Testes de biocompatibilidade in vitro de duas formas comerciais do agregado de trióxido mineral

Recently, regular and white mineral trioxide aggregate (MTA) are being used in Dentistry as retrofilling materials. Genotoxicity and cytotoxicity tests form an important part of cancer research and risk assessment of potential carcinogens. Thus, the goal of this study was to examine the genotoxicity and cytotoxicity of regular and white MTA in vitro by the single cell gel (comet) assay and trypan blue exclusion test, respectively. Mouse lymphoma cells were exposed to two presentation forms of MTA at final concentrations ranging from 1 to 1,000 μg/mL for 3 h at 37°C. The results showed that both compounds tested did not produce genotoxic effects at all concentrations evaluated. Likewise, no statistically significant differences (p > 0.05) were observed in cytotoxicity. Taken together, our results suggest that regular and white MTA are not genotoxins and are not able to interfere in cellular viability as assessed by single cell gel (comet) assay and trypan blue assay, respectively.

Ex vivo biocompatibility tests of regular and white forms of mineral trioxide aggregate

Aim: To examine the genotoxicity and cytotoxicity of regular and white mineral trioxide aggregate (MTA) ex vivo by the single-cell gel (comet) assay and trypan blue exclusion test, respectively. Methodology: Aliquots of 1 × 10 4 Chinese hamster ovary cells were incubated at 37°C for 3 h with grey and white forms of MTA at final concentrations ranging from 1 to 1000 μg mL -1. The negative control group was treated with vehicle control phosphate buffer solution for 3 h at 37°C and the positive control group was treated with methyl metasulfonate (at 1 μg mL -1) for 1 h at 37°C. After incubation, the cells were centrifuged at 180 g for 5 min and washed twice with fresh medium and resuspended with fresh medium. Each individual treatment was repeated three times consecutively to ensure reproducibility. Parameters from single-cell gel (comet) and cytotoxicity assays were assessed by the Kruskal-Wallis nonparametric test. Results: Neither compounds produced genotoxic effects with respect to the single-cell gel (comet) assay in all concentrations evaluated. In the same way, the dose-response relationships of all compounds tested at concentrations ranging from 1 to 1000 μg mL -1 on cell viability assessed by the trypan blue assay displayed no statistically significant differences (P > 0.05) for either endodontic material. Conclusions: Regular (grey) and white MTA are not genotoxins and do not induce cellular death. © 2006 International Endodontic Journal.

Study of DNA damage induced by dental bleaching agents in vitro

Dental bleaching is a simple and conservative procedure for aesthetic restoration of vital and non-vital discolored teeth. Nevertheless, a number of studies have demonstrated the risk of tissue damage from the contact of these agents with the oral mucosa. In the current study, the genotoxic potential associated with exposure to dental bleaching agents was assessed by the single cell gel (comet) assay in vitro. Chinese hamster ovary (CHO) cells in vitro were exposed to six commercial dental bleaching agents (Clarigel Gold - Dentsply; Whitespeed - Discus Dental; Nite White - Discus Dental; Magic Bleaching - Vigodent; Whiteness HP - FGM and Lase Peroxide - DMC). The results pointed out that all dental bleaching agents tested contributed to DNA damage as depicted by the mean tail moment, being the strongest effect observed with the highest dose of hydrogen peroxide (Whiteness HP and Lase peroxide, at a 35% concentration). On the other hand, Magic Bleaching (Vigodent) induced the lowest level of DNA breakage. Negative and positive controls displayed absence and presence of DNA-damaging, respectively. Taken together, these results suggest that dental bleaching agents may be a factor that increases the level of DNA damage. A higher concentration of hydrogen peroxide produced higher noxious activities in the genome as detected by single cell gel (comet) assay.

Antimicrobial endodontic compounds do not modulate alkylation-induced genotoxicity and oxidative stress in vitro

Objective: To investigate if formocresol, paramonochlorophenol, or calcium hydroxide modulate the genotoxic effects induced by the oxidatively damaging agent hydrogen peroxide (H 2O 2) or the alkylating agent methyl methanesulfonate (MMS) in vitro by using single cell gel (comet) assay. Study design: Chinese hamster ovary (CHO) cells in culture were exposed directly to formocresol, paramonochlorophenol, or calcium hydroxide (adjusted to 100 μg/mL) for 1 hour at 37°C. Subsequently the cultures were incubated with increasing concentrations (0-10 μmol/L) of MMS in phosphate-buffered solution (PBS) for 15 minutes at 37°C or of H 2O 2 at increasing concentrations (0-100 μmol/L) in distilled water for 5 minutes on ice. The negative control cells were treated with PBS for 1 hour at 37°C. The parameter from the comet assay (tail moment) was assessed by the Kruskal-Wallis nonparametric test followed by a post hoc analysis (Dunn test). Results: Clear concentration-related effects were observed for the genotoxin-exposed CHO cells. Increase of MMS-induced DNA damage was not significantly altered by the presence of the compounds tested. Similarly, no significant changes were observed when hydrogen peroxide was used with the endodontic compounds evaluated. Conclusion: Formocresol, paramonochlorophenol, and calcium hydroxide are not able to modulate alkylation-induced genotoxicity or oxidative DNA damage as depicted by the single cell gel (comet) assay. © 2006 Mosby, Inc. All rights reserved.

Genotoxicity in primary human peripheral lymphocytes after exposure to regular and white mineral trioxide aggregate

Objective: Taking into consideration that DNA damage plays an important role in carcinogenesis, the purpose of this study was to evaluate whether regular and white mineral trioxide aggregate (MTA) are able to induce genetic damage in primary human cells. Study design: Human peripheral lymphocytes obtained from 10 healthy volunteers were exposed to 2 presentation forms of MTA at final concentrations ranging from 1 to 1000 μg/mL for 1 hour at 37°C. The negative control group was treated with vehicle control (phosphate buffer solution, PBS) for 1 hour at 37°C and the positive control group was treated with hydrogen peroxide (at 100 μM) for 5 minutes on ice. Results were analyzed by the Friedman nonparametric test. Results: The results pointed out that either regular or white MTA in all concentrations tested did not induce DNA breakage in human peripheral lymphocytes as depicted by the mean tail moment. Conclusion: In summary, our results indicate that exposure to MTA may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single cell gel (comet) assay. © 2006 Mosby, Inc. All rights reserved.

MS222 does not induce primary DNA damage in fish

The genotoxicity of the anaesthetic MS222 (tricaine) was analysed in fish under both in vivo and in vitro conditions. Based on results of the single cell gel/Comet assay, MS222 had no direct genotoxic effect on the experimental fish, indicating that MS222 does not induce primary DNA damage. These results suggest that the use of this important anaesthetic in aquaculture can be considered to be safe in terms of genotoxicity. © Springer Science+Business Media, Inc. 2007.

DNA damage and nitric oxide synthesis in experimentally infected Balb/c mice with Trypanosoma cruzi

This study aimed to evaluate whether experimental Chagas disease in acute phase under benznidazole therapy can cause DNA damage in peripheral blood, liver, heart, and spleen cells or induce nitric oxide synthesis in spleen cells. Twenty Balb/c mice were distributed into four groups: control (non-infected animals); Trypanosoma cruzi infected; T. cruzi infected and submitted to benznidazole therapy; and only treated with benznidazole. The results obtained with the single cell gel (comet) assay showed that T. cruzi was able induce DNA damage in heart cells of both benznidazole treated or untreated infected mice. Similarly, T. cruzi infected animals showed an increase of DNA lesions in spleen cells. Regarding nitric oxide synthesis, statistically significant differences (p < 0.05) were observed in all experimental groups compared to negative control, the strongest effect observed in the T. cruzi infected group. Taken together, these results indicate that T. cruzi may increase the level of DNA damage in mice heart and spleen cells. Probably, nitric oxide plays an important role in DNA damaging whereas benznidazole was able to minimize induced T. cruzi genotoxic effects in spleen cells. © 2006 Elsevier Inc. All rights reserved.

First in vivo evaluation of the mutagenic effect of Brazilian green propolis by comet assay and micronucleus test

Propolis is a hive product containing chiefly beeswax and plant-derived substances such as resin and volatile compounds. Propolis has been used in various parts of the world as an antiseptic and wound healer since ancient times, and interest in the product has recently increased. Considering the lack of data concerning the in vivo mutagenicity of green propolis, the capacity of this natural product to cause damage to the DNA was evaluated, using the alkaline single-cell gel electrophoresis (SCGE) and micronucleus test, in the peripheral blood cells of mice. The doses tested by gavage were 1000, 1500 and 2000 mg/kg. Micronucleus and SCGE assays showed that green propolis caused an increase in the damage to DNA in the peripheral blood cells of mice. The polychromatic: normochromatic erythrocytes ratio was not statistically different from the negative control. Considering the doses and the results obtained in this study, the acute consumption of green propolis produced some mutagenic effects on the blood cells of mice. (C) 2008 Elsevier Ltd. All rights reserved.

Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for ∼10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare. © 2002 Elsevier Science B.V. All rights reserved.

Genetic biomonitoring of an urban population exposed to mutagenic airborne pollutants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of endogenous and synthetic female sex hormones on human blood cells in vitro studied with comet assay

The comet assay has been conducted with numerous cell lines to assess in vitro genotoxicity. In order to use the comet assay as part of an in vitro test for evaluating genotoxicity, however, there are cell-specific factors that need to be better understood. In this present study we have evaluated some factors that may impact upon the DNA damage detected in whole blood (WB) cells and lymphocytes (ILs). Experiments were conducted comparing responses of both cells, and investigating the effects of the female hormonal cycle, and oral contraceptive (OC) use on DNA damage detection in the in vitro comet assay, at three sampling time. No significant differences were detected in the basal levels of DNA damage detected in ILs and WB cells from women OC users and non-users and from men. Basal DNA damage in ILs was unaffected by gender and stage of the menstrual cycle or the stage of the treatment schedule. Our results also indicated that the H2O2 induces DNA damage in human lymphocytes independently of gender, low-dose OC use and hormonal fluctuation. However, data showed that in 3rd sampling of menstrual cycle, lymphocytes were more resistant to H2O2-induced DNA damage than those from OC users and men. © 2007 Elsevier Ltd. All rights reserved.

Sevoflurane induces DNA damage whereas isoflurane leads to higher antioxidative status in anesthetized rats

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

Genomic instability in blood cells is able to predict the oral cancer risk: an experimental study in rats

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

Comparison of oxidative stress in ASA physical status I patients scheduled for minimally invasive surgery under balanced or intravenous anesthesia

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

Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)