Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
BMC Immunology
Published in: BMC Immunology 1/2021

Open Access 01-12-2021 | Research article

Lipopolysaccharide- TLR-4 Axis regulates Osteoclastogenesis independent of RANKL/RANK signaling

Authors: Mohammed S. AlQranei, Linda T. Senbanjo, Hanan Aljohani, Therwa Hamza, Meenakshi A. Chellaiah

Published in: BMC Immunology | Issue 1/2021

Login to get access

Abstract

Background

Lipopolysaccharide (LPS) is an endotoxin and a vital component of gram-negative bacteria’s outer membrane. During gram-negative bacterial sepsis, LPS regulates osteoclast differentiation and activity, in addition to increasing inflammation. This study aimed to investigate how LPS regulates osteoclast differentiation of RAW 264.7 cells in vitro.

Results

Herein, we revealed that RAW cells failed to differentiate into mature osteoclasts in vitro in the presence of LPS. However, differentiation occurred in cells primed with receptor activator of nuclear factor-kappa-Β ligand (RANKL) for 24 h and then treated with LPS for 48 h (henceforth, denoted as LPS-treated cells). In cells treated with either RANKL or LPS, an increase in membrane levels of toll-like receptor 4 (TLR4) receptor was observed. Mechanistically, an inhibitor of TLR4 (TAK-242) reduced the number of osteoclasts as well as the secretion of tumor necrosis factor (TNF)-α in LPS-treated cells. RANKL-induced RAW cells secreted a very basal level TNF-α. TAK-242 did not affect RANKL-induced osteoclastogenesis. Increased osteoclast differentiation in LPS-treated osteoclasts was not associated with the RANKL/RANK/OPG axis but connected with the LPS/TLR4/TNF-α tumor necrosis factor receptor (TNFR)-2 axis. We postulate that this is because TAK-242 and a TNF-α antibody suppress osteoclast differentiation. Furthermore, an antibody against TNF-α reduced membrane levels of TNFR-2. Secreted TNF-α appears to function as an autocrine/ paracrine factor in the induction of osteoclastogenesis independent of RANKL.

Conclusion

TNF-α secreted via LPS/TLR4 signaling regulates osteoclastogenesis in macrophages primed with RANKL and then treated with LPS. Our findings suggest that TLR4/TNF-α might be a potential target to suppress bone loss associated with inflammatory bone diseases, including periodontitis, rheumatoid arthritis, and osteoporosis.
Appendix
Available only for authorised users
Literature
1.
Tella SH, Gallagher JC. Biological agents in the management of osteoporosis. Eur J Clin Pharmacol. 2014;70:1291–301.PubMedCrossRef
2.
3.
Azuma Y, Kaji K, Katogi R, Takeshita S, Kudo A. Tumor necrosis factor-alpha induces differentiation of and bone resorption by osteoclasts. J Biol Chem. 2000;275:4858–64.PubMedCrossRef
4.
Yamaguchi R, Yoshimura A, Yoshioka H, Kaneko T, Hara Y. Ability of supragingival plaque to induce toll-like receptor 4-mediated stimulation is associated with cytokine production by peripheral blood mononuclear cells. J Periodontol. 2009;80:512–20.PubMedCrossRef
5.
Watanabe K, Iizuka T, Adeleke A, Pham L, Shlimon AE, Yasin M, et al. Involvement of toll-like receptor 4 in alveolar bone loss and glucose homeostasis in experimental periodontitis. J Periodontal Res. 2011;46:21–30.PubMedCrossRef
6.
Vaananen K. Mechanism of osteoclast mediated bone resorption--rationale for the design of new therapeutics. Adv Drug Deliv Rev. 2005;57:959–71.PubMedCrossRef
7.
Abercrombie M. The crawling movement of metazoan cells. Proc Natl Acad Sci U S A. 1980;207:129–47.
8.
Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337–42.PubMedCrossRef
9.
Asagiri M, Takayanagi H. The molecular understanding of osteoclast differentiation. Bone. 2007;40:251–64.PubMedCrossRef
10.
Kearns AE, Khosla S, Kostenuik PJ. Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulation of bone remodeling in health and disease. Endocr Rev. 2008;29:155–92.PubMedCrossRef
11.
Nagasawa T, Kiji M, Yashiro R, Hormdee D, Lu H, Kunze M, et al. Roles of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin in periodontal health and disease. Periodontol. 2007;43:65–84.CrossRef
12.
Kawai T, Matsuyama T, Hosokawa Y, Makihira S, Seki M, Karimbux NY, et al. B and T lymphocytes are the primary sources of RANKL in the bone resorptive lesion of periodontal disease. Am J Pathol. 2006;169:987–98.PubMedPubMedCentralCrossRef
13.
Algate K, Haynes DR, Bartold PM, Crotti TN, Cantley MD. The effects of tumour necrosis factor-alpha on bone cells involved in periodontal alveolar bone loss; osteoclasts, osteoblasts and osteocytes. J Periodontal Res. 2016;51:549–66.PubMedCrossRef
14.
Kato H, Taguchi Y, Tominaga K, Umeda M, Tanaka A. Porphyromonas gingivalis LPS inhibits osteoblastic differentiation and promotes pro-inflammatory cytokine production in human periodontal ligament stem cells. Arch Oral Biol. 2014;59:167–75.PubMedCrossRef
15.
Kawai T, Akira S. The role of pattern-recognition receptors in innate immunity: update on toll-like receptors. Nat Immunol. 2010;11:373–84.PubMedCrossRef
16.
Ogawa T, Yagi T. Bioactive mechanism of Porphyromonas gingivalis lipid A. Periodontol. 2010;54:71–7.CrossRef
17.
18.
Silva N, Abusleme L, Bravo D, Dutzan N, Garcia-Sesnich J, Vernal R, et al. Host response mechanisms in periodontal diseases. J Appl Oral Sci. 2015;23:329–55.PubMedPubMedCentralCrossRef
19.
20.
Cekici A, Kantarci A, Hasturk H, Van Dyke TE. Inflammatory and immune pathways in the pathogenesis of periodontal disease. Periodontol. 2014;64:57–80.CrossRef
21.
Estornes Y, Bertrand MJ. IAPs, regulators of innate immunity and inflammation. Semin Cell Dev Biol. 2015;39:106–14.PubMedCrossRef
22.
Fujihara M, Muroi M, Tanamoto K, Suzuki T, Azuma H, Ikeda H. Molecular mechanisms of macrophage activation and deactivation by lipopolysaccharide: roles of the receptor complex. Pharmacol Ther. 2003;100:171–94.PubMedCrossRef
23.
Wong M, Ziring D, Korin Y, Desai S, Kim S, Lin J, et al. TNFalpha blockade in human diseases: mechanisms and future directions. Clin Immunol. 2008;126:121–36.PubMedCrossRef
24.
Idriss HT, Naismith JH. TNF alpha and the TNF receptor superfamily: structure-function relationship(s). Microsc Res Tech. 2000;50:184–95.PubMedCrossRef
25.
Xu Y, Chang L, Huang A, Liu X, Liu X, Zhou H, et al. Functional detection of TNF receptor family members by affinity-labeled ligands. Sci Rep. 2017;7:6944.PubMedPubMedCentralCrossRef
26.
Zhang YH, Heulsmann A, Tondravi MM, Mukherjee A, Abu-Amer Y. Tumor necrosis factor-alpha (TNF) stimulates RANKL-induced osteoclastogenesis via coupling of TNF type 1 receptor and RANK signaling pathways. J Biol Chem. 2001;276:563–8.PubMedCrossRef
27.
Kobayashi K, Takahashi N, Jimi E, Udagawa N, Takami M, Kotake S, et al. Tumor necrosis factor alpha stimulates osteoclast differentiation by a mechanism independent of the ODF/RANKL-RANK interaction. J Exp Med. 2000;191:275–86.PubMedPubMedCentralCrossRef
28.
Lam J, Takeshita S, Barker JE, Kanagawa O, Ross FP, Teitelbaum SL. TNF-alpha induces osteoclastogenesis by direct stimulation of macrophages exposed to permissive levels of RANK ligand. J Clin Invest. 2000;106:1481–8.PubMedPubMedCentralCrossRef
29.
Kudo O, Fujikawa Y, Itonaga I, Sabokbar A, Torisu T, Athanasou NA. Pro-inflammatory cytokine (TNFalpha/IL-1alpha) induction of human osteoclast formation. J Pathol. 2002;198:220–7.PubMedCrossRef
30.
Fuller K, Murphy C, Kirstein B, Fox SW, Chambers TJ. TNFalpha potently activates osteoclasts, through a direct action independent of and strongly synergistic with RANKL. Endocrinology. 2002;143:1108–18.PubMedCrossRef
31.
Abu-Amer Y, Ross FP, Edwards J, Teitelbaum SL. Lipopolysaccharide-stimulated osteoclastogenesis is mediated by tumor necrosis factor via its P55 receptor. J Clin Invest. 1997;100:1557–65.PubMedPubMedCentralCrossRef
32.
Nakanishi-Matsui M, Yano S, Matsumoto N, Futai M. Lipopolysaccharide induces multinuclear cell from RAW264.7 line with increased phagocytosis activity. Biochem Biophys Res Commun. 2012;425:144–9.PubMedCrossRef
33.
Hou GQ, Guo C, Song GH, Fang N, Fan WJ, Chen XD, et al. Lipopolysaccharide (LPS) promotes osteoclast differentiation and activation by enhancing the MAPK pathway and COX-2 expression in RAW264.7 cells. Int J Mol Med. 2013;32:503–10.PubMedCrossRef
34.
Chellaiah MA, Majumdar S, Aljohani H. Peptidomimetic inhibitors of L-plastin reduce the resorptive activity of osteoclast but not the bone forming activity of osteoblasts in vitro. PLoS One. 2018;13:e0204209.PubMedPubMedCentralCrossRef
35.
Hayman AR. Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy. Autoimmunity. 2008;41:218–23.PubMedCrossRef
36.
Gao A, Wang X, Yu H, Li N, Hou Y, Yu W. Effect of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of EphA2 in osteoblasts and osteoclasts. In Vitro Cell Dev Biol Anim. 2016;52:228–34.PubMedCrossRef
37.
Nativel B, Couret D, Giraud P, Meilhac O, d’Hellencourt CL, Viranaicken W, et al. Porphyromonas gingivalis lipopolysaccharides act exclusively through TLR4 with a resilience between mouse and human. Sci Rep. 2017;7:15789.PubMedPubMedCentralCrossRef
38.
Darveau RP, Arbabi S, Garcia I, Bainbridge B, Maier RV. Porphyromonas gingivalis lipopolysaccharide is both agonist and antagonist for p38 mitogen-activated protein kinase activation. Infect Immun. 2002;70:1867–73.PubMedPubMedCentralCrossRef
39.
Taubman MA, Valverde P, Han X, Kawai T. Immune response: the key to bone resorption in periodontal disease. J Periodontol. 2005;76(Suppl 11S):2033–41.PubMedCrossRef
40.
41.
Ii M, Matsunaga N, Hazeki K, Nakamura K, Takashima K, Seya T, et al. A novel cyclohexene derivative, ethyl (6R)-6-[N-(2-Chloro-4-fluorophenyl)sulfamoyl] cyclohex-1-ene-1-carboxylate (TAK-242), selectively inhibits toll-like receptor 4-mediated cytokine production through suppression of intracellular signaling. Mol Pharmacol. 2006;69:1288–95.PubMedCrossRef
42.
Ye LL, Wei XS, Zhang M, Niu YR, Zhou Q. The Significance of Tumor Necrosis Factor Receptor Type II in CD8(+) Regulatory T Cells and CD8(+) Effector T Cells. Front Immunol. 2018;9:583.PubMedPubMedCentralCrossRef
43.
44.
Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in subgingival plaque5. J Clin Periodontol. 1998;25:134–44.PubMedCrossRef
45.
Cheng X, Kinosaki M, Murali R, Greene MI. The TNF receptor superfamily: role in immune inflammation and bone formation7. Immunol Res. 2003;27:287–94.PubMedCrossRef
46.
Liu J, Wang S, Zhang P, Said-Al-Naief N, Michalek SM, Feng X. Molecular mechanism of the bifunctional role of lipopolysaccharide in osteoclastogenesis 298. J Biol Chem. 2009;284:12512–23.PubMedPubMedCentralCrossRef
47.
Zhang P, Liu J, Xu Q, Harber G, Feng X, Michalek SM, et al. TLR2-dependent modulation of osteoclastogenesis by Porphyromonas gingivalis through differential induction of NFATc1 and NF-kappaB 44. J Biol Chem. 2011;286:24159–69.PubMedPubMedCentralCrossRef
48.
49.
Sirover MA. Subcellular dynamics of multifunctional protein regulation: mechanisms of GAPDH intracellular translocation. J Cell Biochem. 2012;113:2193–200.PubMedPubMedCentralCrossRef
50.
Terrasse R, Tacnet-Delorme P, Moriscot C, Perard J, Schoehn G, Vernet T, et al. Human and pneumococcal cell surface glyceraldehyde-3-phosphate dehydrogenase (GAPDH) proteins are both ligands of human C1q protein. J Biol Chem. 2012;287:42620–33.PubMedPubMedCentralCrossRef
51.
AlQranei MS, Chellaiah MA. Osteoclastogenesis in periodontal diseases: possible mediators and mechanisms. J Oral Biosci. 2020;62:123–30.PubMedCrossRef
52.
Lee HK, Lee J, Tobias PS. Two lipoproteins extracted from Escherichia coli K-12 LCD25 lipopolysaccharide are the major components responsible for Toll-like receptor 2-mediated signaling. J Immunol. 2002;168:4012–7.PubMedCrossRef
53.
Hirschfeld M, Ma Y, Weis JH, Vogel SN, Weis JJ. Cutting edge: repurification of lipopolysaccharide eliminates signaling through both human and murine toll-like receptor 2. J Immunol. 2000;165:618–22.PubMedCrossRef
54.
Matsunaga N, Tsuchimori N, Matsumoto T, Ii M. TAK-242 (resatorvid), a small-molecule inhibitor of toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol Pharmacol. 2011;79:34–41.PubMedCrossRef
55.
Takami M, Kim N, Rho J, Choi Y. Stimulation by toll-like receptors inhibits osteoclast differentiation. J Immunol. 2002;169:1516–23.PubMedCrossRef
56.
Staab E, Thiele GM, Clarey D, Wyatt TA, Romberger DJ, Wells AD, et al. Toll-like receptor 4 signaling pathway mediates inhalant organic dust-induced bone loss. PLoS ONE. 2016;11:e0158735.PubMedPubMedCentralCrossRef
57.
Itoh K, Udagawa N, Kobayashi K, Suda K, Li X, Takami M, et al. Lipopolysaccharide promotes the survival of osteoclasts via toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophages. J Immunol. 2003;170:3688–95.PubMedCrossRef
58.
Hussain MA, Saito H, Alles N, Shimokawa H, Aoki K, Ohya K. Lipopolysaccharide-induced bone resorption is increased in TNF type 2 receptor-deficient mice in vivo. J Bone Miner Metab. 2008;26:469–77.CrossRef
59.
Gupta A, Lee BS, Khadeer MA, Tang Z, Chellaiah M, Abu-Amer Y, et al. Leupaxin is a critical adaptor protein in the adhesion zone of the osteoclast. J Bone Miner Res. 2003;18:669–85.PubMedCrossRef
60.
61.
AlQranei MS, Aljohani H, Majumdar S, Senbanjo LT, Chellaiah MA. C-phycocyanin attenuates RANKL-induced osteoclastogenesis and bone resorption in vitro through inhibiting ROS levels, NFATc1 and NF-kappaB activation. Sci Rep. 2020;10:2513.PubMedPubMedCentralCrossRef
62.
Aljohani H, Senbanjo LT, Chellaiah MA. Methylsulfonylmethane increases osteogenesis and regulates the mineralization of the matrix by transglutaminase 2 in SHED cells. PLoS ONE. 2019;14:e0225598.PubMedPubMedCentralCrossRef
63.
Chellaiah M, Hruska KA. Osteopontin stimulates gelsolin associated phosphoinositide levels and PtdIns 3-hydroxyl kinase. Mol Biol Cell. 1996;7:743–53.PubMedPubMedCentralCrossRef
64.
Chellaiah MA, Kizer N, Biswas R, Alvarez U, Strauss-Schoenberger J, Rifas L, et al. Osteopontin deficiency produces osteoclast dysfunction due to reduced CD44 surface expression. Mol Biol Cell. 2003;14:173–89.PubMedPubMedCentralCrossRef
65.
Perez-Aso M, Montesinos MC, Mediero A, Wilder T, Schafer PH, Cronstein B. Apremilast, a novel phosphodiesterase 4 (PDE4) inhibitor, regulates inflammation through multiple cAMP downstream effectors. Arthritis Res Ther. 2015;17:249.PubMedPubMedCentralCrossRef
66.
Chellaiah MA, Biswas RS, Rittling SR, Denhardt DT, Hruska KA. Rho-dependent rho kinase activation increases CD44 surface expression and bone resorption in osteoclasts. J Biol Chem. 2003;278:29086–97.PubMedCrossRef
Metadata
Title
Lipopolysaccharide- TLR-4 Axis regulates Osteoclastogenesis independent of RANKL/RANK signaling
Authors
Mohammed S. AlQranei
Linda T. Senbanjo
Hanan Aljohani
Therwa Hamza
Meenakshi A. Chellaiah
Publication date
01-12-2021
Publisher
BioMed Central
Published in
BMC Immunology / Issue 1/2021
Electronic ISSN: 1471-2172
DOI
https://doi.org/10.1186/s12865-021-00409-9

Other articles of this Issue 1/2021

BMC Immunology 1/2021 Go to the issue

Research

Upregulation of PD-1 expression on circulating CD8+ but not CD4+ T cells is associated with tuberculosis infection in health care workers

Research article

Activin a suppresses peripheral CD8+ T lymphocyte activity in acute-phase Kawasaki disease

Research

Highly individual- and tissue-specific expression of glycoprotein group A and B blood antigens in the human kidney and liver

Research article

Identification of active small-molecule modulators targeting the novel immune checkpoint VISTA

Research

Application of imiquimod-induced murine psoriasis model in evaluating interleukin-17A antagonist

Research

The role of β2 integrin in dendritic cell migration during infection
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine
Image Credits