Pro-inflammatory mediators expression by pulp cells following tooth whitening on restored enamel surface.

This paper aimed to assess the influence of adhesive restoration interface on the diffusion of hydrogen peroxide (H2O2), indirect toxicity, and pro-inflammatory mediators expression by odontoblast-like cells, after in-office tooth whitening. Dental cavities prepared in bovine enamel/dentin discs were adhesively restored and subjected or not to hydrolytic degradation (HD). A whitening gel with 35% H2O2 (WG) was applied for 45 min onto restored and non-restored specimens adapted to artificial pulp chambers giving rise to the groups: SD- intact discs (control); SD/HP- whitened intact discs; RT/HP- restored and whitened discs; and RT/HD/HP- restored and whitened discs subjected to HD. The extracts (culture medium + WG components diffused through enamel/dentin/restoration interface) were collected and applied to odontoblast-like MDPC-23 cells. The study evaluated the amount of H2O2 in the extracts, as well as the cell viability (CV), cell morphology (CM), and gene expression of inflammatory mediators (TNF-α and COX-2) by the pulp cells exposed to the extracts (ANOVA and Tukey tests; 5% significance). All whitened groups presented lower CV than SD (control; p<0.05). The highest CV reduction and gene expression of TNF-α and COX-2 was observed in the RT/HD/HP group in comparison with SD/HP and RT/HP (control; p<0.05). CM alterations occurred in all whitened groups. The intensity of these cell side effects was directly related with the amount of H2O2 in the extracts. We concluded that adhesive restoration of dental cavity increases the H2O2 diffusion after in-office whitening, enhancing the indirect toxicity of this therapy and trigger pro-inflammatory overexpression by MDPC-23 cells.

Influence of smoking on oral cells genotoxicity after at-home bleaching using 22% carbamide peroxide: a cohort study.

OBJECTIVE: This study evaluated the influence of smoking on the oral cells genotoxicity before and after at-home bleaching using 22% carbamide peroxide (CP). MATERIALS AND METHODS: This is a prospective observational analytics cohort study which evaluated nonsmokers (NS; n = 24) and smokers (S; n = 16) patients. At-home bleaching was performed using 22% CP gel in individual trays for 1 h per day for 14 days in both groups. Scrapped cells from marginal gums were collected before the bleaching treatment (D0-baseline) and 1 day (D1), 15 days (D15), and 1 month (D30) after its finishing. Cells were stained with Giemsa 10%, and the micronucleus (MN) and metanuclear alterations (MA) were counted by a trained operator in 1000 cells per patient. The collections and data analysis occurred blindly. Data was analyzed by Kruskal-Wallis, Dunn, and Mann-Whitney test (α = 0.05). RESULTS: MN frequency was not influenced by smoking or bleaching. An increase of MA was observed between D0 and D30 for both groups (p < 0.001); however, no statistical difference was found between NS and S (p > 0.05) in the evaluation times. CONCLUSION: Smoking associated with 22% carbamide peroxide gel for at-home bleaching does not show genotoxic potential analyzed by the MN counts. However, a significant increase of MA was found for smokers and nonsmokers. CLINICAL RELEVANCE: Despite of the increase in MA, smoking associated with 22% CP peroxide at-home bleaching showed no important genotoxic potential (MN) for oral cells. Therefore, at-home bleaching treatment is safe for nonsmokers and smokers even with a high carbamide peroxide concentration of 22%.

Increased whitening efficacy and reduced cytotoxicity are achieved by the chemical activation of a highly concentrated hydrogen peroxide bleaching gel

Abstract Objective This study was designed for the chemical activation of a 35% hydrogen peroxide (H2O2) bleaching gel to increase its whitening effectiveness and reduce its toxicity. Methodology First, the bleaching gel - associated or not with ferrous sulfate (FS), manganese chloride (MC), peroxidase (PR), or catalase (CT) - was applied (3x 15 min) to enamel/dentin discs adapted to artificial pulp chambers. Then, odontoblast-like MDPC-23 cells were exposed for 1 h to the extracts (culture medium + components released from the product), for the assessment of viability (MTT assay) and oxidative stress (H2DCFDA). Residual H2O2 and bleaching effectiveness (DE) were also evaluated. Data were analyzed with one-way ANOVA complemented with Tukey's test (n=8. p<0.05). Results All chemically activated groups minimized MDPC-23 oxidative stress generation; however, significantly higher cell viability was detected for MC, PR, and CT than for plain 35% H2O2 gel. Nevertheless, FS, MC, PR, and CT reduced the amount of residual H2O2 and increased bleaching effectiveness. Conclusion Chemical activation of 35% H2O2 gel with MC, PR, and CT minimized residual H2O2 and pulp cell toxicity; but PR duplicated the whitening potential of the bleaching gel after a single 45-minute session.

Análise de citotoxicidade, desmineralização e potencial clareador de géis clareadores experimentais, baseado em peróxido de carbamida 15% adicionados de nano-hidróxiapatita / Analysis of cytotoxicity, demineralization and bleaching potential of experimental bleaching gels, based on 15% carbamide peroxide added with nanohydroxyapatite

O clareamento dental é considerado um procedimento muito seguro, de efeito rápido e agradável, tornando-se um dos procedimentos mais realizados na odontologia. Contudo, o mesmo apresenta alguns efeitos colaterais, sendo o principal deles a sensibilidade dental, que pode ser causada pela citotoxidade, pela desmineralização, pela geração de espécies reativas do oxigênio na polpa dental e pela degradação da região interprismática facilitando o estimulo nos túbulos dentinários. Diante disso, a presente pesquisa avaliou a desmineralização, a citotoxidade e o potencial clareador de géis clareadores experimentais baseados em peróxido de carbamida (PC) 15%, adicionados de 5% e 10% de nano partículas de hidroxiapatita (n-Hap). Para a avaliação cromática foram utilizados 36 incisivos bovinos divididos em 3 grupos: Grupo1 PC 15%, Grupo 2 PC 15% adicionado 5% nHap e grupo 3 PC 15% com 10% n-Hap, mensurando a cor antes e depois do procedimento terapêutico clareador com auxilio de um espectrofotômetro. As coordenadas do CIELAB foram analisadas para verificação da diferença de cor (ΔE) obtida entre os grupos e dentro do próprio grupo (L*) e, após a analise estatística, foi verificado que todos os géis foram capazes de clarear e não houve diferenças entre os grupos. A citotoxidade foi verificada através da absorbância das células viáveis após o tratamento com o MTT e 1:800 das preparações de agente clareador e gel contendo 5% e 10% de n-Hap aplicadas sobre células L929. Nesse caso os grupos foram desiguais estatisticamente, apresentando o PC 15% como o mais tóxico, seguido do PC 15% adicionado de 10% de n-Hap e o melhor resultado foi obtido pelo gel acrecido de 5% de n-Hap. A análise do conteúdo mineral foi realizada pela fluorescência de raios X por dispersão de energia, onde foi necessário o uso de 15 dentes bovinos, que tiveram seus terços médios seccionados, embutidos em resina epóxi e planificados, e foram dividos em grupos de 5 CPs. Cada CP foi aferido em 8 pontos diferentes antes e depois, nos mesmos pontos da terapia clareadora. Foi então verificado que os géis experimentais apresentaram menor perda de Ca e P. No entanto, não houve diferenças estatísticas entre os grupos 2 e 3. Diante das limitações do presente estudo in vitro é possível concluir que os géis experimentais contendo n-Hap foram eficazes em reduzir a citotoxidade e desmineralização sem perder a eficácia da terapia clareadora

Dental whitening is considered a very safe, quick and pleasant procedure, making it one of the most accomplished procedures in dentistry. However, it has some side effects, the main one being sensibility, which can be caused by cytotoxicity, demineralization, oxygen arrival in the dental pulp and the degradation of the interprismatic region facilitating the stimulation in the dentin tubes. Therefore, the present study evaluated the demineralization, cytotoxicity and bleaching potential of 15% carbamide peroxide-based (PC) based bleaching gels added with 5% and 10% nano-hydroxyapatite (n-Hap) particles. For the chromatic evaluation, 36 bovine incisors were divided into 3 groups: Group 1 PC 15%, Group 2 PC 15% added 5% nHap and group 3 PC 15% with 10% n-Hap, measuring the color before and after the the CIELAB coordinates were analyzed to verify the color difference (Δe) obtained between the groups and within the group (L *). After the statistical analysis it was verified that all the gels were able to lighten and there were no differences between the groups. Cytotoxicity was verified by optical density of viable cells after treatment with MTT and 1: 800 of the bleach and gel preparations containing 5% and 10% nHap applied in L929 cells. In this case, the groups were statistically unequal, presenting 15% PC as the most toxic, followed by PC 15% added with 10% n-Hap and the best result was obtained by the 5% n-Hap increased gel. The mineral content analysis was performed by X-ray Fluorescence by energy dispersion, where it was necessary to use 15 bovine teeth, which had their middle sections sectioned, embedded in epoxy resin and planned, and were divided into groups of 5 CPs, each CP was measured at 8 different points before and after, at the same points of the bleaching therapy, then it was verified that the experimental gels presented a lower loss of Ca and P, however there were no statistical differences between groups 2 and 3. On the face of the limitations of the present in vitro study, it is possible to conclude that the experimental gels were effective in reducing the cytotoxicity and demineralization without losing the effectiveness of the bleaching therapy.

Inflammatory response of human dental pulp to at-home and in-office tooth bleaching

ABSTRACT Tooth bleaching is a technique of choice to obtain a harmonious smile, but bleaching agents may damage the dental pulp. Objective: This study evaluated the inflammatory responses of human dental pulp after the use of two bleaching techniques. Material and Methods: Pulp samples were collected from human third molars extracted for orthodontic reasons and divided into three groups: control - no tooth bleaching (CG) (n=7); at-home bleaching with 15% carbamide peroxide (AH) (n = 10), and in-office bleaching with 38% hydrogen peroxide (IO) (n=12). Pulps were removed and stained with hematoxylin-eosin for microscopic analysis of inflammation intensity, collagen degradation, and pulp tissue organization. Immunohistochemistry was used to detect mast cells (tryptase+), blood vessels (CD31+), and macrophages (CD68+). Chi-square, Kruskal-Wallis, and Mann Whitney tests were used for statistical analysis. The level of significance was set at p<.05. Results: The inflammation intensity and the number of macrophages were significantly greater in IO than in AH and CG (p<0.05). The results of CD31+ (blood vessels per mm2) were similar in CG (61.39±20.03), AH (52.29±27.62), and IO (57.43±8.69) groups (p>0.05). No mast cells were found in the pulp samples analyzed. Conclusion: In-office bleaching with 38% hydrogen peroxide resulted in more intense inflammation, higher macrophages migration, and greater pulp damage then at-home bleaching with 15% carbamide peroxide, however, these bleaching techniques did not induce migration of mast cells and increased the number of blood vessels.

Toxicidade do peróxido de carbamida 16% adicionado de nanopartículas de hidroxiapatita / Carbamide peroxide toxicity added 16% hydroxyapatite nanoparticles

O clareamento dental é um dos tratamentos mais realizados nos consultórios odontológicos a fim de melhorar a aparência do sorriso. O procedimento consiste na aplicação de um gel clareador, a base de peróxido de carbamida ou de hidrogênio, sobre os dentes a serem clareados. A sensibilidade dentária é o efeito adverso mais frequentemente relatado no clareamento dentário e é a principal causa de desmotivação dos pacientes. O mecanismo pelo qual se produz a sensibilidade após clareamento dentário ainda não foi completamente elucidado; no entanto, parece estar associado à rápida difusão dos agentes clareadores através do esmalte e dentina que, devido ao seu grau de citotoxicidade, podem agir agredindo as células pulpares, causando sensibilidade. O objetivo deste estudo foi avaliar a biocompatibilidade de nanopartícula de hidroxiapatita adicionada ao gel clareador peróxido de carbamida 16% com flúor e sem flúor e, para tanto, foram realizados testes de citotoxidade empregando MTT. Como resultados verificou-se que as nanopartículas de hidroxiapatita, quando comparadas aos géis de peróxido de carbamida a 16% com e sem flúor, foram as menos citotóxicas (p< 0.05). As diluições citotóxicas convertidas para 70% das amostras testadas também foram comparadas através do teste de Anova com tukey. Foi possível observar que as partículas de nanoHap quando adicionadas nos géis de clareamento com e sem flúor reduziu significativamento a citotoxidade. Concluímos que o novo material proposto nesta investigação apresenta melhor biocompatibilidade do que o gel sem hidroxiapatita, acompanhado da redução da citotoxidade, tais aspectos sugerem que as nanopartículas de hidroxiapatita podem ter aplicações clínicas futuras em tecidos mineralizados em procedimentos de clareamento dental, contribuindo para a redução da sensibilidade dentária.

The dental bleaching is one of the most procedures performed at dental offices to improve the appearance of the smile. The procedure consists of the application of a bleching gel, based on carbamide peroxide or hydrogen over the teeth to be whitened. Tooth sensibility is the frequently reported adverse effect on tooth whitening and is the leading cause of demotivation by the patients. The mechanism that provides the sensibility after tooth whitening has not already been fully elucidated; however, it appears to be associated with the rapid diffusion of bleaching agents through the enamel and dentin that, because of their degree of cytotoxicity, may act attacking pulp cells, causing the sensibility. The aim of this study was to evaluate the biocompatibility of hydroxyapatite nanoparticles added to whitening gel of 16% carbamide peroxide with and without fluoride and, therefore, cytotoxicity tests were performed using MTT. As a result, it was found that the hydroxyapatite nanoparticles, compared to the gels of 16 % carbamide peroxide with and without fluoride, were less cytotoxic (p <0.05). Cytotoxic dilutions converted to 70 % of the tested samples were compared using ANOVA test with Tukey. We concluded that the new material proposed in this research has a better biocompatibility in comparison with the gel without hydroxyapatite, followed by a reduction of cytotoxicity. These aspects suggest that the hydroxyapatite nanoparticles may have clinical future applications in mineralized tissues when dental bleaching procedures are performed, contributing to the reduction of tooth sensitivity. We conclude that the new material proposed in this research has a better biocompatibility of the gel without hydroxyapatite, accompanied by a reduction of cytotoxicity, these aspects suggest that the hydroxyapatite nanoparticles may have future clinical applications in mineralized tissues in dental bleaching procedures, contributing to a reduction tooth sensitivity.

Effects on gastric mucosa induced by dental bleaching--an experimental study with 6% hydrogen peroxide in rats.

UNLABELLED: The value of aesthetic dentistry has precipitated several developments in the investigation of dental materials related to this field. The free marketing of these products is a problem and it is subject to various interpretations regarding its legality. There are several techniques for tooth whitening, the most used one being the external bleaching. It is the later version of such technique that poses the greatest danger of ingesting the product. The present study analysed the systemic effect of these products when they are swallowed. OBJECTIVE: This experimental study aimed to observe the effects of a tooth whitening product, whose active agent is 6% hydrogen peroxide, on the gastric mucosa of healthy and non-tumour gastric pathology animals. MATERIAL AND METHODS: Fifty Wistar-Han rats were used and then distributed into 5 groups, one for control and four test groups in which the bleaching product was administered in animals with and without non-tumour gastric pathology (induced by the administration of 1 sample of 50% ethanol and 5% of drinking water during 6 days) at different times of study by gavage. There was a decrease in body weight in animals of groups handled during the study period, which was most pronounced in IV and VA groups. Changes in spleen weight relative to body weight revealed no statistically significant changes. An analysis of the frequency was performed on the results of macroscopic observation of the gastric mucosa. RESULTS: The gastric mucosa revealed lesions in all manipulated groups, being more frequent in groups III and IV. It appears that there is a synergism when using hydrogen peroxide and 50% ethanol in the same group. CONCLUSION: Therefore, it seems that there are some signs of toxicity 3 to 4 days after administration of 6% hydrogen peroxide. The prescription of these therapies must be controlled by the clinician and the risks must be minimized.

Effects on gastric mucosa induced by dental bleaching – an experimental study with 6% hydrogen peroxide in rats

The value of aesthetic dentistry has precipitated several developments in the investigation of dental materials related to this field. The free marketing of these products is a problem and it is subject to various interpretations regarding its legality. There are several techniques for tooth whitening, the most used one being the external bleaching. It is the later version of such technique that poses the greatest danger of ingesting the product. The present study analysed the systemic effect of these products when they are swallowed.Objective This experimental study aimed to observe the effects of a tooth whitening product, whose active agent is 6% hydrogen peroxide, on the gastric mucosa of healthy and non-tumour gastric pathology animals.Material and Methods Fifty Wistar-Han rats were used and then distributed into 5 groups, one for control and four test groups in which the bleaching product was administered in animals with and without non-tumour gastric pathology (induced by the administration of 1 sample of 50% ethanol and 5% of drinking water during 6 days) at different times of study by gavage. There was a decrease in body weight in animals of groups handled during the study period, which was most pronounced in IV and VA groups. Changes in spleen weight relative to body weight revealed no statistically significant changes. An analysis of the frequency was performed on the results of macroscopic observation of the gastric mucosa.Results The gastric mucosa revealed lesions in all manipulated groups, being more frequent in groups III and IV. It appears that there is a synergism when using hydrogen peroxide and 50% ethanol in the same group.Conclusion Therefore, it seems that there are some signs of toxicity 3 to 4 days after administration of 6% hydrogen peroxide. The prescription of these therapies must be controlled by the clinician and the risks must be minimized.

Evaluation of genotoxicity and efficacy of at-home bleaching in smokers: a single-blind controlled clinical trial.

OBJECTIVE: This single-blind controlled study evaluated the genotoxicity and efficacy of at-home bleaching in smokers and nonsmokers. METHODS: We selected 60 patients with central incisors A2 or darker: 30 smokers (experimental group) and 30 nonsmokers (control group). The bleaching was carried out with 10% carbamide peroxide for three hours a day for three weeks. The color was evaluated using a shade guide, Vita Bleachedguide 3D-Master, at baseline, during bleaching (first, second, and third weeks), and one week and one month after bleaching. Smears were obtained with a moistened wooden spatula from marginal gingiva. All the cytologic smears were stained with Giemsa solution. From each slide, 1000 cells were examined under 40× magnification and where micronuclei (MN) were located, they were examined under 100× magnification. The change in shade guide units at the different assessment periods and the frequency of MN were subjected to a two-way repeated measures analysis of variance and Tukey test (α=0.05). RESULTS: In both groups we detected a whitening of approximately 4 to 5 shade guide units, without color rebound after one month (p>0.05). The frequency of MN was significantly higher in the experimental group than in the control group, regardless of the bleaching treatment (p>0.001). CONCLUSION: The efficacy of bleaching does not appear to be affected by the smoking habit. Additionally, at-home bleaching did not induce DNA damage to the gingival tissue during the bleaching period.

Effect of Fluoride-Treated Enamel on Indirect Cytotoxicity of a 16% Carbamide Peroxide Bleaching Gel to Pulp Cells

The aim of this study was to evaluate the possibility of fluoride solutions applied to enamel to protect pulp cells against the trans-enamel and transdentinal cytotoxicity of a 16% carbamide peroxide (CP) bleaching gel. The CP gel was applied to enamel/dentin discs adapted to aicial pulp chambers (8 h/day) during 1, 7 or 14 days, followed by fluoride (0.05% or 0.2%) application for 1 min. The extracts (culture medium in contact with dentin) were applied to MDPC-23 cells for 1 h, and cell metabolism (MTT assay), alkaline phosphatase (ALP) activity and cell membrane damage (flow cytometry) were analyzed. Knoop microhardness of enamel was also evaluated. Data were analyzed statistically by ANOVA and Kruskal-Wallis tests (α=0.05). For the MTT assay and ALP activity, significant reductions between the control and the bleached groups were observed (p<0.05). No statistically significant difference occurred among bleached groups (p>0.05), regardless of fluoride application or treatment days. Flow cytometry analysis demonstrated 30% of cell membrane damage in all bleached groups. After 14 days of treatment, the fluoride-treated enamel presented significantly higher microhardness values than the bleached-only group (p<0.05). It was concluded that, regardless of the increase in enamel hardness due to the application of fluoride solutions, the treated enamel surface did not prevent the toxic effects caused by the 16% CP gel to odontoblast-like cells.

Resumo O objetivo do presente estudo foi avaliar o possível efeito protetor de soluções fluoretadas aplicadas sobre o esmalte dentário frente à citotoxicidade trans-amelodentinária de um gel clareador com 16% de peróxido de carbamida (PC). O gel de PC foi aplicado sobre discos de esmalte/dentina adaptados a câmaras pulpares aiciais (8 h/dia) durante períodos de 1, 7 ou 14 dias, seguido de aplicação de soluções fluoretadas (0,05% ou 0,2%) durante 1 min. Os extratos (meio de cultura em contato com a dentina) foram aplicados sobre células MDPC-23 durante 1 h, seguido de análise do metabolismo celular (teste do MTT), atividade de fosfatase alcalina (ALP) e danos à membrana celular (citometria de fluxo). A microdureza Knoop do esmalte dental foi avaliada. Os dados foram analisados pelos testes de ANOVA e Kruskal-Wallis. Para o teste do MTT e atividade de ALP, redução significante entre os grupos controle e clareados foram observados (p<0,05). Nenhuma diferença entre os grupos clareados foi observada (p>0,05), independente da aplicação das soluções fluoretadas ou tempo de tratamento. A análise por citometria de fluxo demonstrou lesão à membrana celular em torno de 30% para todos os grupos clareados. Após 14 dias de tratamento, os espécimes clareados e fluoretados apresentaram aumento significante na microdureza do esmalte (p<0,05). Pôde-se concluir que apesar do aumento na dureza do esmalte decorrente da aplicação das soluções fluoretadas, este tratamento não preveniu os efeitos tóxicos causados pelo gel com 16% de PC sobre as células odontoblastóides. .

Soft-tissue alterations following exposure to tooth-whitening agents.

BACKGROUND: Tooth-whitening agents are widely used, either as self-application products or under the supervision of a dentist. These products may be associated with transient gross morphologic changes in oral soft tissues. However, their potential effects on human keratinocytes and fibroblasts in a stratified squamous epithelium have yet to be elucidated. METHODS: In this study, three-dimensional human tissue equivalents are exposed to varying concentrations of tooth-whitening agents for increasing time periods. Tissue alterations are investigated in terms of morphology, proliferation, apoptosis, and protein expression. RESULTS: All whitening agents tested altered tissue morphology, induced proliferation of basal keratinocytes, and caused apoptosis of cells in all epithelial strata. In addition, whitening agents induced alterations in the expression of cytokines that are linked to inflammation. CONCLUSIONS: These results suggest that whitening agents may induce similar changes in vivo and that these products should be used for limited periods of time or under the supervision of a dental professional.