Mutanase from Trichoderma harzianum inductively Produced by Mutan: Short-Term Treatment to Degrade Mature Streptococcus mutans Biofilm.

This study aimed to evaluate the disruptive effect of fungal mutanase against cariogenic biofilm after short-term treatment. For that, mature Streptococcus mutans biofilms (n = 9) were exposed to active or inactivated enzymes produced by Trichoderma harzianum for 1 min, two times per day. Biofilms were analyzed by amount of matrix water-insoluble polysaccharides, bacterial viability, acidogenicity, and morphology by scanning electron microscopy (SEM). The group treated with active enzymes (AE) had a significantly lower amount of insoluble polysaccharides (893.30 ± 293.69) when compared to the negative control group (NaCl, 2192.59 ± 361.96), yet no significant difference was found when comparing to the positive control group (CHX, 436.82 ± 151.07). Also, there was no significant effect on bacteria metabolism and viability (P-value < 0.05). Data generated by the quantitative analysis were confirmed through scanning electron microscopy images. Thus, fungal mutanase degraded the biofilm after a short-term treatment without interfering with bacterial viability and metabolism. Such findings offer insight to the development of routine oral care products containing this input.

Effects of the green tea catechin epigallocatechin-3-gallate on Streptococcus mutans planktonic cultures and biofilms: systematic literature review of in vitro studies.

This study aimed at performing a systematic review of the literature on the effects of epigallocatechin-3-gallate (EGCG) on Streptococcus mutans planktonic cultures and biofilms. The selected references demonstrated that EGCG suppresses S. mutans acid production by inhibiting the activity of enzymes such as lactate dehydrogenase and FIF0-ATPase. Regarding virulence factors, one study reported a reduction in soluble and insoluble polysaccharide synthesis, another demonstrated that EGCG inhibited GTase activity, and another showed effects of EGCG on the expression of gtf B, C, and D. The effects of EGCG on S. mutans biofilms were reported only by 2 of the selected studies. Moreover, high variability in effective concentrations and microbial assessment methods were observed. The literature suggests that EGCG has effects against S. mutans planktonic cells viability and virulence factors. However, the literature lacks studies with appropriate biofilm models to evaluate the precise effectiveness of EGCG against S. mutans biofilms.

High Amount of Organic Matter during Caries Formation Reduces Remineralization and Resin Infiltration of Enamel Caries.

The amount of organic material in the cariogenic environment correlates with the amount of organic material incorporated in carious enamel. The incorporated organic material may be expected to reduce the pore volumes available for remineralization and resin infiltration, but these expected outcomes have not yet been quantified. We tested the effect of the amount of organic content in the cariogenic agent on remineralization and the resin-occluded pore volume in artificial subsurface enamel caries. An acid gel (organic-rich; G1) and an aqueous solution (organic-poor; G2) were used to induce subsurface lesions in human enamel. Undemineralized histological sections were prepared, microradiographed, and then submitted to resin infiltration in vitro. The enamel component volumes (mineral, organic, remineralizable [total water volume], loosely and firmly bound water volumes, and resin-occluded volume) were measured (by microradiography and polarizing microscopy) at histological sites (n = 38, G1; n = 34, G2). The main outcomes were the differences between the experimental and the predicted volumes (Δremineralizable and Δresin-occluded volumes). Resin infiltration was confirmed by confocal scanning laser microscopy. Compared to G2, G1 presented more incorporated organic volume and lower Δremineralizable volume (p = 0.003; Hedges g = 0.66; power = 0.87), a lower increase in loosely bound water volume (p = 0.0013; Hedges g = 0.74; power = 0.93), a lower remineralization volume in the surface layer (p = 0.017; Hedges g = 0.68; power = 0.8), and a lower Δresin-occluded volume (p = 0.0015; Hedges g = 0.73; power = 0.92). In conclusion, the higher amount of organic matter in the cariogenic gel negatively affected remineralization and the resin-occluded volume in subsurface lesions.

Elemental ion release and cytotoxicity of antimicrobial acrylic resins incorporated with nanomaterial.

OBJECTIVES: This study evaluated the release of ions and the cytotoxicity of acrylic resins incorporated with silver vanadate decorated with silver nanoparticles (AgVO3 ). BACKGROUND: The inhibition of the accumulation of microorganisms on the resins is critical in preventing diseases. However, the hypothesis is that the release of ions from the incorporation of AgVO3 may be important in biocompatibility. MATERIALS AND METHODS: Specimens of autopolymerising (AP) and heat-polymerising resin (HP) with AgVO3 were prepared and immersed in culture medium. The release of silver ions (Ag) and vanadium (V) was evaluated by mass spectrometry with inductively coupled plasma (ICP-MS) (n=9) and the cell viability of fibroblasts L929 by MTT (3-[4,5-dimethylthiazol- 2yl]-2,5-diphenyltetrazolium bromide) (n=12). The results were evaluated with analysis of variance (ANOVA), Tukey and Pearson correlation test (α=.05). RESULTS: The groups containing AgVO3 presented a difference in relation to the control (0%) regarding the release of Ag and V (P<.0001). All groups showed a reduction in L929 viability when compared with the cellular control (100%) (P<.0001). In comparison with the control resins for HP, a reduction in the metabolism of cells occurred starting at 2.5% and for AP at 5% (P<.0001). A positive correlation was found between the concentration of AgVO3 and the ion release, and a negative between the ion release and the cell viability. CONCLUSIONS: Significant numbers of Ag and V ions were released from resins with higher concentrations of AgVO3 , presenting cytotoxicity for cells, suggesting that the use of low concentrations is indicated to avoid risks to patients.

Effect of Liquid Crystalline Systems Containing Antimicrobial Compounds on Infectious Skin Bacteria.

This study aimed (i) to prepare liquid crystalline systems (LCS) of glyceryl monooleate (GMO) and water containing antibacterial compounds and (ii) to evaluate their potential as drug delivery systems for topical treatment of bacterial infections. Therefore, LCS containing CPC (cetylpyridinium chloride) (LCS/CPC) and PHMB (poly(hexamethylene biguanide) hydrochloride) (LCS/PHMB) were prepared and the liquid crystalline phases were identified by polarizing light microscopy 24 h and 7 days after preparation. The in vitro drug release profile and in vitro antibacterial activity of the systems were assessed using the double layer agar diffusion method against Staphylococcus aureus, methicillin-resistant S. aureus, Staphylococcus epidermidis, Escherichia coli, and Enterococcus faecalis. The interaction between GMO and the drugs was evaluated by a drug absorption study. Stable liquid crystalline systems containing CPC and PHMB were obtained. LCS/PHMB decreased the PHMB release rate and exerted strong antibacterial activity against all the investigated bacteria. In contrast, CPC interacted with GMO so strongly that it became attached to the system; the amount released was not sufficient to exert antibacterial activity. Therefore, the studied liquid crystalline systems were suitable to deliver PHMB, but not CPC. Accordingly, it was demonstrated that GMO interacts with each drug differently, which may interfere in the final efficiency of GMO/water LCS.

Caries removal in deciduous teeth using an Er:YAG laser: a randomized split-mouth clinical trial.

OBJECTIVES: The aim of the present clinical randomized split-mouth study was to evaluate the effectiveness and efficiency of an Er:YAG laser for caries removal in primary molars, microbiological dentin analysis, and clinical restorations after 1 year in 29 children. MATERIALS AND METHODS: The children's teeth were randomized into two groups: (I) an Er:YAG laser group and (II) a bur preparation group. The efficiency of the treatments (the time necessary for the removal of carious tissue) was evaluated based on the time spent on caries removal in the deciduous molars. The effectiveness (caries removal capacity) of the caries removal was determined by means of a blind test in which the examiner performed a tactile and visual examination of the dentin. Microbiological analysis was performed by counting the Streptococcus mutans and Lactobacillus sp in the remaining dentin. Clinical analysis of restorations was performed using the USPHS method in combination with photographs of restored teeth, 7 days after the restorative procedure and again after 1 year. All cavities were restored with the Adper Single Bond 2/Filtek Z350 system. The obtained data were analyzed with a significance level of 5 %. RESULTS: The Er:YAG laser was less effective and had the same efficacy as bur preparation during caries removal at the pulpal wall of deciduous molars. In the surrounding walls, bur preparation was the more effective method. Regardless of the method employed, the affected dentin in the pulpal wall had similar amounts of S. mutans and Lactobacillus sp. The restorations were clinically accepted by the USPHS method over a 1-year period. CONCLUSION: It can be concluded that caries removal with an Er:YAG laser has no influence on the clinical behavior of restorations. CLINICAL RELEVANCE: Irradiation with an Er:YAG laser is appropriate for caries removal in primary teeth.

Structure of rhamnoglucan, an unexpected alkali-stable polysaccharide extracted from Streptococcus mutans cell wall.

This study identified a rhamnose-containing cell wall polysaccharide (RhaCWP) in an alkaline extract prepared to analyze intracellular polysaccharides (IPS) from Streptococcus mutans biofilm. IPS was an 1,4-α-D-glucan with branchpoints introduced by 1,6-α-glucan while RhaCWP presented 1,2-α-L-and 1,3-α-L rhamnose backbone and side chains connected by 1,2-α-D-glucans, as identified by nuclear magnetic resonance (NMR) spectroscopy and methylation analyses. The MW of IPS and RhaCWP was 11,298 Da, as determined by diffusion-ordered NMR spectroscopy. Therefore, this study analyzed the chemical structure of RhaCWP and IPS from biofilm in a single fraction prepared via a convenient hot-alkali extraction method. This method could be a feasible approach to obtain such molecules and improve the comprehension of the structure-function relationships in polymers from S. mutans in future studies.

Epigallocatechin gallate reduces the virulence of cariogenic Streptococcus mutans biofilm by affecting the synthesis of biofilm matrix components.

INTRODUCTION: There have been reports on the effects of epigallocatechin gallate (EGCG) against Streptococcus mutans viability and acidogenesis. However, the effects of EGCG on the virulence of S. mutans biofilm development have yet to be fully investigated using validated cariogenic biofilm models. OBJECTIVE: Thus, this study aimed to evaluate the effects of EGCG on S. mutans biofilm virulence using a validated cariogenic model and clinically relevant treatment regimens, twice a day for 1.5 min. METHODS: Effects of EGCG on bacterial viability, polyssacharide synthesis and biofilm acidogenesis were evaluated. The morphology and 3D structure of the biofilms were evaluated by scanning electron (SEM) and confocal laser scanning microscopy, respectively. RESULTS: No significant change in S. mutans viability or culture medium pH were observed when comparing EGCG-treated and NaCl-treated biofilms. EGCG significantly reduced the accumulation of soluble and insoluble polysaccharides, resulting in the formation of a biofilm with interspaced exopolysaccharide-microcolony complexes unevenly distributed on enamel. The SEM images of the biofilm treated with EGCG depict multilayers of cells arranged in short chains of microorganisms adhered to an unstructured matrix, which is not continuous and does not enmesh or protect the microorganisms entirely. Importantly, confocal images demonstrated that treatment with EGCG affected the 3D structure and organization of S. mutans biofilm, which presented a biofilm matrix more confined to the location of the microcolonies. CONCLUSION: In conclusion, EGCG lowered the virulence of S. mutans matrix-rich biofilm by reducing the synthesis of biofilm matrix components, altering the biofilm matrix structure, organization, and distribution.

Chitosan nanoparticles loaded with epigallocatechin-3-gallate: synthesis, characterisation, and effects against Streptococcus mutans biofilmEpigallocatechin-loaded chitosan nanoparticles: effects against Streptococcus mutans biofilm.

This study evaluated the effects of chitosan nanoparticles loaded with epigallocatechin-3-gallate (EGCG) against Streptococcus mutans biofilm. EGCG-loaded chitosan (Nchi + EGCG) nanoparticles and Chitosan (Nchi) nanoparticles were prepared by ion gelation process and characterised regarding particle size, polydispersion index, zeta potential, and accelerated stability. S mutans biofilms were treated twice daily with NaCl 0.9% (negative control), Nchi, Nchi + EGCG, and chlorhexidine (CHX) 0.12% (positive control). After 67 h, the biofilms were evaluated for acidogenesis, bacterial viability and dry weight. Biofilm morphology and structure were analysed by scanning electron microscopy. The nanoformulations presented medium to short-term stability, size of 500 nm, and polydispersion index around 0.400. Treatments affected cell morphology and biofilm structure. However, no effects on microbial viability, biofilm dry weight, and acidogenesis were observed. Thus, the nanoformulations disassembled the biofilm matrix without affecting microbial viability, which makes them promising candidates for the development of dental caries preventive and therapeutic agents.

Investigation of the antimicrobial effect of anodic iontophoresis on Gram-positive and Gram-negative bacteria for skin infections treatment.

Iontophoresis, a non-invasive application of a constant low-intensity electric current, is a promising strategy to accelerate wound healing. Although its mechanisms are not yet fully elucidated, part of its action seems related to inhibiting bacteria growth. This work aimed to investigate the antimicrobial effect of iontophoresis using Staphylococcus epidermidis and Escherichia coli strains, Gram-positive and Gram-negative bacteria, respectively. Anodic iontophoresis was applied to each bacterial suspension using Ag/AgCl electrodes, and bacteria viability was evaluated after 24 h incubation by counting colony-forming units. A Quality-by-Design approach was performed to assess the influence of the iontophoresis' intensity and application time on bacterial viability. Cell morphology was evaluated by scanning electron microscopy. Iontophoresis showed antimicrobial effects on the Gram-positive bacteria only at 5 mA and 60 min application. However, a linear relationship was observed between intensity and application time for the Gram-negative one, causing drastic morphological changes and up to 98 % death. The cell wall of Gram-negative bacteria seems more susceptible to disorganization triggered by iontophoresis-induced ion transport than Gram-positive ones. Therefore, anodic iontophoresis can be a powerful ally in controlling Gram-negative bacteria proliferation in wounds.

Influence of glucose supplementation on biofilm formation of Candida albicans and Candida glabrata isolated from diabetic and non-diabetic individuals.

OBJECTIVE: This study evaluated the effect of different glucose concentration on biofilm formation of Candida albicans and Candida glabrata strains isolated from diabetic and non-diabetic individuals. METHODS: The study was divided into two stages: (I) selection and identification of 48 C. albicans and C. glabrata strains by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR/RFLP); (II) evaluation of biofilm formation by means of viability rates (colony-forming units), biofilm dry matter (mg) and biofilm-covered areas (µm2). Statistical comparisons were performed through nonparametric analysis of longitudinal data in factorial experiments with pairwise comparisons using Friedman Conover's test (α = 0.05). RESULTS: All the Candida spp. had their identifications confirmed by PCR/RFLP. C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed higher viability rates than strains from non-diabetic individuals. No difference was observed on viability of C. glabrata biofilm. Regarding biofilm dry matter, C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed lower amount in weight than strains from non-diabetic individuals. In C. glabrata strains, this result was only observed in biofilms cultivated with no glucose supplementation. With regard to biofilm-covered areas, only glucose supplementation and non-diabetic condition showed a positive effect on C. albicans biofilm development, and no condition affected C. glabrata biofilm formation. CONCLUSION: The strain type (C. albicans and C. glabrata) isolated from diabetic and non-diabetic individuals influenced on biofilm formation, but glucose supplementation did not.

Besifloxacin liposomes with positively charged additives for an improved topical ocular delivery.

Topical ophthalmic antibiotics show low efficacy due to the well-known physiological defense mechanisms of the eye, which prevents the penetration of exogenous substances. Here, we aimed to incorporate besifloxacin into liposomes containing amines as positively charged additives and to evaluate the influence of this charge on drug delivery in two situations: (i) iontophoretic and (ii) passive treatments. Hypothesis are (i) charge might enhance the electromigration component upon current application improving penetration efficiency for a burst drug delivery, and (ii) positive charge might prolong formulation residence time, hence drug penetration. Liposomes elaborated with phosphatidylcholine (LP PC) or phosphatidylcholine and spermine (LP PC: SPM) were stable under storage at 6 ºC for 30 days, showed mucoadhesive characteristics, and were non-irritant, according to HET-CAM tests. Electron paramagnetic resonance spectroscopy measurements showed that neither the drug nor spermine incorporations produced evident alterations in the fluidity of the liposome's membranes, which retained their structural stability even under iontophoretic conditions. Mean diameter and zeta potential were 177.2 ± 2.7 nm and - 5.7 ± 0.3 mV, respectively, for LP PC; and 175.4 ± 1.9 nm and + 19.5 ± 1.0 mV, respectively, for LP PC:SPM. The minimal inhibitory concentration (MIC) and the minimal bactericide concentration (MBC) of the liposomes for P. aeruginosa showed values lower than the commercial formulation (Besivance). Nevertheless, both formulations presented a similar increase in permeability upon the electric current application. Hence, liposome charge incorporation did not prove to be additionally advantageous for iontophoretic therapy. Passive drug penetration was evaluated through a novel in vitro ocular model that simulates the lacrimal flow and challenges the formulation resistance in the passive delivery situation. As expected, LP PC: SPM showed higher permeation than the control (Besivance). In conclusion, besifloxacin incorporation into positively charged liposomes improved passive topical delivery and can be a good strategy to improve topical ophthalmic treatments.

The Impact of CO2 Laser Treatment and Acidulated Phosphate Fluoride on Enamel Demineralization and Biofilm Formation.

Introduction: This study evaluated the impact of CO2 laser treatment and acidulated phosphate fluoride (APF) on enamel demineralization and biofilm formation, using in vitro and in situ designs. Methods: Demineralized enamel slabs were distributed among 8 groups: placebo, placebo + continuous CO2 laser, placebo + repeated CO2 laser, placebo + ultrapulsed CO2 laser, 1.23% APF, APF + continuous CO2 laser, APF + repeated CO2 laser and APF + ultrapulsed CO2 laser. In the in vitro study, 15 enamel slabs from each group were subjected to a pH-cycling regimen for 14 days. In the cross over in situ design, 11 volunteers wore palatal appliances with demineralized enamel slabs for 2 periods of 14 days each. Drops of sucrose solution were dripped onto enamel slabs 8×/day. Biofilms formed on slabs were collected and the colony-forming units (CFU) of Streptococcus mutans and Lactobacillus were determined. Results: For both in vitro and in situ studies, there was no significant difference between treatments (P>0.05). However, all treatments increased microhardness of demineralized enamel (P<0.05). After a further in situ cariogenic challenge, with the exception of the placebo, all treatments maintained microhardness values (P<0.05). Microbiological analysis showed no difference in Streptococcus mutans (P>0.05) or Lactobacillus (P>0.05) counts between groups. Conclusion: The results suggest that APF gel combined with the CO2 laser, regardless of the pulse emission mode used, was effective in controlling enamel demineralization, but none of the tested treatments was able to prevent bacterial colonization.

Mango peel as a potential enzyme inducer in Trichoderma harzianum: a strategy for cariogenic biofilm degradation and reuse of industrial waste

Water-insoluble exopolysaccharides (I-EPS) are a virulence factor for dental biofilms. It has already been demonstrated that mango pulp induces the secretion of glucan-hydrolytic enzymes in the fungus Trichoderma harzianum, and that they have an effect on I-EPS from young biofilms. Aim: Evaluate the effect of mango peel as an enzyme inducer in T. harzianum, and the effect of enzymes secreted on mature biofilms. Methods: Fractions of the peel (PL) and ethanol-precipitated pulp (PP) of Tommy Atkins mangoes were sterilized and added to a culture medium containing T. harzianum for induction of hydrolytic enzymes. After 192 h, the culture medium was centrifuged and the supernatant (enzyme extract) was used as treatment on S. mutans biofilms (n=9): a) NaCl 0.9 %; b) 0.12 % chlorhexidine digluconate; and c) extract of enzymes induced by PL or PP. Acidogenicity, bacterial viability, quantification of insoluble polysaccharides, and three-dimensional analysis of the biofilm by scanning electron microscopy (SEM) was performed. Data were analyzed by ANOVA followed by the Tukey test (α=5 %). Results: The hydrolytic enzymes did not alter the metabolism or bacterial viability of the biofilm (p<0.05). Although the images obtained by SEM suggest some degree of matrix degradation, the quantification of I-EPS for the PL and PP groups did not differ from the control group (p>0.05), suggesting a slight effect on the disorganization of the mature S. mutans biofilm. Conclusion: The results suggest that mango peel fraction can induce secretion of mutanase by T. harzianum, however in an insufficient amount to generate significant degradation on cariogenic biofilm.

Effect of methylene blue-induced photodynamic therapy on a Streptococcus mutans biofilm model.

BACKGROUND: Several studies have reported the use of antimicrobial photodynamic therapy (aPDT) to control biofilm but its efficacy depends on several factors, such as biofilm model used. This study aims to examine whether exposure to diode laser combined with methylene blue affects the bacterial viability and polysaccharide content in a Streptococcus mutans cariogenic biofilm model, which simulated 'feast-famine' episodes of exposure to sucrose that occur in the oral cavity. MATERIALS AND METHODS: S. mutans biofilms were formed on acrylic resin discs and exposed to a 10% sucrose solution for 1min, eight times/day. After growing for 48h, the biofilms were submitted to the following treatments, twice daily (n=4): (i) 0.9% NaCl (NaCl) as the negative control; (ii) 0.12% chlorhexidine digluconate (CHX) as the positive antibacterial control; (iii) diode laser combined with methylene blue, using an energy density of 320J/cm2 (aPDT). After 120h of growth, the biofilm formed on each disc was collected to determine the viable bacterial counts and concentration of insoluble exopolysaccharides (IEPS) and intracellular polysaccharides (IPS). RESULTS: Bacterial counts in the biofilms formed differed among the treatments. Compared with NaCl, aPDT significantly destabilized biofilm (p<0.0001). aPDT and CHX equally lowered the concentration of IEPS and IPS in biofilms. CONCLUSION: Under the experimental conditions assessed, our findings indicate that a twice-daily treatment with diode laser combined with methylene blue effectively decreased bacterial viability and the intra- and extracellular polysaccharide concentration in biofilms of S. mutans, a cariogenic bacterium.

Quantification of Streptococcus mutans in Different Types of Ligature Wires and Elastomeric Chains.

This study aimed to test the hypothesis that Streptococcus mutans contamination levels differ according to the type of the orthodontic ligature. Thirteen patients were selected. Each quadrant was randomly subjected to one of the following ligature-use protocols: I) elastomeric chain, II) steel ligature crossed over the archwire, III) steel ligature crossed under the archwire, and IV) steel ligature in a figure-eight pattern under the archwire. After seven days, the devices were removed and the Streptococcus mutans colony-forming unit count per mg of biofilm weight was determined. Twelve specimens (n=3) were also processed for scanning electron microscopy analysis. ANOVA and Tukey-Kramer test were used for comparisons to assess S. mutans differences between groups at a 5% significance level. There was no statistical difference in detectable levels of S. mutans among the groups (p=0.294). Scanning electron microscopy results showed abundant biofilms and microbial contamination in all groups. In conclusion, S. mutans contamination levels are similar in the different orthodontic ligatures.

The short-term in situ model to evaluate the anticariogenic potential of ionomeric materials.

OBJECTIVES: Aiming to contribute to the study of mechanisms involved in the anticariogenic properties of dental materials, this study assessed the suitability of a short-term in situ model to evaluate the anticariogenic potential of ionomeric materials. METHODS: The study used a 3-phase crossover, double blind design, and in each phase eight volunteers wore palatal appliances containing four enamel blocks restored with one of the following materials: composite resin (CR-Z250) (negative control), a conventional glass ionomer cement (GIC-Ketac-Fil) or a resin-modified GIC (RM-GIC-Vitremer). The restored blocks were covered with a "test plaque" of S. mutans, placed in palatal appliances and a cariogenic challenge was made during 1 min with 20% sucrose solution. After 45 min, test plaque was collected for fluoride (F) analysis. Enamel surface microhardness was previously determined at one side of the restoration and the percentage of surface microhardness change (%SMC) in relation to baseline (other side) was calculated. F concentration in enamel was also evaluated. RESULTS: Split-plot ANOVA showed a statistically lower %SMC on enamel around the ionomeric materials than around the CR (p<0.05). This result was supported by a statistically higher F concentration in test plaque (P<0.001) and in enamel (P<0.001) restored with the ionomeric materials when compared to the CR. CONCLUSIONS: The results suggest that the short-term in situ model tested is useful for studying the anticariogenic potential of dental materials that release fluoride.

Quantification of Streptococcus mutans in Different Types of Ligature Wires and Elastomeric Chains

Abstract This study aimed to test the hypothesis that Streptococcus mutans contamination levels differ according to the type of the orthodontic ligature. Thirteen patients were selected. Each quadrant was randomly subjected to one of the following ligature-use protocols: I) elastomeric chain, II) steel ligature crossed over the archwire, III) steel ligature crossed under the archwire, and IV) steel ligature in a figure-eight pattern under the archwire. After seven days, the devices were removed and the Streptococcus mutans colony-forming unit count per mg of biofilm weight was determined. Twelve specimens (n=3) were also processed for scanning electron microscopy analysis. ANOVA and Tukey-Kramer test were used for comparisons to assess S. mutans differences between groups at a 5% significance level. There was no statistical difference in detectable levels of S. mutans among the groups (p=0.294). Scanning electron microscopy results showed abundant biofilms and microbial contamination in all groups. In conclusion, S. mutans contamination levels are similar in the different orthodontic ligatures.

Resumo Este estudo teve como objetivo testar a hipótese de que os níveis de contaminação de Streptococcus mutans diferem de acordo com o tipo de ligadura ortodôntica. Treze pacientes foram selecionados. Cada quadrante foi submetido aleatoriamente a um dos seguintes protocolos de uso de ligadura: I) ligadura elastomérica, II) ligadura de aço trançada sobre o arco, III) ligadura de aço trançada sob o arco e IV) ligadura de aço em um padrão de "oito" sob o arco. Após sete dias, os dispositivos foram removidos e a contagem das unidades formadoras de colônia de S. mutans por mg de peso de biofilme foi determinada. Doze espécimes (n = 3) também foram processados para análise por microscopia eletrônica de varredura. Análise de variância e teste de Tukey-Kramer foram utilizados para comparações a fim de avaliar as diferenças de níveis de S. mutans entre os grupos com significância de 5%. Não houve diferença estatisticamente significante em níveis detectáveis de S. mutans entre os grupos (p = 0,294). Os resultados da microscopia eletrônica de varredura mostraram biofilmes abundantes e contaminação microbiana em todos os grupos. Em conclusão, os níveis de contaminação de S. mutans são semelhantes nas diferentes ligaduras ortodônticas.

Teaching of bioethics in dental graduate programs in Brazil.

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.