TNF-alpha stimulation increases dental pulp stem cell migration in vitro through integrin alpha-6 subunit upregulation

Arch Oral Biol. 2017 Mar:75:48-54. doi: 10.1016/j.archoralbio.2016.12.005. Epub 2016 Dec 15.

Abstract

Objective: The dissemination of stem cells into tissues requiring inflammatory and reparative response is fundamentally dependent upon their chemotactic migration. Expression of TNF-α is up regulated in inflamed pulps. Dental pulp cells are also known to express integrin α6 subunit. Expression of integrin subunit α6 has been linked to the acquisition of migratory potential in a wide variety of cell types in both pathological and physiological capacities. Therefore, in this study we examined the effects of a pleiotropic cytokine TNF-α on the migration of hDPSCs and investigated its relationship with expression of integrin α6 in hDPSCs during chemotactic migration.

Design: hDPSC cultures were established. Protein expression profile of α6 integrin subunit was determined. Effect of exogenous TNF-α (50ng/mL) on hDPSCs' migration potential was evaluated by transwell inserts and in vitro scratch assay. Upregulation/downregulation of TNF-α mediated migration was assayed in presence/absence of integrin α6 respectively. To suppress integrin α6 expression, cells were transfected with integrin α6 siRNA and then cell migration and cytoskeletal changes were evaluated.

Results: Our results showed significant increase of hDPSCs' migration after stimulation with TNF-α. By knockdown of integrin α6, which is upregulated by TNF-α, we observed a decrease in the TNF-α directed chemotaxis of hDPSCs.

Conclusion: In this study, we show that activation of integrin α6 brought about by TNF-α led to an increase in migratory activity in DPSCs in vitro thus describing a novel association between a cytokine TNF-α and α6 chain of an adhesion receptor integrin in regulating migration of hDPSCs.

Keywords: Cytokine; Dental pulp stem cell; Integrin α6; Migration; Regeneration; TNF-α.

MeSH terms

  • Adult
  • Cell Movement / drug effects*
  • Chemotaxis
  • Cytokines / metabolism
  • Cytoskeleton / drug effects
  • Dental Pulp / cytology
  • Dental Pulp / drug effects*
  • Down-Regulation
  • Gene Knockdown Techniques
  • Humans
  • Integrin alpha6 / genetics
  • Integrin alpha6 / metabolism*
  • Molar, Third
  • RNA, Small Interfering / genetics
  • Regeneration / drug effects
  • Stem Cells / cytology
  • Stem Cells / drug effects*
  • Tumor Necrosis Factor-alpha / pharmacology*
  • Up-Regulation / drug effects*

Substances

  • Cytokines
  • Integrin alpha6
  • RNA, Small Interfering
  • Tumor Necrosis Factor-alpha