salicylates and thiazolyl-blue

The purpose of this study was to compare ISO standards versus a new technique for in vitro evaluation of cytotoxicity of root canal sealers. The cytotoxicity of AH Plus, Cortisomol, and Sealapex was first recorded according to ISO standards on L 929 fibroblasts by the MTT assay. In parallel, 30 single-rooted teeth were cut at the cementum enamel junction (CEJ), and the roots were prepared and sterilized before filling with the lateral condensation using one of three sealers (n = 10). The apexes of the roots were dipped into 1 ml of minimum essential medium for 1, 2, and 30 days renewing the medium every other day. After 24-h contact between the medium and the filled roots, the medium was used to measure the cytotoxicity on L 929 with the MTT assay. ISO standards always gave a statistically higher cytotoxicity than the root-dipping technique (p < 0.0001), whatever the sealer and the exposure time. The ISO standards showed statistically significant differences among the sealers (p < 0.0001). AH Plus was noncytotoxic, Cortisomol showed a high cytotoxicity decreasing over time (p < 0.001), and Sealapex displayed a high cytotoxicity that did not decrease over time (NS). The new technique showed statistically significant differences among the sealers (p = 0.001), but the differences were so small that they were likely not clinically relevant. The high cytotoxicity of Sealapex decreased over time but the cytotoxicity of AH Plus and Cortisomol did not. The results show that the ISO standards may strongly over-evaluate the cytotoxicity of the endodontic sealers, emphasize the difference among the sealers, and may clinically correspond to a large overfilling. The new technique reduces the discrimination of the test and may clinically correspond to a classical filling. Therefore, both methods might be considered as clinically relevant, corresponding to classical and overfilling conditions.

Topics: Analysis of Variance; Animals; Biocompatible Materials; Calcium Hydroxide; Cell Line; Coloring Agents; Epoxy Resins; Fibroblasts; Gutta-Percha; Humans; Mice; Root Canal Filling Materials; Root Canal Obturation; Root Canal Preparation; Salicylates; Tetrazolium Salts; Thiazoles; Zinc Oxide-Eugenol Cement

The objective of this study was to compare the cytotoxicity, inflammatory response, osteogenic effect, and the signaling mechanism of these biologic activities of 4 calcium compound-based root canal sealers (ie, Sealapex [Sybron Kerr, WA], apatite root sealer [ARS; Dentsply Sankin, Tokyo, Japan], MTA Fillapex [Angelus Indústria de Produtos Odontológicos S/A, Londrina, PR, Brazil], and iRoot SP [Innovative BioCreamix Inc, Vancouver, Canada]) in human periodontal ligament cells.. Cytotoxicity was assessed using the 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide assay. Levels of inflammatory mediators were measured by enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and Western blot analysis. Osteogenic potential was evaluated by alkaline phosphatase activity, alizarin red staining, and marker genes by reverse-transcription polymerase chain reaction. The signal transduction pathways were examined by Western blotting.. None of the sealers were cytotoxic. ARS, MTA Fillapex, and iRoot SP induced a lower expression of proinflammatory mediators than Sealapex. All sealers increased ALP activity and the formation of mineralized nodules and up-regulated the expression of osteoblastic marker messenger RNA. ARS, MTA Fillapex, and iRoot SP showed superior osteogenic potential compared with Sealapex. The expression and/or activation of integrin receptors and downstream signaling molecules, including focal adhesion kinase, paxillin, Akt, mitogen-activated protein kinase, and nuclear factor κB, was induced by ARS, MTA Fillapex, and iRoot SP treatment but not by Sealapex treatment.. We show for the first time that ARS, MTA Fillapex, and iRoot SP induce a lower expression of inflammatory mediators and enhance osteoblastic differentiation of PDLCs via the integrin-mediated signaling pathway compared with Sealapex.

Topics: Alkaline Phosphatase; Aluminum Compounds; Anthraquinones; Biocompatible Materials; Calcium Compounds; Calcium Hydroxide; Cell Differentiation; Cell Line; Cell Survival; Coloring Agents; Drug Combinations; Durapatite; Focal Adhesion Kinase 1; Humans; Inflammation Mediators; Materials Testing; Mitogen-Activated Protein Kinases; Nanoparticles; NF-kappa B; Osteoblasts; Osteogenesis; Oxides; Paxillin; Periodontal Ligament; Proto-Oncogene Proteins c-akt; Root Canal Filling Materials; Salicylates; Signal Transduction; Silicates; Tetrazolium Salts; Thiazoles

A series of salicylanilide derivatives (compounds 1-32) were synthesised by reacting substituted salicylic acids and anilines. The chemical structures of these compounds were determined by (1)H-NMR, electrospray ionisation mass spectrometry (ESI-MS) and elemental analysis. The compounds were assayed for their antiproliferative activities against the Hep-G2 cell line by the 3-(4,5-dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Among the compounds tested, 22 and 28 showed the most favouable antiproliferative activities with 50% inhibitory concentration (IC(50)) values of 1.7 and 1.3 μM, respectively, which were comparable to the positive control of 5-fluorouracil (IC(50)=1.8 μM). A solid-phase ELISA assay was also performed to evaluate the ability of compounds 1-32 to inhibit the autophosphorylation of the epidermal growth factor receptor tyrosine kinase (EGFR TK). Docking simulations of 22 and 28 were carried out to illustrate the binding mode of the molecule into the EGFR active site, and the result suggested that both compounds 22 and 28 could bind the EGFR kinase well.

Topics: Aniline Compounds; Antineoplastic Agents; ErbB Receptors; Fluorouracil; Hep G2 Cells; Humans; Inhibitory Concentration 50; Molecular Dynamics Simulation; Nuclear Magnetic Resonance, Biomolecular; Phosphorylation; Protein Binding; Salicylanilides; Salicylates; Structure-Activity Relationship; Tetrazolium Salts; Thiazoles

The biologic safety profile of oral health care products is often assumed on the basis of simplistic test models such as monolayer cell culture systems. We developed and characterized a tissue-engineered human oral mucosal model, which was proven to represent a potentially more informative and more clinically relevant alternative for the biologic assessment of mouthwashes. The aim of this study was to evaluate the biologic effects of alcohol-containing mouthwashes on an engineered human oral mucosal model.. Three-dimensional (3D) models were engineered by the air/liquid interface culture technique using human oral fibroblasts and keratinocytes. The models were exposed to phosphate buffered saline (negative control), triethylene glycol dimethacrylate (positive control), cola, and three types of alcohol-containing mouthwashes. The biologic response was recorded using basic histology; a cell proliferation assay; 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tissue-viability assay; transmission electron microscopy (TEM) analysis; and the measurement of release of interleukin (IL)-1beta by enzyme-linked immunosorbent assay.. Statistical analysis showed that there was no significant difference in tissue viability among the mouthwashes, cola, and negative control groups. However, exposure to the positive control significantly reduced the tissue viability and caused severe cytotoxic epithelial damage as confirmed by histology and TEM analysis. A significant increase of IL-1beta release was observed with the positive control and, to a lesser extent, with two of the tested mouthrinses.. The 3D human oral mucosal model can be a suitable model for the biologic testing of mouthwashes. The alcohol-containing mouthwashes tested in this study do not cause significant cytotoxic damage and may slightly stimulate IL-1beta release.

Topics: Cell Proliferation; Coloring Agents; Drug Combinations; Ethanol; Humans; Interleukin-1beta; Microscopy, Electron, Transmission; Models, Biological; Mouth Mucosa; Mouthwashes; Salicylates; Terpenes; Tetrazolium Salts; Thiazoles; Tissue Engineering; Tissue Survival

Recently, a new sealer composed of Portland cement named Endo-CPM-Sealer was developed. The aim of this study was to investigate the effects of Endo-CPM-Sealer (EGEO SRL, Buenos Aires, Argentina), Sealapex (Sybron Endo, Glendora, CA), and Angelus MTA (Angelus, Londrina, Brazil) on cell viability and cytokine (interleukin [IL]-1beta and IL-6) production by mouse fibroblasts.. Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts. Cells cultured with only empty polyethylene tubes were used as the control. After 24 hours, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to evaluate the cell viability. For cytokine assay, mouse fibroblasts were incubated in 24-well flat-bottom plates with set material disks at the bottom. Cells cultured without the material disks served as the negative control. After 24 hours of incubation, culture media were collected for cytokine evaluation by using an enzyme-linked immunosorbent assay. The data were statistically analyzed by analysis of variance and Bonferroni correction.. Endo-CPM-Sealer, Sealapex, and Angelus MTA did not inhibit the cell viability. All materials induced IL-6 releasing, but the amount was not statistically significant compared with the control group. Angelus MTA induced IL-1beta releasing significantly more than the control.. All materials were not considered cytotoxic in fibroblast culture.

Topics: Aluminum Compounds; Animals; Biocompatible Materials; Calcium Compounds; Calcium Hydroxide; Cell Line; Cell Survival; Coloring Agents; Drug Combinations; Fibroblasts; Interleukin-1beta; Interleukin-6; Interleukins; Materials Testing; Mice; Oxides; Root Canal Filling Materials; Salicylates; Silicates; Tetrazolium Salts; Thiazoles; Time Factors

Salicylates are novel biologically active compounds that exhibit multiple therapeutic activities. The anti-cancer effectiveness of calcium salicylate has been investigated on human HT-1080 fibrosarcoma cell lines at relatively low concentrations (predominantly 0.4 mM) compared to those previously reported. Although low calcium salicylate concentrations did not retard tumour growth progression significantly, as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and time-lapse assays, its cytotoxic characteristics were proven to be prominent by various morphological and immunocytological techniques. The results here demonstrate evidence for approximately 25% apoptosis after treatment with calcium salicylate, which up-regulatd the expression of p53, p21 and Bax, and down-regulated Bcl-2 in HT-1080 cells.

Topics: Antineoplastic Agents; Apoptosis; bcl-2-Associated X Protein; Cell Cycle; Cell Line, Tumor; Cell Survival; Fibrosarcoma; Gene Expression Regulation, Neoplastic; Humans; Indicators and Reagents; p21-Activated Kinases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-bcl-2; Salicylates; Tetrazolium Salts; Thiazoles; Tumor Suppressor Protein p53

We determined the roles of reactive oxygen species (ROS) in the expression of cyclooxygenase-2 (COX-2) and the production of prostaglandin E2 (PGE2) in lipopolysaccharide (LPS)-activated microglia. LPS treatment increased intracellular ROS in rat microglia dose-dependently. Pre-treatment with superoxide dismutase (SOD)/catalase, or SOD/catalase mimetics that can scavenge intracellular ROS, significantly attenuated LPS-induced release in PGE2. Diphenylene iodonium (DPI), a non-specific NADPH oxidase inhibitor, decreased LPS-induced PGE2 production. In addition, microglia from NADPH oxidase-deficient mice produced less PGE2 than those from wild-type mice following LPS treatment. Furthermore, LPS-stimulated expression of COX-2 (determined by RT-PCR analysis of COX-2 mRNA and western blot for its protein) was significantly reduced by pre-treatment with SOD/catalase or SOD/catalase mimetics. SOD/catalase mimetics were more potent than SOD/catalase in reducing COX-2 expression and PGE2 production. As a comparison, scavenging ROS had no effect on LPS-induced nitric oxide production in microglia. These results suggest that ROS play a regulatory role in the expression of COX-2 and the subsequent production of PGE2 during the activation process of microglia. Thus, inhibiting NADPH oxidase activity and subsequent ROS generation in microglia can reduce COX-2 expression and PGE2 production. These findings suggest a potential therapeutic intervention strategy for the treatment of inflammation-mediated neurodegenerative diseases.

Topics: Animals; Animals, Newborn; Blotting, Western; Brain; Catalase; Catecholamines; Cell Count; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Dose-Response Relationship, Drug; Drug Interactions; Extracellular Space; Female; Fluoresceins; Imidazolines; Intracellular Space; Isoenzymes; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Mice, Knockout; Microglia; NADPH Oxidases; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Organometallic Compounds; Pregnancy; Prostaglandin-Endoperoxide Synthases; Rats; Rats, Inbred F344; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Salicylates; Superoxide Dismutase; Tetrazolium Salts; Thiazoles

In HeLa cells evidence is provided that active oxygen species such as hydrogen peroxide and superoxide at low levels are important growth regulatory signals. They may constitute a novel regulatory redox system of control superimposed upon the established cell growth signal transduction pathways. Whilst for example hydrogen peroxide can be added exogenously to elicit growth responses in these cells, it is clear that cellularly generated superoxide and hydrogen peroxide are important. Experiments with superoxide dismutase, superoxide dismutase mimics and inhibitors of both superoxide dismutase and xanthine oxidase suggest that superoxide generated intracellularly and superoxide released extracellularly are both relevant to growth control in HeLa cells.

Topics: Allopurinol; Catalase; Cell Division; Ditiocarb; HeLa Cells; Humans; Hydrogen Peroxide; Oxidation-Reduction; Oxypurinol; Reactive Oxygen Species; Salicylates; Signal Transduction; Superoxide Dismutase; Superoxides; Tetrazolium Salts; Thiazoles; Xanthine Oxidase