Single Cell Gel Research Articles

Longitudinal follow-up of oxidative stress and DNA damage parameters in detergent workers.

Background:The aim of this study was the follow-up of work place enzyme and detergent dust exposure effects and smoking habit on DNA damage parameters of workers and the evaluation of their antioxidant enzyme activities and lipid peroxidation with regard to bag-filter installation in the work place.Material and Methods:All investigated parameters were studied in a group of 153 workers of enzyme-free detergent production plant (E-free) and a group of 138 workers of enzyme-plus detergent plant (E-plus) and compared with 45 controls 7.2 years before and 3.1 years after filter system installation. The following methods were used: antioxidant enzymes by an ultraviolet–visibles spectrophotometer, malondialdehyde (MDA), 8-hydroxy-2'deoxyguanosine (8OH-2'dG) by high-performance liquid chromatography, trace elements by atomic absorption spectroscopy, and comet assay by single cell gel electrophoresis.Results:Compared with controls, significant increases were observed in both detergent-exposed groups with respect to the levels of MDA, antioxidant enzyme activities, and DNA damage parameters, including 8OH-2'dG, endonuclease III-sensitive sites, and DNA strand breaks, with enhancement effect of smoking before filter system installation. After filter installation, besides significant decrease in the detergent and enzyme dust of airborne and oxidative stress indicators, there was improvement in all DNA damage investigated parameters at the end of this study. The levels of cumulative exposure index of detergent dusts decreased significantly after airborne improvement and showed positive correlation with internal biochemical parameters.Conclusions:We concluded that high levels of enzyme and detergent contents of work place dusts had a cumulative effect and smoking had a synergistic effect on the imbalance of antioxidant status and lipid peroxidation, suggesting that oxidation stress is important in the occurrence and progression of DNA damage over this study. Detergent and enzyme contents in respirable and total dust had the main role and sufficient potential in their genotoxicity.

Evaluation of radioinduced damage and repair capacity in human breast cancer cells, MCF-7 and T-47D

Ionising radiation is an established etiological agent for breast cancer, but is also a therapeutic modality used in cancer treatment. The accumulation of deoxyribonucleic acid (DNA) damage and deficient DNA repair are considered susceptibility factors that predispose individuals to breast cancer development. In the present study, the genetic damage induced by gamma radiation and repair capacity in the target cells, i.e., cell lines originating from breast cancer, were analysed using a micronucleus (MN) test and comet assay (single-cell alkaline gel electrophoresis). Two breast tumour cell lines, MCF-7 and T-47D, were irradiated in a 60Co source (0.722 Gy/min) with various doses (0.5, 1.0, 2.0, 4.0 and 5.0 Gy). Cytogenetic data showed similar spontaneous damage of two cell lines; the radioinduced damage, however, was higher in T-47D starting from 2 Gy, with a more accelerated proliferation rate than MCF-7 at all analysed doses. Both tumour cell lines were able to repair a considerable part of the radioinduced damage within 1 h after exposure, indicating a relative radioresistance of these cell lines to the genotoxic action of ionising radiation.

Environmental and occupational biomonitoring using the Comet assay

Biomonitoring of human populations exposed to potential mutagens or carcinogens can provide an early detection system for the initiation of cell disregulation in the development of cancer. In recent years, the Comet assay, also known as a "single cell gel" (SCG) electrophoresis assay, has become an important tool for assessing DNA damage in exposed populations. This is the method of choice for population-based studies of environmental and occupational exposure to air pollutants, metals, pesticides, radiation, and other xenobiotics as we show in this review. To appreciate the role of the Comet assay in the field of biomonitoring, we review data from 122 studies that employed the assay. These studies evaluated environmental versus occupational exposures and the levels of DNA damage in cells of individuals exposed in each case. Our review of the literature reveals the importance of the need to establish standard methodological conditions that affect unwinding and electrophoresis times and tail values (tail length, tail DNA, tail moment), with the goal of being able to compare data collected in different laboratories throughout the world. The Comet assay is susceptible to subtle artifacts of manipulation depending on the type and timing of sampling performed. Therefore, in the reporting of DNA damage detected by the Comet assay, the context of how the DNA damage was created also needs to be reported and considered in the interpretation of Comet assay results. The success of the Comet assay is reflected by its use over the past 20 years in the field of biomonitoring, and by the increasing number of studies that continue to report its use. As the shortcomings of the assay are identified and considered in the interpretation of DNA damage detection, the Comet assay will continue to provide improved reliability as a biomarker in human biomonitoring studies.

Studies of the Toxicological Potential of Capsinoids: V. Genotoxicity Studies of Dihydrocapsiate

A series of studies was performed to evaluate the safety of dihydrocapsiate (4-hydroxy-3-methoxybenzyl 8-methylnonanoate; CAS no. 205687-03-2). This study evaluated the potential genotoxicity of this compound using a variety of in vitro and in vivo test systems, including bacterial reverse mutation test, chromosomal aberration test, micronucleus test, gene mutation assay with transgenic rats, and single-cell gel (SCG) assay (Comet assay). In vitro tests (bacterial reverse mutation test and chromosomal aberration test) produced positive results in the absence of metabolic activation, but negative results in the presence of metabolic activation. The in vivo gene mutation assay (with transgenic rats) produced negative results, as did the in vivo mouse micronucleus assay, which failed to induce micronucleated polychromatic erythrocytes. Although the rat SCG assay produced statistically significant increases in the Olive tail moment and % tail DNA of the liver and intestine in the 2000 mg/kg group (compared with the negative-control group), a number of factors caused the authors to question the validity of these findings. Taken together, these results suggest that dihydrocapsiate has a low or extremely low likelihood of inducing genotoxicity.

Genotoxicity and cytotoxicity in multiple organs induced by titanium miniplates in Wistar rats

Internal fixture materials are currently used as metallic biomaterials in rehabilitation of human body. Nevertheless, metal release due to corrosion phenomena appears to play a crucial role for human health. Thus, the goal of the present study was to evaluate whether liver, kidney, and lung are particularly sensitive organs for DNA damaging and cytotoxicity following implantation of internal fixture materials composed by titanium alloy in vivo. A total of 18 rats underwent surgical titanium miniplates in their tibias, being randomly distributed into three groups: 30 days, 90 days, and 180 days after implantation. A total of six animals served as negative control (animals that not received any implant). After experimental design, the lung, liver, and kidney were removed for histopatological and genotoxic analysis as depicted by H.E. stain and single cell gel (comet) assay, respectively. No significant statistically differences (p > 0.05) for DNA damaging were found to all experimental groups when compared to negative control for all organs evaluated. In addition, no remarkable morphological alterations were detected under histopathological analysis. Taken together, such results suggest that titanium miniplates are neither able to induce DNA damage in multiple organs nor to cause some abnormalities in lung, liver, and kidney.

Single-Cell Gel (Comet) Assay as a Promising Tool for the Detection of DNA Damage Induced by Compounds Used in Dental Practice: The Oral Cancer Risk Assessment

Taking into consideration genetic damage plays an important role in oral carcinogenesis, the purpose of this article is to provide an overview on the application of single-cell gel (comet) assay as a promising tool for the detection of DNA damage induced by some compounds used in dental practice as far as to assess the oral cancer risk. The single-cell gel (comet) assay is a microgel electrophoresis technique able to detect DNA damage at the level of single cells. In the alkaline version, which is mainly used, DNA single-strand breaks, DNA double-strand breaks, alkali-labile sites, and single-strand breaks containing incomplete excision repair sites cause increased DNA migration. Therefore, this interesting methodology estimates the risk of these substances with respect to genotoxicity, and will be added to those used for regulatory purposes in improving oral health and preventing oral carcinogenesis.

Paracoccidioidomycosis: no genetic damage in human peripheral blood cells of patients assessed by single-cell gel (comet) assay

Paracoccidioidomycosis is a systemic fungal infection caused by Paracoccidioides brasiliensis. As infectious diseases can cause DNA damage, the authors aimed at analyzing DNA breakage in peripheral blood cells of patients with paracoccidioidomycosis by using the comet assay. The results suggested that paracoccidioidomycosis does not cause genotoxicity.

Genotoxicity in primary human peripheral lymphocytes after exposure to radiopacifiers in vitro

Taking into consideration that DNA damage plays an important role in carcinogenesis, the purpose of this study was to evaluate whether some radiopacifiers widely used in clinical practice are able to induce genetic damage in primary human cells in vitro. Human peripheral lymphocytes obtained from 10 healthy volunteers were exposed to barium sulphate (BaSO(4)), zirconium oxide (ZnO(2)) and bismuth oxide (Bi(2)O(3)) at final concentrations ranging from 1 to 1000 microg/mL for 1 h at 37 degrees C. The negative control group was treated with vehicle control (phosphate buffer solution) for 1 h at 37 degrees C and the positive control group was treated with hydrogen peroxide (at 100 microM) for 5 min on ice. Results were analyzed by the Friedman non-parametric test. The results pointed all compounds tested out did not induce DNA breakage in human peripheral lymphocytes as depicted by the mean tail moment and tail intensity in all concentrations tested. In summary, our results indicate that exposure to these radiopacifiers may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single cell gel (comet) assay.

Genetic damage in human peripheral lymphocytes exposed to antimicrobial endodontic agents

Formocresol, paramonochlorophenol, or calcium hydroxide have been widely used in dental practice to eradicate bacteria and consequently to produce root canal disinfection. Taking into consideration strong evidence for a relationship between DNA damage and carcinogenesis, the purpose of the present study was to evaluate the genotoxic effects of antimicrobial endodontic compounds in human peripheral lymphocytes by single-cell gel (comet) assay. This technique detects DNA strand breaks in individual cells. A total of 10 microL of the tested substance solution (formocreso1, paramonochlorofeno1, and calcium hydroxide at 100-microg/mL concentration) was added to human peripheral lymphocytes from 10 volunteers for 1 hour at 37 degrees C. The negative control group was treated with vehicle control (PBS) for 1 hour at 37 degrees C, as well. For the positive control group, lymphocytes were exposed to hydrogen peroxide at 100 microM during 5 minutes on ice. No DNA breakage was detected after a treatment of peripheral lymphocytes by formocresol, paramonochlorophenol, or calcium hydroxide at 100 microg/mL. In summary, our results indicate that exposure to formocresol, paramonochlorophenol, or calcium hydroxide may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single-cell gel (comet) assay.

Absence of DNA damage in multiple organs (blood, liver, kidney, thyroid gland and urinary bladder) after acute fluoride exposure in rats

Fluoride has been widely used in dentistry as a caries prophylactic agent. However, there has been some speculation that excess fluoride could cause an impact on genome integrity. In the current study, the potential DNA damage associated with exposure to fluoride was assessed in cells of blood, liver, kidney, thyroid gland and urinary bladder by the single cell gel (comet) assay. Male Wistar rats aging 75 days were distributed into seven groups: Groups 1 (control), 2, 3, 4, 5, 6 and 7 received 0 (deionized water), 10, 20, 40, 60, 80 and 100 mgF/Kg body weight from sodium fluoride (NaF), respectively, by gastrogavage. These groups were killed at 2 h after the administration of the fluoride doses. The level of DNA strand breaks did not increase in all organs evaluated and at all doses of NaF tested, as depicted by the mean tail moment. Taken together, our results suggest that oral exposure to NaF did not result in systemic genotoxic effect in multiple organs related to fluoride toxicity. Since DNA damage is an important step in events leading to carcinogenesis, this study represents a relevant contribution to the correct evaluation of the potential health risk associated with chemical exposure.

Lack of Effect of Prior Treatment with Fluoride on Genotoxicity of Two Chemical Agents in vitro

The goal of this study was to investigate the ability of fluoride to modulate the genotoxic effects induced by the oxidative agent hydrogen peroxide (H₂O₂) and the alkylating agent methyl methanesulfonate (MMS) in vitro by the single-cell gel (comet) assay. Chinese hamster ovary cells were exposed in culture for 1 h at 37°C to sodium fluoride at 7–100 µg/ml. NaF-treated and control cells were then incubated with 0–10 µM MMS in phosphate-buffered saline (PBS) for 15 min at 37°C, or 0–100 µM H₂O₂ in distilled water for 5 min on ice. Negative control cells were treated with PBS for 1 h at 37°C. Clear concentration-related effects were observed for the two genotoxins. Increase of DNA damage induced by either MMS or H₂O₂was not significantly altered by pretreatment with NaF. The data indicate that NaF does not modulate alkylation-induced genotoxicity or oxidative DNA damage as measured by the single-cell gel (comet) assay.

Genotoxicity of Corrosion Eluates Obtained From Endosseous Implants

Commercially pure titanium alloys are currently used as metallic biomaterials in implantology. Corrosion phenomena appear to play a decisive role in metallic implant long-term behavior. Thus, the goal of this study was to examine the genotoxic potential of corrosion eluates obtained from dental implants using Chinese ovary hamster cells in vitro by the single-cell gel (comet) assay. This technique detects deoxyribonucleic acid strand breaks in individual cells in alkaline conditions. The materials tested included 3 dental implants commercially available. Each of the tested materials was corroded in a solution consisting of equal amounts of acetic acid and sodium chloride (0.1 M) for 1, 3, 7, 14, and 21 days. The Chinese ovary hamster cultures were then exposed to all corrosion eluates obtained from endosseous dental implants for 30 minutes at 37 degrees C. None of the eluates was found to exhibit genotoxicity, regardless of the type of dental implant used. The results suggest that all dental implants tested in this study did not induce deoxyribonucleic acid breakage as depicted by the single-cell gel (comet) assay.

Biocompatibility of gutta-percha solvents using in vitro mammalian test-system

Taking into consideration that DNA damage and cellular death play important roles during carcinogenesis, the purpose of the present study was to evaluate in vitro genotoxic or cytotoxic effects of chloroform and eucalyptol by single cell gel (comet) assay and trypan blue exclusion test, respectively. Chloroform and eucalyptol were exposed to Chinese hamster ovary cells in culture directly for 3 hours at 37 degrees C at final concentrations ranging from 1.25 to 10 microL/mL. The negative control group was treated with vehicle control (phosphate-buffered solution), and the positive control group was treated with methyl metasulfonate (MMS, at 1 microg/mL concentration). All data were analyzed by the Kruskal-Wallis nonparametric test followed by the Dunn test. The results showed that both gutta-percha solvents were cytotoxic at concentrations of 2.5, 5, and 10 microL/mL (P < .05). On the other hand, both solvents did not induce DNA breakage at 1.25 microL/mL concentration. These results suggest that both chloroform or eucalyptol are strong cytotoxicants, but they may not be a factor that increases the level of DNA lesions in mammalian cells.

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.

Biocompatibility of glass–ionomer cements using mouse lymphoma cells in vitro

Glass-ionomer cements are widely used in dentistry as restorative materials and adhesives for composite restorations. A number of genotoxicity studies have been conducted using these materials with results conflicting so far. Thus, the approach was aimed to look at the genotoxic and cytotoxic potential of three different glass-ionomer cements available commercially (Ketac Cem, Ketac Molar and Vitrebond) by the single cell gel (comet) assay and trypan blue exclusion test, respectively. For this, such materials were exposed to mouse lymphoma cells in vitro for 1 h at 37 degrees C. Data were assessed by Kruskall-Wallis non-parametric test. The results showed that all powders assayed did not show genotoxic effects. On the other hand, the liquid from Vitrebond at 0.1% dilution caused an increase of DNA injury. Significant statistically differences (P < 0.05) in cytotoxicity provoked by all powders tested were observed for exposure at 1,000 micro g mL(-1) concentration and 100 micro g mL(-1) for Ketac Molar. With respect to liquids of glass-ionomer cements evaluated, the major toxic effect on cell viability was produced at 1%, beginning at the dilution of 0.5% for Vitrebond. Taken together, these results support the notion that some components of glass-ionomer cements show both genotoxic and cytotoxic effects in higher concentrations.

DNA damage in multiple organs after exposure to chlorhexidine in Wistar rats

Since chlorhexidine is effective against microorganisms, it is widely recommended in dentistry. However, studies have provided evidence that chlorhexidine is toxic for a variety of cell types. In order to identify potential genotoxins in different cell types, the purpose of this study was to investigate whether chlorhexidine digluconate is able to cause, in terms of DNA damage, alterations in leukocytes, liver, kidney and urinary bladder by the single cell gel (comet) assay. Ten male Wistar rats were divided into two groups: a negative control and the experimental group treated with 3 ml of 0.12% chlorhexidine digluconate by gavage once a day for 8 days. Statistically significant increases of DNA damage was observed in leukocytes and kidney cells of the chlorhexidine digluconate treated group as depicted by the mean tail moment. Taken together, the data indicate that leukocytes and kidney cells are potential targets for primary DNA damage following oral exposure to chlorhexidine digluconate as detected by single cell gel (comet) assay.

유청단백질 및 Lactobacillus spp. 추출물이 전립선 세포 내 항산화 활성에 미치는 영향

본 실험은 가수분해 시킨 유청단백질과 Lb. casei HY2782의 세포 추출물을 인간 전립선 세포에처리하여 세포 내 glutathione 농도 상승효과와 산화제 tbutyl hydroperoxide (TBHP)에 의한핵산 손상 방지 효과를 알아보기 위한 실험으로 전립선 세포 RWPE1 cells와 PC3MMM2 cells에 hydrolyzed WPI(500g/m)를 48시간 동안 처리 시, 가수분해 시키지 않은 WPI 보다 각각 28.2%, 38.4%씩 GSH 농도가 증가하는 것을 유의적으로 확인할 수 있었고(p<0.05), 카제인의 경우 RWPE1 cells and PC3MMM2 cells에 처리 시 가수분해 시킨 카제인과의 유의차가 보이지 않았으며 (p<0.05), glutathione 합성저해제인 Buthionine sulfoximin (BSO) 처리 시 Glutathione 농도가 현저하게 낮아지는 것을 확인할 수가 있었다(Anderson, 1998). 전립선 세포 RWPE1 cells 와 PC3MMM2 cells에 Lb. casei HY2782 세포 추출물 처리 시 PC3MMM2 cell에서는 GSH의 농도가 유의적으로 증가하는 것을 확인 할 수 있었고(p<0.05), RWPE1 cell에서는 증가 성향을 확인 할 수가 있었다.가수분해 시킨 유청단백질과 Lb. casei cell 세포추출물 처리 시 산화제에 의한 세포의 DNA 손상 억제정도와 사멸율의 변화를 확인 한 결과, 가수분해 WPI를 처리 후 Oxidant tbutyl hydroperoxide (TBHP)(500mM)를 이용하여 PC3MMM2 세포에 산화적인 스트레스를 주었을 시, TBHP만 처리한 구에서는 62.0%의 생존율을 나타내었고, glutathione 합성저해제인 BSO를 첨가시킨 처리구에서는 33.7%의 생존율을 나타내었다. 가수분해 시킨 WPI와 카제인 처리구에서는 각각 86.7%, 63.6%의 생존율을 나타내었고, WPI와 BSO를 함께 처리한 구에서는 71.5%의 생존율을 나타내었다. 이는 BSO에 의해 GSH의 생성이 저해되어 나타난 결과로 보여지며(Anderson, 1988), 가수분해시킨 WPI 가 가수분해 시킨 카제인보다는 높은 생존율을 나타내는 것을 유의적으로 확인할 수가 있었다. (p<0.05) Lb. casei 세포 추출물처리에 의한 생존율에서는 가수분해시킨 WPI와 유사한 수준의 82.4%의 생존율을 보였다. 가수분해시킨 유청단백질과 Lb. casei cell 세포추출물에 의한 세포에서의 glutathione 농도 증가에 의해 산화제에 의한 DNA의 손상이 적었던 것으로 사료되어지고, 이로 인해 생존율이 높았던 것으로 판단된다. Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

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

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. Human peripheral lymphocytes obtained from 10 healthy volunteers were exposed to 2 presentation forms of MTA at final concentrations ranging from 1 to 1000 microg/mL for 1 hour at 37 degrees C. The negative control group was treated with vehicle control (phosphate buffer solution, PBS) for 1 hour at 37 degrees C and the positive control group was treated with hydrogen peroxide (at 100 microM) for 5 minutes on ice. Results were analyzed by the Friedman nonparametric test. 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. 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.

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

To investigate if formocresol, paramonochlorophenol, or calcium hydroxide modulate the genotoxic effects induced by the oxidatively damaging agent hydrogen peroxide (H2O2) or the alkylating agent methyl methanesulfonate (MMS) in vitro by using single cell gel (comet) assay. Chinese hamster ovary (CHO) cells in culture were exposed directly to formocresol, paramonochlorophenol, or calcium hydroxide (adjusted to 100 microg/mL) for 1 hour at 37 degrees C. Subsequently the cultures were incubated with increasing concentrations (0-10 micromol/L) of MMS in phosphate-buffered solution (PBS) for 15 minutes at 37 degrees C or of H2O2 at increasing concentrations (0-100 micromol/L) in distilled water for 5 minutes on ice. The negative control cells were treated with PBS for 1 hour at 37 degrees 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). 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. 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.

Genotoxicity and cytotoxicity of glass ionomer cements on Chinese hamster ovary (CHO) cells

Glass ionomer cements are widely used in dentistry as restorative materials and adhesives for composite restorations. However, the results of genotoxicity studies using these materials are inconclusive in literature. The goal of this study was to examine the genotoxic and cytotoxic potential of three different glass ionomer cements available commercially (Ketac Cem, Ketac Molar and Vitrebond) by the single cell gel (comet) assay and trypan blue exclusion test, respectively. For this, such materials were exposed to Chinese hamster ovary (CHO) cells in vitro for 1 h at 37( composite function)C. Data were assessed by Kruskall-Wallis nonparametric test. The results showed that the powder from Ketac Molar displayed genotoxicity only in the maximum concentration evaluated (100 microg/mL). In the same way, the liquid from Vitrebond at 0.1% dilution caused an increase of DNA injury. Significant differences (P<0.05) in cytotoxicity provoked by all powders tested of glass ionomer cements were observed for exposure at 1,000 microg/mL concentration. With respect to liquids of glass ionomer cements evaluated, the major toxic effect on cell viability was produced at 10%, beginning at the dilution of 0.5% for Vitrebond. Taken together, we conclude that some components of glass ionomer cements show both genotoxic and cytotoxic effects.