Effect of triethylene glycol dimethacrylate on the cytotoxicity, cyclooxygenase-2 expression and prostanoids production in human dental pulp cells

Int Endod J. 2012 Sep;45(9):848-58. doi: 10.1111/j.1365-2591.2012.02042.x. Epub 2012 Apr 6.

Abstract

Aim: To evaluate the effect of TEGDMA on cell cycle progression as well as alterations of cell cycle-related gene and protein expression.

Methodology: Human dental pulp cells were exposed to 0-5 mmol L(-1) TEGDMA for 24 h. Cytotoxicity was evaluated by 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. Cell cycle progression was analysed by propidium iodide (PI) flow cytometry. Cell death pathway was surveyed by annexin V/PI dual-staining flow cytometry. The mRNA expression of cell cycle-related genes (cdc2, cyclinB1 and p21) and COX-2 was evaluated by reverse transcriptase-polymerase chain reaction, and their protein expression was evaluated by Western blotting. The production of PGE(2) and PGF(2α) in the culture medium was determined by enzyme-linked immunosorbent assay.

Results: Triethylene glycol dimethacrylate inhibited cellular growth and induced cell cycle deregulation in dental pulp cells. High-dose exposure provoked both necrotic and apoptotic cell death. The gene and protein expression of cdc2, cyclin B1 and cdc25C declined obviously whilst cells treated with 2.5 mmol L(-1) TEGDMA concurrent with the elevated expression of p21. The mRNA and protein expression of COX-2, along with production of PGE(2) and PGF(2α), are drastically raised by 2.5-5 mmol L(-1) TEGDMA.

Conclusions: Triethylene glycol dimethacrylate induced cytotoxicity, cell cycle arrest and apoptosis in dental pulp cells, which was associated with the decline of cdc2, cyclin B1, cdc25C expression and elevation of p21 expression. Concomitantly, COX-2 expression, PGE(2) and PGF(2α) production increased. These effects may contribute to explain the pulpal damage and inflammation induced by TEGDMA after operative procedures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Annexin A5 / pharmacology
  • Apoptosis / drug effects
  • CDC2 Protein Kinase
  • Cell Culture Techniques
  • Cell Cycle / drug effects
  • Cell Death / drug effects
  • Cell Proliferation / drug effects
  • Cell Shape / drug effects
  • Coloring Agents
  • Cyclin B / drug effects
  • Cyclin B1 / drug effects
  • Cyclin-Dependent Kinase Inhibitor p21 / drug effects
  • Cyclin-Dependent Kinases
  • Cyclooxygenase 2 / drug effects*
  • Dental Materials / toxicity*
  • Dental Pulp / cytology
  • Dental Pulp / drug effects*
  • Dinoprost / analysis
  • Dinoprostone / analysis
  • Enzyme Inhibitors / pharmacology
  • Flow Cytometry / methods
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • Humans
  • Necrosis
  • Polyethylene Glycols / toxicity*
  • Polymethacrylic Acids / toxicity*
  • Propidium
  • Prostaglandins / biosynthesis*
  • Tetrazolium Salts
  • Thiazoles
  • Time Factors
  • cdc25 Phosphatases / drug effects

Substances

  • Annexin A5
  • CCNB1 protein, human
  • CDKN1A protein, human
  • Coloring Agents
  • Cyclin B
  • Cyclin B1
  • Cyclin-Dependent Kinase Inhibitor p21
  • Dental Materials
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Polymethacrylic Acids
  • Prostaglandins
  • Tetrazolium Salts
  • Thiazoles
  • triethylene glycol dimethacrylate
  • Propidium
  • Polyethylene Glycols
  • Dinoprost
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • CDC2 Protein Kinase
  • CDK1 protein, human
  • Cyclin-Dependent Kinases
  • CDC25C protein, human
  • cdc25 Phosphatases
  • thiazolyl blue
  • Fluorescein-5-isothiocyanate
  • Dinoprostone