RESUMO
BACKGROUND: Dental composites have become the standard filling material to restore teeth, but during the placement of these restorations, high amounts of respirable composite dust (<5 µm) including many nano-sized particles may be released in the breathing zone of the patient and dental operator. Here we tested the respirable fraction of several composite particles for their cytotoxic effect using an alveolar macrophage model system. âMETHODS: Composite dust was generated following a clinical protocol, and the dust particles were collected under sterile circumstances. Dust was dispersed in fluid, and 5-µm-filtered to enrich the respirable fractions. Quartz DQ12 and corundum were used as positive and negative control, respectively. Four concentrations (22.5 µg/ml, 45 µg/ml, 90 µg/ml and 180 µg/ml) were applied to NR8383 alveolar macrophages. Light and electron microscopy were used for subcellular localization of particles. Culture supernatants were tested for release of lactate dehydrogenase, glucuronidase, TNF-α, and H2O2. RESULTS: Characterization of the suspended particles revealed numerous nano-sized particles but also many high volume particles, most of which could be removed by filtering. Even at the highest concentration (180 µg/ml), cells completely cleared settled particles from the bottom of the culture vessel. Accordingly, a mixture of nano- and micron-scaled particles was observed inside cells where they were confined to phagolysosomes. The filtered particle fractions elicited largely uniform dose-dependent responses, which were elevated compared to the control only at the highest concentration, which equaled a mean cellular dose of 120 pg/cell. A low inflammatory potential was identified due to dose-dependent release of H2O2 and TNF-α. However, compared to the positive control, the released levels of H2O2 and TNF-α were still moderate, but their release profiles depended on the type of composite. CONCLUSIONS: Alveolar macrophages are able to phagocytize respirable composite dust particle inclusive nanoparticles. Since NR8383 cells tolerate a comparatively high cell burden (60 pg/cell) of each of the five materials with minimal signs of cytotoxicity or inflammation, the toxic potential of respirable composite dust seems to be low. These results are reassuring for dental personnel, but more research is needed to characterize the actual exposure and uptake especially of the pure nano fraction.
Assuntos
Resinas Compostas , Poeira , Macrófagos Alveolares/metabolismo , Animais , Células Cultivadas , Estresse Oxidativo , RatosRESUMO
AIM: To investigate the cytotoxicity and genotoxicity of a new silicate-based BioRoot RCS® sealer in comparison with contemporary sealers. METHODOLOGY: A periodontal ligament cell line using lentiviral gene transfer of human telomerase reverse transcriptase (hTERT) was used and exposed to subtoxic concentrations of 24-h eluates from two epoxy resin-based (AH Plus Jet® and Acroseal® ), four various methacrylate-based endodontic sealers (EndoREZ® , RealSeal® , RealSeal SE® and Hybrid Root SEAL® ) and three silicate-based sealers (BioRoot RCS® , iRootSP® and MTA Fillapex® ). The XTT-based cell viability assay was used for cytotoxicity screening of materials. The γ-H2AX assay was used for genotoxicity screening. In the γ-H2AX immunofluorescence assay, PDL-hTERT cells were exposed to eluates of the substances for 6 h and DNA double-strand breaks (DSB) were detected microscopically. Induced foci represented DSBs, which can induce ATM-dependent phosphorylation of the histone H2AX. The statistical significance of the differences between the experimental groups was compared using the Student's t-test (P < 0.05). RESULTS: The cytotoxicity of the 24-h eluates could be ranked in the following order: Hybrid Root SEAL® >RealSeal® >Acroseal® >RealSeal SE® ≥ AH Plus Jet® > EndoREZ® >MTA Fillapex® > iRoot SP® >BioRoot RCS® . In negative controls (cells which received medium only) 4.08 ± 0.53 DSB foci (mean ± SEM) whilst in positive controls 10.76 ± 4.05 DSB foci/cell were found. BioRoot RCS® and RealSeal SE® exhibited significant differences in foci formation at 1/3 EC50 compared with their 1/10 EC50 concentration (P < 0.05). Both concentrations (1/10 and 1/3 of EC50) of AH Plus Jet® , Acroseal® , RealSeal® and MTA Fillapex® sealers were not significantly different when compared with the medium control (P < 0.05). CONCLUSIONS: New BioRoot RCS® was not toxic whilst Hybrid Root SEAL® demonstrated more toxicity and DNA double-strand breaks when compared with other resin- and silicate-based root canal sealers.
Assuntos
Quebras de DNA de Cadeia Dupla/efeitos dos fármacos , Selantes de Fossas e Fissuras/toxicidade , Materiais Restauradores do Canal Radicular/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Resinas Compostas/toxicidade , Resinas Epóxi/toxicidade , Humanos , Testes de Mutagenicidade , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacosRESUMO
Phenol has been traditionally used in dental treatment as a sedative for the pulp or as disinfectant for carious cavity and root canal. However, phenol is regarded as a mutagenic and carcinogenic agent and its use in dental practice is now therefore restricted. Monochlorophenols are derivatives of phenol, which are still used clinically as root canal disinfectants, they are even more active antiseptics/disinfectants than phenol, and the so-called Walkhoff (ChKM) solution makes use of monochlorophenol for root canal disinfection. Ingredients in the ChKM solution are the monochlorophenol compound 4-chlorophenol (4-CP), camphor, and menthol. In literature, the use of the ChKM solution is controversial because of a possible DNA toxicity of the ingredient 4-CP. However, it is unknown whether ChKM can really induce DNA damage in human oral cells. In this study, the induction of DNA double-strand breaks (DSBs) by ChKM and monochlorophenol compounds (2-chlorophenol, 2-CP; 3-chlorophenol, 3-CP; and 4-chlorophenol, 4-CP) was tested in human gingival fibroblasts (HGFs). DNA DSBs (foci) induced in HGFs unexposed and exposed to monochlorophenols or ChKM solution were investigated using the γ-H2AX DNA focus assay, which is a direct marker for DSBs. DSBs result in the ATM-dependent phosphorylation of the histone H2AX. When cells were exposed to medium or medium + DMSO (1 %) (negative controls), an average of 3 foci per cell were found. In positive control cells (H2O2 + medium, or H2O2 + medium + DMSO (1 %), an average of 35 foci each were found. About 20 DSB foci per cell were found, when HGFs were exposed to 2-CP (4 mM), 3-CP (2.3 mM), 4-CP (2.1 mM), or ChKM (corresponding to 1.5 mM 4-CP). Our results show increasing DNA toxicities in the order of 2-CP < 3-CP < 4-CP < ChKM solution. An additive DNA toxicity was found for 4-CP in combination with camphor in the ChKM solution, compared to the 4-CP alone. No significant differences regarding multi-foci cells (cells that contain more than 40 foci) were found when HGFs were exposed to the EC50 concentrations (given in parenthesis) of ChKM (1.5 mM), 4-CP (2.1 mM), or 2-CP (4 mM). Significantly fewer multi-foci cells were found when HGFs were exposed to the EC50 concentration (given in parenthesis) of 3-CP (2.3 mM), compared to the EC50 concentrations of ChKM, 4-CP, or 2-CP. Monochlorophenol compounds and/or ChKM solution can induce DSBs in primary human oral (cavity) cells, which underscores their genotoxic capacity.
Assuntos
Cânfora/farmacologia , Clorofenóis/farmacologia , Quebras de DNA de Cadeia Dupla/efeitos dos fármacos , Desinfetantes de Equipamento Odontológico/farmacologia , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Mentol/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Clorofenóis/química , Desinfetantes de Equipamento Odontológico/química , Combinação de Medicamentos , Fibroblastos/citologia , Fibroblastos/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Gengiva/citologia , Gengiva/metabolismo , Histonas/genética , Histonas/metabolismo , Humanos , Concentração Inibidora 50 , Isomerismo , Testes de Mutagenicidade , Fosforilação/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacosRESUMO
Bisphenol-A-glycidyldimethacrylate (BisGMA) is used in many resin-based dental materials. It was shown in vitro that BisGMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of [(14)C]BisGMA applied via gastric and intravenous administration (at dose levels well above those encountered in dental care) were examined in vivo in guinea pigs to test the hypothesis that BisGMA reaches cytotoxic levels in mammalian tissues. [(14)C]BisGMA was taken up rapidly from the stomach and intestine after gastric administration and was widely distributed in the body following administration by each route. Most [(14)C] was excreted within one day as (14)CO(2). The peak equivalent BisGMA levels in guinea pig tissues examined were at least 1000-fold less than known toxic levels. The peak urine level in guinea pigs that received well in excess of the body-weight-adjusted dose expected in humans was also below known toxic levels. The study therefore did not support the hypothesis.
Assuntos
Bis-Fenol A-Glicidil Metacrilato/farmacocinética , Materiais Dentários/farmacocinética , Animais , Bile/química , Bis-Fenol A-Glicidil Metacrilato/administração & dosagem , Bis-Fenol A-Glicidil Metacrilato/análise , Sangue , Dióxido de Carbono/análise , Radioisótopos de Carbono , Ducto Cístico , Materiais Dentários/análise , Fezes/química , Cobaias , Injeções Intravenosas , Instilação de Medicamentos , Veias Jugulares , Masculino , Compostos Radiofarmacêuticos , Distribuição Aleatória , Fatores de Tempo , Distribuição Tecidual , UrinaRESUMO
OBJECTIVES: Resin (co)monomers may be released from restorative dental materials and can diffuse into the tooth pulp or the gingiva, and can reach the saliva and the circulating blood. Genotoxic potential of some dental composite components has been clearly documented. The genotoxic effects of xenobiotics can represent a possible step in tumor initiation and/or embryotoxicity/teratogenesis. A modified fluorescent mouse embryonic stem cell test (R.E.Tox) was used to test the embryotoxic potential of following dental restorative materials: Bisphenol A glycidylmethacrylate (BisGMA), urethanedimethacrylate (UDMA), hydroxyethylmethacrylate (HEMA), and triethyleneglycoldimethacrylate (TEGDMA), as well as some of their metabolic intermediates 2,3-epoxy-2-methyl-propionicacid-methylester (EMPME), methacrylic acid (MA), and 2,3-epoxy-2-methylpropionic acid (EMPA). METHODS: Mouse embryonic stem (ES) cells stably transfected with a vector containing the gene for the green fluorescent protein under control of the cardiac alpha-myosin heavy chain promoter were differentiated in the presence of various concentrations of the test compounds for 12 days. Fluorescence was measured using the TECAN Safire and values were expressed as percent of control values. To distinguish between cytotoxic and embryotoxic effects, all compounds were tested in a standard MTT assay. RESULTS: HEMA, TEGDMA and EMPME did not influence the differentiation process of ES cells towards cardiac myocytes. No cytotoxic effects were observed at any of the concentration levels tested. Exposure to BisGMA resulted in a 50% decrease in cell survival and a very strong inhibition of cell differentiation at 10(-5)M (p<0.01). Embryotoxic effects were also present at 10(-6) and 10(-7)M (p<0.05). EMPA induced a decrease in ES cell differentiation at 10(-5)M (p<0.01) without cytotoxic effects. No embryotoxic effects were induced at lower concentrations. Exposure to UDMA resulted in a slight decrease of cell differentiation at 10(-5)M (p<0.05). Exposure of cells to MA resulted in an increase of cardiac differentiation up to 150% (p<0.05) at 10(-5)M without cytotoxic effects. CONCLUSIONS: BisGMA induced a significant high embryotoxic/teratogenic effect over a large range of concentration. Therefore attention should be focused on this dental monomer, which should be investigated further by in vivo experiments.
Assuntos
Resinas Compostas/toxicidade , Restauração Dentária Permanente/efeitos adversos , Embrião de Mamíferos/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Animais , Bis-Fenol A-Glicidil Metacrilato/toxicidade , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Compostos de Epóxi/toxicidade , Coração/efeitos dos fármacos , Coração/embriologia , Teste de Materiais , Metacrilatos/toxicidade , Camundongos , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/toxicidade , Poliuretanos/toxicidade , Propionatos/toxicidade , Testes de ToxicidadeRESUMO
The cytotoxic potentials of the dental composite components triethyleneglycoldimethacrylate (TEGDMA) and 2-hydroxy-ethylmethacrylate (HEMA) as well as mercuric chloride (HgCl2) and methyl mercury chloride (MeHgCl) were investigated. Proliferating A549 and L2 cell monolayers were cultured in the absence or presence of composite components or mercurials. Twenty-four hours later the tetrazolium salt XTT (sodium 3'-[1-phenyl-aminocarbonyl)-3,4-tetrazolium]bis(4-methoxy-6-nitro)benzenesulphonic acid) was added. Formazan formation was quantified using a microtiter plate reader. EC50 values were obtained as half-maximum-effect concentrations from fitted curves. EC50 values were in A549 cells (mean values +/- standard deviation; n = 12; micromol/l); HEMA 8854+/-1882; TEGDMA 1821+/-529; HgCl2 41+/-7 and MeHgCl 27+/-3. EC50 values in L2 cells were: HEMA 191+/-28; TEGDMA 112+/-16; HgCl2 25+/-6 and MeHgCl 8+/-6. All tested substances induced a dose-dependent loss of viability in A549 and L2 cells after 24 h. The EC50 values of both mercurials were significantly (p < 0.05) lower compared to the values of both composite components. TEGDMA was about 5-fold (A549 cells) and about 2-fold (L2 cells) more toxic compared to HEMA. It is to be assumed that the risk of lung cell damage by dental composite components is even more unlikely.
Assuntos
Resinas Acrílicas/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Resinas Compostas/farmacologia , Pulmão/efeitos dos fármacos , Compostos de Mercúrio/farmacologia , Metacrilatos/farmacologia , Polietilenoglicóis/farmacologia , Ácidos Polimetacrílicos/farmacologia , Poliuretanos/farmacologia , Animais , Linhagem Celular , Humanos , Pulmão/citologia , Ratos , Células Tumorais CultivadasRESUMO
No toxicokinetic data are available about the dental composite component 2-hydroxyethylmethacrylate (HEMA) in vivo in the literature. Therefore, the excretion of HEMA in feces and urine in vivo and, using the pendular perfusion technique with segments of jejunum and colon, in the biliary and enteric excretion in situ were investigated in anesthetized guinea pigs. In the in situ experiments, guinea pigs (n = 4) received HEMA (0.02 mmol/kgbw labelled with a tracer dose 14C-HEMA 0.3 kBq/gbw) injected into the jugular vein. In the in vivo experiments, guinea pigs (n = 4) received HEMA (+ 14C-HEMA, same dose as above) via gastric tube. Urine and feces were collected for 24h. In the in situ experiments, organs from guinea pigs were removed 60 min after the beginning of the experiment, and then the 14C-radioactivity was measured. During the 60 min perfusion period the calculated amount of 14C-activity excreted into the total jejunum and colon was 6.0 +/- 1.0% and 2.7 +/- 0.7% of the dose administered, respectively (mean +/- sem). Of the 14C-HEMA dose, 5.3 +/- 0.3% was found in the bile. Significantly (p < 0.05) higher bile/blood concentration ratios were found at 10-40 min after the injection of HEMA, as compared to the ratio at 60 min. The total 14C-recovery in all organs tested was 20.0 +/- 2.6%. During 24h the amounts of 14C-activity excreted in the feces and urine were 1.1 +/- 0.1% or 17.1 +/- 1.50% of the dose administered, respectively (mean +/- sem). The total 14C-recovery in all organs tested was 11.6 +/- 0.6%. In a second series of in vivo experiments, exhaled air from the animals was captured during the 24h experimental period. 14C was exhaled to 63.6 +/- 2.11% of the administered 14C-HEMA dose (mean +/- sem; n = 4) as 14C-carbondioxide. The results indicate a rapid clearance of 14C-HEMA and/or 14C-HEMA metabolite(s) from the organism, exhalation being the major route of elimination.
Assuntos
Materiais Biocompatíveis/farmacocinética , Metacrilatos/farmacocinética , Animais , Materiais Biocompatíveis/administração & dosagem , Radioisótopos de Carbono/farmacocinética , Relação Dose-Resposta a Droga , Cobaias , Masculino , Metacrilatos/administração & dosagem , Fatores de Tempo , Distribuição TecidualRESUMO
The monomer triethyleneglycoldimethacrylate (TEGDMA) is used as a diluent in many resin-based dental materials. It was previously shown in vitro that TEGDMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of 14C-TEGDMA applied via gastric, intradermal, and intravenous administration at dose levels well above those encountered in dental care were examined in vivo in guinea pigs and mice as a test of the hypothesis that TEGDMA reaches cytotoxic levels in mammalian tissues. 14C-TEGDMA was taken up rapidly from the stomach and small intestine after gastric administration in both species and was widely distributed in the body following administration by each route. Most 14C was excreted within one day as 14CO2. The peak equivalent TEGDMA levels in all mouse and guinea pig tissues examined were at least 1000-fold less than known toxic levels. The study therefore did not support the hypothesis.
Assuntos
Resinas Compostas/metabolismo , Resinas Compostas/toxicidade , Polietilenoglicóis/metabolismo , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/metabolismo , Ácidos Polimetacrílicos/toxicidade , Animais , Resinas Compostas/administração & dosagem , Feminino , Cobaias , Injeções Intravenosas , Injeções Subcutâneas , Intubação Gastrointestinal , Masculino , Taxa de Depuração Metabólica , Camundongos , Camundongos Endogâmicos BALB C , Polietilenoglicóis/administração & dosagem , Ácidos Polimetacrílicos/administração & dosagem , Distribuição TecidualRESUMO
Lewisite is a highly toxic arsenic compound which can cause skin damage. In the present study effects of Lewisite on cell membrane integrity and energy metabolism as well as antidotal effects of DL-2,3-dimercaptopropanesulfonate (DMPS), and meso-2,3-dimercaptosuccinic acid (m-DMSA) were investigated in a keratinocyte derived cell line (SCL II) and primary human keratinocytes (HK). Cells were incubated in Lewisite (60 microM) containing medium for 5 min. During the following 6 h lactate dehydrogenase (LDH) activity in the supernatant, intracellular ATP content, tetrazolium reduction, glucose consumption and lactate formation were measured. Glucose consumption and lactate production were decreased in both cell lines after Lewisite exposure. In SCL II cells an increase of LDH activity in the supernatant, a decrease of ATP content, and an impaired ability to reduce tetrazolium was found 3 h after Lewisite exposure. In HK cultures tetrazolium reduction was significantly decreased already after 2 h, whereas LDH increase in the supernatant and ATP content decrease occurred only at 6 h after Lewisite exposure. When DMPS or m-DMSA was added directly after Lewisite exposure to SCL II cells, glucose consumption and lactate formation were restored and LDH leakage was prevented. SCL II cells might be more prone to membrane damage whereas in keratinocytes mitochondrial impairment seems to be the predominant effect of Lewisite.
Assuntos
Arsenicais/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Queratinócitos/efeitos dos fármacos , Trifosfato de Adenosina/análise , Arsenicais/antagonistas & inibidores , Sobrevivência Celular/efeitos dos fármacos , Quelantes/farmacologia , Formazans/análise , Glucose/metabolismo , Humanos , Queratinócitos/metabolismo , L-Lactato Desidrogenase/análise , Ácido Láctico/análise , Succímero/farmacologia , Sais de Tetrazólio/análise , Fatores de Tempo , Células Tumorais Cultivadas , Unitiol/farmacologiaRESUMO
OBJECTIVES AND METHODS: In a previous study it was postulated that toxicity of 2-hydroxyethylmethacrylate (HEMA) and triethleneglycoldimethacrylate (TEGDMA) is based on oxidative metabolites. In this study the influence of antioxidative vitamins (including uric acid) on the toxicity of HEMA or TEGDMA was tested. Toxicity of HEMA and TEGDMA was determined in rat alveolar epithelial L2, human malignant A549, and human fibroblast-like 11Lu cells by inhibition of methionine incorporation (as a marker of protein synthesis inhibition) and by determination of glutathione depletion, as well as by measurement of GSSG increase. RESULTS: Toxicity of the composite components HEMA and TEGDMA was demonstrated by GSH depletion as the most sensitive method. Five hundred micromoles per litre Vitamin C or 250 micromol/l Vitamin E were mostly able to decrease toxicity of HEMA and TEGDMA in the cell lines tested. In addition, 250 micromol/l Vitamin A was only effective in L2 cells impairing HEMA toxicity and 250 micromol/l uric acid impairing TEGDMA toxicity as assessed by decreased GSH depletion. In A549 cells only methionine incorporation inhibition but not GSH depletion was significantly affected. By contrast, in 11Lu cells methionine incorporation inhibition was not significantly changed, but GSH depletion was. CONCLUSIONS: The postulated mechanism of HEMA or TEGDMA toxicity based on radical metabolites is supported by the effectivity of the antioxidative substances tested in mitigating toxicity and by the greater susceptibility of the glutathione redox system as compared to protein synthesis inhibition in assessing toxicity.
Assuntos
Antioxidantes/farmacologia , Resinas Compostas/toxicidade , Metacrilatos/toxicidade , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/toxicidade , Vitaminas/farmacologia , Animais , Ácido Ascórbico/farmacologia , Materiais Biocompatíveis/toxicidade , Linhagem Celular , Glutationa/análise , Dissulfeto de Glutationa/análise , Glutationa Redutase , Humanos , Metionina/metabolismo , NADH NADPH Oxirredutases/metabolismo , Ratos , Tiorredoxina Dissulfeto Redutase , Ácido Úrico/farmacologia , Vitamina E/farmacologiaRESUMO
OBJECTIVE: The effect of dental composite components triethyleneglycoldimethacrylate (TEGDMA) and hydroxyethylmethacrylate (HEMA), as well as mercuric chloride (HgCl2) and methylmercury chloride (MeHgCl) was investigated on the release of lactatedehydrogenase (LDH) from alveolar epithelial lung cell lines in vitro. METHODS: The confluent cell layers from the A549 (human, malignant) and the L2 cells (rat) were incubated with various concentrations of HEMA, TEGDMA, MeHgCl and HgCl2 at 37 degrees C in 2% (v/v) CO2 atmosphere for 8h. In further experiments the L2 cells were incubated with the same compounds for 6-48 h. LDH release was measured and the values were expressed as percentage of the LDH content. The values were plotted on a concentration log-scale and the substance concentration at the maximum slope was assessed as effective concentration (EC50). RESULTS: A significant (p<0.05) increase in the LDH release was found in the L2 cells after 8-h incubation with HEMA (4 mmol/l), TEGDMA (2 mmol/l), MeHgCl (0.01 mmol/l) and HgCl2 (0.015 mmol/l), and in A549 cells with HEMA (14 mmol/l), TEGDMA (15 mmol/l), MeHgCl (0.15 mmol/l) and HgCl2 (0.05 mmol/l), compared to controls. The EC50 values from compounds in the L2 cells are shown in the following table (mean; sem in parentheses; n=3-6; #n=1): [see text]. SIGNIFICANCE: The toxic effect of HgCl2 and MeHgCl from the L2 cells was about 100-700-fold higher than of the dental composite components. A significant (p<0.05) time dependent increase of toxicity was observed with TEGDMA, HEMA and MeHgCl.
Assuntos
Materiais Biocompatíveis/toxicidade , Resinas Compostas/toxicidade , Materiais Dentários/toxicidade , Pulmão/efeitos dos fármacos , Compostos de Mercúrio/toxicidade , Animais , Anti-Infecciosos Locais/toxicidade , Linhagem Celular , Células Epiteliais/efeitos dos fármacos , Humanos , L-Lactato Desidrogenase/efeitos dos fármacos , Modelos Lineares , Pulmão/citologia , Neoplasias Pulmonares/patologia , Cloreto de Mercúrio/toxicidade , Metacrilatos/toxicidade , Compostos de Metilmercúrio/toxicidade , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/toxicidade , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/efeitos dos fármacos , Ratos , Estatística como Assunto , Temperatura , Fatores de Tempo , Células Tumorais CultivadasRESUMO
OBJECTIVE: Unconverted 2-hydroxyethylmethacrylate (HEMA) can be released from dental resin materials and can enter the body in humans. In the present study the uptake, distribution and excretion of 14C-HEMA applied via different routes were examined in vivo in guinea pigs. METHODS: HEMA (0.02 mmol/kg bw labelled with a tracer dose 14C-HEMA 0.3 Bq/g bw) was administered by gastric tube or by subcutaneous injection. Urine, feces, and exhaled carbon dioxide were collected for 24 h after administration. Guinea pigs were killed 24 h after the beginning of the experiment and various organs removed and 14C radioactivity measured. RESULTS: Low fecal 14C levels (about 2% of the dose) and urinary levels of about 15% after 24 h were noted with either route of administration. Direct measurement of exhaled CO(2) showed that about 70% of the dose left the body via the lungs. Two pathways for the metabolism of 14C-HEMA can be described. It is likely that 14C-pyruvate is formed in vivo resulting in the formation of toxic 14C-HEMA intermediates. 14C-HEMA was taken up rapidly from the stomach and small intestine after gastric administration and was widely distributed in the body following administration by each of the routes. CONCLUSIONS: Clearance from most tissues following gastric and intradermal administration was essentially complete within one day. The peak HEMA levels in all tissues examined after 24 h were at least onemillion-fold less than known toxic levels.
Assuntos
Metacrilatos/farmacocinética , Metacrilatos/toxicidade , Resinas Acrílicas/farmacocinética , Resinas Acrílicas/toxicidade , Animais , Testes Respiratórios , Radioisótopos de Carbono/farmacocinética , Radioisótopos de Carbono/urina , Resinas Compostas/farmacocinética , Resinas Compostas/toxicidade , Materiais Dentários/farmacocinética , Materiais Dentários/toxicidade , Relação Dose-Resposta a Droga , Fezes , Cobaias , Absorção Intestinal , Masculino , Taxa de Depuração Metabólica , Distribuição Tecidual , UrinaRESUMO
1 Dimercaprol (BAL), 2,3-dimercaptopropanesulphonate sodium (DMPS) and meso-2,3-dimercaptosuccinic acid (DMSA) are effective arsenic antidotes, but the question which one is preferable for optimal therapy of arsenic poisoning is still open to discussion. Major drawbacks of BAL include (a) its low therapeutic index, (b) its tendency to redistribute arsenic to brain and testes, for example, (c) the need for (painful) intramuscular injection and (d) its unpleasant odour. 2 The newer antidotes DMPS and DMSA feature low toxicity and high therapeutic index. They can be given orally or intravenously due to their high water solubility. While these advantages make it likely that DMPS and DMSA will replace BAL for the treatment of chronic arsenic poisoning, acute intoxication-especially with lipophilic organoarsenicals-may pose a problem for the hydrophilic antidotes, because their ionic nature can adversely affect intracellular availability. 3 This article focuses on aspects dealing with the power of BAL, DMPS, and DMSA to mobilize tissue-bound arsenic in various experimental models, such as monolayers of MDCK (= Madin-Darby canine kidney) cells from dog kidney, isolated perfused liver from guinea-pigs, and perfused jejunal segments from rat small intestine. 4 The results show that hydrophilic DMPS and DMSA may fail to rapidly and completely remove arsenic that has escaped from the extracellular space across tight epithelial barriers. However, owing to their low toxicity, which allows larger doses to be applied, and the potential modification of their pharmacokinetics by means of inert oral anion-exchange resins, DMPS and DMSA may advantageously replace BAL whenever intervention time is not critical. With severe intoxication by organic arsenicals, when the point-of-no-return is a limiting factor, BAL may still have a place as an arsenic antidote.
Assuntos
Antídotos/uso terapêutico , Intoxicação por Arsênico , Dimercaprol/uso terapêutico , Intoxicação/tratamento farmacológico , Animais , Antídotos/farmacocinética , Antídotos/toxicidade , Dimercaprol/farmacocinética , Dimercaprol/toxicidade , Cães , Cobaias , Humanos , Camundongos , Ratos , Succímero/farmacocinética , Succímero/uso terapêutico , Succímero/toxicidade , Unitiol/farmacocinética , Unitiol/uso terapêutico , Unitiol/toxicidadeRESUMO
OBJECTIVES: Previous studies have shown that resin composites may cause persistent inflammation of oral or pulpal tissues as well as cell death through eluted substances. The aim of this study was to investigate the leaching of ingredients from commercial dental fissure sealers as well as their cytotoxic effects on human gingival fibroblast (HGF). METHODS: The sealers tested were: Helioseal(®) F, Helioseal(®) Refill, Fissurit(®) F, Grandio(®) Seal, Ultraseal XT(®) plus and Delton(®) FS. Ten discs of each sealer were respectively immersed in methanol or water and incubated at 37°C. The eluates were analysed by gas chromatography/mass spectrometry at day 1, 3 and 7. In the XTT-test, eight discs of each fissure sealer were immersed into medium. The eluates of the respective sealer were mixed and used undiluted and diluted with medium. HGF were incubated with the dilutions at 37°C for 24h. Then XTT-salt was added and the XTT-formazan formation was quantified. RESULTS: In eluates from polymerized sealers, comonomers (triethylene glycol dimethacrylate (TEGDMA)) and additives were found (e.g. camphorquinone (CQ), butylated hydroxytoluene, triphenylstibane). 7 d after the beginning of the experiments the highest amount of TEGDMA was found in the aqueous eluate from Grandio(®) Seal (9944.31 (2250.56) µmol/l). The most cytotoxic eluate found in the XTT-test was from Fissurit(®) F (EC50 value at 27.13 (7.04)%; (mean(SD)). SIGNIFICANCE: Because of the use of sealers in preventative dental medicine it should be taken into account that substances like TEGDMA or CQ, that are often causing allergic reactions, are elutable. Before using the sealers patients should be asked for allergic reactions to these substances.
Assuntos
Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Metacrilatos/química , Selantes de Fossas e Fissuras/química , Bis-Fenol A-Glicidil Metacrilato/química , Bis-Fenol A-Glicidil Metacrilato/toxicidade , Hidroxitolueno Butilado/química , Hidroxitolueno Butilado/toxicidade , Cânfora/análogos & derivados , Cânfora/química , Cânfora/toxicidade , Linhagem Celular , Corantes , Resinas Compostas/química , Resinas Compostas/toxicidade , Meios de Cultura , Formazans , Cromatografia Gasosa-Espectrometria de Massas , Gengiva/citologia , Humanos , Metacrilatos/toxicidade , Metanol/química , Compostos Organometálicos/química , Compostos Organometálicos/toxicidade , Selantes de Fossas e Fissuras/toxicidade , Polietilenoglicóis/química , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/química , Ácidos Polimetacrílicos/toxicidade , Solventes/química , Fatores de Tempo , Água/químicaRESUMO
OBJECTIVES: The toxicity of monomers like bisphenol-A-glycidyldimethacrylate (BisGMA) and urethane-dimethacrylate (UDMA) to cells is well studied. In these studies solubilizers, which have a toxic potential, are used to dissolve the basic monomers in the aqueous medium. In these experiments it is not possible to confidently exclude a synergistic effect of basic monomers and solubilizers in cells. Moreover, less is known about the synergistic interaction between basic- and comonomers (triethyleneglycoldimethacrylate (TEGDMA); 2-hydroxyethylmethacrylate (HEMA)) in cells. We dissolved the basic monomers in the comonomers and incubated human gingival fibroblasts (HGFs) with these binary mixtures in different concentrations. METHODS: Proliferating HGFs monolayers were cultured in the absence or presence of mixtures of BisGMA/TEGDMA, BisGMA/HEMA, UDMA/TEGDMA and UDMA/HEMA. Twenty-four hours later XTT was added and the formazan formation was quantified. EC(50) values were obtained at half-maximum-effect concentrations from fitted curves. RESULTS: EC(50) values were (mmol/l; mean±sem; n=5): 0.01 BisGMA/0.48 TEGDMA; 0.04 BisGMA/4.99 HEMA; 0.04 UDMA/1.60 TEGDMA and 0.02 UDMA/2.26 HEMA. All tested mixtures induced a dose-dependent loss of viability in HGFs after 24h. SIGNIFICANCE: The EC(50) values of binary mixtures were significantly (p<0.05) lower compared to the EC(50) values of the pure substances indicating a synergistic interaction of the mixtures on the HGFs. The widely used (co)monomers BisGMA and TEGDMA have the lowest EC(50) values. The highest decrease of EC(50) values, compared with the pure substances, were found in the mixture UDMA/HEMA. Worst case calculations show that the EC(50) values from binary mixtures are at least 6 fold lower compared with known amounts of elutable (co)monomers from polymerized composites.
Assuntos
Gengiva/efeitos dos fármacos , Metacrilatos/toxicidade , Bis-Fenol A-Glicidil Metacrilato/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Fibroblastos/efeitos dos fármacos , Gengiva/citologia , Humanos , Polietilenoglicóis/toxicidade , Ácidos Polimetacrílicos/toxicidade , Poliuretanos/toxicidadeRESUMO
OBJECTIVES: Silorane-based dental monomers contain an epoxy functional group. Less is known about the toxicological and inflammatory potential of silorane-based composites. Therefore we compared the release of 24 cytokines from human leukocytes after incubation with silorane-based Filtek™ Silorane (Silo) and methacrylate-based TetricEvo Flow® (TC). METHODS: Leukocytes from nine healthy test persons (P) were incubated with Silo or TC for up to 72h. All 24h cytokines were quantified with a magnetic bead assay. RESULTS: Silo stimulates the leukocytes to higher release of cytokines when compared to TC. 72h after beginning the experiment, leukocytes from P6 incubated with Silo secreted more than an 18-fold amount of interleukin (IL)-6 when compared with leukocytes incubated with TC (771.8 vs 42.1pg/ml). Only leukocytes from P8 incubated with Silo release up to 14.4pg/ml IL-2 after 72h. SIGNIFICANCE: The significantly higher induction of cytokines with Silo in comparison to TC is test person independent. This indicates a higher sensitization potential for Silo. Because of the cytokine release pattern (especially the release of T-cell dependent IL-2) from leukocytes from P8 after incubation with Silo it is likely that P8 can develop an allergic Type IV sensitization to Silo. Therefore the cytokine release assay is a helpful tool for providing information about possible immunological reactions to dental resins in individual cases as well as for a general risk assessment and comparison between different dental materials.
Assuntos
Citocinas/biossíntese , Hipersensibilidade a Drogas/imunologia , Leucócitos/efeitos dos fármacos , Metacrilatos/farmacologia , Siloxanas/farmacologia , Humanos , Inflamação/imunologia , Interleucina-2/biossíntese , Interleucina-6/biossíntese , Leucócitos/metabolismo , Metacrilatos/efeitos adversos , Siloxanas/efeitos adversosRESUMO
The aim of this study was to investigate the triethylene glycol (TEGDMA) elution kinetics from light-cured composite with and without chewing simulation over a time period of 86 h. An experimental composite with TEGDMA labeled with a tracer dose of 14C-TEGDMA was used. The material parameters were in the range of commercially available composites. The mastification was simulated with the Fatigue-machine and the MUC-3 chewing simulator. 14C was eluted to 2.55% of the applied 14C-TEGDMA dose within 86 h after chewing simulation with the Fatigue-machine and to 2.60% after chewing simulation with the MUC-3. Similar 14C-kinetic data were found for 14C-elution with and without chewing simulation with the Fatigue-machine and with MUC-3. During the first 26 h after the beginning of the experiments a linear 14C-elution kinetic was observed, followed by a second linear 14C-elution kinetic with a lower slope up to 86 h in both apparatus. It could be shown that chewing simulation has no significant (p<0.05) effect on the release of 14C-TEGDMA (and/or 14C-degradation products) from polymerized composites.
Assuntos
Resinas Compostas/química , Mastigação , Polietilenoglicóis/análise , Ácidos Polimetacrílicos/análise , Radioisótopos de Carbono/análise , Análise do Estresse Dentário/instrumentação , Humanos , Cinética , Cura Luminosa de Adesivos Dentários , Teste de MateriaisRESUMO
The relative effectiveness of British Anti-Lewisite (BAL), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), and a new metal binding agent 2,3-bis(acetylthio)propanesulfonamide (BAPSA) was compared by determining their effect on pyruvate metabolism in perfused livers of guinea pigs after repeated treatment with As2O3. Guinea pigs received As2O3, 2.5 mg/kg s.c. twice daily on 5 consecutive days (total dose 25 mg/kg). Sixteen hours after the last dose the livers were perfused (2.5 ml/min/g liver) with Krebs-Henseleit buffer and glucose (10 mmol/l) as substrate for 80 min. After 50 min of perfusion 0.1 or 0.7 mmol/l BAL, DMPS, DMSA, or BAPSA were added to the perfusate for 30 min. Samples of the effluent were collected every 10 min; lactate and pyruvate were determined enzymatically. As compared to controls, a significant decrease in the pyruvate and lactate efflux was observed in perfused livers of guinea pigs treated with As2O3. After influx of BAL (0.1 mmol/l), DMSA (0.7 mmol/l), and BAPSA (0.1 and 0.7 mmol/l) respectively, the pyruvate and lactate efflux and the oxygen consumption (exception BAL 0.1 mmol/l) increased and reached control values without arsenic treatment. On the other hand, the pyruvate and lactate efflux and the oxygen consumption was further significantly decreased after influx of 0.7 mmol/l BAL.
Assuntos
Arsênio/toxicidade , Arsenicais , Quelantes/farmacologia , Lactatos/metabolismo , Fígado/efeitos dos fármacos , Óxidos , Piruvatos/metabolismo , Animais , Antídotos/farmacologia , Trióxido de Arsênio , Dimercaprol/farmacologia , Cobaias , Técnicas In Vitro , Ácido Láctico , Fígado/metabolismo , Masculino , Consumo de Oxigênio/efeitos dos fármacos , Perfusão , Ácido Pirúvico , Succímero/farmacologiaRESUMO
Divergent pattern in pyruvate efflux from livers perfused with As2O3 and livers of animals previously repeatedly treated with the toxicant was observed in earlier experiments (Reichl et al. 1987, 1988). Further studies of the effect of As2O3 on carbohydrate metabolism were therefore performed. Male guinea pigs received either a single dose of As2O3 10 mg.kg-1 s.c. or repeated doses of 2.5 mg.kg-1 bis in die (b.i.d.) on 5 consecutive days. One hour after the single dose or 1 h and 16 h after the last injection in the repeated treatment group the animals were sacrificed under anaesthesia. The livers were removed by a freeze-stop technique and the contents of glycogen and glycolysis intermediates were measured. In the single dose group a decrease in fructose-1,6-diphosphate and glycerol-aldehyde-3-phosphate and an increase in phosphoenol-pyruvate and pyruvate was observed. In the repeat dose, 1-h group a significant decrease in glycogen, glucose-6-phosphate, fructose-6-phosphate, glycerolaldehyde-3-phosphate, dihydroxyacetonephosphate, 2-phosphoglycerate and pyruvate was found. In the repeat dose, 16-h group the contents of glycogen, glucose-6-phosphate, pyruvate and lactate were diminished. The most prominent finding after repeated As2O3 administration was a marked depletion in total carbohydrate content. This was due mainly to depletion of glycogen.
Assuntos
Arsênio/toxicidade , Metabolismo dos Carboidratos , Animais , Arsênio/administração & dosagem , Glicólise/efeitos dos fármacos , Cobaias , Injeções , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/metabolismo , MasculinoRESUMO
The effect of the dithiols British Anti-Kewisite (BAL), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA) and a new metal binding agent 2,3-bis-(acetylthio)- propanesulfonamide (BAPSA) on the biliary excretion of arsenic in perfused livers of guinea pigs after acute experimental poisoning with As2O3 was investigated. Guinea pigs received As2O3, 10.0 mg/kg subcutaneously at 9 a.m. as a single injection. One hour after the injection the livers were perfused (2.5 ml x min.-1 x g-1 liver) with Krebs-Henseleit buffer and glucose for 80 min. After 40 min. of saline perfusion (control) 0.1 or 0.7 mmol/l BAL, DMSA, DMPS, or BAPSA were added to the perfusate and arsenic elimination in the bile and effluent perfusate was measured. The biliary excretion of arsenic in control livers between 40 and 80 min. was 0.7% of the total arsenic liver content before perfusion (= arsenic liver content after perfusion + portion excreted in the bile+perfusate). After antidote addition (0.1 mmol/l) the excretion was 0.2% for livers perfused with BAL, 6.8% for DMSA, 10.6% for DMPS, and 11.1% for BAPSA, respectively. After 0.7 mmol/l antidote the excretion of arsenic was 0.1% in livers perfused with BAL, 9.6% for DMSA, 12.3% for DMPS, and 13.3% for BAPSA, respectively. Except BAL, all compounds and most effectively BAPSA increased biliary excretion of arsenic. This indicates that excretion of arsenic which normally is mainly renal is shifted towards faecal excretion by the dithiols.