Abstract Schinus terebinthifolia Raddi green fruits essential oil (EO) was evaluated regarding its phytochemical profile, antimicrobial and cytotoxic activities, and toxicity. Gas chromatography with mass spectrometry was applied to identify its constituents, thereafter the minimum inhibitory concentration, minimum bactericidal and fungicidal concentrations, and its antibiofilm activity were evaluated. The EO cytotoxicity was assessed in tumor and non-tumor human cells, and in vivo toxicity was evaluated in a Galleria mellonella model. The major constituents of S. terebinthifolia EO were alpha-phellandrene and beta-phellandrene. The EO had a weak activity against all strains of Candida albicans (MIC 1000μg/mL) and had no activity against non-albicans strains, bacteria, and C. albicans biofilm. Cytostatic activity against all tumor cell lines was shown. Additionally, cell viability remained at EO concentrations up to 62.5 μg/mL. At 16 mg/mL, 50% hemolysis was observed, and it had low toxicity in vivo. Overall, the S. terebinthifolia EO was characterized by low antimicrobial and antibiofilm activities, with no evidence of toxicity to human tumor and non-tumor cells.

Abstract: This study developed experimental gels containing titanium tetrafluoride (TiF4) combined with commercial 35% hydrogen peroxide (HP), and evaluated bleaching efficacy and pH of the gels, and mineral content and morphology of enamel submitted to these treatments. In phase-1, different stock gels mixed with TiF4 were combined with HP. In phase-2, the selected gels were tested on enamel/dentin specimens (n=8): HP; HP and Natrosol+TiF4 (HPnT); HP and Natrosol+Chemygel+TiF4 (HPncT); HP and Aristoflex+TiF4 (HPaT). Bleaching was performed in four sessions (3x15min-application/session). Color (CIEL*a*b*) and whiteness index (WID) were measured after each session, whereas whiteness index differences (ΔWID), color alteration (CIELab–ΔE, CIEDE2000-ΔE00), enamel morphology and pH, at end of bleaching therapy. The change in Knoop microhardness (ΔKHN) was compared before and after bleaching. Data were analyzed by two-way repeated measures ANOVA and Bonferroni (CIEL*, a*, b*), one-way ANOVA and Tukey (ΔWID, ΔE, ΔE00), and LSD (ΔKHN) tests (α=5%). SEM and pH measurements were submitted to descriptive analysis. No differences were observed in lightness (L*) or WID among the groups (p > 0.05), but HP exhibited lower b* values (p<0.05), higher ΔWID than HPnT, and the highest ΔE among the groups (p < 0.05). No differences in ΔE00 were observed between HP and HPncT (p > 0.05), and HPncT showed higher ΔKHN than HP (p < 0.05). HP presented pH values closer to neutral (6.9), whereas experimental agents showed acidic pH values (2.3–3.9). No morphological changes were observed in HP or HPncT groups. HPncT was able to bleach the enamel and maintain enamel microhardness and surface integrity, even at low pH.

BACKGROUND Essential oils (EO) extracted from Cinnamomum verum has been used as an antimicrobial agents for centuries. The effects of C. verum leaf oil against virulence of microorganisms is not well studied yet. OBJECTIVES This study evaluates the effect of C. verum leaf oil against three virulence factors of Candida albicans, C. tropicalis and C. dubliniensis and its in-vivo toxicity. METHODS Chemical composition of EO was determined using gas chromatography-mass spectrometry (GC-MS). Minimum inhibitory concentration (MIC) was determined using clinical and laboratory standards institute (CLSI) M27-A3 broth microdilution. Effect of EO on initial adhesion was quantified using XTT assay after allowing Candida cells to adhere to the polystyrene surface for 2 h. Biofilm formation of Candida in the presence of EO was quantified using XTT viability assay. Efficacy on reduction of germ tube formation was evaluated using standard protocol. Visualisation of biofilm formation and progression under the EO treatment were done using scanning electron microscope (SEM) and Time lapses microscope respectively. In-vivo toxicity of EO was determined using Galleria mellonella larvae. Chlorhexidine digluconate: positive control. RESULTS Eugenol was the main compound of EO. MIC was 1.0 mg/mL. 50% reduction in initial adhesion was achieved by C. albicans, C. tropicalis and C. dubliniensis with 1.0, > 2.0 and 0.34 mg/mL respectively. 0.5 and 1.0 mg/mL significantly inhibit the germ tube formation. MBIC50 for forming biofilms were ≤ 0.35 mg/mL. 1.0 mg/mL prevent biofilm progression of Candida. SEM images exhibited cell wall damages, cellular shrinkages and decreased hyphal formation. No lethal effect was noted with in-vivo experiment model at any concentration tested. CONCLUSION C. verum leaf oil acts against virulence factors of Candida and does not show any toxicity.

Abstract: We evaluated the antifungal and antibiofilm potential of the hydroalcoholic extract of bark from Anadenanthera colubrina (vell.) Brenan, known as Angico, against Candida spp. Antifungal activity was evaluated using the microdilution technique through the Minimum Inhibitory and Fungicide Concentrations (MIC and MFC). The antibiofilm potential was tested in mature biofilms formed by Candida species and analyzed through the counting of CFU/mL and scanning electron micrograph (SEM). In vivo toxicity and therapeutic action was evaluated in the Galleria mellonella model. The treatment with the extract, in low doses, was able to reduce the growth of planktonic cells of Candida species. MIC values range between 19.5 and 39 µg/mL and MFC values range between 79 and 625 µg/mL. In addition was able to reduce the number of CFU/mL in biofilms and to cause structural alteration and cellular destruction, observed via SEM. A. colubrina showed low toxicity in the in vivo assay, having not affected the viability of the larvae at doses below 100mg/kg and high potential in the treatment of C. albicans infection. Considering its high antifungal potential, its low toxicity and potential to treatment of infections in in vivo model, A. colubrina extract is a strong candidate for development of a new agent for the treatment of oral candidiasis.

Abstract: The aim of this study was to evaluate in vitro the antifungal, antibiofilm and antiproliferative activities of the extract from the leaves of Guapira graciliflora Mart. The phytochemical characterization of the extract was performed using electrospray ionization mass spectrometry (ESI-MS). The antimicrobial activity of the extract and its fractions was evaluated using the broth microdilution method against species of Candida. The inhibition of C. albicans biofilm was evaluated based on the number of colony-forming units (CFU) and metabolic activity (MTT). The antiproliferative activity of the extract and its fraction was evaluated in the presence of human tumor and non-tumor cells, and the cytotoxicity of the extract was determined on the RAW 264.7 macrophage line – both using the sulforhodamine B method. The phytochemical characterization indicated the presence of the flavonoids rutin and kaempferol. The extract and the methanol fraction exhibited moderate antifungal activity against C. albicans, C. krusei, and C. glabrata, and strong activity against C. dubliniensis. In the biofilms at 24 and 48 hours, the concentration of 12500 µg/mL of the extract was the most effective at reducing the number of CFU s/mL (44.4% and 42.9%, respectively) and the metabolic activity of C. albicans cells (34.6% and 52%, respectively). The extract and its fractions had no antiproliferative effect on the tumor lines tested, with mean activity (log GI50) equal to or greater than 1.71 µg/mL. Macrophage cell viability remained higher than 80% for concentrations of the extract of up to 62.5 µg/mL. G. graciliflora has flavonoids in its chemical composition and demonstrates potential antifungal and antibiofilm activity, with no evidence of a significant change in the viability of human tumor and non-tumor cell lines.

Abstract We investigated the anti-caries effects of an experimental propolis varnish in vivo, and further tested its toxicity against fibroblasts. Fifty-six SPF female Wistar rats were infected with Streptococcus mutans UA159 (SM) and allocated into four groups (n = 14/group): G1, propolis varnish (15%/PV); G2, chitosan varnish (CV/vehicle); G3, gold standard (GS/Duraphat®); and G4, untreated. The animals received a single varnish application on their molars and were submitted to a high cariogenic challenge (Diet-2000, 56% sucrose, and 5% sucrose-added water, ad libitum) for 4 weeks. Total cultivable microbiota and SM were counted, and smooth-surface and sulcal caries were scored. PV, CV and GS cytotoxic effects were tested against fibroblasts. The data were analyzed using ANOVA with the Tukey-Kramer test (p ≤ 0.05). Total microbiota and SM counts did not differ among the treatments (p = 0.78), or in relation to the untreated group (p = 0.52). PV reduced development of smooth-surface enamel caries compared with the untreated group (p = 0.0018), with no significant difference from GS (p = 0.92); however, the PV effects were no longer observed when the dentin was affected. Neither PV nor GS prevented enamel sulcal lesion onset, but GS significantly reduced the severity of dentinal sulcal lesions (p < 0.0001). No significant difference was observed in fibroblast viability between PV and GS (p < 0.0001). In conclusion, PV prevented smooth-surface enamel caries and showed low cell toxicity. Nevertheless, due to the high cariogenic challenge, its effects were not sustained throughout the experiment. Further studies are encouraged to establish a protocol to sustain the long-term anti-caries activity of PV in the oral cavity.

Abstract The effect of a 4% titanium tetrafluoride (TiF 4 ) varnish on enamel demineralization was evaluated. Twelve volunteers participated in this double-blind, randomized crossover study. Six enamel specimens were positioned in intraoral appliances throughout four treatment stages: 4% TiF 4 varnish (experimental varnish), 5% sodium fluoride (NaF) varnish (Duraphat ® ), placebo varnish, and negative control (deionized water). After 24 h, the varnishes were removed and plaques were allowed to accumulate. A 20% sucrose solution was dripped onto enamel blocks (10x/day). Enamel alterations were analyzed by surface microhardness (SMH), percentage of surface loss (%SML), cross-sectional microhardness (CSMH), scanning electron microscopy (SEM), and energy dispersive X-ray spectrometry (EDS). Student’s paired t-test was used for SMH analysis and repeated-measures analysis of variance (ANOVA) for %SML and CSMH (∆Z) analyses (p-value=0.05). The TiF 4 varnish group had lower %SML than the placebo and control groups (p=0.044 and p=0.003, respectively), thus showing its capacity to inhibit surface demineralization. TiF 4 and NaF varnishes demonstrated a protective effect against mineral loss on the enamel subsurface. Both were statistically different from the control group when CSMH was analyzed (p=0.000). A titanium dioxide film was observed on enamel surfaces of the TiF 4 group SEM images. EDS confirmed the presence of titanium in all TiF 4 samples. The 4% TiF 4 varnish is a promising compound capable of reacting with enamel to protect it against surface and subsurface demineralization.

Ensaios biológicos realizados com diferentes tipos de própolis brasileira têm mostrado que a própolis do tipo 6 (G6) tem forte atividade antimicrobiana e menor teor de flavonóides. Este trabalho teve como objetivo avaliar a correlação entre a composição fenólica e atividades biológicas apresentadas pela própolis G6, do Estado da Bahia, e a própolis verde, do tipo 12 (G12). Os teores de flavonóides e fenólicos totais na própolis G6 foram diferentes dos teores da própolis G12. Apesar da G12 apresentar maior atividade antioxidante, a própolis G6 apresenta maior atividade antimicrobiana e citotóxica. Os resultados mostraram que os compostos fenólicos não são os únicos compostos responsáveis pela atividade biológica da própolis. Estudos mais específicos da composição química, em adição à avaliação das atividades biológicas, são requeridos para determinar a qualidade da própolis.

Biological assays that have been performed on different types of Brazilian propolis have shown that type 6 propolis (G6) has a strong antimicrobial activity and a low flavonoid content. This study aimed to evaluate the correlation between the phenolic composition and the biological activities displayed by propolis G6 from the state of Bahia and green propolis, also known as type 12 (G12). The values of the flavonoids and the total phenolics in propolis G6 were different than those in propolis G12. Although the G12 variety presented greater antioxidant activity, propolis G6 proved to have greater antimicrobial and cytotoxic activities. The results showed that the phenolic compounds may not be the only compounds responsible for the biological activity. More detailed studies of the chemical composition and an assessment of the biological activity are required to establish the quality of propolis.

A cavidade oral apresenta vários genótipos de Streptococcus mutans, que podem possuir diferentes capacidades de virulência. Entretanto, pouco se sabe sobre a diversidade e virulência de genótipos de S. mutans isolados in vivo sob uma condição controlada de alto desafio cariogênico. Este estudo avaliou a diversidade genotípica de S. mutans identificados no biofilme dental formado in vivo na presença de sacarose, assim como a acidogenicidade e aciduricidade desses genótipos. Para possibilitar formação de biofilme, voluntários bochecharam com água destilada ou solução de sacarose 8x/dia durante 3 dias. S. mutans isolados da saliva e do biofilme dental foram genotipados por PCR com primers-arbitrários. Genótipos isolados do biofilme foram avaliados em relação à habilidade de reduzir o pH da suspensão devido à glicólise, em relação à susceptibilidade a ácidos e também atividade F-ATPase. A maioria dos voluntários apresentou apenas 1 genótipo na saliva, que foram detectados em quase todas as amostras de biofilme em altas proporções. Genótipos isolados somente na presença de sacarose apresentaram maior acidogenicidade do que aqueles genótipos isolados apenas na presença de água. Genótipos de biofilmes formados na presença de sacarose foram mais acidúricos após 30 e 60 min de incubação em pH 2,8 e 5,0, respectivamente. Os resultados do presente estudo sugerem que biofilmes formados sob condição de alto desafio cariogênico podem apresentar genótipos de S. mutans mais acidúricos e mais acidogênicos.

The oral cavity harbors several Streptococcus mutans genotypes, which could present distinct virulence properties. However, little is known about the diversity and virulence traits of S. mutans genotypes isolated in vivo under controlled conditions of high cariogenic challenge. This study evaluated the genotypic diversity of S. mutans isolated from dental biofilms formed in vivo under sucrose exposure, as well as their acidogenicity and aciduricity. To form biofilms, subjects rinsed their mouths with distilled water or sucrose solution 8 times/day for 3 days. S. mutans collected from saliva and biofilms were genotyped by arbitrarily-primed PCR. Genotypes identified in the biofilms were evaluated regarding their ability to lower the suspension pH through glycolysis and their acid susceptibility and F-ATPase activity. Most subjects harbored only one genotype in saliva, which was detected in almost all biofilm samples at high proportions. Genotypes isolated only in the presence of sucrose had higher acidogenicity than those isolated only in the presence of water. Genotypes from biofilms formed with sucrose were more aciduric after 30 and 60 min of incubation at pH 2.8 and 5.0, respectively. The present results suggest that biofilms formed under high cariogenic conditions may harbor more aciduric and acidogenic S. mutans genotypes.

Propolis is a resinous hive product collected by honeybees from various plant sources. It has a complex chemical composition, constituted by various phenolic compounds. Extracts of increasing polarity (n-hexane, chloroform, and ethanol) were obtained from a sample of red propolis from the state of Alagoas. Assays were carried out for determination of contents of phenolics, along with antibacterial and antioxidant activities. The EEP, fractions and sub-fractions showed strong biological activities and were related with phenolic the content compounds contents. The sub-fractions were more bioactive than the EEP and fractions, demonstrating that the antioxidant and antibacterial activities are not a result of synergistic effect between the various chemical compounds in propolis.

The composition and biological activities of propolis, a resinous hive product collected by honeybees from various plant sources, depends on various factors such as season and vegetation of the area. The aim of this study was to evaluate the influence of the seasonal effect on the ethanolic extracts of Brazilian propolis (EEP) type 6 and type 12, collected during 6 months in terms of antibacterial activity and phenolic composition. The antimicrobial properties were evaluated by MIC and MBC on S. mutans Ingbritt 1600 and the profile of chemical composition by UV-visible spectrophotometry, HPLC-RF and GC-MS. The results demonstrated that the season in which propolis is collected influences its chemical composition, resulting in modifications in its antibacterial activity.

O desenvolvimento de tecnologias aplicadas à saúde traz dilemas morais inesperados aos profissionais de pesquisa, em relação às práticas que envolvem seres humanos. Devido à relevância deste assunto, o objetivo deste estudo foi avaliar o ensino de bioética na educação de estudantes de programas de pós-graduação em Odontologia do Brasil. Oitenta e sete programas de pós-graduação em Odontologia, avaliados pela CAPES (2001-2003), foram estudados. Os dados foram extraídos, de maneira independente, por 2 avaliadores, por meio de consulta ao sítio da CAPES e aos sítios eletrônicos dos programas, diretamente ou via links dos programas disponíveis no sítio da CAPES. Dos 87 programas, 48 apresentaram disciplina de Ética/Bioética. Trinta e oito porcento dos programas com conceitos 5, 6 e 7 da CAPES mantêm disciplinas de Bioética, enquanto 62% dos programas com conceitos 3 e 4 apresentaram conteúdos de Bioética. Desta forma, os resultados deste estudo representam um alerta para os educadores envolvidos em ensino da pesquisa odontológica. Estes resultados também mostraram que a instrução em bioética ainda é incipiente nos programas de pós-graduação em Odontologia no Brasil, ainda que a resolução 196/96 do Conselho Nacional de Saúde tenha sido publicada há dez anos. Por este motivo, seria necessário assegurar uma pedagogia ética na formação do jovem pesquisador.

In the field of human research, researchers are faced with unexpected moral dilemmas, as a result of the development of technologies applied to health. Due to the great importance of this issue, our objective was to evaluate bioethics instruction in the education of researchers in Brazilian graduate programs in dentistry. Eighty-seven graduate programs in dentistry, recognized by CAPES (Coordination for the Improvement of Higher Education Personnel) were evaluated in this study. Data were extracted independently by two researchers from the CAPES website, and from the websites of the graduate programs, directly or via links to the programs available at the CAPES website. Forty-eight out of 87 programs had an ethics/bioethics course as part of their curricula. Of the graduation programs graded 5, 6 or 7 by CAPES, 38% included bioethics courses, while 62% of the programs graded 3 or 4 by CAPES had bioethics courses as part of their curricula. These findings are an alert to those involved in dental research education, as they showed that, although resolution 196/96 by the National Council of Health regulating human research in Brazil was published ten years ago, bioethics instruction in Brazilian graduate programs in dentistry is still at an incipient stage. This situation indicates a need for ethics pedagogy in the education of young researchers.

O objetivo deste trabalho foi investigar in vitro a atividade antimicrobiana do hidróxido de cálcio em combinação com vários veículos contra alguns microrganismos mais frequentemente isolados dos canais radiculares. A atividade antimicrobiana foi determinada pelo método de difusão no ágar. Cilindros de aço inoxidável foram colocados sobre as placas de agar já inoculadas, dentro dos quais os medicamentos testados e seus controles foram introduzidos. As zonas de inibição do crescimento microbiano foram medidas e anotadas após o período de incubação de cada placa e os resultados foram analisados estatisticamente. A atividade antimicrobiana in vitro dos medicamentos testados, em ordem decrescente, foi: Ca(OH)2 + PMCF + glicerina, Ca(OH)2 + PMCF, Ca(OH)2 + glicerina, Ca(OH)2 + anestésico, Ca(OH)2 + salina, Ca(OH)2 + H2O, Ca(OH)2 + polietilenoglicol. Pastas com veículos oleosos produziram as maiores zonas de inibição, quando comparadas com as com veículos aquosos ou oleosos. Concluimos que a habilidade de difusão e a atividade antimicrobiana do hidróxido de cálcio estão relacionadas com os tipos de veículos utilizados.

The aim of this study was to investigate in vitro the antimicrobial activity of calcium hydroxide in combination with several vehicles against some microorganisms commonly isolated from root canals. Antimicrobial activity was determined by the agar diffusion method. Stainless-steel cylinders were placed on each inoculated agar medium. The test medicaments and their controls were placed inside the cylinders. The zones of growth inhibition were measured and recorded after incubation for each plate and the results were analyzed statistically (ANOVA). The in vitro antimicrobial effects of the medications were ranked from strongest to weakest as follows: Ca(OH)2 + CMCP + glycerine, Ca(OH)2 + CMCP, Ca(OH)2 + glycerine, Ca(OH)2 + anesthetic, Ca(OH)2 + saline, Ca(OH)2 + H2O, Ca(OH)2 + polyethyleneglycol. The pastes with oily vehicles showed significantly larger mean zones of inhibition compared to those with aqueous or viscous vehicles. It was concluded that diffusion and antimicrobial activity of calcium hydroxide were affected by the type of vehicle used.

Cariogenicity and cariostatic properties of cow, human and infant formula milk-review. The purpose of this study is to introduce some information about local and systemic effects of different kinds of rnilk in oral health, through the explicitness of its cariogenicity and cariostatic properties. Different kinds of milk and milk products are consumed most commonly by the worldwide population, raising the interest of researchers in its influence in the oral health for some decades. Several studies have been conducted to associate the consumption of bovine-, human- and infant formula- milk with caries development and other dental defect, but controversial results have not been able to define the cariogenic and cariostatic potential of different kinds of milk. Bovine milk has some cariostatic components as casein, lipids and antibacterial enzyme, though it has 4% lactose, supposedly cariogenic sugar. Human milk has been related to a sort of caries which is like bottle caries, even though some studies have demonstrated its non cariogenicity. The infant formula milk, which is elaborated for specific period in the childhood, has received low control on its potential for developing caries. It could be of great value to elucidate the controversy surrounding the cariogenicity and cariostatic properties of different kinds of milk, concerning caries prevention during infant and adult life.

A proposta desse trabalho é apresentar uma revisão da literatura sobre o efeito tópico e sistêmico dos diferentes tipos de leite na saúde dental, através da explicitação de sua cariogenicidade e propriedades cariostáticas. O leite em suas diferentes fórmulas bem como seus derivados são produtos consumidos em larga escala pela população mundial, merecendo há algumas décadas a atenção de pesquisadores sobre suas influências na cavidade oral. Diversos estudos têm sido conduzidos com objetivo de relacionar o consumo de leite bovino, materno e em fórmula infantil com o desenvolvimento de cáries e outros defeitos dentais. Mas, muitos resultados controversos ainda não conseguiram definir com exatidão os potenciais cariogênicos e cariostáticos dos diferentes tipos de leite. O leite bovino possui alguns componentes como a caseína, lipídios e enzimas antibacterianas que apresentam potencial cariostático, embora contenha também 4% de lactose, açúcar sugestivamente cariogênico. O leite humano, por sua vez, tem sido relatado como responsável pelo aparecimento de um tipo de cárie similar a cárie de mamadeira, apesar de alguns estudos defenderem sua não cariogenicidade. Já o leite em fórmula infantil, que é elaborado para determinadas épocas da infância, tem recebido baixo controle sobre sua capacidade de desenvolver cáries. Desvendar a controvérsia entre a cariogenicidade e propriedades cariostáticas dos diferentes tipos de leite, ou simplesmente instruirse sobre esse assunto, poderia ter grande valor na prevenção de cáries durante a infância e na vida adulta.