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Glutathione (GSH) is important for the self-protection of cells against oxidative stress and toxic xenobiotics, whereas reactive oxygen species (ROS) at elevated concentrations may cause detrimental alterations of cell membranes, DNA, and other cellular structures. The present investigation addressed the effects of triethylene-glycoldimethacrylate (TEGDMA) and camphorquinone (CQ) on glutathione metabolism and the formation of ROS in oral cells. Primary human pulp fibroblasts were exposed to various concentrations of TEGDMA and CQ (0.1-5 mM). Subsequently, GSH concentration and ROS formation were analyzed with the use of the monobromobimane assay (GSH) and 2',7'-dichlorofluorescein diacetate (DCFH-DA) (ROS). The endogenous ROS hydrogen peroxide (H2O2) was used as a positive control (0.02-2 mM). TEGDMA significantly decreased GSH at concentrations between 0.5 and 5 mM (p<0.05), but did not elevate ROS levels. Contrary, CQ increased ROS formation at concentrations>or=1 mM, but had only a moderate effect on GSH at the highest test concentration. Hydrogen peroxide increased ROS and simultaneously decreased GSH at concentrations of >or=0.2 mM. These data show that the investigated substances may cause cell damage due to various mechanisms, GSH decrease and/or ROS increase. As a consequence, TEGDMA and CQ released into an aqueous environment from resinous materials might interact, thus generating significant cytotoxic effects even at low concentrations.
Glutathione (GSH) is important for the self-protection of cells against oxidative stress and toxic xenobiotics, whereas reactive oxygen species (ROS) at elevated concentrations may cause detrimental alterations of cell membranes, DNA, and other cellular structures. The present investigation addressed the effects of triethylene-glycoldimethacrylate (TEGDMA) and camphorquinone (CQ) on glutathione metabolism and the formation of ROS in oral cells. Primary human pulp fibroblasts were exposed to various concentrations of TEGDMA and CQ (0.1-5 mM). Subsequently, GSH concentration and ROS formation were analyzed with the use of the monobromobimane assay (GSH) and 2',7'-dichlorofluorescein diacetate (DCFH-DA) (ROS). The endogenous ROS hydrogen peroxide (H2O2) was used as a positive control (0.02-2 mM). TEGDMA significantly decreased GSH at concentrations between 0.5 and 5 mM (p<0.05), but did not elevate ROS levels. Contrary, CQ increased ROS formation at concentrations>or=1 mM, but had only a moderate effect on GSH at the highest test concentration. Hydrogen peroxide increased ROS and simultaneously decreased GSH at concentrations of >or=0.2 mM. These data show that the investigated substances may cause cell damage due to various mechanisms, GSH decrease and/or ROS increase. As a consequence, TEGDMA and CQ released into an aqueous environment from resinous materials might interact, thus generating significant cytotoxic effects even at low concentrations.
The objective of this investigation was to analyze whether various combinations of the ROS scavengers glutathione (GSH), N-acetyl-cysteine (NAC), and vitamins C and E decrease DNA damage due to visible-light-irradiated (VL-irradiated) camphorquinone/N,N-dimethyl-p-toluidine (CQ/DMT) compared with individual vitamin C or E. PhiX-174 RF plasmid DNA was used to determine single and double strand breaks as parameters of DNA damage. Individual ROS scavengers and combinations of the antioxidants were added to plasmid DNA treated with VL-irradiated CQ/DMT/Cu (II). After incubation, DNA was loaded into a 1% agarose gel. Following electrophoresis, gels stained with 0.5 microg/mL ethidium bromide were photographed under ultraviolet illumination and analyzed with NIH ImageJ software. Results were evaluated between groups for statistical significance using Student's paired t-test (p < 0.05). Glutathione significantly reduced oxidative DNA damage at all test concentrations when combined with vitamin C or vitamin E. The concentration of damaged DNA observed in the presence of combinations of GSH with vitamin C or vitamin E was significantly lower compared with all other combinations of antioxidants investigated in our study (p < 0.05). In contrast to GSH, NAC was not able to compensate the pro-oxidative effects of vitamin C and vitamin E. Only at a concentration of 2 mM, NAC combined with vitamin C efficiently prevented CQ/DMT/Cu (II)-associated DNA damage. Our data indicate that solely the combinations of GSH with vitamin C or vitamin E significantly reduce the severity of oxidative DNA damage caused by CQ/DMT, whereas NAC may even increase the pro-oxidant activity of vitamin C and vitamin E.
Toxicity potentiation of two monomers [bisphenol-A-glycidyldimethacrylate (BisGMA) and urethanedimethacrylate (UDMA)] as well as two comonomers [triethyleneglycoldimethacrylate (TEGDMA) and 2-hydroxyethylmethacrylate (HEMA)], each in combination with H(2)O(2), was investigated on the viability on human gingival fibroblasts (HGF) and human pulpal fibroblasts (HPF). The applied concentration of H(2)O(2) was 0.06 or 0.1 mmol/l, respectively, corresponding to the EC(0) of H(2)O(2) in HGF or HPF. The cell viability was assessed by the XTT test. From this test the half maximum effect concentrations (EC(50)) were calculated from fitted sigmoidale curves. EC(50) values were (HGF; mmol/l; mean +/- s.e.m.; n = 5): HEMA 11.9 +/- 0.9, TEGDMA 3.7 +/- 0.3, H(2)O(2) 0.36 +/- 0.04, UDMA 0.27 +/- 0.08, and BisGMA 0.11 +/- 0.03. No significant (P < 0.05) differences in the EC(50) values were observed when HGF was exposed to substances, as compared to HPF. No significant decrease of the EC(50) values was found when HGF or HPF, respectively, was exposed to HEMA or BisGMA in addition with H(2)O(2) up to the concentration of 0.1 mmol/l, as compared to those EC(50) values of each compound without H(2)O(2) addition. A significant decrease of the TEGDMA EC(50) value from 3.7 to 2.1 or 0.4 mmol/l, respectively, was found when cells were exposed to TEGDMA in combination with H(2)O(2) (0.06 or 0.1 mmol/l), as compared to that TEGDMA EC(50) value without H(2)O(2) addition. A significant decrease of the UDMA EC(50) value from 0.27 to 0.11 or 0.08 mmol/l, respectively, was found when HGF or HPF was exposed to UDMA in combination with H(2)O(2) (0.06 or 0.1 mmol/l), as compared to that UDMA EC(50) value without H(2)O(2) addition. The addition of H(2)O(2) (0.06 or 0.1 mmol/l) resulted in a toxicity potentiation of TEGDMA and UDMA, but not of HEMA and BisGMA, on HGF or HPF.
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