PPARγ inhibits inflammation and RANKL expression in epoxy resin-based sealer-induced osteoblast precursor cells E1 cells

Arch Oral Biol. 2013 Jan;58(1):28-34. doi: 10.1016/j.archoralbio.2012.04.015. Epub 2012 May 14.

Abstract

Objectives: The AH26 of epoxy resin-based sealer is used widely owing to its excellent physical characteristics but it induces oxidative stress and cytotoxicity at the periapical tissues. AH26 exhibited cytotoxicity towards MC-3T3-E1 cells, which resulted in mitochondria-mediated apoptosis. Peroxisome proliferator-activated receptor (PPARγ) has an anti-inflammatory effect in several tissue and cells, but its action of AH26-related inflammation is not completely understood. The aim of this study is to investigate the anti-inflammatory and anti-osteoclastic mechanisms of PPARγ in AH26-induced MC-3T3 E1 cells.

Methods: AH26 was prepared according to the manufacturer's instructions. The 1-day extraction sample, which was diluted by 30%, was tested in this experiment. Recombinant deficiency adenoviral PPARγ (Ad/PPARγ) was used to examine PPARγ over-expression in MC-3T3 E1 cells. AH26-induced reactive oxygen species (ROS) formation was analysed using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) with fluorescence-activated cell sorting (FACS), and the expression of receptor activator of nuclear factor-κB ligand (RANKL) and inflammatory molecules was determined by immunoblotting. The anti-inflammatory and anti-osteoclastic mechanisms of the PPARγ-involved signal pathway was examined by immunoblotting.

Results: The AH26 elutes induced inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), RANKL expression and ROS formation. In addition, the AH26 elutes suppressed the expression of PPARγ. However, the recovery of PPARγ expression with Ad/PPARγ resulted in the inhibition of iNOS, COX-2, RANKL and ROS formation despite the AH26 treatment in MC-3T3 E1 cells. The mechanism of PPARγ was confirmed by the blocking of nuclear factor kappa B (NF-κB) translocation to the nucleus after the suppression of ERK1/2, SAPK/JNK and AP-1 in AH26-induced MC-3T3 E1 cells.

Conclusion: From this result, PPARγ acts to inhibit bone destruction in AH26-induced bone cells. Therefore, the anti-inflammatory and anti-osteoclastic character of PPARγ might be applicable for healing periapical lesions more rapidly or reducing the induction of cellular inflammation caused by some endodontic sealers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Adenoviridae / genetics
  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Bismuth / pharmacology*
  • Blotting, Western
  • Cell Separation
  • Cyclooxygenase 2 / drug effects
  • Epoxy Resins / pharmacology*
  • Extracellular Signal-Regulated MAP Kinases / drug effects
  • Flow Cytometry
  • Fluoresceins
  • Fluorescent Dyes
  • Genetic Vectors / genetics
  • Inflammation Mediators / metabolism
  • MAP Kinase Kinase 4 / drug effects
  • MAP Kinase Signaling System / drug effects
  • Mice
  • NF-kappa B / drug effects
  • Nitric Oxide Synthase Type II / drug effects
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism
  • Osteoclasts / drug effects*
  • PPAR gamma / analysis
  • PPAR gamma / antagonists & inhibitors
  • PPAR gamma / pharmacology*
  • RANK Ligand / drug effects*
  • Reactive Oxygen Species / analysis
  • Root Canal Filling Materials / pharmacology*
  • Signal Transduction / drug effects
  • Silver / pharmacology*
  • Titanium / pharmacology*
  • Transcription Factor AP-1 / drug effects
  • Transfection

Substances

  • Anti-Inflammatory Agents
  • Epoxy Resins
  • Fluoresceins
  • Fluorescent Dyes
  • Inflammation Mediators
  • NF-kappa B
  • PPAR gamma
  • RANK Ligand
  • Reactive Oxygen Species
  • Root Canal Filling Materials
  • Tnfsf11 protein, mouse
  • Transcription Factor AP-1
  • diacetyldichlorofluorescein
  • Silver
  • epoxy resin AH-26
  • Titanium
  • Nitric Oxide Synthase Type II
  • Nos2 protein, mouse
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2
  • Extracellular Signal-Regulated MAP Kinases
  • MAP Kinase Kinase 4
  • Bismuth