Skip to main content

Advertisement

SpringerLink
Log in

High dose of TNF-α suppressed osteogenic differentiation of human dental pulp stem cells by activating the Wnt/β-catenin signaling

  • Original Paper
  • Published:
Journal of Molecular Histology Aims and scope Submit manuscript

Abstract

Dental pulp stem cells (DPSCs) were a clonogenic, highly proliferative cells capable of self-renewal and multi-lineage differentiation including chondrocytes, adipocytes, neural cells and osteoblasts, which make it an attractive choice for bone regeneration and repair of craniofacial defects. Recent studies showed that tumor necrosis factor α (TNF-α) may affect osteoclastogenesis and bone formation. However, the effect and mechanism of TNF-α on DPSCs is not clear. In this study, we found that low dose TNF-α promoted mineralization and high dose TNF-α suppressed osteogenic differentiation of DPSCs. Levels of ALP, Osteopontin, Osteocalcin, Osterix and Runx2 were up-regulated in DPSCs treated with TNF-α at low concentration, while down-regulated in DPSCs treated with TNF-α at high concentration. Blockade of Wnt/β-catenin signaling reversed the inhibitory effect observed on osteogenic differentiation of DPSCs treated with TNF-α at high concentration. In addition, we did not detect any proliferative effect of TNF-α on DPSCs by cell cycle and cell counts analysis. In summary, our data suggested that high concentration TNF-α suppressed mineralization and mineralization-related gene expressions through the Wnt/β-catenin signaling in DPSCs. Our findings may provide a foundation for autologous transplantation of DPSCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

TNF-α:

Tumor necrosis factor α

DPSCs:

Dental pulp stem cell

MSCs:

Mesenchymal stem cells

ALP:

Alkaline phosphatase

BMP-2:

Bone morphogenic protein 2

DMEM:

Dulbecco modified eagle medium

FBS:

Fetal bovine serum

PFA:

Paraformaldehyde

References

  • Bennett CN, Ouyang H, Ma YL, Zeng Q, Gerin I, Sousa KM, Lane TF, Krishnan V, Hankenson KD, MacDougald OA (2007) Wnt10b increases postnatal bone formation by enhancing osteoblast differentiation. J Bone Miner Res 22:1924–1932

    Article  CAS  PubMed  Google Scholar 

  • Bocker W, Docheva D, Prall WC, Egea V, Pappou E, Rossmann O, Popov C, Mutschler W, Ries C, Schieker M (2008) IKK-2 is required for TNF-alpha-induced invasion and proliferation of human mesenchymal stem cells. J Mol Med 86:1183–1192

    Article  PubMed  Google Scholar 

  • Cho HH, Kim YJ, Kim SJ, Kim JH, Bae YC, Ba B, Jung JS (2006) Endogenous Wnt signaling promotes proliferation and suppresses osteogenic differentiation in human adipose derived stromal cells. Tissue Eng 12:111–121

    Article  CAS  PubMed  Google Scholar 

  • Cho HH, Shin KK, Kim YJ, Song JS, Kim JM, Bae YC, Kim CD, Jung JS (2010) NF-kappaB activation stimulates osteogenic differentiation of mesenchymal stem cells derived from human adipose tissue by increasing TAZ expression. J Cell Physiol 223:168–177

    CAS  PubMed  Google Scholar 

  • Choung HW, Lee JH, Lee DS, Choung PH, Park JC (2013) The role of preameloblast-conditioned medium in dental pulp regeneration. J Mol Histol 44:715–721

    Article  CAS  PubMed  Google Scholar 

  • Daoussis D, Andonopoulos AP, Liossis SN (2010) Wnt pathway and IL-17: novel regulators of joint remodeling in rheumatic diseases. Looking beyond the RANK-RANKL-OPG axis. Semin Arthritis Rheum 39:369–383

    Article  CAS  PubMed  Google Scholar 

  • De Boer J, Wang HJ, Van Blitterswijk C (2004) Effects of Wnt signaling on proliferation and differentiation of human mesenchymal stem cells. Tissue Eng 10:393–401

    Article  PubMed  Google Scholar 

  • de Mendonca CA, Bueno DF, Martins MT, Kerkis I, Kerkis A, Fanganiello RD, Cerruti H, Alonso N, Passos-Bueno MR (2008) Reconstruction of large cranial defects in nonimmunosuppressed experimental design with human dental pulp stem cells. J Craniofa Surg 19:204–210

    Google Scholar 

  • Doble BW, Woodgett JR (2003) GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci 116:1175–1186

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Gilbert L, He X, Farmer P, Rubin J, Drissi H, van Wijnen AJ, Lian JB, Stein GS, Nanes MS (2002) Expression of the osteoblast differentiation factor RUNX2 (Cbfa1/AML3/Pebp2alpha A) is inhibited by tumor necrosis factor-alpha. J Biol Chem 277:2695–2701

    Article  CAS  PubMed  Google Scholar 

  • Gronthos S, Mankani M, Brahim J, Robey PG, Shi S (2000) Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci USA 97:13625–13630

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, DenBesten P, Robey PG, Shi S (2002) Stem cell properties of human dental pulp stem cells. J Dent Res 81:531–535

    Article  CAS  PubMed  Google Scholar 

  • Guo AJ, Choi RC, Cheung AW, Chen VP, Xu SL, Dong TT, Chen JJ, Tsim KW (2011) Baicalin, a flavone, induces the differentiation of cultured osteoblasts: an action via the Wnt/beta-catenin signaling pathway. J Biol Chem 286:27882–27893

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hess K, Ushmorov A, Fiedler J, Brenner RE, Wirth T (2009) TNFα promotes osteogenic differentiation of human mesenchymal stem cells by triggering the NF-kappaβ signaling pathway. Bone 45:367–376

    Article  CAS  PubMed  Google Scholar 

  • Hill PA, Tumber A, Meikle MC (1997) Multiple extracellular signals promote osteoblast survival and apoptosis. Endocrinology 138:3849–3858

    CAS  PubMed  Google Scholar 

  • Huang H, Zhao N, Xu X, Xu Y, Li S, Zhang J, Yang P (2011) Dose-specific effects of tumor necrosis factor alpha on osteogenic differentiation of mesenchymal stem cells. Cell Prolif 44:420–427

    Article  CAS  PubMed  Google Scholar 

  • Kim YJ, Lee MH, Wozney JM, Cho JY, Ryoo HM (2004) Bone morphogenetic protein-2-induced alkaline phosphatase expression is stimulated by Dlx5 and repressed by Msx2. J Biol Chem 279:50773–50780

    Article  CAS  PubMed  Google Scholar 

  • Kjeldsen M, Holmstrup P, Bendtzen K (1993) Marginal periodontitis and cytokines: a review of the literature. J Periodontol 64:1013–1022

    Article  CAS  PubMed  Google Scholar 

  • Kong X, Liu Y, Ye R, Zhu B, Zhu Y, Liu X, Hu C, Luo H, Zhang Y, Ding Y, Jin Y (2013) GSK3beta is a checkpoint for TNF-alpha-mediated impaired osteogenic differentiation of mesenchymal stem cells in inflammatory microenvironments. Biochim Biophys Acta 1830:5119–5129

    Article  CAS  PubMed  Google Scholar 

  • Lacey DC, Simmons PJ, Graves SE, Hamilton JA (2009) Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation. Osteoarthritis and Cartilage/OARS, Osteoarthritis Research Society 17:735–742

    Article  CAS  PubMed  Google Scholar 

  • Lee S, An S, Kang TH, Kim KH, Chang NH, Kang S, Kwak CK, Park HS (2011) Comparison of mesenchymal-like stem/progenitor cells derived from supernumerary teeth with stem cells from human exfoliated deciduous teeth. Regen Med 6:689–699

    Article  PubMed  Google Scholar 

  • Lencel P, Hardouin P, Magne D (2010) Do cytokines induce vascular calcification by the mere stimulation of TNAP activity? Med Hypotheses 75:517–521

    Article  CAS  PubMed  Google Scholar 

  • Li B, Shi M, Li J, Zhang H, Chen B, Chen L, Gao W, Giuliani N, Zhao RC (2007) Elevated tumor necrosis factor-alpha suppresses TAZ expression and impairs osteogenic potential of Flk-1 + mesenchymal stem cells in patients with multiple myeloma. Stem Cells Dev 16:921–930

    Article  CAS  PubMed  Google Scholar 

  • Liu N, Shi S, Deng M, Tang L, Zhang G, Liu N, Ding B, Liu W, Liu Y, Shi H, Liu L, Jin Y (2011) High levels of beta-catenin signaling reduce osteogenic differentiation of stem cells in inflammatory microenvironments through inhibition of the noncanonical Wnt pathway. J Bone Miner Res 26:2082–2095

    Article  CAS  PubMed  Google Scholar 

  • Liu W, Liu Y, Guo T, Hu C, Luo H, Zhang L, Shi S, Cai T, Ding Y, Jin Y (2013) TCF3, a novel positive regulator of osteogenesis, plays a crucial role in miR-17 modulating the diverse effect of canonical Wnt signaling in different microenvironments. Cell Death Dis 4:e539

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Liu W, Konermann A, Guo T, Jager A, Zhang L, Jin Y (2014) Canonical Wnt signaling differently modulates osteogenic differentiation of mesenchymal stem cells derived from bone marrow and from periodontal ligament under inflammatory conditions. Biochim Biophys Acta 1840:1125–1134

    Article  CAS  PubMed  Google Scholar 

  • Lu Z, Wang G, Dunstan CR, Zreiqat H (2012) Short-term exposure to tumor necrosis factor-alpha enables human osteoblasts to direct adipose tissue-derived mesenchymal stem cells into osteogenic differentiation. Stem Cells Dev 21:2420–2429

    Article  CAS  PubMed  Google Scholar 

  • Maeda K, Takahashi N, Kobayashi Y (2013) Roles of Wnt signals in bone resorption during physiological and pathological states. J Mol Med 91:15–23

    Article  CAS  PubMed  Google Scholar 

  • Orsini G, Mazzoni A, Orciani M, Putignano A, Procaccini M, Falconi M, Pashley DH, Tay FR, Breschi L (2011) Matrix metalloproteinase-2 expression induced by two different adhesive systems on human pulp fibroblasts. J Endod 37:1663–1667

    Article  PubMed  Google Scholar 

  • Park SJ, Bae HS, Park JC (2015) Osteogenic differentiation and gene expression profile of human dental follicle cells induced by human dental pulp cells. J Mol Histol 46:93–106

    Article  CAS  PubMed  Google Scholar 

  • Scheller EL, Chang J, Wang CY (2008) Wnt/beta-catenin inhibits dental pulp stem cell differentiation. J Dent Res 87:126–130

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Yang X, Zhang S, Pang X, Fan M (2012) Pro-inflammatory cytokines induce odontogenic differentiation of dental pulp-derived stem cells. J Cell Biochem 113:669–677

    Article  CAS  PubMed  Google Scholar 

  • Zhou FH, Foster BK, Zhou XF, Cowin AJ, Xian CJ (2006) TNF-alpha mediates p38 MAP kinase activation and negatively regulates bone formation at the injured growth plate in rats. J Bone Miner Res 21:1075–1088

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors deny any conflicts of interest related to this study.

Author information

Author notes

    Authors and Affiliations

    1. Department of Stomatology, Zoucheng People’s Hospital, Zoucheng, 273500, Shandong, People’s Republic of China

      Zhenjie Qin, Zhixiu Fang, Lei Zhao & Jing Chen

    2. Department of Pharmacy, Zoucheng People’s Hospital, Zoucheng, 273500, Shandong, People’s Republic of China

      Yuanteng Li

    3. Department of Obstetrics and Gynecology, Zoucheng People’s Hospital, Zoucheng, 273500, Shandong, People’s Republic of China

      Guangyun Liu

    Authors
    1. Zhenjie Qin
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Zhixiu Fang
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Lei Zhao
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Jing Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Yuanteng Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Guangyun Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Guangyun Liu.

    Additional information

    Zhenjie Qin and Zhixiu Fang contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Qin, Z., Fang, Z., Zhao, L. et al. High dose of TNF-α suppressed osteogenic differentiation of human dental pulp stem cells by activating the Wnt/β-catenin signaling. J Mol Hist 46, 409–420 (2015). https://doi.org/10.1007/s10735-015-9630-7

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s10735-015-9630-7

    Keywords

    Search

    Navigation