Cell toxicity of methacrylate monomers-the role of glutathione adduct formation.

Polymer-based dental restorative materials are designed to polymerize in situ. However, the conversion of methacrylate monomer to polymer is never complete, and leakage of the monomer occurs. It has been shown that these monomers are toxic in vitro; hence concerns regarding exposure of patients and dental personnel have been raised. Different monomer methacrylates are thought to cause toxicity through similar mechanisms, and the sequestration of cellular glutathione (GSH) may be a key event. In this study we examined the commonly used monomer methacrylates, 2-hydroxyethylmethacrylate (HEMA), triethylenglycol-dimethacrylate (TEGDMA), bisphenol-A-glycidyl-dimethacrylate (BisGMA), glycerol-dimethacrylate (GDMA) and methyl-methacrylate (MMA). The study aimed to establish monomers' ability to complex with GSH, and relate this to cellular toxicity endpoints. Except for BisGMA, all the monomer methacrylates decreased the GSH levels both in cells and in a cell-free system. The spontaneous formation of methacrylate-GSH adducts were observed for all methacrylate monomers except BisGMA. However, we were not able to correlate GSH depletion and toxic response measured as SDH activity and changes in cell growth pattern. Together, the current study indicates mechanisms other than GSH-binding to be involved in the toxicity of methacrylate monomers.

HEMA reduces cell proliferation and induces apoptosis in vitro.

OBJECTIVES: Methacrylate monomers have been identified in aqueous extracts of freshly cured compomers. Both cells in the pulpal cavity and various cells of the oral mucosa can potentially be exposed to these leachables. Short-term exposure to dental monomers at relatively high concentrations induces adverse biological effects in vitro. The mechanisms involved have not been fully elucidated although involvement of various signaling pathways including ROS formation, activation of MAP-kinases and caspases has been suggested. The aim of this study was to investigate potential cellular responses following long-term exposure to relatively low and potentially more clinical relevant HEMA concentrations. METHODS: A submandibular gland cell line was exposed to HEMA (20-600 microM) for up to 72h. The impact on cell proliferation, apoptosis, and possible underlying mechanisms was assessed by flow cytometry, microscopy and western blotting. RESULTS: Exposure to HEMA (600 microM) resulted in reduced cell proliferation after 24h and increased apoptosis after 60h. Further, we observed ATM dependent phosphorylation of p53, advocating an initial DNA damage in the HEMA exposed cells. SIGNIFICANCE: In conclusion, we show that exposure to relatively low concentration of HEMA for a prolonged time result in cell death, possibly as a consequence of DNA damage.

Toxicity evaluation of root canal sealers in vitro.

AIM: To compare the toxicity of methacrylate resin-based root canal sealers with sealers based on epoxy resin and silicone by two-well established cell culture methods. METHODOLOGY: Specimens of AH Plus, EndoREZ, RoekoSeal and Epiphany were prepared for direct contact in the Millipore filter diffusion test and as extracts in the MTT assay. Mouse fibroblasts (L929) were used as toxicity targets. Differences in cytotoxicity between fresh and set specimens and between the extracts of root canal sealers were determined by t-test (P < 0.05). RESULTS: In the filter diffusion test, freshly mixed Epiphany and AH Plus were rated severely toxic and RoekoSeal and EndoREZ nontoxic. When set, Epiphany was moderately toxic, whereas AH Plus, RoekoSeal and EndoREZ were nontoxic. Epiphany was significantly more toxic than RoekoSeal and EndoREZ (P < 0.05). In the MTT assay with set specimens, Epiphany was rated severely toxic; AH Plus and RoekoSeal slightly toxic; and EndoREZ nontoxic. Epiphany was significantly more toxic than the other three materials in this test (P < 0.001). CONCLUSION: The multi-methacrylate resin-based (Epiphany) root canal sealer was significantly more toxic to L-929 cells than the silicone-based Roeko Seal and the single methacrylate-based EndoREZ root canal sealers. AH Plus showed intermediate toxicity.

Corrosion and biocompatibility testing of palladium alloy castings.

OBJECTIVE: The biocompatibility of palladium-copper alloys has been questioned in the literature. The intention of the present work was to study: (a) the release of ions in an immersion test from a copper-containing alloy, Option((R)) (79% Pd, 10% Cu, 9% Ga, 2% Au), compared with an alloy without Cu, IS85 (82% Pd, 6% Ga, 3.5% Sn, 3.5% In, 2.5% Ag, 2.5% Au); (b) the effect of oxide films produced by preoxidation on corrosion resistance; and (c) the possibility of biologically synergetic effects of ions released. METHODS: Specimens of both alloys were cast, rubbed and ultrasonically cleaned. Metallographic specimens were prepared after (a) casting and (b) preoxidizing treatment at approximately 1000 degrees C and studied by SEM and EDS. Immersion tests were carried out in a solution of 0.1mol/l of NaCl and 0.1mol/l of lactic acid at 37 degrees C for 7 days. The alloy specimens were tested in the following three steps: (1) as preoxidized; (2) after subsequent removal of a 0.1mm thick layer by grinding; and (3) after an additional removal of approximately 0.1mm by grinding. The test solutions were analyzed by means of ICP to record the amounts of ions that had leached out from the alloy specimens. The biocompatibility was studied by cell culture tests and the HET-CAM method. Test solutions were prepared by dissolving PdCl(2) and CuCl(2) to appropriate concentrations. RESULTS: The metallographic investigations revealed moderate segregations in interdendritic regions and grain boundaries. After preoxidation in air a zone of oxidation from 25 up to 200 microm thickness for Option and from 5 to 10 microm for IS85 was found. Oxidation was observed along a rim for both alloys and for Option also along interdendritic positions. The oxides were seen as small, dark spots <1 microm in a metallic matrix. These results indicate that: (1) the oxidation of the copper-containing palladium alloy is far more severe than that of the alloy with no copper; and (2) the elemental release from these oxides is substantially larger than that from the corrosion of the metallic structure. The results of the cell culture testing showed that Cu was most toxic, followed by Cu(2+)+Pd(2+) (1:2), based on the determination of the concentration that caused 50% cytotoxicity. The HET-CAM testing showed Cu(2+)+Pd(2+) (1:2) to have the highest irritation score. SIGNIFICANCE: The copper-containing Pd alloy developed a 0.1mm thick rim with small oxide particles in a metallic matrix during preoxidation. If this rim is not completely removed, significantly more Cu, Ga and Pd ions have been shown to leach into the test solution than from the alloy structure. No synergetic effect of Cu and Pd ions was observed in cultured cells, while the mixture Pd(2+)+Cu(2+) (1:2) was most irritating to mucous membrane as evaluated by the HET-CAM method.