Abstract
A member of the tumor necrosis factor (TNF) family, receptor activator of nuclear factor kappaB ligand (RANKL; also known as ODF, OPGL, and TRANCE), plays critical roles in osteoclast differentiation and activation in the presence of macrophage colony-stimulating factor (M-CSF). Recently, TNF-alpha has also been shown to induce the formation of multinucleated osteoclast-like cells (MNCs) in the presence of M-CSF from mouse macrophages. We demonstrated that mononuclear preosteoclast-like cells (POCs) were formed in the presence of conditioned medium of osteoblastic cells in a rat bone marrow culture depleted of stromal cells. Using this culture system, in this study we examined whether TNF-alpha affects differentiation into POCs from hematopoietic progenitor cells. Human TNF-alpha (hTNF-alpha) markedly stimulated the formation of POCs. Moreover, a concentration as low as 0.005 ng/mL of hTNF-alpha increased the level of mRNA for calcitonin receptor (CTR) and cathepsin-K of POCs. The POCs induced by hTNF-alpha formed MNCs, which showed dentine-resorbing activity after coculture with primary osteoblasts. Stimulation was observed after 24 h of treatment with hTNF-alpha only on day 1 or day 2 of the culture. After 24 h of hTNF-alpha treatment, expression of the receptor activator of nuclear factor kappaB (RANK) mRNA was markedly increased. The addition of soluble RANKL (sRANKL) to the preformed POCs efficiently induced MNCs. Interestingly, treatment of bone marrow cells with hTNF-alpha and sRANKL synergistically augmented the formation of MNCs. This formation was abolished by the addition of human osteoprotegerin (hOPG). These results suggest that cooperation of TNF-alpha and RANKL is important for osteoclastogenesis.
MeSH terms
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Animals
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Bone Marrow Cells / cytology
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Bone Marrow Cells / drug effects
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Bone Marrow Cells / metabolism
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Bone Remodeling / drug effects
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Bone Remodeling / physiology*
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Bone and Bones / cytology
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Bone and Bones / drug effects
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Bone and Bones / metabolism
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Carrier Proteins / pharmacology
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Cathepsins / genetics
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Cell Differentiation / drug effects
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Cell Differentiation / physiology*
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Cells, Cultured / cytology
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Cells, Cultured / drug effects
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Cells, Cultured / metabolism
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Coculture Techniques
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Culture Media, Conditioned / pharmacology
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Drug Interactions / physiology*
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Glycoproteins / metabolism
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Glycoproteins / pharmacology
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Hematopoietic Stem Cells / cytology
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Hematopoietic Stem Cells / drug effects
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Hematopoietic Stem Cells / metabolism*
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Male
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism*
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Membrane Glycoproteins / pharmacology
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Osteoblasts / cytology
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Osteoblasts / drug effects
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Osteoblasts / metabolism
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Osteoclasts / cytology
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Osteoclasts / drug effects
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Osteoclasts / metabolism*
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Osteoprotegerin
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RANK Ligand
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RNA, Messenger / drug effects
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RNA, Messenger / metabolism
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Rats
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Rats, Sprague-Dawley
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Receptor Activator of Nuclear Factor-kappa B
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Receptors, Calcitonin / genetics
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Receptors, Cytoplasmic and Nuclear / metabolism
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Receptors, Tumor Necrosis Factor
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Stromal Cells / cytology
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Stromal Cells / metabolism
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Tumor Necrosis Factor-alpha / metabolism*
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Tumor Necrosis Factor-alpha / pharmacology
Substances
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Carrier Proteins
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Culture Media, Conditioned
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Glycoproteins
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Membrane Glycoproteins
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Osteoprotegerin
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RANK Ligand
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RNA, Messenger
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Receptor Activator of Nuclear Factor-kappa B
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Receptors, Calcitonin
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Receptors, Cytoplasmic and Nuclear
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Receptors, Tumor Necrosis Factor
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TNFRSF11A protein, human
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TNFRSF11B protein, human
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TNFSF11 protein, human
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Tnfrsf11a protein, mouse
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Tnfrsf11b protein, mouse
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Tnfrsf11b protein, rat
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Tnfsf11 protein, mouse
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Tumor Necrosis Factor-alpha
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Cathepsins