Molecular and Cellular Biochemistry

, Volume 455, Issue 1–2, pp 21–28 | Cite as

WNT5A supports viability of senescent human dental follicle cells

  • Christian MorsczeckEmail author
  • Anja Reck
  • Torsten E. Reichert


The osteogenic differentiation of dental follicle cells (DFCs) is inhibited by the onset of cellular senescence, but the cause for this is largely unknown. Recently it was shown that WNT5a, which is an inductor of the non-canonical WNT pathway, stimulates both cellular senescence and osteogenic differentiation of different cell types. In this study, we investigated the role of WNT5a for viability and osteogenic differentiation in human DFCs after the induction of cellular senescence. DFCs were cultivated until the induction of cellular senescence. The induction of cellular senescence was confirmed by β-galactosidase staining, estimation of population doubling time, and slightly telomere length shortening. After induction of cellular senescence, the expression of WNT5A and the potential to induce the osteogenic differentiation decreased. Inhibition of WNT5A by specific siRNAs had significant effect on the viability of DFCs. Cell proliferation was reduced, whereas both cellular senescence and cell death were increased in DFCs. However, an inhibition of WNT5A did only slightly effect the osteogenic differentiation of DFCs. Our results suggest that WNT5A supports viability during both cell proliferation and osteogenic differentiation of DFCs.


Dental follicle cells Cellular senescence Osteogenic differentiation WNT5A 



This work was supported by grant of the Deutsche Forschungsgemeinschaft (DFG MO1875/10-1).

Supplementary material

11010_2018_3467_MOESM1_ESM.jpg (723 kb)
Figure S1: Western Blot membrane for estimation of WNT5A expression in DFCs before and after induction of senescence. Supplementary material 1 (JPG 722 KB)
11010_2018_3467_MOESM2_ESM.jpg (379 kb)
Figure S2: Gene expression of WNT5A in DFCs after inhibition with siRNAs WNT5A#2 and WNT5A#2. For control DFCs were transfected with ALLSTAR siRNA. Supplementary material 2 (JPG 379 KB)
11010_2018_3467_MOESM3_ESM.jpg (963 kb)
Figure S3: Inhibition of WNT5A during the osteogenic differentiation of DFCs at passage 9 after transfection with WNT5A specific siRNAs or with Allstar siRNA for control. After 7 days of osteogenic differentiation, the gene expression of osteogenic differentiation markers SPONDIN-1, CEMP-1 and RUNX2 (A) and the alkaline phosphatase (ALP) activity (B) were estimated. (C) Alizarin red staining after 28 days of the osteogenic differentiation with ODM. (D) DFCs at passage 9 were transfected either with specific WNT5A siRNAs or control siRNAs (Allstar) and the gene expression of osteogenic differentiation markers ALP, CEMP-1 and RUNX2 were estimated. Columns represent the mean + SD (n = 3). Supplementary material 3 (JPG 963 KB)
11010_2018_3467_MOESM4_ESM.jpg (800 kb)
Figure S4: Induction of cell death in DFCs after 24 h of the osteogenic differentiation with the osteogenic differentiation medium (ODM). For control DFCs were cultured in standard medium (DMEM). Supplementary material 4 (JPG 799 KB)


  1. 1.
    Morsczeck C, Gotz W, Schierholz J, Zeilhofer F, Kuhn U, Mohl C, Sippel C, Hoffmann KH (2005) Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth. Matrix Biol 24:155–165CrossRefGoogle Scholar
  2. 2.
    Diekwisch TG (2001) The developmental biology of cementum. Int J Dev Biol 45:695–706Google Scholar
  3. 3.
    Honda MJ, Imaizumi M, Tsuchiya S, Morsczeck C (2010) Dental follicle stem cells and tissue engineering. J Oral Sci 52:541–552CrossRefGoogle Scholar
  4. 4.
    Morsczeck C (2015) Molecular mechanisms in dental follicle precursor cells during the osteogenic differentiation. Histol Histopathol 30:1161–1169Google Scholar
  5. 5.
    Morsczeck C, Reichert TE (2018) Dental stem cells in tooth regeneration and repair in the future. Expert Opin Biol Ther 18:187–196. CrossRefGoogle Scholar
  6. 6.
    Morsczeck C, Gresser J, Ettl T (2016) The induction of cellular senescence in dental follicle cells inhibits the osteogenic differentiation. Mol Cell Biochem 417:334–339CrossRefGoogle Scholar
  7. 7.
    Morsczeck C, Hullmann M, Reck A, Reichert TE (2018) The cell cycle regulator protein P16 and the cellular senescence of dental follicle cells. Mol Cell Biochem 439:45–52. CrossRefGoogle Scholar
  8. 8.
    Morsczeck C, Schmalz G (2010) Transcriptomes and proteomes of dental follicle cells. J Dent Res 89:445–456CrossRefGoogle Scholar
  9. 9.
    Xiang L, Chen M, He L, Cai B, Du Y, Zhang X, Zhou C, Wang C, Mao JJ, Ling J (2014) Wnt5a regulates dental follicle stem/progenitor cells of the periodontium. Stem Cell Res Ther 5:135CrossRefGoogle Scholar
  10. 10.
    Baschant U, Rauner M, Balaian E, Weidner H, Roetto A, Platzbecker U, Hofbauer LC (2016) Wnt5a is a key target for the pro-osteogenic effects of iron chelation on osteoblast progenitors. Haematologica 101:1499–1507. CrossRefGoogle Scholar
  11. 11.
    Viale-Bouroncle S, Klingelhöffer C, Ettl T, Reichert TE, Morsczeck C (2015) A protein kinase A (PKA)/β-catenin pathway sustains the BMP2/DLX3-induced osteogenic differentiation in dental follicle cells (DFCs). Cell Signal 27(3):598–605CrossRefGoogle Scholar
  12. 12.
    Asem MS, Buechler S, Wates RB, Miller DL, Stack MS (2016) Wnt5a signaling in cancer. Cancers (Basel). Google Scholar
  13. 13.
    Cai J, Mutoh N, Shin JO, Tani-Ishii N, Ohshima H, Cho SW, Jung HS (2011) Wnt5a plays a crucial role in determining tooth size during murine tooth development. Cell Tissue Res 345:367–377. CrossRefGoogle Scholar
  14. 14.
    Lin M, Li L, Liu C, Liu H, He F, Yan F, Zhang Y, Chen Y (2011) Wnt5a regulates growth, patterning, and odontoblast differentiation of developing mouse tooth. Dev Dyn 240:432–440CrossRefGoogle Scholar
  15. 15.
    Wu X, Hu L, Li Y, Li Y, Wang F, Ma P, Wang J, Zhang C, Jiang C, Wang S (2018) SCAPs regulate differentiation of DFSCs During tooth root development in swine. Int J Med Sci 15:291–299. CrossRefGoogle Scholar
  16. 16.
    Bitler BG, Nicodemus JP, Li H, Cai Q, Wu H, Hua X, Li T, Birrer MJ, Godwin AK, Cairns P, Zhang R (2011) Wnt5a suppresses epithelial ovarian cancer by promoting cellular senescence. Cancer Res 71:6184–6194. CrossRefGoogle Scholar
  17. 17.
    Webster MR, Xu M, Kinzler KA, Kaur A, Appleton J, O’Connell MP, Marchbank K, Valiga A, Dang VM, Perego M, Zhang G, Slipicevic A, Keeney F, Lehrmann E, Wood W, Becker KG, Kossenkov AV, Frederick DT, Flaherty KT, Xu X, Herlyn M, Murphy ME, Weeraratna AT (2015) Wnt5A promotes an adaptive, senescent-like stress response, while continuing to drive invasion in melanoma cells. Pigment Cell Melanoma Res 28:184–195. CrossRefGoogle Scholar
  18. 18.
    Winer J, Jung CK, Shackel I, Williams PM (1999) Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem 270:41–49CrossRefGoogle Scholar
  19. 19.
    Gil ME, Coetzer TL (2004) Real-time quantitative PCR of telomere length. Mol Biotechnol 27:169–172CrossRefGoogle Scholar
  20. 20.
    Abukawa H, Zhang W, Young CS, Asrican R, Vacanti JP, Kaban LB, Troulis MJ, Yelick PC (2009) Reconstructing mandibular defects using autologous tissue-engineered tooth and bone constructs. J Oral Maxillofac Surg 67:335–347CrossRefGoogle Scholar
  21. 21.
    Griesmann H, Ripka S, Pralle M, Ellenrieder V, Baumgart S, Buchholz M, Pilarsky C, Aust D, Gress TM, Michl P (2013) WNT5A-NFAT signaling mediates resistance to apoptosis in pancreatic cancer. Neoplasia 15:11–22CrossRefGoogle Scholar
  22. 22.
    Miura M, Chen X-D, Allen MR, Bi Y, Gronthos S, Seo B-M, Lakhani S, Flavell RA, Feng X-H, Robey PG, Young M, Shi S (2004) A crucial role of caspase-3 in osteogenic differentiation of bone marrow stromal stem cells. J Clin Investig 114:1704–1713CrossRefGoogle Scholar
  23. 23.
    Nemoto E, Sakisaka Y, Tsuchiya M, Tamura M, Nakamura T, Kanaya S, Shimonishi M, Shimauchi H (2016) Wnt3a signaling induces murine dental follicle cells to differentiate into cementoblastic/osteoblastic cells via an osterix-dependent pathway. J Periodontal Res 51(2):164–174CrossRefGoogle Scholar
  24. 24.
    Hasegawa D, Wada N, Yoshida S, Mitarai H, Arima M, Tomokiyo A, Hamano S, Sugii H, Maeda H (2018) Wnt5a suppresses osteoblastic differentiation of human periodontal ligament stem cell-like cells via Ror2/JNK signaling. J Cell Physiol 233:1752–1762. CrossRefGoogle Scholar
  25. 25.
    Zhou Y, Zheng L, Li F, Wan M, Fan Y, Zhou X, Du W, Pi C, Cui D, Zhang B, Sun J, Zhou X (2018) Bivalent histone codes on WNT5A during odontogenic differentiation. J Dent Res 97:99–107. CrossRefGoogle Scholar
  26. 26.
    Sakisaka Y, Tsuchiya M, Nakamura T, Tamura M, Shimauchi H, Nemoto E (2015) Wnt5a attenuates Wnt3a-induced alkaline phosphatase expression in dental follicle cells. Exp Cell Res 336(1):85–93CrossRefGoogle Scholar
  27. 27.
    Viale-Bouroncle S, Buergers R, Morsczeck C, Gosau M (2013) β-Tricalcium phosphate induces apoptosis on dental follicle cells. Calcif Tissue Int 92:412–417CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Oral and Maxillofacial SurgeryUniversity Hospital RegensburgRegensburgGermany

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