyword(s):

Bone Resorption ◽

Mrna Expression ◽

Bone Marrow Cells ◽

Osteoclast Differentiation ◽

Cathepsin K ◽

Active Role ◽

Osteoclast Formation ◽

Calcium Deposition ◽

Tartrate Resistant Acid Phosphatase

Abstract Previous studies have found that 8-prenylflavonoids have a higher osteogenic activity than do flavonoids, which suggested that the 8-prenyl group may play an active role in bone-protective properties. To address this hypothesis, activities of 8-prenylnaringenin (PNG) and naringenin (NG) in osteoblast and osteoclast differentiation and function were compared in vitro. PNG was found to have a stronger ability than NG to improve osteoblast differentiation and osteogenic function in cultured rat calvarial osteoblasts, as demonstrated by levels of alkaline phosphatase activity, osteocalcin, calcium deposition, and the number and area of mineralized bone nodules, as well as mRNA expression of osteogenesis-related genes Bmp-2, OSX, and Runx-2. In addition, although expression of osteoclastogenic inducer receptor activator of nuclear factor kappa-B ligand (RANKL) was not affected, that of osteoclastogenesis inhibitor osteoprotegerin (OPG) and consequently the OPG/RANKL ratio were increased, more potently by PNG than NG. PNG was also found to have a higher potency than NG in inhibiting the osteoclast formation in rabbit bone marrow cells and their resorptive activity, as revealed by lower numbers of osteoclasts formed, lower numbers and areas of bone resorption pits, and lower mRNA expression levels of tartrate-resistant acid phosphatase and cathepsin K. Furthermore, PNG induced apoptosis of mature osteoclasts at a higher degree and at an earlier time than did NG. These results indicate that the 8-prenyl group plays an important role and contributes to the higher bone-protective activity of PNG in comparison with NG.

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Amaranthus mangostanus Inhibits the Differentiation of Osteoclasts and Prevents Ovariectomy-Induced Bone Loss

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/1927017 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11 ◽

Cited By ~ 2

Author(s):

Yeon-Hui Jeong ◽

Haeng Jeon Hur ◽

Ae Sin Lee ◽

Sang Hee Lee ◽

Mi Jeong Sung

Keyword(s):

Bone Loss ◽

Biological Activities ◽

Osteoclast Differentiation ◽

Bone Destruction ◽

Cathepsin K ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Potential Candidate ◽

Bone Homeostasis ◽

Activated T Cells

Bone homeostasis is dynamically balanced between bone forming osteoblasts and bone resorbing osteoclasts. Osteoclasts play an important role in bone destruction and osteoporosis, and they are derived from monocyte/macrophages in response to macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor κB (NF-κB) ligand (RANKL). Amaranthus mangostanus L. (AM) is a plant with powerful antioxidant and other biological activities including anti-inflammatory, antidiabetic, and antihyperlipidemic effects. However, its effects on bone health are unknown. In this study, we explored whether AM could affect RANK-mediated osteoclastogenesis. AM significantly suppressed RANKL-induced osteoclast differentiation and expression of osteoclast-specific genes, TRAP, cathepsin K, NF-activated T-cells (NFATc1), and Dc-stamp in RAW 264.7 cells. Moreover, AM significantly inhibited extracellular signal-regulated kinase (ERK), Akt, and NF-κB signaling pathways in RAW 264.7 cells. In addition, AM preserved ovariectomy-induced bone loss in mice. Taken together, our results suggest that AM might be a potential candidate for the treatment of postmenopausal osteoporosis.

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Polymorphonuclear Neutrophils in Osteomyelitis: Link to Osteoclast Generation and Bone Resorption

European Journal of Inflammation ◽

10.1177/1721727x1201000317 ◽

2012 ◽

Vol 10 (3) ◽

pp. 413-426 ◽

Cited By ~ 13

Author(s):

M.M. Gaida ◽

B. Mayer ◽

S. Stegmaier ◽

P. Schirmacher ◽

C. Wagner ◽

...

Keyword(s):

Mhc Class Ii ◽

Bone Erosion ◽

Ex Vivo ◽

Bone Destruction ◽

Cathepsin K ◽

Class Ii ◽

Osteoclast Formation ◽

Polymorphonuclear Neutrophils ◽

Tartrate Resistant Acid Phosphatase

Chronic and persistent inflammatory processes in bones may lead to severe erosions with consequent functional impairment sometimes requiring amputation of the limb. To explore the relationship between inflammation and bone erosion, biopsies of patients with osteomyelitis due to arterial occlusive disease or to diabetes mellitus were examined (n=31). Histologically, inflammation and bone erosion were confirmed. In the eroded bones the number of osteoclasts correlated with the abundance of infiltrated polymorphonuclear neutrophils (PMN), which were highly activated as shown by expression of MHC class II. For functional characterisation of the infiltrating PMN, patients with implant-associated osteomyelitis, a condition associated with persistent bacterial infection and bone destruction, were recruited. The cells were recovered from infected sites and examined ex vivo. These PMN expressed MHC class II and produced interleukin (IL)-8, a further indication of PMN activation. To assess a possible link between infiltrating PMN and bone erosion, we tested the effect of IL-8 on osteoclast generation in vitro. CD14+ monocytes derived from the peripheral blood of healthy individuals were cultivated with monocyte colony stimulating factor (M-CSF) and IL-8. Within 3 days, a translocation of the transcription factor NFATcl into the nucleus was seen, and by 10 to 20 days multinucleated cells with typical osteoclast morphology appeared that expressed tartrate-resistant acid phosphatase (TRAP) and cathepsin K. Moreover, the cells were able to resorb bone, proving that IL-8 was able to induce the differentiation of monocytes to osteoclasts. Because IL-8 is a major cytokine produced by activated PMN, we propose that in the course of persistent infection infiltrating PMN contribute to induction of osteoclast formation, thus providing a link between inflammation and bone erosion.

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Kalkitoxin Reduces Osteoclast Formation and Resorption and Protects against Inflammatory Bone Loss

International Journal of Molecular Sciences ◽

10.3390/ijms22052303 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2303

Author(s):

Liang Li ◽

Ming Yang ◽

Saroj Kumar Shrestha ◽

Hyoungsu Kim ◽

William H. Gerwick ◽

...

Keyword(s):

Bone Loss ◽

Transmembrane Protein ◽

Osteoclast Differentiation ◽

Mapk Signaling ◽

Osteoclast Formation ◽

Bone Diseases ◽

Tartrate Resistant Acid Phosphatase ◽

Nuclear Factor ◽

Mineral Density ◽

Multinucleated Cells

Osteoclasts, bone-specified multinucleated cells produced by monocyte/macrophage, are involved in numerous bone destructive diseases such as arthritis, osteoporosis, and inflammation-induced bone loss. The osteoclast differentiation mechanism suggests a possible strategy to treat bone diseases. In this regard, we recently examined the in vivo impact of kalkitoxin (KT), a marine product obtained from the marine cyanobacterium Moorena producens (previously Lyngbya majuscula), on the macrophage colony-stimulating factor (M-CSF) and on the receptor activator of nuclear factor κB ligand (RANKL)-stimulated in vitro osteoclastogenesis and inflammation-mediated bone loss. We have now examined the molecular mechanism of KT in greater detail. KT decreased RANKL-induced bone marrow-derived macrophages (BMMs) tartrate-resistant acid phosphatase (TRAP)-multinucleated cells at a late stage. Likewise, KT suppressed RANKL-induced pit area and actin ring formation in BMM cells. Additionally, KT inhibited several RANKL-induced genes such as cathepsin K, matrix metalloproteinase (MMP-9), TRAP, and dendritic cell-specific transmembrane protein (DC-STAMP). In line with these results, RANKL stimulated both genes and protein expression of c-Fos and nuclear factor of activated T cells (NFATc1), and this was also suppressed by KT. Moreover, KT markedly decreased RANKL-induced p-ERK1/2 and p-JNK pathways at different time points. As a result, KT prevented inflammatory bone loss in mice, such as bone mineral density (BMD) and osteoclast differentiation markers. These experiments demonstrated that KT markedly inhibited osteoclast formation and inflammatory bone loss through NFATc1 and mitogen-activated protein kinase (MAPK) signaling pathways. Therefore, KT may have potential as a treatment for destructive bone diseases.

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Cells in regenerating deer antler cartilage provide a microenvironment that supports osteoclast differentiation

Journal of Experimental Biology ◽

10.1242/jeb.204.3.443 ◽

2001 ◽

Vol 204 (3) ◽

pp. 443-455

Author(s):

C. Faucheux ◽

S. Nesbitt ◽

M. Horton ◽

J. Price

Keyword(s):

Type I Collagen ◽

Mononuclear Cells ◽

Osteoclast Differentiation ◽

Cathepsin K ◽

Collagen Matrix ◽

Tartrate Resistant Acid Phosphatase ◽

Type I ◽

Deer Antler

Deer antlers are a rare example of mammalian epimorphic regeneration. Each year, the antlers re-grow by a modified endochondral ossification process that involves extensive remodelling of cartilage by osteoclasts. This study identified regenerating antler cartilage as a site of osteoclastogenesis in vivo. An in vitro model was then developed to study antler osteoclast differentiation. Cultured as a high-density micromass, cells from non-mineralised cartilage supported the differentiation of large numbers of osteoclast-like multinucleated cells (MNCs) in the absence of factors normally required for osteoclastogenesis. After 48 h of culture, tartrate-resistant acid phosphatase (TRAP)-positive mononuclear cells (osteoclast precursors) were visible, and by day 14 a large number of TRAP-positive MNCs had formed (783+/−200 per well, mean +/− s.e.m., N=4). Reverse transcriptase/polymerase chain reaction (RT-PCR) showed that receptor activator of NF κ B ligand (RANKL) and macrophage colony stimulating factor (M-CSF) mRNAs were expressed in micromass cultures. Antler MNCs have the phenotype of osteoclasts from mammalian bone; they expressed TRAP, vitronectin and calcitonin receptors and, when cultured on dentine, formed F-actin rings and large resorption pits. When cultured on glass, antler MNCs appeared to digest the matrix of the micromass and endocytose type I collagen. Matrix metalloproteinase-9 (MMP-9) may play a role in the resorption of this non-mineralised matrix since it is highly expressed in 100 % of MNCs. In contrast, cathepsin K, another enzyme expressed in osteoclasts from bone, is only highly expressed in resorbing MNCs cultured on dentine. This study identifies the deer antler as a valuable model that can be used to study the differentiation and function of osteoclasts in adult regenerating mineralised tissues.

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Edgeworthia papyrifera Regulates Osteoblast and Osteoclast Differentiation In Vitro and Exhibits Anti-osteoporosis Activity in Animal Models of Osteoporosis

Planta Medica ◽

10.1055/a-0942-1960 ◽

2019 ◽

Vol 85 (09/10) ◽

pp. 766-773 ◽

Cited By ~ 1

Author(s):

Pansoo Kim ◽

Yeon-Ju Nam ◽

Woo Jung Kim ◽

Jin Kyu Kim ◽

Gyeongbeen Lee ◽

...

Keyword(s):

Bone Resorption ◽

Animal Models ◽

Osteoclast Differentiation ◽

Raw 264.7 Cells ◽

Tartrate Resistant Acid Phosphatase ◽

Raw 264.7 ◽

Treatment Of Osteoporosis ◽

Increased Risk ◽

Interesting Candidate

AbstractOsteoporosis is a clinical condition characterized by low bone strength that leads to an increased risk of fracture. Strategies for the treatment of osteoporosis involve inhibition of bone resorption by osteoclasts and an increase of bone formation by osteoblasts. Here, we identified the extract derived from the stem part of Edgeworthia papyrifera that enhanced differentiation of MC3T3-E1 cells to osteoblast-like cells and inhibited osteoclast differentiation of RAW 264.7 cells in vitro. In support of our observation, rutin and daphnoretin, which were previously reported to inhibit osteoclast differentiation, were identified in E. papyrifera extract. In an animal model of osteoporosis, the ovariectomy-induced increases in bone resorption biomarkers such as pyridinoline and tartrate-resistant acid phosphatase were significantly reduced by E. papyrifera extract administration at 25.6 and 48.1%, respectively. Furthermore, the ovariectomy-induced bone loss in animal models of osteoporosis was significantly prevented by the administration of E. papyrifera in our study. Taking these observations into account, we suggest that E. papyrifera is an interesting candidate for further exploration as an anti-osteoporotic agent.

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Chitosan oligosaccharide inhibits skull resorption induced by lipopolysaccharides in mice

BMC Oral Health ◽

10.1186/s12903-019-0946-7 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 1

Author(s):

Ke Guo ◽

Zong Lin Liu ◽

Wen Chao Wang ◽

Wei Feng Xu ◽

Shi Qi Yu ◽

...

Keyword(s):

Bone Resorption ◽

Therapeutic Potential ◽

Bone Destruction ◽

Osteoclast Formation ◽

Water Soluble ◽

Tartrate Resistant Acid Phosphatase ◽

Chitosan Oligosaccharide ◽

Micro Computed Tomography ◽

Bone Damage ◽

Dose Dependent Decrease

Abstract Background Low-molecular-weight chitosan oligosaccharide (LMCOS), a chitosan degradation product, is water-soluble and easily absorbable, rendering it a popular biomaterial to study. However, its effect on bone remodelling remains unknown. Therefore, we evaluated the effect of LMCOS on lipopolysaccharide (LPS)-induced bone resorption in mice. Methods Six-week-old male C57BL/6 mice (n = five per group) were randomly divided into five groups: PBS, LPS, LPS + 0.005% LMCOS, LPS + 0.05% LMCOS, and LPS + 0.5% LMCOS. Then, the corresponding reagents (300 μL) were injected into the skull of the mice. To induce bone resorption, LPS was administered at 10 mg/kg per injection. The mice were injected three times a week with PBS alone or LPS without or with LMCOS and sacrificed 2 weeks later. The skull was removed for micro-computed tomography, haematoxylin-eosin staining, and tartrate-resistant acid phosphatase staining. The area of bone damage and osteoclast formation were evaluated and recorded. Results LMCOS treatment during LPS-induced skull resorption led to a notable reduction in the area of bone destruction; we observed a dose-dependent decrease in the area of bone destruction and number of osteoclasts with increasing LMCOS concentration. Conclusions Our findings showed that LMCOS could inhibit skull bone damage induced by LPS in mice, further research to investigate its therapeutic potential for treating osteolytic diseases is required.

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Inhibition of osteoclast differentiation and bone resorption by polyunsaturated fatty acids in RAW 264.7 murine macrophages

Suid-Afrikaanse Tydskrif vir Natuurwetenskap en Tegnologie ◽

10.4102/satnt.v31i1.318 ◽

2012 ◽

Vol 31 (1) ◽

Author(s):

J. C. Boeyens ◽

W-H. Chua ◽

M.C. Kruger ◽

A.M. Joubert ◽

M. Coetzee

Keyword(s):

Fatty Acids ◽

Bone Resorption ◽

Polyunsaturated Fatty Acids ◽

Cell Line ◽

Osteoclast Differentiation ◽

Inhibitory Effect ◽

Osteoclast Formation ◽

Raw 264.7 ◽

Murine Macrophages

This study investigated the effects of polyunsaturated fatty acids on osteoclast formation and bone resorption in RAW 264.7 murine pre-osteoclasts. Data obtained suggests an inhibitory effect of these compounds on osteoclastogenesis and bone resorption in the cell line tested.

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Cell-cell fusion: human multinucleated osteoclasts

Open Life Sciences ◽

10.2478/s11535-009-0058-5 ◽

2009 ◽

Vol 4 (4) ◽

pp. 543-548 ◽

Cited By ~ 2

Author(s):

Zhi-Yong Zeng ◽

Jun-Min Chen

Keyword(s):

Cell Fusion ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Transmembrane Protein ◽

Osteoclast Differentiation ◽

Osteoclast Formation ◽

Tartrate Resistant Acid Phosphatase ◽

Peripheral Blood Mononuclear ◽

Healthy Donors ◽

Cell Cell

AbstractOsteoclasts are known to be formed by fusion of circulating mononuclear precursor cells which originate from haematopoietic stem cells. The precise mechanisms regulating the cell-cell fusion of these circulating cells to multinucleated osteoclasts remain unclear. In the present study, human peripheral blood mononuclear cells (PBMNCs) from healthy donors were treated with the macrophagecolony stimulating factor (M-CSF) and receptor activator of nuclear factor (NF)-κB ligand (RANKL) to induce osteoclast differentiation. Osteoclast formation and resorption activity were investigated through the use of tartrate-resistant acid phosphatase (TRAP) staining and lacunar resorption on dentine slices respectively. Real-time reverse-transcription polymerase chain reaction (PCR) was used to detect expression of dendritic cell-specific transmembrane protein (DC-STAMP) in these cells. The results showed that under the treatment of M-CSF and RANKL, PBMNCs differentiated into multinucleated osteoclasts through cell-cell fusion of mononucleated cells. These osteoclasts were TRAP positive and capable of resorbing the bone. Expression of DC-STAMP was much higher in the cells treated with both M-CSF and RANKL than those treated with M-CSF alone. We concluded that human PBMNCs might differentiate into active osteoclasts under certain conditions and the DC-STAMP, which is believed critical for osteoclast development, will be a possible therapeutic target for osteoclast related diseases in future.

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