Dose-response and time-course of α-tocoferol mediating the cytoprotection of dental pulp cells against hydrogen peroxide.

This in vitro study evaluated the potential protective effect of vitamin E alpha-tocopherol (α-T) isomer against the toxicity of hydrogen peroxide (HP) applied on dental pulp cells. Odontoblast-like MDPC-23 cells were seeded on 96-well plates for 72 h, treated with different concentrations of α-T (1, 3, 5, and 10 mM) for different times (1, 4, 8, and 24 h) and then exposed or not to a 0.018% HP solution for 30 min. In positive and negative control groups, cells were exposed to HP or culture medium (DMEM containing 5% DMSO), respectively. Cell viability was assessed by the MTT assay and the absorbance numeric data, expressed as percentage values, were subjected to the statistical analysis by Kruskal-Wallis and Mann-Whitney tests (α=5%). Considering the cells in the negative control as having 100% of cell viability, all combinations of α-T concentrations and pretreatment times showed a protective effect against HP cytotoxicity. Significant reduction of cell viability (59%) was observed in the positive control compared with the negative control. The highest values of pulp cell viability were obtained after pretreatment with 1 and 3 mM α-T concentrations for 24 h followed by exposure to HP (126% and 97% of cell viability, respectively). Under the tested conditions, the most effective cell protection against the cytotoxic effects of HP was provided by the lowest concentrations of α-T (1 and 3 mM) applied for 24 h.

Dose-Response and Time-Course of a-Tocoferol Mediating the Cytoprotection Of Dental Pulp Cells Against Hydrogen Peroxide

This in vitro study evaluated the potential protective effect of vitamin E alpha-tocopherol (α-T) isomer against the toxicity of hydrogen peroxide (HP) applied on dental pulp cells. Odontoblast-like MDPC-23 cells were seeded on 96-well plates for 72 h, treated with different concentrations of α-T (1, 3, 5, and 10 mM) for different times (1, 4, 8, and 24 h) and then exposed or not to a 0.018% HP solution for 30 min. In positive and negative control groups, cells were exposed to HP or culture medium (DMEM containing 5% DMSO), respectively. Cell viability was assessed by the MTT assay and the absorbance numeric data, expressed as percentage values, were subjected to the statistical analysis by Kruskal-Wallis and Mann-Whitney tests (α=5%). Considering the cells in the negative control as having 100% of cell viability, all combinations of α-T concentrations and pretreatment times showed a protective effect against HP cytotoxicity. Significant reduction of cell viability (59%) was observed in the positive control compared with the negative control. The highest values of pulp cell viability were obtained after pretreatment with 1 and 3 mM α-T concentrations for 24 h followed by exposure to HP (126% and 97% of cell viability, respectively). Under the tested conditions, the most effective cell protection against the cytotoxic effects of HP was provided by the lowest concentrations of α-T (1 and 3 mM) applied for 24 h.

Neste estudo, foi avaliado o potencial protetor do isômero alfa-tocoferol da vitamina E (-T) contra a ação tóxica do peróxido de hidrogênio (PH) aplicado sobre células pulpares. Células odontoblastóides MDPC-23 foram semeadas em placas de 96 compartimentos por 72 h, tratadas com diferentes concentrações de α-T (1, 3, 5 e 10 mM) por diferentes períodos (1, 4, 8 e 24 h) e, então, expostas ou não a uma solução com 0,0018% de PH por 30 min. Nos controles positivo e negativo, as células foram expostas ao PH ou meio de cultura (DMEM contendo 5% de DMSO), respectivamente. A viabilidade celular foi avaliada pelo teste do MTT e os valores numéricos de absorbância, expressos em porcentagem, foram submetidos a análise estatística pelos testes de Kruskal-Wallis e Mann-Whitney (α=5%). Considerando as células do grupo controle negativo como apresentando 100% de viabilidade celular, todas as combinações de α-T, nas diferentes concentrações, e tempos de pré-tratamento demonstraram um efeito protetor contra a citotoxicidade do PH. Redução significativa da viabilidade (59%) foi observada para o grupo controle positivo comparado ao controle negativo. Os maiores valores de viabilidade celular foram obtidos após pré-tratamento com 1 e 3 mM de α-T por 24 h seguido de exposição ao PH (126% e 97% de viabilidade celular, respectivamente). Assim, de acordo com as condições experimentais, o efeito protetor mais efetivo contra os efeitos tóxicos do PH foi observado para as menores concentrações de α-T (1 e 3 mM) aplicado por 24 h.

Efeito protetor da vitamina E (α-Tocoferol) contra a atividade citotóxica do peróxido de hidrogênio / Protective effect of vitamin E (α-tocopherol) against the cytotoxicity of hydrogen peroxide

O clareamento dental de consultório, realizado com géis que apresentam elevada concentração de peróxido de hidrogênio (PH), pode causar danos intensos para as células da polpa dentária. Dentro deste contexto, a administração de agentes antioxidantes previamente ao clareamento dental, tem sido considerada uma terapia promissora, pois pode eliminar ou pelo menos minimizar os efeitos adversos deste procedimento clínico estético. Assim, o objetivo do presente estudo foi avaliar o possível efeito protetor da Vitamina E (alfa-Tocoferol / α-T) contra à ação citotóxica do PH sobre células odontoblastóides (MDPC-23). Para isto, células foram semeadas em placas de 96 wells durante 72 horas e então submetidas a diferentes tempos de pré-tratamento (1, 4, 8 e 24 horas) com variadas concentrações de α-T (1, 3, 5 e 10 mM). Após os períodos de pré-tratamento, as células foram expostas ou não ao PH (0,018%) durante 30 minutos. Nos grupos controle positivo e negativo, as células foram somente expostas a uma solução de PH (0,018%) ou meio de cultura, respectivamente. O metabolismo celular foi avaliado pelo teste de MTT. A absorbância foi transformada em porcentagem e analisada pelo Teste de Kruskal-Wallis, complementado por Mann-Whitney (α=0,05%). Todas as concentrações de αT e tempos de pré-tratamento propostos neste estudo apresentaram proteção das células contra os efeitos citotóxicos do PH. Porém, os melhores resultados foram obtidos com as concentrações de 1 e 3 mM (126% e 97%, respectivamente) por 24 horas em relação a 41% de metabolismo celular do grupo controle positivo (p<0,05). Portanto, as células pré-tratadas com α-T por 24 horas, em concentrações baixas (1 e 3 mM), apresentaram os melhores resultados de viabilidade celular, ou seja, maior efeito protetor frente à agressão direta do PH

The in-office tooth bleaching using gels with high concentrations of hydrogen peroxide (HP) may cause irreversible damage to pulp tissue. Therefore, the administration of antioxidazing agents previously to the tooth bleaching procedures has been considered as a promising therapy, mainly due the capacity of these agents to prevent, or at last reduce, the negative side-effects caused by toxic products used in this esthetic treatment. Therefore, the aim of the present in vitro study was to evaluate the protective activity of vitamin E (alfa-Tocoferol / α-T) against the toxic effects of HP applied to cultured odontoblast-like cells (MDPC-23). Cells were seeded in wells of 96-well plates for 72 hs and then pre-treated with different concentrations of α-T (1, 3, 5, and 10 mM) for variable periods (1, 4, 8, and 24 hs). Following, the cells were exposed to a HP solution (0,018%) for 30 min. In positive and negative control groups, the cells were exposed only to HP solution (0,018%) or culture medium, respectively. The cell metabolism was assessed by MTT assay and the absorbance numeric data, expressed as percentage values, were subjected to the statistical analysis of Kruskal-Wallis complemented by Mann-Whitney test (α=0.05%). All α-T concentrations and pre-treatments evaluated in this study showed cell protection against the cytotoxic effects of HP. However, considering the MDPC23 cells in the negative control group as presenting 100% metabolism, it was observed that the most relevant data occurred when the cells were pre-treated with α-T at 1 and 3 mM (126% and 97% of cell metabolism, respectively) for 24hs compared to the positive control group (41%) (p<0.05). Based upon the methodology used in the present investigation, it can be concluded that low concentrations of α-T (1 and 3 mM) applied for 24 hs to the cultured odontoblast-like MDPC-23 cells provide the best protective effects against the cytotoxicity caused by hydrogen peroxide

Effectiveness of chlorhexidine on the disinfection of complete dentures colonised with fluconazole-resistant Candida albicans: in vitro study.

This in vitro study evaluated different concentrations of chlorhexidine (CHX) solution on the disinfection of dentures colonised with a reference (ATCC 90028) and azole-resistant (R1, R2 e R3) strains of Candida albicans. Sterile dentures were individually inoculated with one of the strains and incubated at 37 °C for 24 h. Then, each denture was immersed in sterile saline (control) or CHX (2%, 1% or 0.2%) for 10 min. Samples of serial dilutions were spread on Agar Sabouraud Dextrose and incubated at 37 °C for 48 h. The colonies were counted and the values of log(cfu ml(-1)) were analysed by Kruskal-Wallis test (P < 0.05). Dentures immersed in CHX were incubated for 7 days. For all strains, the cfu ml(-1) values of 0.2% CHX were significantly higher than those of 2% and 1% CHX. There was no difference between the cfu ml(-1) values of 2% and 1% CHX. For dentures immersed in CHX, ATCC 90028 strain showed lower cfu ml(-1) values than R2 and R3 strains. For control dentures, cfu ml(-1) values of ATCC 90028 strain were higher than those of R strains. Immersion in 2% CHX resulted in the highest number of dentures without fungal growth after 7 days. For denture disinfection, 2% CHX was the most effective concentration, and R strains were less susceptible to disinfection. Chlorhexidine is effective in disinfection of dentures contaminated with azole-resistant C. albicans.

Direct and transdentinal antibacterial activity of chlorhexidine.

PURPOSE: To evaluate the antibacterial effect of different chlorhexidine (CHX) concentrations against Streptococcus mutans using the agar-diffusion method with and without human dentin discs placed between the bacteria and the test substances. METHODS: For the direct application (agar-well technique), a base layer containing 15 mL of BHI agar and 300 microL of S. mutans inoculum (10(9) cfu/mL) was prepared in Petri dishes. Six wells per dish were made at equidistant points and immediately filled with CHX gels (0.12%, 0.2%, 1% and 2%), 35% phosphoric acid and pure natrosol (n = 6 wells/substance). Paper discs soaked in sterile distilled water served as control group (n = 6). For the indirect application (transdentinal diffusion), 0.2 mm- and 0.5 mm-thick human dentin discs (36 discs/thickness) had the hydraulic conductance determined, which allowed the homogeneous allocation of them to the experimental and control groups. The discs were placed at equidistant points on the Petri dishes containing BHI with the S. mutans inoculum (six discs per dish; one per substance) with the pulpal side in contact with the bacteria. In the discs treated with CHX gels, dentin surface was etched with H3PO4 and rinsed with distilled water before CHX gel application for 1 minute. After both direct and indirect application, the dishes were incubated for 24 hours and the bacterial growth inhibition zones formed around the wells and dentin discs were measured. Data were analyzed statistically by the non-parametric Kruskal-Wallis and Mann-Whitney tests at 5% significance level. RESULTS: In the direct test, all CHX concentrations presented a dose-dependent antibacterial activity against S. mutans. In the indirect test, there were statistically significant differences (P < 0.05) among all groups and the largest microbial growth inhibition zones were observed when 2% CHX was applied on 0.2 mm-thick discs (P < 0.05). It was concluded that all evaluated CHX gels exhibited both direct and transdentinal antibacterial activity against S. mutans. This effect of CHX was strongly influenced by the CHX concentration as well as the dentin barrier thickness.