Mineral trioxide aggregate enhances the osteogenic capacity of periodontal ligament stem cells via NF-κB and MAPK signaling pathways

J Cell Physiol. 2018 Mar;233(3):2386-2397. doi: 10.1002/jcp.26110. Epub 2017 Aug 25.

Abstract

Mineral trioxide aggregate (MTA), as a bioactive material, has a widespread application in clinical practice. To date, the effects of MTA on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs) remain unclear. hPDLSCs were isolated from human periodontal ligament tissues and cultured with MTA conditioned media. Cell counting kit-8 (CCK-8) assay was performed to assess the proliferation capacity of MTA-treated hPDLSCs. Immunofluorescence assay, alkaline phosphatase (ALP) activity, alizarin red staining, real-time RT-PCR, and western blot analyses were used to investigate the odonto/osteogenic capacity of hPDLSCs as well as the involvement of NF-κB and MAPK pathways. ALP activity assay revealed that 2 mg/ml was the optimal concentration for the induction of hPDLSCs by MTA. The protein expression of DSP, RUNX2, OCN, OSX, OPN, DMP1, ALP, and COL-I in MTA-treated hPDLSCs was significantly higher than those in control group (p < 0.01). When hPDLSCs were treated with the inhibitors of NF-κB and MAPK pathways (U0126, SP600125, SB203580, and BMS345541), the effects of MTA on the differentiation of hPDLSCs were suppressed. Mechanistically, P65 was detected to transfer from cytoplasm to nuclei, as indicated by western blot and immunofluorescence assay. Moreover, MAPK-related proteins and its downstream transcription factors were also upregulated in MTA-treated hPDLSCs. Together, mineral trioxide aggregate can promote the odonto/osteogenic capacity of hPDLSCs via activating the NF-κB and MAPK pathways.

Keywords: differentiation; mineral trioxide aggregate; mitogen-activated protein kinase; nuclear factor kappa B; periodontal ligament stem cells.

MeSH terms

  • Aluminum Compounds / pharmacology*
  • Calcium Compounds / pharmacology*
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Drug Combinations
  • Gene Expression Regulation
  • Humans
  • Mitogen-Activated Protein Kinases / metabolism*
  • NF-kappa B / metabolism*
  • Osteogenesis / drug effects*
  • Oxides / pharmacology*
  • Periodontal Ligament / drug effects*
  • Periodontal Ligament / enzymology
  • Signal Transduction / drug effects*
  • Silicates / pharmacology*
  • Stem Cells / drug effects*
  • Stem Cells / enzymology
  • Time Factors
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Aluminum Compounds
  • Calcium Compounds
  • Drug Combinations
  • NF-kappa B
  • Oxides
  • Silicates
  • Transcription Factors
  • mineral trioxide aggregate
  • Mitogen-Activated Protein Kinases