Biotechnology Letters

, Volume 38, Issue 9, pp 1443–1448 | Cite as

Enterococcus faecalis promotes osteoclast differentiation within an osteoblast/osteoclast co-culture system

  • Zuhui Deng
  • Shuai Wang
  • Boon Chin Heng
  • Changyong Yuan
  • Chengfei Zhang
Original Research Paper



To investigate the effects of heat-killed Enterococcus faecalis ATCC 29212 and P25RC clinical strain (derived from an obturated root canal with apical periodontitis) on osteoclast differentiation within an osteoblast/osteoclast co-culture system.


Heat-killed E. faecalis significantly increased the proportion of multinucleated osteoclastic cells (MNCs) within the co-culture system. The IL-6 level was significantly increased upon exposure to heat-killed E. faecalis. Gene expression levels of NFATc1 and cathepsin K were significantly up-regulated compared to the untreated control. EphrinB2 and EphB4 expressions at both the mRNA and protein levels were also significantly upregulated compared to the untreated control.


Heat-killed E. faecalis can induce osteoclast differentiation within the osteoblast/osteoclast co-culture system in vitro, possibly through ephrinB2-EphB4 bidirectional signaling.


Enterococcus faecalis EphrinB2 EphB4 Multinucleated osteoclastic cells Osteoblasts Osteoclasts Osteoclast differentiation 



This work was supported by RFCID/HHSRF HMRF Grant No. 12110772 and Shenzhen Key Laboratory of ENT (No. ZDSYS 201506050935272).

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. Baker PJ, Dixon M, Evans RT, Dufour L, Johnson E, Roopenian DC (1999) CD4(+) T cells and the proinflammatory cytokines gamma interferon and interleukin-6 contribute to alveolar bone loss in mice. Infect Immun 67:2804–2809PubMedPubMedCentralGoogle Scholar
  2. Boyce BF (2013) Advances in the regulation of osteoclasts and osteoclast functions. J Dent Res 92:860–867CrossRefPubMedPubMedCentralGoogle Scholar
  3. Edwards CM, Mundy GR (2008) Eph receptors and ephrin signaling pathways: a role in bone homeostasis. Int J Med Sci 5:263–272CrossRefPubMedPubMedCentralGoogle Scholar
  4. Feng X, McDonald JM (2011) Disorders of bone remodeling. Annu Rev Pathol 6:121–145CrossRefPubMedPubMedCentralGoogle Scholar
  5. Fujisaki K, Tanabe N, Suzuki N, Kawato T, Takeichi O, Tsuzukibashi O, Makimura M, Ito K, Maeno M (2007) Receptor activator of NF-kappa B ligand induces the expression of carbonic anhydrase II, cathepsin K, and matrix metalloproteinase-9 in osteoclast precursor RAW264.7 cells. Life Sci 80:1311–1318CrossRefPubMedGoogle Scholar
  6. Gomes BP, Pinheiro ET, Gade-Neto CR, Sousa EL, Ferraz CC, Zaia AA, Teixeira FB, Souza-Filho FJ (2004) Microbiological examination of infected dental root canals. Oral Microbiol Immunol 19:71–76CrossRefPubMedGoogle Scholar
  7. Graves DT, Oates T, Garlet GP (2011) Review of osteoimmunology and the host response in endodontic and periodontal lesions. J Oral Microbiol 3:5304. doi: 10.3402/jom.v3i0.5304 CrossRefGoogle Scholar
  8. Hienz SA, Paliwal S, Ivanovski S (2015) Mechanisms of bone resorption in periodontitis. J Immunol Res 2015:615486CrossRefPubMedPubMedCentralGoogle Scholar
  9. Kurihara N, Bertolini D, Suda T, Akiyama Y, Roodman GD (1990) IL-6 stimulates osteoclast-like multinucleated cell-formation in long-term human marrow cultures by inducing IL-1 release. J Immunol 144:4226–4230PubMedGoogle Scholar
  10. Pfeilschifter J, Chenu C, Bird A, Mundy GR, Roodman GD (1989) Interleukin-1 and tumor necrosis factor stimulate the formation of human osteoclastlike cells-invitro. J Bone Miner Res 4:113–118CrossRefPubMedGoogle Scholar
  11. Rocas IN, Siqueira JF, Santos KRN (2004) Association of Enterococcus faecalis with different forms of periradicular diseases. J Endod 30:315–320CrossRefPubMedGoogle Scholar
  12. Takayanagi H (2007) Osteoimmunology: shared mechanisms and crosstalk between the immune and bone systems. Nat Rev Immunol 7:292–304CrossRefPubMedGoogle Scholar
  13. Wang S, Deng Z, Seneviratne CJ, Cheung GS, Jin L, Zhao B, Zhang C (2015a) Enterococcus faecalis promotes osteoclastogenesis and semaphorin 4D expression. Innate Immun 21:726–735CrossRefPubMedGoogle Scholar
  14. Wang S, Deng Z, Ye X, Geng X, Zhang C (2015b) Enterococcus faecalis attenuates osteogenesis through activation of p38 and ERK1/2 pathways in MC3T3-E1 cells. Int Endod J. doi: 10.1111/iej.12578 Google Scholar
  15. Yamashita T, Takahashi N, Udagawa N (2012) New roles of osteoblasts involved in osteoclast differentiation. World J Orthop 3:175–181CrossRefPubMedPubMedCentralGoogle Scholar
  16. Zhan X, Zhang C, Dissanayaka WL, Cheung GS, Jin L, Yang Y, Yan F, Tong EH (2013) Storage media enhance osteoclastogenic potential of human periodontal ligament cells via RANKL-independent signaling. Dent Traumatol 29:59–65CrossRefPubMedGoogle Scholar
  17. Zhao C, Irie N, Takada Y, Shimoda K, Miyamoto T, Nishiwaki T, Suda T, Matsuo K (2006) Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis. Cell Metab 4:111–121CrossRefPubMedGoogle Scholar
  18. Zhu XF, Wang QQ, Zhang CF, Cheung GSP, Shen Y (2010) Prevalence, phenotype, and genotype of Enterococcus faecalis isolated from saliva and root canals in patients with persistent apical periodontitis. J Endod 36:1950–1955CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Zuhui Deng
    • 1
    • 2
  • Shuai Wang
    • 1
  • Boon Chin Heng
    • 1
  • Changyong Yuan
    • 1
  • Chengfei Zhang
    • 1
  1. 1.Endodontics, Comprehensive Dental Care, Faculty of DentistryThe University of Hong KongHong Kong SARChina
  2. 2.Shenzhen ENT InstituteShenzhen Longgang ENT HospitalShenzhenChina

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