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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
Article
  • 72 Downloads

Abstract

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.

Keywords

Dental follicle cells Cellular senescence Osteogenic differentiation WNT5A 

Notes

Acknowledgements

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)

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Copyright information

© 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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