Cytotoxicity and DNA double-strand breaks in human gingival fibroblasts exposed to eluates of dental composites

Yang Yang; Franz-Xaver Reichl; Jianwei Shi; Xiuli He; Reinhard Hickel; Christof Högg

doi:10.1016/j.dental.2017.10.002

Cytotoxicity and DNA double-strand breaks in human gingival fibroblasts exposed to eluates of dental composites

Dent Mater. 2018 Feb;34(2):201-208. doi: 10.1016/j.dental.2017.10.002. Epub 2017 Oct 15.

Authors

Yang Yang¹, Franz-Xaver Reichl¹, Jianwei Shi², Xiuli He¹, Reinhard Hickel³, Christof Högg⁴

Affiliations

¹ Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, Germany; Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-University of Munich, Nußbaumstr. 26, 80336 Munich, Germany.
² Department of Orthodontics, Ludwig-Maximilians-University of Munich, Goethestr. 70, 80336 Munich, Germany.
³ Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, Germany.
⁴ Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, Germany; Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-University of Munich, Nußbaumstr. 26, 80336 Munich, Germany. Electronic address: christof.hoegg@lrz.uni-muenchen.de.

PMID: 29042079
DOI: 10.1016/j.dental.2017.10.002

Abstract

Objective: Previously, single composite components were used to study cytotoxicity and induction of DNA double-strand breaks (DNA-DSBs) of dental composite resins. In the present study, cytotoxicity and induction of DNA-DSBs in human gingival fibroblasts (HGFs) were investigated with dental composite eluates consisting of multiple components. The eluates were qualified and quantified.

Methods: The composites Esthet.X^® HD, Venus^®, X-tra fil^®, CLEARFIL™ AP-X, Admira^® Fusion and QuiXfil^® were polymerized and immersed into Dulbecco's modified Eagle's medium (DMEM) for 72h. Subsequently, HGFs were incubated with the corresponding composite eluates. The cell viability of HGFs was obtained from an XTT assay. DNA-DSBs were determined using a γ-H2AX assay. The qualification and quantification of eluates were performed by gas chromatography/mass spectrometry (GC/MS).

Results: HGFs exposed to the eluates of all investigated composites showed no significant loss of cell viability, compared to negative control. Significant DNA-DSBs induction could be found in HGFs exposed to the eluates of Esthet.X^® HD (0.43±0.05 foci/cell) and Venus^® (0.39±0.04 foci/cell), compared to control (0.22±0.03 foci/cell). A total of 12 substances were detected from the investigated composite eluates. Five of them were methacrylates: tetraethyleneglycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA), hydroxypropyl methacrylate (HPMA), ethyleneglycol dimethacrylate (EGDMA) and trimethylolpropane trimethacrylate (TMPTMA). The highest concentration of HEMA (110.5μM), HPMA (86.08μM) and TMPTMA (4.50μM) was detected in the eluates of QuiXfil^®. The highest concentration of TEGDMA was 1080μM in Venus^® eluates and the highest concentration of EGDMA was 3.18μM in Esthet.X^® HD eluates.

Significance: Significant DNA-DSBs induction can be found in HGFs exposed to the eluates of Esthet.X^® HD and Venus^®. The interactive effects among released (co)monomers and additives may influence the cytotoxicity and induction of DNA-DSBs, compared to exposure with single composite component.

Keywords: Culture medium; Cytotoxicity; DMEM; DNA double-strand breaks; Elution; Genotoxicity; Multiple components; TEGDMA; γ-H2AX.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Cell Survival / drug effects
Composite Resins / chemistry
Composite Resins / toxicity*
DNA Breaks, Double-Stranded*
Fibroblasts / drug effects*
Gas Chromatography-Mass Spectrometry
Gingiva / cytology*
Humans
In Vitro Techniques
Methacrylates
Siloxanes

Substances

Clearfil AP-X
Composite Resins
Esthet-X
Methacrylates
QuiXfil
Siloxanes
Venus (composite resin)
X-tra fil composite resin
admira