Cytotoxicity and induction of DNA double-strand breaks by components leached from dental composites in primary human gingival fibroblasts.

Abstract

INTRODUCTION:

The public interest steadily increases in the biological adverse effects caused by components released from resin-based dental restorations.

OBJECTIVE:

In this study, the cytotoxicity and the genotoxicity were investigated of following released components from dental resin restorations in human gingival fibroblasts (HGF): tetraethyleneglycol dimethacrylate (TEEGDMA), neopentylglycol dimethacrylate (Neopen), diphenyliodoniumchloride (DPIC), triphenyl-stibane (TPSB) and triphenylphosphane (TPP).

METHODS:

XTT based cell viability assay was used for cytotoxicity screening of substances. γ-H2AX assay was used for genotoxicity screening. In the γ-H2AX assay, HGFs were exposed to the substances for 6h. Induced foci represent double DNA strand breaks (DSBs), which can induce ATM-dependent phosphorylation of the histone H2AX. Cell death effects (apoptosis and necrosis), induced by the substances were visually tested by the same investigator using the fluorescent microscope.

RESULTS:

All tested substances induced a dose-dependent loss of viability in HGFs. Following toxicity ranking among the substances at EC50-concentration were found in the XTT assay (mM, mean±SEM; n=5): DPIC>Neopen>TPSB>TPP>TEEGDMA. DSB-foci per HGF-cell were obtained, when HGFs were exposed to the EC50-concentration of each substance in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP>TEEGDMA. Multi-foci cells (cells that contain more than 40 foci each) in 80 HGF-cells at EC50-concentration of each substance were found as follow (mean±SEM; n=3): DPIC>Neopen>TPP>TPSB>TEEGDMA. Cell apoptosis contained in each substance at EC50-concentration in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP >TEEGDMA. Cell necrosis contained in each substance at EC50-concentration in the following order (mean±SEM; n=3): DPIC>Neopen>TPSB>TPP>TEEGDMA.

CONCLUSION:

Leached components from dental resin restorations can induce DNA DSBs and cell death effects in HGFs.

• Authors: Mohamed Shehata, Jürgen Durner, Ayce Eldenez, Kirsten Van Landuyt, Panorea Styllou, Lena Rothmund, Reinhard Hickel, Harry Scherthan, Werner Geurtsen, Bernd Kaina, Thomas Carell, Franz X Reichl
• Journal: Dental materials : official publication of the Academy of Dental Materials (Dent Mater) Vol. 29 Issue 9 Pg. 971-9 (Sep 2013) ISSN: 1879-0097 [Electronic] England
• PMID: 23915819 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Chemical References

• Biphenyl Compounds
• Composite Resins
• Dental Materials
• H2AX protein, human
• Histones
• Indicators and Reagents
• Methacrylates
• Mutagens
• Onium Compounds
• Organophosphorus Compounds
• Polymethacrylic Acids
• Stilbenes
• Tetrazolium Salts
• diphenyliodonium
• 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide
• triethylene glycol dimethacrylate
• neopentyl methacrylate
• triphenylphosphine
• Polyethylene Glycols
• estrobin

Topics

• Apoptosis (drug effects)
• Biphenyl Compounds (toxicity)
• Cell Culture Techniques
• Cell Line
• Cell Survival (drug effects)
• Composite Resins (toxicity)
• DNA Breaks, Double-Stranded (drug effects)
• Dental Materials (toxicity)
• Dose-Response Relationship, Drug
• Fibroblasts (drug effects)
• Gingiva (cytology, drug effects)
• Histones (drug effects)
• Humans
• Indicators and Reagents
• Materials Testing
• Methacrylates (toxicity)
• Microscopy, Fluorescence
• Mutagens (toxicity)
• Necrosis
• Onium Compounds (toxicity)
• Organophosphorus Compounds (toxicity)
• Polyethylene Glycols (toxicity)
• Polymethacrylic Acids (toxicity)
• Stilbenes (toxicity)
• Tetrazolium Salts