Effects of intrapulpal temperature change induced by visible light units on the metabolism of odontoblast-like cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytotoxicity of resin-based light-cured liners

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Transdentinal diffusion and cytotoxicity of self-etching adhesive systems

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Toxicity of chlorhexidine on odontoblast-like cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on cultured odontoblast cell lines after consecutive applications

To evaluate the trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on an odontoblast-like cell lines (MDPC-23) after consecutive applications.Fifteen enamel/dentine discs were obtained from bovine central incisor teeth and placed individually in artificial pulp chambers. Three groups (n = 5 discs) were formed according to the following enamel treatments: G1: 35% H2O2 bleaching gel (15 min); G2: 35% H2O2 bleaching gel (15 min) + halogen light (20 s); G3: control (no treatment). After repeating the treatments three consecutive times, the extracts (culture medium + gel components that had diffused through enamel/dentine discs) in contact with the dentine were collected and applied to previously cultured MDPC-23 cells (50 000 cells cm(-2)) for 24 h. Cell metabolism was evaluated by the MTT assay and data were analysed statistically (alpha = 5%; Kruskal-Wallis and Mann-Whitney U-test). Cell morphology was analysed by scanning electron microscopy.Cell metabolism decreased by 92.03% and 82.47% in G1 and G2 respectively. G1 and G2 differed significantly (P < 0.05) from G3. Regardless of halogen light activation, the application of the bleaching gel on the cultured odontoblast-like cells caused significantly more severe cytotoxic effects than those observed in the nontreated control group. In addition, significant morphological cell alterations were observed in G1 and G2.After three consecutive applications of a 35% H2O2 bleaching agent, the diffusion of the gel components through enamel and dentine caused severe toxic effects to cultured pulp cells.

Increased Viability of Odontoblast-Like Cells Subjected to Low-Level Laser Irradiation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of low-level laser irradiation on odontoblast-like cells

Low-level laser therapy (LLLT), also referred to as therapeutic laser, has been recommended for a wide array of clinical procedures, among which the treatment of dentinal hypersensitivity. However, the mechanism that guides this process remains unknown. Therefore, the objective of this study was to evaluate in vitro the effects of LLL irradiation on cell metabolism (MTT assay), alkaline phosphatase (ALP) expression and total protein synthesis. The expression of genes that encode for collagen type-1 (Col-1) and fibronectin (FN) was analyzed by RT-PCR. For such purposes, oclontoblast-like cell line (MDPC-23) was previously cultured in Petri dishes (15000 cells/cm(2)) and submitted to stress conditions during 12 h. Thereafter, 6 applications with a monochromatic near infrared radiation (GaAlAs) set at predetermined parameters were performed at 12-h intervals. Non-irradiated cells served as a control group. Neither the MTT values nor the total protein levels of the irradiated group differed significantly from those of the control group (Mann-Whitney test; p > 0.05). on the other hand, the irradiated cells showed a decrease in ALP activity (Mann-Whitney test; p < 0.05). RT-PCR results demonstrated a trend to a specific reduction in gene expression after cell irradiation, though not significant statistically (Mann-Whitney test; p > 0.05). It may be concluded that, under the tested conditions, the LLLT parameters used in the present study did not influence cell metabolism, but reduced slightly the expression of some specific proteins.

Mineral Trioxide Aggregate-based Endodontic Sealer Stimulates Hydroxyapatite Nucleation in Human Osteoblast-like Cell Culture

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

In vitro cytotoxicity and in vivo-biocompatibility of contemporary resin-modified glass-ionomer cements

Objectives. To evaluate the effects of current resin-modified glass-ionomer cements (RMGICs) applied on culture of cells or implanted into subcutaneous tissue of rats.Methods. Experiment 1 - Thirty round-shaped samples of every RMGICs: Rely X Luting Cement (RL), Vitremer (VM), and Vitrebond (VB) were placed into wells with 1.1 mL of culture medium (DMEM), and incubated for 24,48 or 72 h. The extracts from every sample were applied on the MDPC-23 cells. Fresh DMEM was used as control group. The MTT assay was carried out for mitochondrial respiration. Experiment 2 - Fifty-four polyethylene tubes filled with the experimental materials were implanted into the dorsal subcutaneous tissue of rats. At 7, 30, and 90 days the animals were killed and the biopsies were processed for histological evaluation.Results. Experiment 1 - Both time of elution and material significantly influenced cell respiratory activity. in general, the extracts obtained at 24 h were less cytotoxic than 48 and 72 h incubation. The cytotoxic effect of VM and RL were not statistically different (P < 0.05) for the 24-hour period. VB showed the highest cytotoxic effect. Experiment 2 - All RMGICs elicited at 7 days a moderate to intense inflammatory reaction which decreased over time. However, connective healing occurred for most of samples at 90-day evaluation.Significance. Glass-ionomer cements may cause noticeable inflammatory response when in direct contact to connective tissue. The toxic effects of this kind of soluble material depend on the amount of components released in the aqueous environment. (C) 2005 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Antimicrobial Properties and Cytotoxicity of an Antimicrobial Monomer for Application in Prosthodontics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Photodynamic therapy associating Photogem (R) and blue LED on L929 and MDPC-23 cell culture

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nutritional stress enhances cell viability of odontoblast-like cells subjected to low level laser irradiation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytotoxicity of monomers, plasticizer and degradation by-products released from dental hard chairside reline resins

Objectives. The aim of this study was to evaluate the cytotoxic effect of the monomers isobutyl methacrylate (IBMA) and 1,6-hexanediol dimethacrylate (1,6-HDMA), the plasticizer di-n-butyl phthalate (DBP), and the degradation by-products methacrylic acid (MA) and benzoic acid (BA) on L929 cells. Based on previous investigations on the release of these compounds from hard chairside reline resins, a range of concentrations (mu mol/L) were selected for the cytotoxicity tests (IBMA, 5.491406.57; 1,6-HDMA, 1.2239.32; DBP, 1.12143.8; MA, 9.07581; BA, 3.19409).Methods. Cytotoxic effects were assessed using MTT and 3H-thymidine assays after the cells had been exposed to the test compounds at the given concentrations for 24h. Cytotoxicity was rated based on cell viability relative to controls (cells exposed to medium without test substances).Results. DNA synthesis activity was inhibited by all compounds. Mitochondrial dehydrogenase activity decreased in cells treated with monomers, plasticizer and MA by-product, whereas no cytotoxic effect was observed on contact with BA at the majority of concentrations tested. The ranges of suppression for 3H-thymidine assay were: IBMA, 2595%; 1,6-HDMA, 9598%; DBP, 4098%; MA, 9799%; BA, 5471%. For MTT assay, the ranges of suppression were: IBMA, 096%; 1,6-HDMA, 2689%; DBP, 1780%; MA, 5266%; BA, 027%. The 3H-thymidine assay was more sensitive than the MTT assay.Significance. This study evaluated the cytotoxicity of a wide range of concentrations of monomers (IBMA and 1,6-HDMA), plasticizer (DBP) and degradation by-products (MA and BA), including those expected to be released from hard chairside reline resins. The differences observed in the cytotoxicity of these compounds, along with other properties, may assist the dental practitioners in the selection of reline materials with improved service life performance and low risk of adverse reactions in patients who wear relined dentures.

Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

Introduction: Endodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinamica, Ibipora, PR, Brazil). Methods: Macrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls. Results: Analysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01). Conclusion: The tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid-based solutions induce lower No release than EDTA-based irrigants. (J Endod 2009;35:824-828)

Antiulcerogenic effect and cytotoxic activity of semi-synthetic crotonin obtained from Croton cajucara Benth.

Trans-dehydrocrotonin, the major diterpene isolated from the bark of Croton cajucara, has good antiulcerogenic activity which, however, is accompanied by toxic effects. on the basis of these results, a semi-synthetic crotonin, named 4SRC, was prepared to determine whether this substance has similar antiulcerogenic activity with lower or no toxicity. The natural crotonin was also isolated from the bark of C. cajucara but was not used due to the small amount obtained. The cytotoxic effect of semi-synthetic crotonin, expressed as cell viability, was assessed in (a) lung fibroblast cell line (V79) derived from Chinese hamsters, a system commonly used for cytotoxicity studies, and (b) rat hepatocytes isolated from male Wistar rats. After treatment, cell viability was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction (MTT reduction), total acid content and neutral red uptake assays. To evaluate V79 cell viability, different concentrations of semi-synthetic crotonin were incubated with the cells. To evaluate the antiulcerogenic effects of semi-synthetic crotonin (50, 100 and 200 mg/kg), we used the models of gastric ulcer induced by ethanol/HCl, stress, indomethacin/bethanechol, and ethanol in male Swiss mice and male Wistar rats. The substance had an IC50 = 500 muM in the neutral red uptake and MTT reduction tests and an IC50 = 200 muM in the nucleic acid content test. With regard to hepatocyte viability after treatment with semi-synthetic crotonin at different concentrations, semi-synthetic crotonin had an IC50 = 10-500 muM in the nucleic acid content and MTT reduction tests and an IC50 = 120 muM in the neutral red uptake test. In another experiment, V79 cells were incubated with the metabolites produced by hepatocytes treated with different concentrations of semi-synthetic crotonin. After a 4-h incubation, semi-synthetic crotonin had an IC50 = 500 muM in the MTT reduction and neutral red uptake tests and an IC50 = 370 muM in nucleic acid content test. The substance had significant antiulcerogenic activity in all models studied, suggesting the presence of a possible antisecretory effect combined with a cytoprotective effect. For this reason, the effect of semi-synthetic crotonin was also evaluated on biochemical parameters of gastric juice and gastric wall mucus, both obtained from pylorus-ligated mice. No significant differences were observed in these parameters between semi-synthetic crotonin-treated and control animals. The results obtained with semi-synthetic crotonin are promising, with a significant preventive effect against gastric ulcer induced by different agents. Our data also show that semi-synthetic crotonin was less toxic than dehydrocrotonin and that the cytotoxic effects decreases with the time that isolated hepatocytes were in culture. (C) 2003 Elsevier B.V. B.V. All rights reserved.