Effects of transforming growth factor-β1 on plasminogen activation in stem cells from the apical papilla: role of activating receptor-like kinase 5/Smad2 and mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signalling

Int Endod J. 2020 May;53(5):647-659. doi: 10.1111/iej.13266. Epub 2020 Feb 7.

Abstract

Aim: To study the effects of TGF-β1 on the plasminogen activation (PA) system of stem cells from the apical papilla (SCAP) and its signalling.

Methodology: SCAP cells were isolated from the apical papilla of immature permanent teeth extracted for orthodontic reasons. They were exposed to various concentration of TGF-β1 with/without pretreatment and coincubation by SB431542 (ALK/Smad2/3 inhibitor), or U0126 (MEK/ERK inhibitor). MTT assay, Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to detect their effects on cell viability, and the protein expression of plasminogen activator inhibitor-1 (PAI-1), urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and their secretion. The paired Student's t-test was used for statistical analysis.

Results: TGF-β1 significantly stimulated PAI-1 and soluble uPAR (suPAR) secretion of SCAP cells (P < 0.05), whereas uPA secretion was inhibited. Accordingly, TGF-β1 induced both PAI-1 and uPAR protein expression of SCAP cells. SB431542 (an ALK5/Smad2/3 inhibitor) pretreatment and coincubation prevented the TGF-β1-induced PAI-1 and uPAR of SCAP. U0126 attenuated the TGF-β1-induced expression/secretion of uPAR, but not PAI-1 in SCAP. SB431542 reversed the TGF-β1-induced decline of uPA.

Conclusions: TGF-β1 may affect the repair/regeneration activities of SCAP via differential increase or decrease of PAI-1, uPA and uPAR. These effects induced by TGF-β1 are associated with ALK5/Smad2/3 and MEK/ERK activation. Elucidation the signalling pathways and effects of TGF-β1 is useful for treatment of immature teeth with open apex by revascularization/revitalization procedures and tissue repair/regeneration.

Keywords: PAI-1; TGF-β1; regeneration; repair; signal transduction; stem cells from apical papilla; urokinase plasminogen.

MeSH terms

  • Extracellular Signal-Regulated MAP Kinases*
  • Humans
  • Mitogen-Activated Protein Kinase Kinases
  • Plasminogen
  • Smad2 Protein
  • Stem Cells
  • Transforming Growth Factor beta1*
  • Transforming Growth Factors

Substances

  • SMAD2 protein, human
  • Smad2 Protein
  • Transforming Growth Factor beta1
  • Transforming Growth Factors
  • Plasminogen
  • Extracellular Signal-Regulated MAP Kinases
  • Mitogen-Activated Protein Kinase Kinases