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Bone Resorption Activity Assay Kit, Fluorescence Detection

Here's why pit assays are passé.

Cosmo Bio's Bone Resorption Activity Assay Kit is a multiwell solution for culture, differentiation and quantitative evaluation of bone resorption activity of osteoclasts. The assay reports the fluorescence signal arising from the accumulation of fluoresceinamine-labeled chondroitin sulfate released from plate-bound calcium phosphate (carbonate apatite) into culture well media.

Bone Resorption Assay Kit leverages fluoresceinated calcium phosphate (CaP) as a substrate for cellular bone resorption activity. Culture plates (included in kits and also sold separately) are CaP pre-coated with a synthetic crystalline carbonate apatite, a uniform and defined alternative to natural apatite found in dentin discs. Prior to use, CaP-coated plates are freshly bound to fluoresceinamine-labeled chondroitin sulfate (included in kits and also sold separately). Osteoclastic resorption activity causes labeled chondroitin sulfate to be released from plate-bound calcium phosphate. Released labeled chondroitin sulfate is quantitated by simple fluorometric evaluation of fluorescence intensity. This kit also permits parallel performance of the slower, more labor intensive, but traditional pit assay. It should be noted that CosmoBio CaP-pre-coated plates have been used with trans-well inserts in cellular co-culture experimental designs.

(Heo, S. C. et al. Int J Mol Sci 22, 695 (2021) and Steller, D., Scheibert, A., Sturmheit, T. & Hakim, S. G. Sci Rep-uk 10, 16861 (2020)).


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Bone Resorption Assay KIT

■Features

  • ●Time course of bone resorption activity easily obtained by measuring fluorescence of culture medium sample.
  • ●Well-established method utilized by researchers worldwide. Imaging not required.
  • ●Convenient CaP (hydroxyapatite) pre-coated wells; will not move within well.
  • ●Ready to use sterile components.
  • ●Fluorescence excitation and emission wavelengths identical to FITC. NOTE: use culture media lacking Phenol Red.
  • ●Permits microscopic observation of cell morphology.
  • ●Permits pit area assay after cell removal * using image analysis software.
  • ●Suitable for evaluating osteoclast response to drugs such as bisphosphonates.
BONE RESORPTION ASSAY KIT 24

■Data

The data shown below are the results using RAW264 cells. (Miyazaki T., et al., Anal Biochem, 410:7-12, 2011)

Measurement Principle
A) A schematic of the mechanism for measuring bone resorption activity.
B) RAW264 cells differentiated into osteoclastic cells over days in culture.

The fluorescence intensity of the conditioned medium was increased by the addition of RANKL (100 ng/mL).（*: p<0.05, **: p<0.001). ○: RANKL(-), ●: RANKL(+)

Bone resorption activity and RANKL concentration

RANKL-dependent increases of the fluorescence intensity (C) and pit area (D) (mean ± S.D., n = 3, **: p<0.001).

Evaluation of drugs for treating osteoporosis

The inhibitory effects of Pamidronate and β-Estradiol on the resorption of CaP induced by RANKL (100 ng/mL) were evaluated by fluorescence intensity (E) and pit area (F) (mean ± S.D., n = 3, *:p<0.05,**: p<0.001).

Pit Formation

A microscopic photograph of a CaP-coated plate (on day 6). G: without RANKL; H: with RANKL (100 ng/mL)

■Specification

 DescriptionCatalog Number
Bone Resorption Assay Kit 24
  • ●1 X 24 well calcium phosphate (CaP) coated plate
  • ●FACS (fluorosceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-24KIT
Bone Resorption Assay Kit 24 X 2
  • ●2 X 24 well calcium phosphate (CaP) coated plate
  • ●FACS (fluorosceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-24X2KIT
Bone Resorption Assay Kit 48
  • ●1 x 48 well calcium phosphate (CaP) coated plate
  • ●FACS (fluorosceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-48KIT
Bone Resorption Assay Kit 48 X 2
  • ●2 x 48 well calcium phosphate (CaP) coated plate
  • ●FACS (fluorosceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-48X2KIT
Bone Resorption Assay Kit 96
  • ●1 x 96 well calcium phosphate (CaP) coated plates
  • ●FACS (fluoresceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-96KIT
Bone Resorption Assay Kit 96 X 2
  • ●2 x 96 well calcium phosphate (CaP) coated plates
  • ●FACS (fluoresceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-96X2KIT
Bone Resorption Assay Kit 96 stripwell
  • ●1 x 96 well stripwell calcium phosphate (CaP) coated plates
  • ●FACS (fluoresceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-S96KIT
Bone Resorption Assay Kit 96 stripwell X 2
  • ●2 x 96 well stripwell calcium phosphate (CaP) coated plates
  • ●FACS (fluoresceinamine labeled chondroitin sulfate)
  • ●Bone Resorption Assay Buffer
 CSR-BRA-S96X2KIT

Bone Resorption Assay KIT

Product nameCatalog Number
Bone Resorption Assay Kit 24 CSR-BRA-24KIT
Bone Resorption Assay Kit 24x2 CSR-BRA-24X2KIT
Bone Resorption Assay Kit 48 CSR-BRA-48KIT
Bone Resorption Assay Kit 48x2 CSR-BRA-48X2KIT
Bone Resorption Assay Kit 96 CSR-BRA-96KIT
Bone Resorption Assay Kit 96x2 CSR-BRA-96X2KIT
Bone Resorption Assay Kit 96 stripwell CSR-BRA-S96KIT
Bone Resorption Assay Kit 96 stripwell x2 CSR-BRA-S96X2KIT

Bone Resorption Assay PLATE

■Features

  • ●Observation of cell morphology is possible using a microscope.
  • ●Analysis of pit area is also possible using image analyzing software after removing cells.
  • ●Plate is easy to handle because it is pre-coated with CaP.
  • ●Plate is sterilized and ready to use

■Data

Figure A. Phase-contrast micrograph of RAW264 cells cultured with RANKL (100 ng/mL) for 6 days on CaP-coated plates. OC indicates where osteoclast-like cells were observed.
Figure B. Photograph of the plate after removing cells. Pits can be observed macroscopically (Left: without RANKL; Right: with RANKL).
Figure C. Micrograph of the pits in a CaP-coated plate (with RANKL).
Figure D. RANKL-dependent increase of the pit area (mean ± S.D., n = 3).
Figure E.The inhibitory effect of the bisphosphonate, Pamidronate, on CaP resorption induced by RANKL (100 ng/mL).

Bone Resorption Assay PLATE

4-well Plate
24-well Plate
48-well Plate
96-well Plate
96-well stripwell Plate
Product nameCatalog Number
Bone Resorption Assay Plate 4 well CSR-BRA-4P
Bone Resorption Assay Plate 24 well CSR-BRA-24P
Bone Resorption Assay Plate 24 well X 2 CSR-BRA-24X2P
Bone Resorption Assay Plate 48 well CSR-BRA-48P
Bone Resorption Assay Plate 48 well X 2 CSR-BRA-48X2P
Bone Resorption Assay Plate 96 well CSR-BRA-96P
Bone Resorption Assay Plate 96 well X 2 CSR-BRA-96X2P
Bone Resorption Assay Plate 96 stripwell CSR-BRA-S96P
Bone Resorption Assay Plate 96 stripwell X 2 CSR-BRA-S96X2P

Related products

Product nameCatalog Number
Bone Resorption Assay FACS CSR-BRA-FACS1
Bone Resorption Assay Buffer CSR-BRA-B1

References

  1. Miyazaki, T. and Suzuki, O.Using Fluorescent Polyanions to Assay for Osteoclastic Calcium-Resorption Activity in Calcium: Chemistry, Analysis, Function and Effects (ed. Preedy, V. R.) vol. 10 111–125 (The Royal Society of Chemistry, 2016).
  2. Miyazaki, T., Miyauchi, S., Anada, T., Imaizumi, H. and Suzuki, O.Evaluation of osteoclastic resorption activity using calcium phosphate coating combined with labeled polyanion Anal Biochem 410, 7–12 (2011).
  3. Miyazaki, T., Miyauchi, S., Tawada, A., Anada, T. and Suzuki, O.Effect of chondroitin sulfate-E on the osteoclastic differentiation of RAW264 cells Dent Mater J 29, 403–410 (2010).

Citations

in 2023

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  2. Urabe, Fumihiko, et al. "Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein." Journal of Extracellular Vesicles 12.3 (2023): 12312.
  3. Min, Hong Ki, et al. "Dasatinib, a selective tyrosine kinase inhibitor, prevents joint destruction in rheumatoid arthritis animal model." International Journal of Rheumatic Diseases 26.4 (2023): 718-726.
  4. Rabjohns, Emily M., et al. "Aged G Protein-Coupled Receptor Kinase 3 (Grk3)-Deficient Mice Exhibit Enhanced Osteoclastogenesis and Develop Bone Lesions Analogous to Human Paget’s Disease of Bone." Cells 12.7 (2023): 981.
  5. Kim, Seong-Kyu, et al. "Histone Deacetylase 6 Inhibitor CKD-WID Suppressed Monosodium Urate-Induced Osteoclast Formation by Blocking Calcineurin-NFAT Pathway in RAW 264.7 Cells." Pharmaceuticals 16.3 (2023): 446.
  6. Ding, Mina, et al. "Crucial Role of Lysine-Specific Histone Demethylase 1 in RANKL-Mediated Osteoclast Differentiation." International Journal of Molecular Sciences 24.4 (2023): 3605.
  7. Whitlock, Jarred M., et al. "Cell surface-bound La protein regulates the cell fusion stage of osteoclastogenesis." Nature Communications 14.1 (2023): 616.
  8. Chen, Qi, et al. "Aberrant activation of TGF-β1 induces high bone turnover via Rho GTPases-mediated cytoskeletal remodeling in Camurati-Engelmann disease." Bone cell differentiation in health and disease 16648714 (2023).
  9. Whitlock, Jarred M., et al. "An inducible explant model of osteoclast-osteoprogenitor coordination in exacerbated osteoclastogenesis." Iscience 26.4 (2023).
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in 2022

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    Aster saponin A2 inhibits osteoclastogenesis through mitogen-activated protein kinase-c-Fos-NFATc1 signaling pathway J Vet Sci. 2022 Jul; 23(4): e47.
  2. HK Min, et al.
    DJ-1 controls T cell differentiation and osteoclastogenesis in rheumatoid arthritisScientific Reports volume 12, Article number: 12767 (2022)
  3. C Tortora, et al.
    Effects of CB2 and TRPV1 Stimulation on Osteoclast Overactivity Induced by Iron in Pediatric Inflammatory Bowel Disease Inflammatory Bowel Diseases, Volume 28, Issue 8, August 2022, Pages 1244–1253, https://doi.org/10.1093/ibd/izac073
  4. F Rossi, et al.
    Alteration of ost0eoclast activity in childhood cancer survivors: Role of iron and of CB2/TRPV1 receptors PLUS ONE July 21, 2022 https://doi.org/10.1371/journal.pone.0271730
  5. E Cho, et al.
    Identification of Novel Genes for Cell Fusion during Osteoclast Formation Int. J. Mol. Sci. 2022, 23(12), 6421; https://doi.org/10.3390/ijms23126421
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    An inducible explant model for dissecting osteoclast-osteoblast coordination in health and disease. Jarred M. Whitlock, Luis F. de Castro, Michael T. Collins, Leonid V. Chernomordik, Alison M. Boyce doi: https://doi.org/10.1101/2022.10.27.514052
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    Exercise-linked skeletal irisin ameliorates diabetes-associated osteoporosis by inhibiting the oxidative damage–dependent miR-150-FNDC5/pyroptosis axis Diabetes 2022;71(12):2777–2792 https://doi.org/10.2337/db21-0573
  8. ON Austah, et al.
    Trigeminal neurons control immune-bone cell interaction and metabolism in apical periodontitis Cellular and Molecular Life Sciences volume 79, Article number: 330 (2022)
  9. SH Mun, et al.
    THOC5 regulates human osteoclastogenesis Volume 101, Issue 3, June–August 2022, 151248 https://doi.org/10.1016/j.ejcb.2022.151248
  10. Q Chen, et al.
    Aberrant activation of TGF-β1 induces high bone turnover via Rho GTPases-mediated cytoskeletal remodeling in Camurati-Engelmann disease Front Endocrinol (Lausanne). 2022 Oct 17;13:913979. doi: 10.3389/fendo.2022.913979. eCollection 2022.
  11. H Okawa, et al.
    Fluorescent risedronate analogue 800CW-pRIS improves tooth extraction-associated abnormal wound healing in zoledronate-treated mice Communications Medicine volume 2, Article number: 112 (2022)

in 2021

  1. Zang, L. et al.10-Gingerol Suppresses Osteoclastogenesis in RAW264.7 Cells and Zebrafish Osteoporotic ScalesFrontiers Cell Dev Biology 9, 588093 (2021).
  2. Chen, Z. et al.2-NPPA Mitigates Osteoclastogenesis via Reducing TRAF6-Mediated c-fos ExpressionFront Pharmacol 11, 599081 (2021).
  3. Watahiki, A. et al.Deficiency of Lipin2 Results in Enhanced NF-κB Signaling and Osteoclast Formation in RAW-D Murine MacrophagesInt J Mol Sci 22, 2893 (2021).
  4. Tamura, H. et al.Effects of Erythromycin on Osteoclasts and Bone Resorption via DEL-1 Induction in MiceAntibiotics10, 312 (2021).
  5. Heo, S. C. et al.Elevated Expression of Cathepsin K in Periodontal Ligament Fibroblast by Inflammatory Cytokines Accelerates Osteoclastogenesis via Paracrine Mechanism in Periodontal DiseaseInt J Mol Sci 22, 695 (2021).
  6. Ha, Y.-J. et al.Fucoxanthin Suppresses Osteoclastogenesis via Modulation of MAP Kinase and Nrf2 SignalingMar Drugs 19, 132 (2021).
  7. Lee, E.-J. et al.Hydroxycoumarin Scopoletin Inhibits Bone Loss through Enhancing Induction of Bone Turnover Markers in a Mouse Model of Type 2 DiabetesBiomed 9, 648 (2021).
  8. Chen, Z. et al.N‐[2‐(4‐benzoyl‐1‐piperazinyl)phenyl]‐2‐(4‐chlorophenoxy) acetamide is a novel inhibitor of resorptive bone loss in miceJ Cell Mol Med 25, 1425–1438 (2021).
  9. Lee, J. et al.PDK2 Deficiency Prevents Ovariectomy‐Induced Bone Loss in Mice by Regulating the RANKL‐NFATc1 Pathway During OsteoclastogenesisJ Bone Miner Res 36, 553–566 (2021).
  10. Nakane, S., Imamura, K., Hisanaga, R., Ishihara, K. & Saito, A.Systemic administration of cytotoxic T lymphocyte‐associated antigen 4 (CTLA‐4)‐Ig abrogates alveolar bone resorption in induced periodontitis through inhibition of osteoclast differentiation and activation: An experimental investigationJ Periodontal Res (2021) doi:10.1111/jre.12909.
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    Development of a Cause-based Treatment for NBP-induced MRONJ Journal of Oral and Maxillofacial Surgery Volume 79, Issue 10, Supplement, October 2021, Pages e51-e52
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    Demethylbelamcandaquinone B from Marantodes pumilum var. alata (Blume) Kuntze inhibits osteoclast differentiation in RAW264. 7 cells Asian Pacific Journal of Tropical Biomedicine Year : 2021 | Volume : 11 | Issue : 12 | Page : 535-542 DOI: 10.4103/2221-1691.331269
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    Role of the Endocannabinoid/Endovanilloid System in the Modulation of Osteoclast Activity in Paget's Disease of Bone Int. J. Mol. Sci. 2021, 22(18), 10158; https://doi.org/10.3390/ijms221810158
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    LOXG473A induces the formation of osteoclasts in RAW264. 7 cells via IL-6/JAK2/STAT3 signaling Experimental Cell Research Volume 409, Issue 1, 1 December 2021, 112890
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    Suppressive effects of an apoptotic mimicry prepared from jumbo-flying squid-skin phospholipids on the osteoclastogenesis in receptor activator of nuclear …Journal of the Chinese Medical Association 84(1):p 51-60, January 2021. | DOI: 10.1097/JCMA.0000000000000447
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    Kalkitoxin reduces osteoclast formation and resorption and protects against inflammatory bone loss Int. J. Mol. Sci. 2021, 22(5), 2303; https://doi.org/10.3390/ijms22052303
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    Aqueous extract of Chrysanthemum morifolium Ramat. inhibits RANKL-induced osteoclast differentiation by suppressing the c-fos/NFATc1 pathway Archives of Oral Biology Volume 122, February 2021, 105029
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    Novel Majeed Syndrome–Causing LPIN2 Mutations Link Bone Inflammation to Inflammatory M2 Macrophages and Accelerated Osteoclastogenesis Arthritis & Rheumatology Volume 73, Issue 6 p. 1021-1032 https://doi.org/10.1002/art.41624

in 2020

  1. Na, W. et al.Aesculetin Inhibits Osteoclastic Bone Resorption through Blocking Ruffled Border Formation and Lysosomal TraffickingInt J Mol Sci 21, 8581 (2020).
  2. Wada, S. et al.Bach1 Inhibition Suppresses Osteoclastogenesis via Reduction of the Signaling via Reactive Oxygen Species by Reinforced AntioxidationFrontiers Cell Dev Biology 8, 740 (2020).
  3. Zhou, Y. et al.BMP9 Reduces Bone Loss in Ovariectomized Mice by Dual Regulation of Bone RemodelingJ Bone Miner Res 35, 978–993 (2020).
  4. Lee, E.-J. et al.Coumarin Ameliorates Impaired Bone Turnover by Inhibiting the Formation of Advanced Glycation End Products in Diabetic Osteoblasts and OsteoclastsBiomol 10, 1052 (2020).
  5. Nishioku, T. et al.Dimethyl fumarate prevents osteoclastogenesis by decreasing NFATc1 expression, inhibiting of erk and p38 MAPK phosphorylation, and suppressing of HMGB1 releaseBiochem Bioph Res Co 530, 455–461 (2020).
  6. Shang, N. & Wu, J.Egg-Derived Tripeptide IRW Attenuates LPS-Induced Osteoclastogenesis in RAW 264.7 Macrophages via Inhibition of Inflammatory Responses and NF-κB/MAPK ActivationJ Agr Food Chem 68, 6132–6141 (2020).
  7. Steller, D., Scheibert, A., Sturmheit, T. & Hakim, S. G.Establishment and validation of an in vitro co-culture model for oral cell lines using human PBMC-derived osteoclasts, osteoblasts, fibroblasts and keratinocytesSci Rep-uk 10, 16861 (2020).
  8. Kumagai, M. et al.Fluorinated Kavalactone Inhibited RANKL-Induced Osteoclast Differentiation of RAW264 CellsBiological Pharm Bulletin 43, 898–903 (2020).
  9. Choi, Y. et al.Gastrin-Releasing Peptide (GRP) Stimulates Osteoclastogenesis in PeriodontitisCells 10, 50 (2020).
  10. Korea, D. of O. M. and I., Wonkwang University College of Dentistry, Iksan 54538, Republic of, Park, G. H., Gu, D. R. & Lee, S. H.Inhibitory effect of Chaenomelis Fructus ethanol extract on receptor activator of nuclear factor-kappa B ligand-mediated osteoclastogenesisInt J Oral Biology 45, 15–24 (2020).
  11. Park, K. H., Gu, D. R., Kim, M. S. & Lee, S. H.Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca2+-Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 ExpressionJ Bone Metabolism 27, 53–63 (2020).
  12. Jeong, S. et al.Isoliquiritigenin Derivatives Inhibit RANKL-Induced Osteoclastogenesis by Regulating p38 and NF-κB Activation in RAW 264.7 CellsMolecules 25, 3908 (2020).
  13. Kim, S.-I. et al.Linarin and its aglycone acacetin abrogate actin ring formation and focal contact to bone matrix of bone-resorbing osteoclasts through inhibition of αvβ3 integrin and core-linked CD44Phytomedicine 79, 153351 (2020).
  14. Lee, J. et al.N-[2-(4-Acetyl-1-Piperazinyl)Phenyl]-2-(3-Methylphenoxy)Acetamide (NAPMA) Inhibits Osteoclast Differentiation and Protects against Ovariectomy-Induced OsteoporosisMolecules 25, 4855 (2020).
  15. Matsubara, T. et al.Plectin stabilizes microtubules during osteoclastic bone resorption by acting as a scaffold for Src and Pyk2Bone 132, 115209 (2020).
  16. Cho, E. et al.PMSA prevents osteoclastogenesis and estrogen-dependent bone loss in miceBone 142, 115707 (2020).
  17. Kao, Y.-F., Tu, M.-C., Chai, H.-J., Lin, Y.-L. & Chen, Y.-C.Suppressive effects of an apoptotic mimicry prepared from jumbo-flying squid-skin phospholipids on the osteoclastogenesis in receptor activator of nuclear factor kappa B ligand/macrophage colony-stimulating factor-induced RAW 264.7 cellsJ Chin Med Assoc 84, 51–60 (2020).
  18. Tortora et al.The Role of Cannabinoid Receptor Type 2 in the Bone Loss Associated With Pediatric Celiac DiseaseJournal of Pediatric Gastroenterology and Nutrition 71, 633–640 (2020).

in 2019

  1. Yeh, I.-J. et al.6-Shogaol Suppresses 2-Amino-1-Methyl-6-Phenylimidazo [4,5-b] Pyridine (PhIP)-Induced Human 786-O Renal Cell Carcinoma Osteoclastogenic Activity and Metastatic PotentialNutrients 11, 2306 (2019).
  2. Li, J. et al.Artemisinin inhibits breast cancer‐induced osteolysis by inhibiting osteoclast formation and breast cancer cell proliferationJ Cell Physiol 234, 12663–12675 (2019).
  3. Kim, H. Y. et al.Auranofin Inhibits RANKL-Induced Osteoclastogenesis by Suppressing Inhibitors of κB Kinase and Inflammasome-Mediated Interleukin-1β SecretionOxid Med Cell Longev 2019, 1–12 (2019).
  4. Matsuike, R. et al.Continuous Compressive Force Induces Differentiation of Osteoclasts with High Levels of Inorganic DissolutionMed Sci Monitor 25, 3902–3909 (2019).
  5. Angireddy, R. et al.Cytochrome c oxidase dysfunction enhances phagocytic function and osteoclast formation in macrophagesFaseb J 33, 9167–9181 (2019).
  6. Kim, S., Kang, S.-S., Choi, S.-I., Kim, G.-H. & Imm, J.-Y.Ecklonia cava Extract Containing Dieckol Suppresses RANKL-Induced Osteoclastogenesis via MAP Kinase/NF-kappa B Pathway Inhibition and Heme Oxygenase-1 InductionJ Microbiol Biotechn 29, 11–20 (2019).
  7. Li, J. et al.eIF2α signaling regulates autophagy of osteoblasts and the development of osteoclasts in OVX miceCell Death Dis 10, 921 (2019).
  8. Chen, Z., Cho, E., Lee, J., Lee, S. & Lee, T.-H.Inhibitory Effects of N-[2-(4-acetyl-1-piperazinyl) phenyl]-2-(2-chlorophenoxy) acetamide on Osteoclast Differentiation In Vitro via the Downregulation of TRAF6Int J Mol Sci 20, 5196 (2019).
  9. Kasagi, S. et al.MicroRNA-124 inhibits TNF-α- and IL-6-induced osteoclastogenesisRheumatology International39, 689–695 (2019).
  10. Nishiguchi, A. & Taguchi, T.Osteoclast-Responsive, Injectable Bone of Bisphosphonated-Nanocellulose that Regulates Osteoclast/Osteoblast Activity for Bone RegenerationBiomacromolecules 20, 1385–1393 (2019).
  11. Ma, Y. et al.Osthole inhibits osteoclasts formation and bone resorption by regulating NF‐κB signaling and NFATc1 activations stimulated by RANKLJ Cell Biochem 120, 16052–16061 (2019).
  12. Lee, K.-A. et al.Promotion of osteoclastogenesis by IL-26 in rheumatoid arthritisArthritis Res Ther 21, 283 (2019).
  13. Cho, E., Chen, Z., Lee, J., Lee, S. & Lee, T.-H.PSTP-3,5-Me Inhibits Osteoclast Differentiation and Bone ResorptionMolecules 24, 3346 (2019).
  14. Hsu, L.-C., Reddy, S. V., Yilmaz, Ö. & Yu, H.Sphingosine-1-Phosphate Receptor 2 Controls Podosome Components Induced by RANKL Affecting Osteoclastogenesis and Bone ResorptionCells 8, 17 (2019).
  15. Abdallah, B. M. et al.The Coumarin Derivative 5′-Hydroxy Auraptene Suppresses Osteoclast Differentiation via Inhibiting MAPK and c-Fos/NFATc1 PathwaysBiomed Res Int 2019, 1–10 (2019).

in 2018

  1. Jin, S. H. et al.Actin-binding LIM protein 1 regulates receptor activator of NF-κB ligand-mediated osteoclast differentiation and motilityBmb Rep 51, 356–361 (2018).
  2. Kwon, Y.-B., Wang, F.-F. & Jang, H.-D.Anti-osteoclastic effect of caffeic acid phenethyl ester in murine macrophages depends upon the suppression of superoxide anion production through the prevention of an active-Nox1 complex formationJ Nutritional Biochem 58, 158–168 (2018).
  3. Kittaka, M. et al.Cherubism Mice Also Deficient in c‐Fos Exhibit Inflammatory Bone Destruction Executed by Macrophages That Express MMP14 Despite the Absence of TRAP+ OsteoclastsJ Bone Miner Res 33, 167–181 (2018).
  4. Na, S. et al.Dose analysis of photobiomodulation therapy on osteoblast, osteoclast, and osteocyteJ Biomed Opt 23, 075008 (2018).<>
  5. Ahn, S.-H. et al.Inhibitory Effects of 2N1HIA (2-(3-(2-Fluoro-4-Methoxyphenyl)-6-Oxo-1(6H)-Pyridazinyl)-N-1H-Indol-5-Ylacetamide) on Osteoclast Differentiation via Suppressing Cathepsin K ExpressionMolecules 23, 3139 (2018).
  6. Wijekoon, H. M. S. et al.Inhibitory effects of sodium pentosan polysulfate on formation and function of osteoclasts derived from canine bone marrowBmc Vet Res 14, 152 (2018).
  7. Wik, H. M. S.Pentosan Polysulfate: an Effective Treatment for Model of Rheumatoid Arthritis and a Novel Candidate for Inhibition of Cytokines-Induced Osteoclastogenesis(2018).
  8. Chu, G., Zhang, W., Chen, M., Yang, H. & Yuan, Z.Punicalagin inhibits RANKL-induced osteoclastogenesis by suppressing NF-KB and MAPK signaling pathwaysInt J Clin Exp Med 11, 6571–6582 (2018).
  9. Lin, B., Ke, Q., Leaman, D. W., Goel, V. & Agarwal, A.Regulation of RANKL‐induced osteoclastogenesis by RING finger protein RNF114J Orthopaed Res 36, 159–166 (2018).
  10. Funaki, Y. et al.Resolvin E1 Inhibits Osteoclastogenesis and Bone Resorption by Suppressing IL-17-induced RANKL Expression in Osteoblasts and RANKL-induced Osteoclast DifferentiationYonago Acta Med 61, 008–018 (2018).
  11. Nishida, M., Saegusa, J., Tanaka, S. & Morinobu, A.S100A12 facilitates osteoclast differentiation from human monocytesPlos One 13, e0204140 (2018).
  12. Fessler, J. et al.Senescent T-Cells Promote Bone Loss in Rheumatoid ArthritisFront Immunol 9, 95 (2018).
  13. Sapkota, M., Li, L., Kim, S.-W. & Soh, Y.Thymol inhibits RANKL-induced osteoclastogenesis in RAW264.7 and BMM cells and LPS-induced bone loss in miceFood Chem Toxicol 120, 418–429 (2018).

in 2017

  1. Lee, S.-H., Lee, J.-Y., Kwon, Y.-I. & Jang, H.-D.Anti-Osteoclastic Activity of Artemisia capillaris Thunb. Extract Depends upon Attenuation of Osteoclast Differentiation and Bone Resorption-Associated Acidification Due to Chlorogenic Acid, Hyperoside, and ScoparoneInt J Mol Sci 18, 322 (2017).
  2. Wijekoon, S. et al.Chronological differential effects of pro-inflammatory cytokines on RANKL-induced osteoclast differentiation of canine bone marrow-derived macrophagesJ Vet Med Sci 17–0393 (2017) doi:10.1292/jvms.17-0393.
  3. Kim, H. et al.Comprehensive study on the roles of released ions from biodegradable Mg–5 wt% Ca–1 wt% Zn alloy in bone regenerationJ Tissue Eng Regen M 11, 2710–2724 (2017).
  4. Cong, F. et al.Ginsenoside Rb2 inhibits osteoclast differentiation through nuclear factor-kappaB and signal transducer and activator of transcription protein 3 signaling pathwayBiomed Pharmacother 92, 927–934 (2017).
  5. Maruyama, K. et al.Nociceptors Boost the Resolution of Fungal Osteoinflammation via the TRP Channel-CGRP-Jdp2 AxisCell Reports 19, 2730–2742 (2017).
  6. Lee, D. et al.Nutrikinetics of Isoflavone Metabolites After Fermented Soybean Product (Cheonggukjang) Ingestion in Ovariectomized MiceMol Nutr Food Res 61, 1700322 (2017).
  7. Bellini, G. et al.PKCβII-mediated cross-talk of TRPV1/CB2 modulates the glucocorticoid-induced osteoclast overactivityPharmacol Res 115, 267–274 (2017).
  8. Park, K. H. et al.Pueraria lobate Inhibits RANKL-Mediated Osteoclastogenesis Via Downregulation of CREB/PGC1β/c-Fos/NFATc1 SignalingThe American Journal of Chinese Medicine 45, (2017).
  9. Hirai, T. et al.The effectiveness of new triple combination therapy using synthetic disease-modifying anti-rheumatic drugs with different pharmacological function against rheumatoid arthritis: the verification by an in vitro and clinical studyClin Rheumatol 36, 51–58 (2017).
  10. 66. Gu, D. R. et al.The inhibitory effect of beta-lapachone on RANKL-induced osteoclastogenesisBiochem Bioph Res Co 482, 1073–1079 (2017).

in 2016

  1. Nishioku, T., Terasawa, M., Baba, M., Yamauchi, A. & Kataoka, Y.CD147 promotes the formation of functional osteoclasts through NFATc1 signallingBiochem Bioph Res Co 473, 620–624 (2016).
  2. Antika, L. D. et al.Dietary compound gossypetin inhibits bone resorption through down-regulating lysosomal cathepsin K activity and autophagy-related protein induction in actin ring-bearing osteoclastsJ Funct Food 24, 390–402 (2016).
  3. Qiu, Z.-C. et al.Discovery of a New Class of Cathepsin K Inhibitors in Rhizoma Drynariae as Potential Candidates for the Treatment of OsteoporosisInt J Mol Sci 17, 2116 (2016).
  4. Feres, M. F. N., Kucharski, C., Diar-Bakirly, S. & El-Bialy, T.Effect of low-intensity pulsed ultrasound on the activity of osteoclasts: An in vitro studyArch Oral Biol 70, 73–78 (2016).
  5. Li, L., Sapkota, M., Kim, S. & Soh, Y.Herbacetin inhibits RANKL-mediated osteoclastogenesis in vitro and prevents inflammatory bone loss in vivoEur J Pharmacol 777, 17–25 (2016).
  6. Shiota, J., Murao, H., Miura, A., Mikami, M. & Tanaka, K.Iguratimod, a Disease-Modifying Anti-Rheumatic Drug, Inhibits Osteoclastogenesis and Bone Resorption through Suppression of the Nuclear Factor of Activated T Cells Signaling PathwayOpen J Rheumatology Autoimmune Dis 06, 106–119 (2016).
  7. Ishikawa, S. et al.Inductive Effect of Palmatine on Apoptosis in RAW 264.7 CellsEvid-based Compl Alt 2016, 1–9 (2016).
  8. Joung, Y. H. et al.Methylsulfonylmethane Inhibits RANKL-Induced Osteoclastogenesis in BMMs by Suppressing NF-κB and STAT3 ActivitiesPlos One 11, e0159891 (2016).
  9. Cheng, T.-L. et al.Myeloid thrombomodulin lectin-like domain inhibits osteoclastogenesis and inflammatory bone lossSci Rep-uk 6, 28340 (2016).
  10. Pengjam, Y. et al.NF-κB pathway inhibition by anthrocyclic glycoside aloin is key event in preventing osteoclastogenesis in RAW264.7 cellsPhytomedicine 23, 417–428 (2016).
  11. Kaneki, K. et al.Palmatine’s Suppression on NFATc1 for RANKL‒treated RAW264.7 CellsJapanese Pharmacology & Therapeutics 44, 997–1004 (2016).
  12. Takigawa, S. et al.Role of miR-222-3p in c-Src-Mediated Regulation of OsteoclastogenesisInt J Mol Sci 17, 240 (2016).
  13. Yu, H.Sphingosine-1-Phosphate Receptor 2 Regulates Proinflammatory Cytokine Production and OsteoclastogenesisPlos One 11, e0156303 (2016).

in 2015

  1. Maruyama, K. et al.5-Azacytidine-induced Protein 2 (AZI2) Regulates Bone Mass by Fine-tuning Osteoclast SurvivalJ Biol Chem 290, 9377–9386 (2015).
  2. Davis, J. W. E. O.-D. A. R., Olmsted-Davis, E., Davis, A. R. & Sevick-Muraca, E. AnInjectable Method for Posterior Lateral Spine Fusion(2015).
  3. Sapkota, M., Li, L., Choi, H., Gerwick, W. H. & Soh, Y.Bromo-honaucin A inhibits osteoclastogenic differentiation in RAW 264.7 cells via Akt and ERK signaling pathwaysEur J Pharmacol 769, 100–109 (2015).
  4. Miyazaki, T., Miyauchi, S., Anada, T., Tawada, A. & Suzuki, O.Chondroitin Sulfate‐E Binds to Both Osteoactivin and Integrin αVβ3 and Inhibits Osteoclast DifferentiationJ Cell Biochem 116, 2247–2257 (2015).
  5. Gohda, J. et al.HIV-1 replicates in human osteoclasts and enhances their differentiation in vitroRetrovirology 12, 12 (2015).
  6. Mishima, K. et al.Lansoprazole Upregulates Polyubiquitination of the TNF Receptor-Associated Factor 6 and Facilitates Runx2-mediated OsteoblastogenesisEbiomedicine 2, 2046–2061 (2015).
  7. Park, S. et al.Osteonecrosis of the Jaw Developed in Mice: DISEASE VARIANTS REGULATED BY γδ T CELLS IN ORAL MUCOSAL BARRIER IMMUNITYJ Biol Chem 290, 17349–17366 (2015).
  8. Kloos, B. et al.Pasteurella multocida toxin- induced osteoclastogenesis requires mTOR activationCell Commun Signal 13, 40 (2015).
  9. Yamashita, Y. et al.RANKL pretreatment plays an important role in the differentiation of pit-forming osteoclasts induced by TNF-α on murine bone marrow macrophagesArch Oral Biol 60, 1273–1282 (2015).
  10. Kim, K.-W., Kim, B.-M., Moon, H.-W., Lee, S.-H. & Kim, H.-R.Role of C-reactive protein in osteoclastogenesis in rheumatoid arthritisArthritis Res Ther 17, 41 (2015).
  11. Lee, S.-H. & Jang, H.-D.Scoparone attenuates RANKL-induced osteoclastic differentiation through controlling reactive oxygen species production and scavengingExp Cell Res 331, 267–277 (2015).
  12. TSAI, Y.-M. et al.Syringetin suppresses osteoclastogenesis mediated by osteoblasts in human lung adenocarcinomaOncol Rep 34, 617–626 (2015).
  13. Kim, K.-W., Kim, H.-R., Kim, B.-M., Cho, M.-L. & Lee, S.-H.Th17 Cytokines Regulate Osteoclastogenesis in Rheumatoid ArthritisAm J Pathology 185, 3011–3024 (2015).
  14. Hong, S. H. et al.The antibody atliximab attenuates collagen-induced arthritis by neutralizing AIMP1, an inflammatory cytokine that enhances osteoclastogenesisBiomaterials 44, 45–54 (2015).
  15. Choe, C.-H. et al.Transmembrane protein 173 inhibits RANKL‐induced osteoclast differentiationFebs Lett 589, 836–841 (2015).
  16. Hung, J.-Y. et al.Tricetin, a dietary flavonoid, suppresses benzo(a)pyrene-induced human non-small cell lung cancer bone metastasisInt J Oncol 46, 1985–1993 (2015).

in 2014

  1. Eda, H. et al.A novel Bruton’s tyrosine kinase inhibitor CC-292 in combination with the proteasome inhibitor carfilzomib impacts the bone microenvironment in a multiple myeloma model with resultant antimyeloma activityLeukemia 28, 1892–1901 (2014).
  2. Ohba, T. et al.Bisphosphonates Inhibit Osteosarcoma‐Mediated Osteolysis Via Attenuation of Tumor Expression of MCP‐1 and RANKLJ Bone Miner Res 29, 1431–1445 (2014).
  3. Nishida, H. et al.Blockade of CD26 Signaling Inhibits Human Osteoclast DevelopmentJ Bone Miner Res 29, 2439–2455 (2014).
  4. Sharma, R. et al.Caspase-2 Maintains Bone Homeostasis by Inducing Apoptosis of Oxidatively-Damaged OsteoclastsPlos One 9, e93696 (2014).
  5. Kim, Y.-H. et al.Fisetin antagonizes cell fusion, cytoskeletal organization and bone resorption in RANKL-differentiated murine macrophagesJ Nutritional Biochem 25, 295–303 (2014).
  6. Kanzaki, H. et al.Nuclear Nrf2 Induction by Protein Transduction Attenuates OsteoclastogenesisFree Radical Bio Med 77, 239–248 (2014).
  7. Hsu, Y.-L. et al.Obtusifolin Suppresses Phthalate Esters-Induced Breast Cancer Bone Metastasis by Targeting Parathyroid Hormone-Related ProteinJ Agr Food Chem 62, 11933–11940 (2014).
  8. Kang, J. H. et al.Osteoprotegerin Expressed by Osteoclasts: An Autoregulator of OsteoclastogenesisJ Dent Res 93, 1116–1123 (2014).
  9. Lee, C. W. et al.Potential anti-osteoporotic activity of low-molecular weight hyaluronan by attenuation of osteoclast cell differentiation and function in vitroBiochem Bioph Res Co 449, 438–443 (2014).

in 2013

  1. Hokugo, A., Sun, S., Park, S., McKenna, C. E. & Nishimura, I.Equilibrium-dependent bisphosphonate interaction with crystalline bone mineral explains anti-resorptive pharmacokinetics and prevalence of osteonecrosis of the jaw in ratsBone 53, 59–68 (2013).
  2. Choi, Y. S. et al.Joint-protective effects of compound K, a major ginsenoside metabolite, in rheumatoid arthritis: in vitro evidenceRheumatol Int 33, 1981–1990 (2013).
  3. Kuo, C.-H. et al.Lung tumor-associated dendritic cell-derived resistin promoted cancer progression by increasing Wolf–Hirschhorn syndrome candidate 1/Twist pathwayCarcinogenesis 34, 2600–2609 (2013).
  4. Orosa, B. et al.Lysophosphatidic acid receptor inhibition as a new multipronged treatment for rheumatoid arthritisAnn Rheum Dis 73, 298 (2013).
  5. Tanaka, H. et al.Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in OsteoclastsPlos One 8, e59402 (2013).
  6. Harada, K. et al.Polyphosphate-Mediated Inhibition of Tartrate-Resistant Acid Phosphatase and Suppression of Bone Resorption of OsteoclastsPlos One 8, e78612 (2013).
  7. Moon, S.-J. et al.Temporal differential effects of proinflammatory cytokines on osteoclastogenesisInt J Mol Med31, 769–777 (2013).
  8. Rossi, F. et al.The 17-β-oestradiol inhibits osteoclast activity by increasing the cannabinoid CB2 receptor expressionPharmacol Res 68, 7–15 (2013).
  9. Kanzaki, H., Shinohara, F., Kajiya, M. & Kodama, T.The Keap1/Nrf2 Protein Axis Plays a Role in Osteoclast Differentiation by Regulating Intracellular Reactive Oxygen Species SignalingJ Biol Chem 288, 23009–23020 (2013).

in 2012

  1. Fabre, C. et al.Dual Inhibition of Canonical and Noncanonical NF-κB Pathways Demonstrates Significant Antitumor Activities in Multiple MyelomaAm Assoc Cancer Res 18, 4669–4681 (2012).
  2. Kim, J. et al.Novel antiosteoclastogenic activity of phloretin antagonizing RANKL‐induced osteoclast differentiation of murine macrophagesMol Nutr Food Res 56, 1223–1233 (2012).
  3. Kim, J. et al.Osteoblastogenesis and osteoprotection enhanced by flavonolignan silibinin in osteoblasts and osteoclastsJ Cell Biochem 113, 247–259 (2012).
  4. JL, K. et al.Osteogenic activity of yellow flag iris (Iris pseudacorus) extract modulating differentiation of osteoblasts and osteoclastsAm J Chin Med 40, 1289–305 (2012).