title: Modulation of osteoclastogenesis with macrophage M1- and M2-inducing stimuli.
abstract: Macrophages are generated through the differentiation of monocytes in tissues and they have important functions in innate and adaptive immunity. In addition to their roles as phagocytes, macrophages can be further differentiated, in the presence of receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF), into osteoclasts (multinucleated giant cells that are responsible for bone resorption). In this work, we set out to characterize whether various inflammatory stimuli, known to induce macrophage polarization, can alter the type of multinucleated giant cell obtained from RANKL differentiation. Following a four-day differentiation protocol, along with lipopolysaccharide (LPS)/interferon gamma (IFNγ) as one stimulus, and interleukin-4 (IL-4) as the other, three types of multinucleated cells were generated. Using various microscopy techniques (bright field, epifluorescence and scanning electron), functional assays, and western blotting for osteoclast markers, we found that, as expected, RANKL treatment alone resulted in osteoclasts, whereas the addition of LPS/IFNγ to RANKL pre-treated macrophages generated Langhans-type giant cells, while IL-4 led to giant cells resembling foreign body giant cells with osteoclast-like characteristics. Finally, to gain insight into the modulation of osteoclastogenesis, we characterized the formation and morphology of RANKL and LPS/IFNγ-induced multinucleated giant cells.
authors.affiliation: Ontario Cancer Institute and Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.
authors.forename: Sujeeve
authors.initials: S
authors.lastname: Jeganathan
authors.affiliation: Department of Biological Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada.
authors.forename: Cara
authors.initials: C
authors.lastname: Fiorino
authors.affiliation: Department of Biological Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada.
authors.forename: Urja
authors.initials: U
authors.lastname: Naik
authors.affiliation: Department of Biological Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada.
authors.forename: He Song
authors.initials: HS
authors.lastname: Sun
authors.affiliation: Department of Biological Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada.
authors.forename: Rene E
authors.initials: RE
authors.lastname: Harrison
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D015744:Giant Cells, Langhans / Q000166:cytology / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D010010:Osteoclasts / Q000166:cytology / Q000378:metabolism*; D053245:RANK Ligand / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D053245:RANK Ligand; C505689:Tnfsf11 protein, mouse; D015847:Interleukin-4; D007371:Interferon-gamma
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