Abstract
Osteoclasts are multinucleated giant cells that originate from a monocyte/macrophage lineage, and are involved in the inflammatory bone destruction accompanied by periodontitis. Recent studies have shown that osteoclast precursors reside not only in the bone marrow, but also in the peripheral blood and spleen, though the precise characteristics of each precursor have not been analyzed. We hypothesized that the number of osteoclast precursors in those tissues may increase under pathological conditions and contribute to osteoclast formation in vivo in a mouse model. To test this hypothesis, we attempted to identify cell populations that possess osteoclast differentiation potential in the bone marrow, spleen, and blood by analyzing macrophage/monocyte-related cell surface markers such as CD11b, CD14, and colony-stimulating factor-1 receptor (c-Fms). In the bone marrow, the CD11b− cell population, but not the CD11b+ cell population, differentiated into osteoclasts in the presence of receptor activator of nuclear factor-κB ligand and macrophage colony-stimulating factor. On the other hand, in the spleen and blood, CD11b+ cells differentiated into osteoclasts. Interestingly, lipopolysaccharide (LPS) administration to the mice dramatically increased the proportion of CD11b+ c-Fms+ CD14+ cells, which differentiated into osteoclasts, in the bone marrow and spleen. These results suggest that LPS administration increases the proportion of a distinct cell population expressing CD11b+, c-Fms+, and CD14+ in the bone marrow and spleen. Thus, these cell populations are considered to contribute to the increase in osteoclast number during inflammatory bone destruction such as periodontitis.
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Abbreviations
- IL-1α/β:
-
Interleukin-1α/β
- LPS:
-
Lipopolysaccharide
- M-CSF:
-
Macrophage colony-stimulating factor
- OPG:
-
Osteoprotegerin
- RANK:
-
Receptor activator of nuclear factor-κB
- RANKL:
-
Receptor activator of nuclear factor-κB ligand
- TLR4:
-
Toll-like receptor 4
- TNFα:
-
Tumor necrosis factor α
References
Ash P, Loutit JF, Townsend KM (1980) Osteoclasts derived from haematopoietic stem cells. Nature 283:669–670
Charles JF, Hsu LY, Niemi EC, Weiss A, Aliprantis AO, Nakamura MC (2012) Inflammatory arthritis increases mouse osteoclast precursors with myeloid suppressor function. J Clin Investig 122:4592–4605
Fujikawa Y, Quinn JM, Sabokbar A, McGee JO, Athanasou NA (1996) The human osteoclast precursor circulates in the monocyte fraction. Endocrinology 137:4058–4060
Han C, Jin J, Xu S, Liu H, Li N, Cao X (2010) Integrin CD11b negatively regulates TLR-triggered inflammatory responses by activating Syk and promoting degradation of MyD88 and TRIF via Cbl-b. Nat Immunol 11:734–742
Ishii M, Egen JG, Klauschen F, Meier-Schellersheim M, Saeki Y, Vacher J, Proia RL, Germain RN (2009) Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis. Nature 458:524–528
Jacome-Galarza CE, Lee SK, Lorenzo JA, Aguila HL (2013) Identification, characterization, and isolation of a common progenitor for osteoclasts, macrophages, and dendritic cells from murine bone marrow and periphery. J Bone Miner Res 28:1203–1213
Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, Khoo W, Wakeham A, Dunstan CR, Lacey DL, Mak TW, Boyle WJ, Penninger JM (1999) OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature 397:315–323
Miyamoto T, Arai F, Ohneda O, Takagi K, Anderson DM, Suda T (2000) An adherent condition is required for formation of multinuclear osteoclasts in the presence of macrophage colony-stimulating factor and receptor activator of nuclear factor κB ligand. Blood 96:4335–4343
Nair SP, Meghji S, Wilson M, Reddi K, White P, Henderson B (1996) Bacterially induced bone destruction: mechanisms and misconceptions. Infect Immun 64:2371–2380
Nakamichi Y, Mizoguchi T, Arai A, Kobayashi Y, Sato M, Penninger JM, Yasuda H, Kato S, DeLuca HF, Suda T, Udagawa N, Takahashi N (2012) Spleen serves as a reservoir of osteoclast precursors through vitamin D-induced IL-34 expression in osteopetrotic op/op mice. Proc Natl Acad Sci USA 109:10006–10011
Poltorak A, He X, Smirnova I, Liu MY, Van Huffel C, Du X, Birdwell D, Alejos E, Silva M, Galanos C, Freudenberg M, Ricciardi-Castagnoli P, Layton B, Beutler B (1998) Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282:2085–2088
Shi C, Pamer EG (2011) Monocyte recruitment during infection and inflammation. Nat Rev Immunol 11:762–774
Suda T, Takahashi N, Udagawa N, Jimi E, Gillespie MT, Martin TJ (1999) Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev 20:345–357
Takahashi N, Udagawa N, Akatsu T, Tanaka H, Isogai Y, Suda T (1991) Deficiency of osteoclasts in osteopetrotic mice is due to a defect in the local microenvironment provided by osteoblastic cells. Endocrinology 128:1792–1796
Takahashi N, Udagawa N, Suda T (1999) A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function. Biochem Biophys Res Commun 256:449–455
Takeshita S, Kaji K, Kudo A (2000) Identification and characterization of the new osteoclast progenitor with macrophage phenotypes being able to differentiate into mature osteoclasts. J Bone Miner Res 15:1477–1488
Taubman MA, Valverde P, Han X, Kawai T (2005) Immune response: the key to bone resorption in periodontal disease. J Periodontol 76:2033–2041
Udagawa N, Takahashi N, Yasuda H, Mizuno A, Itoh K, Ueno Y, Shinki T, Gillespie MT, Martin TJ, Higashio K, Suda T (2000) Osteoprotegerin produced by osteoblasts is an important regulator in osteoclast development and function. Endocrinology 141:3478–3484
Walker DG (1975a) Bone resorption restored in osteopetrotic mice by transplants of normal bone marrow and spleen cells. Science 190:784–785
Walker DG (1975b) Control of bone resorption by hematopoietic tissue. The induction and reversal of congenital osteopetrosis in mice through use of bone marrow and splenic transplants. J Exp Med 142:651–663
Wong BR, Rho J, Arron J, Robinson E, Orlinick J, Chao M, Kalachikov S, Cayani E, Bartlett FS 3rd, Frankel WN, Lee SY, Choi Y (1997) TRANCE is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells. J Biol Chem 272:25190–25194
Xing L, Schwarz EM, Boyce BF (2005) Osteoclast precursors, RANKL/RANK, and immunology. Immunol Rev 208:19–29
Yasuda H, Shima N, Nakagawa N, Mochizuki SI, Yano K, Fujise N, Sato Y, Goto M, Yamaguchi K, Kuriyama M, Kanno T, Murakami A, Tsuda E, Morinaga T, Higashio K (1998a) Identity of osteoclastogenesis inhibitory factor (OCIF) and osteoprotegerin (OPG): a mechanism by which OPG/OCIF inhibits osteoclastogenesis in vitro. Endocrinology 139:1329–1337
Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N, Suda T (1998b) Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA 95:3597–3602
Yoshida H, Hayashi S, Kunisada T, Ogawa M, Nishikawa S, Okamura H, Sudo T, Shultz LD (1990) The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature 345:442–444
Acknowledgements
We thank Yoichi Miyamoto, Ayako Mochizuki, Akihiro Matsunaga, and Akifumi Matsumoto at Showa University, School of Dentistry for technical support and constructive advice for this study.
Funding
This work was supported by JSPS KAKENHI (Grant Numbers 26293398, 24659830), Industry to Support Private Universities Building up Their Foundations of Strategic Research (S1411009, S1201014, S0801016), and Private University Research Branding Project by MEXT Japan.
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Supplemental Fig. 1
Gating strategy for FACS analysis and cell sorting. After doublets and dead cells were gated out using 7-AAD, the single living cells were classified into each cell populations using antibodies. (TIFF 13995 kb)
Supplemental Fig. 2
Effect of LPS administration on the osteoclast differentiation potential of CD11b+ cells in the bone marrow and spleen of C3H/HeN and C3H/HeJ mice. C3H/HeN and C3H/HeJ mice were injected intraperitoneally with LPS (500 μL/kg) or saline. After 24 h, CD11b+cells from the bone marrow and spleen were isolated using magnetic-labeled anti-CD11b antibodies. The CD11b+cells (1 × 105) were then cultured in the presence of RANKL (100 ng/mL) and M-CSF (50 ng/mL) in 96-well culture plates. After 5 days of culture, cells were fixed and stained for TRAP, an osteoclast marker enzyme and TRAP-positive multinucleated cells were counted as osteoclasts. Data are presented as the mean values of four independent experiments. The error bars represent the SD. **P < 0.01 N.S., not significant. (TIFF 13995 kb)
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Enomoto, T., Takami, M., Yamamoto, M. et al. LPS administration increases CD11b+ c-Fms+ CD14+ cell population that possesses osteoclast differentiation potential in mice. Cytotechnology 69, 529–537 (2017). https://doi.org/10.1007/s10616-017-0094-3
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DOI: https://doi.org/10.1007/s10616-017-0094-3