Synergistic antimicrobial potential of EGCG and fosfomycin against biofilms associated with endodontic infections

Objective: This study aimed to evaluate the cytotoxicity and synergistic effect of epigallocatechin gallate (EGCG) and fosfomycin (FOSFO) on biofilms of oral bacteria associated with endodontic infections. Methodology: This study determined minimum inhibitory and bactericidal concentration (MIC/MBC) and fractionated inhibitory concentration (FIC) of EGCG and FOSFO against Enterococcus faecalis, Actinomyces israelii, Streptococcus mutans, and Fusobacterium nucleatum. Monospecies and multispecies biofilms with those bacteria formed in polystyrene microplates and in radicular dentin blocks of bovine teeth were treated with the compounds and control chlorhexidine (CHX) and evaluated by bacterial counts and microscopy analysis. Toxicity effect of the compounds was determined on fibroblasts culture by methyl tetrazolium assays. Results: The combination of EGCG + FOSFO demonstrated synergism against all bacterial species, with an FIC index ranging from 0.35 to 0.5. At the MIC/FIC concentrations, EGCG, FOSFO, and EGCG+FOSFO were not toxic to fibroblasts. EGCG+FOSFO significantly reduced monospecies biofilms of E. faecalis and A. israelli, whereas S. mutans and F. nucleatum biofilms were eliminated by all compounds. Scanning electron microscopy of multispecies biofilms treated with EGCG, EGCG+FOSFO, and CHX at 100x MIC showed evident biofilm disorganization and substantial reduction of extracellular matrix. Confocal microscopy observed a significant reduction of multispecies biofilms formed in dentin tubules with 84.85%, 78.49%, and 50.6% of dead cells for EGCG+FOSFO, EGCG, and CHX at 100x MIC, respectively. Conclusion: EGCG and fosfomycin showed a synergistic effect against biofilms of oral pathogens related to root canal infections without causing cytotoxicity

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

Objectives: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. Material and Methods: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (

Aferição do pH de diferentes agentes clareadores indicados para o clareamento intracoronário

Objective: to measure pH values of bleaching agents that are indicated to intracoronal bleaching technique in different time intervals. Methods: Each group (G) received five samples (n=5): G1 – distilled water (AD); G2 – hydrogen peroxide (H2O2) 30%; G3 – sodium perborate (PbS) + AD; G4 – PbS + H2O2 30%; G5 – sodium percarbonate (PcS) + AD; and G6 – PcS + H2O2 30%. pH values were stated using a digital pHmeter, in different time intervals: immediately after handling (T0), 24 hours (T1) and 168 hours after handling (T2). The results were submitted to statistical analysis through Kruskal-Wallis and Mann Whitney tests, in this order, allowing multiple comparisons among the groups. To verify the effect of time in each group, Friedman test was applied. Results: In the evaluation of the effect of time in each group, it was observed that G2 presented acid behavior, while the other groups exhibited values close to neutrality or alkaline. Conclusions: H2O2 30% was the only agent that showed acidic behavior in every evaluation time. Meanwhile, PcS + H2O had the highest pH values.Objetivo: Aferir os valores de pH de diferentes produtos clareadores indicados para uso na técnica intracoronária em diferentes intervalos de tempo. Materiais e métodos: Para cada grupo experimental (G), foram preparadas cinco amostras (n=5): G1 – água destilada (AD); G2 – peróxido de hidrogênio (H2O2) a 30%; G3 – perborato de sódio (PbS) + AD; G4 – PbS + H2O2 a 30%; G5 – percarbonato de sódio (PcS) + AD; e G6 – PcS + H2O2 a 30%. O pH foi determinado por um pHmetro digital, em diferentes intervalos de tempo: imediatamente após a manipulação (T0), 24 horas (T1) e 168 horas após a manipulação (T2). Os dados obtidos foram submetidos à análise estatística por meio do teste de Kruskal-Wallis, seguido pelo teste de Mann Whitney para comparações múltiplas entre os grupos. Para verificar o efeito do tempo em cada grupo, foi aplicado o teste de Friedman. Resultados: Na avaliação do efeito do tempo em cada grupo, observou-se que G2 apresentou comportamento ácido, enquanto os demais grupos exibiram valores de pH próximos da neutralidade ou alcalinos. Conclusão: H2O2 30% foi o único agente que apresentou comportamento ácido em todos os tempos de avaliação, e a mistura PcS + H2O foi a que apresentou os maiores valores de pH

Influência da atenção odontológica precoce na experiência de cárie em adultos jovens

Os programas preventivos para crianças em idade pré-escolar são eficazes para prevenção e manutenção da saúde bucal. No entanto, a efetividade dessas ações preventivas após o término do programa até a idade adulta ainda não foi investigada. O objetivo do trabalho foi avaliar a influência da atenção odontológica precoce durante a idade pré-escolar na experiência de cárie em adultos jovens. Foram avaliados 72 indivíduos (18 a 25 anos, 41,7% gênero masculino) que participaram do programa preventivo da Bebê-Clínica/UEL durante seus cinco primeiros anos de vida, e 36 indivíduos (18 a 25 anos, 41,7% gênero masculino) que não participaram de programas preventivos (grupo controle). Aplicou-se questionário estruturado com questões englobando aspectos sociodemográficos, hábitos dietéticos e hábitos de higiene bucal. Em seguida realizou-se exame clínico para diagnóstico de placa visível e índices CPOD (número de dentes cariados, perdidos e obturados) e CPOS (número de superfícies cariadas, perdidas e obturadas), além de coleta de saliva para quantificar SIgA salivar e anticorpos SIgA anti-S. mutans por ensaio imunoenzimático. As médias dos índices CPOD e CPOS encontrados foram de 2,72 ± 2,8 e 3,07± 3,3 para o grupo de estudo (n=72), e 3,06 ± 2,9 e 3,92 ± 4,6 para o grupo controle (n=36), sem diferença estatisticamente significativa. No grupo de estudo, o gênero masculino apresentou prevalência de cárie significativamente menor que o gênero feminino. Níveis menores de anticorpos SIgA anti-S. mutans foram observados no grupo de estudo. Conclui-se que a atenção odontológica precoce não influenciou significativamente a prevalência de cárie em adultos jovens, porém, prode ter promovido modulação negativa na resposta imunológica humoral bucal contra os S. mutans.Oral health programs for preschool children are effective for protection and maintaining oral health. However, the effectiveness of these preventive actions after the end of the program up to adulthood has not been investigated yet. The objective of this study was to investigate the influence of early dental care during childhood in the dental caries experience in young adults. 72 individuals, who participated in the prevention program of Bebê-Clínica/UEL during their first five years of life, and 36 subjects, who did not participate in the preventive program (control group), were evaluated. A structured questionnaire was applied addressing socio-demographic characteristics, dietary and oral hygiene habits. Afterwards, a clinical diagnosis of visible dental plaque and DMFT (Decayed, Missing or Filled Teeth) and DMFS (Decayed, Missing or Filled Surface) indexes were held. In addition, a saliva sample was collected to quantify salivary SIgA anti-S. mutans and anti-S. mutans SIgA antibodies by immunoenzymatic assay. The mean DMFT and DMFS indexes found were 2,72 ± 2,8 and 3,07 ± 3,3 for the study group (n = 72), and 3,06 ± 2,9 and 3,92 ± 4,6 for the control group (n = 36) (p>0,05) . In the study group, males showed significantly lower prevalence of dental caries compared to the female gender. Lower levels of anti-S. mutans SIgA were also observed in the study group. We conclude that early dental care did not significantly influence the prevalence of dental caries in young adults. However, it may induce a modulation of oral immune humoral response against S. mutans

Anti-dentine Salivary SIgA in young adults with a history of dental trauma in deciduous teeth

Anti-dentin autoantibodies are associated with inflammatory root resorption in permanent teeth and are modulated by dental trauma and orthodontic force. However, it is not known whether deciduous tooth trauma can stimulate the development of a humoral immune response against dentin. The aim of this study was to evaluate the levels of salivary SIgA reactivity against human dentin extract in young adults with a history of trauma in the primary dentition. A sample of 78 patients, aged 18 to 25, who had completed an early childhood (0 to 5 years old) caries prevention program years earlier at the Universidade Estadual de LondrinaPediatric Clinic, underwent radiographic examination and salivary sampling. Anti-dentin SIgA levels were analyzed by immunoenzymatic assay and Western blotting. Although dental trauma to deciduous teeth had occurred in 34 (43.6%) of the patients, no differences in SIgA levels were detected between individuals who had experienced trauma and those who had not (p > 0.05). Multivariate regression analysis showed no association between dental trauma and SIgA levels (p > 0.05). Patients with a history of deciduous trauma presented low levels of anti-dentin antibodies, associated with orthodontic root resorption (p < 0.05). Western blot analysis showed that salivary antibodies recognized a single band of approximately 45 kDa in dentin extract. We concluded that salivary SIgA recognizes a specific component of the dentin matrix and that anti-dentin antibodies were not triggered by trauma to primary teeth. However, trauma to deciduous teeth may down-modulate SIgA in response to orthodontic root response.</p

Highlights in peptide nanoparticle carriers intended to oral diseases

New biocompatible nanomaterials are leading to a range of emerging health treatments. Thus, peptide drugs present in oral diseases, such as caries, periodontal diseases and oral cancer, are highlighting into clinical practice because the peptides can have an influence the growth of tumor cells or microorganisms and its can exhibit direct cytotoxic activity towards cancer cells or microbial cells. Therefore, it is interesting to speculate what are the natural or synthetic peptides that can be used to develop novel strategies to fight cancer diseases or microbial cells, and so, to represent a novel family of anticancer or antimicrobial agents. The use of buccal drug delivery systems, especially nanoparticles, to carrier peptides shows to be very interesting, because these systems can protect the peptide against enzymatic degradation, in addition to target inaccessible sites. However, this peptide delivery system seems to be unexplored by researchers. On the hand, the application of drug delivery systems for oral diseases could be a proposed treatment for these diseases, including the treatment or co-treatment with other therapies, such as photodynamic therapy, of antimicrobial, periodontal diseases and cancer, or even in the early diagnosis of cancer. The objective of this study is to present a systematic review of nanotechnology-based peptides delivery systems intended to oral diseases.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

Abstract Objectives: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. Material and Methods: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05). Results: Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment. Conclusions: The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations