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Involvement of hydrogen peroxide in the differentiation of clonal HD-11EM cells into osteoclast-like cells.
- Source :
-
Journal of cellular physiology [J Cell Physiol] 1998 Sep; Vol. 176 (3), pp. 574-87. - Publication Year :
- 1998
-
Abstract
- The present study uses the osteoclast precursor clonal line, HD-11EM, to study the potential of hydrogen peroxide (H2O2) in mediating the differentiation of HD-11EM into osteoclast-like cells. HD-11EM cells are a newly established clonal cell line that, in response to 1alpha,25-(OH)2D3, differentiate into osteoclast-like cells that are multinucleated (more than three nuclei), express tartrate-resistant acid phosphatase (TRAP), and excavate resorption pits when cultured on dentin slices in the presence of osteoblasts (Hsia et al., 1995, J. Bone Miner. Res., 10(Suppl 1):S424; Hsia, and Hauschka, 1997, unpublished data). Here we demonstrate that HD-11EM express the reduced nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase specific cytochrome b558 subunits, and that stimulation of HD-11EM with 1 or 10 nM 1alpha,25-(OH)2D3 increases the extracellular release of H2O2 within 5-10 min. Ours is the first report that stimulation of a cell with 1alpha,25-(OH)2D3 enhances the activation of NADPH-oxidase and increases the basal release of superoxide and the formation of its dismutation product, H2O2. To determine the possible involvement of H2O2 in the differentiation of HD-11EM, these cells were exposed to glucose/glucose oxidase. This enzyme system was used to deliver a pure and continuous source of H2O2 in nanomole amounts consistent with quantities produced by HD-11EM in response to 1alpha,25-(OH)2D3. Both 1alpha,25-(OH)2D3 and the exogenously generated H2O2 stimulated a dose- and time-dependent increase in TRAP activity/cell and the number of multinucleated cells 24-48 hr after treatment. Northern analysis confirmed an increase in expression of TRAP mRNA in response to either 1alpha,25-(OH)2D3 or H2O2. Decreases in cell proliferation and v-myc mRNA were also observed in response to these agents. Taken together, our findings indicate that production of H2O2 by HD-11EM is an important local factor involved in differentiation of HD-11EM into osteoclast-like cells, and suggest that H2O2 may play a role in native osteoclast differentiation.
- Subjects :
- Acid Phosphatase genetics
Acid Phosphatase metabolism
Blotting, Northern
Calcitriol pharmacology
Cell Count
Cell Differentiation drug effects
Cell Differentiation physiology
Cell Nucleus physiology
Clone Cells cytology
Clone Cells metabolism
Cytochrome b Group metabolism
Gene Expression Regulation, Enzymologic drug effects
Genes, myc drug effects
Glucose Oxidase metabolism
Glucose Oxidase pharmacology
Humans
Isoenzymes genetics
Isoenzymes metabolism
NADPH Oxidases metabolism
RNA, Messenger analysis
Stem Cells cytology
Stem Cells metabolism
Superoxides metabolism
Tartrate-Resistant Acid Phosphatase
Hydrogen Peroxide metabolism
Osteoclasts cytology
Osteoclasts enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9541
- Volume :
- 176
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Journal of cellular physiology
- Publication Type :
- Academic Journal
- Accession number :
- 9699510
- Full Text :
- https://doi.org/10.1002/(SICI)1097-4652(199809)176:3<574::AID-JCP14>3.0.CO;2-#