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LPS administration increases CD11b+ c-Fms+ CD14+ cell population that possesses osteoclast differentiation potential in mice

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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

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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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Correspondence to Masamichi Takami.

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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

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