Involvement of oxidative stress in mutagenicity and apoptosis caused by dental resin monomers in cell cultures

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Lee, Dong Hee; Lim, Bum-Soon; Lee, Yong-Keun; Ahn, Sug-Joon; Yang, Hyeong-Cheol

Issue Date
Dental Materials 2006;22:1086-1092
resin monomersGMATEGDMAHEMAPulp cellscytotoxicitymicronucleiDNA fragmentationapoptosisROSGSH
Objective. This investigation studied the possibility that apoptosis as well as mutagenicity
induced by resin monomers are mediated by oxidative stress.
Methods. A range of dilutions of three resin monomers (GMA, TEGDMA, and HEMA) was
added to culture medium (DMEM/10% FBS), of V79-4 fibroblasts and RPC-C2A pulp cells
for 24 h. Their cytotoxic effects were measured by a colorimetric functional assay (MTT).
Chromosomal aberration induced by the resin monomers was investigated by counting
micronuclei in V79-4 cells. The effects of the resin monomers on DNA fragmentation were
viewed by agarose gel electrophoresis of DNA, isolated from RPC-C2A pulp cells that were
treated by resin compounds. Resin monomer-induced apoptosis was further confirmed by
flow cytometry (staining with both annexin V-FITC and PI).
Results. All monomers exhibited a dose-dependent cytotoxic effect, and the ranking of the
cytotoxicity based on TC50 was GMA>TEGDMA> HEMA. The resin monomer-induced cytotoxicity
was significantly decreased by co-treatment with N-acetylcystein (NAC), an antioxidant.
The authors also confirmed a dose-dependent genotoxicity of the resin monomers
that had induced micronucleated cells in V79-4 fibroblasts. Similar to the effects on cytotoxicity,
NAC reduced the numbers of micronuclei in comparison with those generated by the
resin monomers. The preventive effects of NAC were also observed in monomer-induced
apoptosis in RPC-C2A cells. A DNA ladder pattern, characteristic of apoptosis, was shown
at cytotoxic concentrations, but NAC blocked the resin monomer-mediated DNA fragmentation.
The preventive effects of NAC on apoptosis were confirmed by Annexin V staining.
Cells exposed to 300 M GMA, 7mM TEGDMA, or 14mM HEMA for 24h showed a significant
increase in apoptotic cells, while NAC co-treatment caused a reduction in apoptotic cells
compared to controls.
Significance. These findings suggest that glutathione depletion and oxidative stress are
responsible for GMA, TEGDMA, and HEMA-induced mutagenicity and apoptosis.
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College of Dentistry/School of Dentistry (치과대학/치의학대학원)Dept. of Dentistry (치의학과)Journal Papers (저널논문_치의학과)
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