Resumo O biofilme subgengival periodontopatogênico é o principal agente etiológico da periodontite. Assim, a pesquisa de antimicrobianos como adjuvantes para o tratamento periodontal na literatura é intensa. Cloreto de cetilpiridínio (CPC) é um agente antimicrobiano comumente usado em enxaguatórios bucais. No entanto não foram encontrados na literatura estudos avaliando os efeitos do CPC em um modelo complexo de biofilme. Portanto, o objetivo deste artigo é avaliar as propriedades antimicrobianas do cloreto de cetilpiridinio 0,075% em um modelo de biofilme subgengival multiespécie in vitro. O biofilme subgengival composto por 31 espécies relacionadas à periodontite foi formado por 7 dias, utilizando o dispositivo calgary. Os tratamentos com CPC e clorexidina (CHX) 0,12% (controle positivo) foram realizados 2x/dia, por 1 min, do dia 3 até o final do período experimental, totalizando 8 tratamentos. Após 7 dias de formação do biofilme, a atividade metabólica do biofilme foi avaliada por reação colorimétrica e a composição microbiana dos biofilmes por hibridização DNA-DNA. A análise estatística foi realizada usando ANOVA com dados transformados via BOX-COX seguido do teste de Dunnett. Tanto o CPC como a CHX reduziram a atividade metabólica do biofilme em aproximadamente 60% e apresentaram atividade antimicrobiana contra 13 espécies diferentes. Especificamente, apenas os níveis de F.n. Vicentii e P. gingivalis foram reduzidos somente pelo tratamento com a CHX enquanto apenas o CPC reduziu A. odontolyticus e A. israelli. O CPC foi tão eficaz quanto o CHX como antimicrobiano através de biofilme subgengival complexo multiespecífico in vitro. No entanto, futuros estudos usando modelos in vivo de doença periodontal experimental devem ser realizados para comprovar tal efeito.

Abstract Periodontopathogenic subgingival biofilm is the main etiological agent of periodontitis. Thus, a search for antimicrobials as adjuvant for periodontal treatment in the literature is intense. Cetylpyridinium chloride (CPC) is a well-known antimicrobial agent commonly used in mouthrinses. However, CPC effects on a complex biofilm model were not found over the literature. Therefore, the aim of this manuscript is to evaluate 0.075% CPC antimicrobial properties in a multispecies subgingival biofilm model in vitro. The subgingival biofilm composed by 31 species related to periodontitis was formed for 7 days, using the calgary device. The treatments with CPC and chlorhexidine (CHX) 0.12% (as positive control) were performed 2x/day, for 1 min, from day 3 until the end of experimental period, totaling 8 treatments. After 7 days of biofilm formation, biofilm metabolic activity was evaluated by a colorimetric reaction and biofilms microbial composition by DNA-DNA hybridization. Statistical analysis was performed using ANOVA with data transformed via BOX-COX followed by Dunnett post-hoc. Both CPC and CHX reduced biofilm metabolic activity in 60% and presented antimicrobial activity against 13 different species. Specifically, only CHX reduced levels of F.n. vicentii and P. gingivalis while only CPC reduced A. odontolyticus and A. israelli. CPC was as effective as CHX as antimicrobial through in vitro complex multispecies subgingival biofilm. However, future studies using in vivo models of experimental periodontal disease should be performed to prove such effect.

Abstract The aim of this study was to evaluate the effects of adjunct systemic antibiotic treatment with metronidazole (MTZ) and amoxicillin (AMX) in patients receiving non-surgical subgingival debridement (NSD) for peri-implantitis. Forty subjects presenting with at least one implant with severe peri-implantitis were randomized into an experimental group [treated with NSD plus MTZ (400 mg) and AMX (500 mg) three times a day for 14 days] and a control group treated with NSD plus placebo. Clinical parameters and submucosal biofilm profiles were evaluated up to 1 year post-treatment. Overall, both treatments improved clinical parameters over time. At 1 year, mean probing depth (PD), mean clinical attachment (CA) level and proportions of red complex pathogens did not differ significantly between the two groups. In addition, mean PD and CA changes to 1-year posttreatment did not differ significantly between the two groups between baseline and 1-year post-treatment. These results suggest that the addition of MTZ and AMX to the treatment protocol of patients undergoing NSD for with severe peri-implantitis does not improve the clinical and microbiological outcomes of NSD. The fact that half of the implants in both groups did not achieve clinical success (PD < 5 mm, no BoP, no bone loss) suggest that neither of the tested protocols were effective for treating severe peri-implantitis.

Abstract The aim of this randomized, single blinded clinical trial was to evaluate the effect of a pre-procedural mouthwash containing cetylpyridinium chloride (CPC), zinc lactate (Zn) and sodium fluoride (F) in the reduction of viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler. Sixty systemically healthy volunteers receiving dental prophylaxis were randomly assigned to one of the following experimental groups (15 per group): (i) rinsing with 0.075% CPC, 0.28% Zn and 0.05% F (CPC+Zn+F), (ii) water or (iii) 0.12% chlorhexidine digluconate (CHX), and (iv) no rinsing. Viable bacteria were collected from different locations in the dental office on enriched TSA plates and anaerobically incubated for 72 hours. The colonies were counted and species were then identified by Checkerboard DNA–DNA Hybridization. The total number of colony-forming units (CFUs) detected in the aerosols from volunteers who rinsed with CPC+Zn+F or CHX was statistically significantly (p<0.05) lower than of those subjects who did not rinse or who rinsed with water. When all locations were considered together, the aerosols from the CPC+Zn+F and CHX groups showed, respectively, 70% and 77% fewer CFUs than those from the No Rinsing group and 61% and 70% than those from the Water group. The mean proportions of bacterial species from the orange complex were statistically significantly (p<0.05) lower in aerosols from the CPC+Zn+F and CHX groups compared with the others two groups. In conclusion, the mouthwash containing CPC+Zn+F, is effective in reducing viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler.

ABSTRACT Objective The aim of this study was to evaluate the association of Porphyromonas endodontalis, Filifactor alocis and Dialister pneumosintes with the occurrence of periodontitis. Material and Methods Thirty subjects with chronic periodontitis (ChP) and 10 with periodontal health (PH) were included in the study. Nine subgingival biofilm samples were collected as follows: i) PH group - from the mesial/buccal aspect of each tooth in two randomly chosen contralateral quadrants; ii) ChP group - from three sites in each of the following probing depth (PD) categories: shallow (≤3 mm), moderate (4-6 mm) and deep (≥7 mm). Checkerboard DNA-DNA hybridization was used to analyze the samples. Results We found the three species evaluated in a higher percentage of sites and at higher levels in the group with ChP than in the PH group (p<0.05, Mann-Whitney test). We also observed these differences when the samples from sites with PD≤4 mm or ≥5 mm of subjects with ChP were compared with those from subjects with PH (p<0.05, Mann-Whitney test). In addition, the prevalence and levels of D. pneumosintes, and especially of F. alocis were very low in healthy subjects (0.12x105 and 0.01x105, respectively). Conclusion F. alocis and D. pneumosintes might be associated with the etiology of ChP, and their role in the onset and progression of this infection should be further investigated. The role of P. endodontalis was less evident, since this species was found in relatively high levels and prevalence in the PH group.

Introducción. Las bacteriemias de origen oral pueden ocurrir después de procedimientos odontológicos y de otros actos cotidianos. Algunas condiciones de la cavidad oral favorecen las bacteriemias como en el caso de pacientes con diabetes mellitus y periodontitis que presentan inflamación gingival exacerbada. Objetivo. Comparar la eficacia de un método independiente de cultivo (PCR cuantitativa) y otro dependiente (BacT-ALERT 3D ® ) en la detección de la bacteriemia. Materiales y métodos. Se tomaron muestras de sangre de individuos con diabetes mellitus de tipo II y periodontitis, antes y después de la masticación de manzana. Una alícuota se procesó por el sistema automatizado de hemocultivo (BacT-ALERT 3D ® ) y se monitorizó durante 15 días; la otra alícuota fue tratada para la extracción del ADN y procesada por RT-PCR usando un conjunto de cebadores de 16S rDNA exclusivos para bacterias. Resultados. En las muestras tomadas antes de masticar se confirmó la ausencia de bacterias mediante los dos métodos. En las muestras tomadas después de masticar la presencia de bacterias se evidenció únicamente en dos hemocultivos y en ninguna de las muestras se detectó la presencia de bacterias con el método de RT-PCR. Conclusiones. La PCR cuantitativa no mostró mayor eficacia que el BacT-ALERT 3D ® en la detección de la bacteriemia de origen oral.

Introduction: Oral-derived bacteremia may occur after several dental procedures and routine daily activities. Some conditions of the oral cavity may favor episodes of bacteremia. This would be the case of patients with diabetes mellitus and periodontitis, who exhibit exacerbated gingival inflammation and may be more prone to developing oral-derived bacteremia. Objective: To compare the effectiveness of an independent culture method (quantitative real-time PCR- qCR) and the most commonly used method (BacT-ALERT 3D ® ) for the diagnosis of bacteremia. Materials and methods: Blood samples were drawn from subjects with type 2 diabetes mellitus and chronic periodontitis before and after apple chewing. Samples were processed by an automated blood culture system (BacT-ALERT 3D ® ) monitored for 15 days with suitable subculture of positive cultures. In parallel, whole DNA from blood samples was purified using a commercial kit and screened by qPCR using a universal primer set of 16S rDNA for bacteria detection. Results: Blood cultures taken before apple chewing were shown to be negative by the two diagnostic methods. After chewing, two samples (11%) showed bacterial growth by BacT-ALERT 3D ® whereas qPCR did not detect the presence of bacteria in any sample. Conclusions: qPCR did not show greater effectiveness than the BacT-ALERT 3D ® in the detection of bacteremia of oral origin.

Objective The aim of this systematic review was to compare the clinical effectiveness of systemic antibiotics administered in the active stage of periodontal treatment or after the healing phase. Material and Methods An electronic search was performed in the databases EMBASE, MEDLINE and Cochrane Central Register of Controlled Trials (CENTRAL), in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) statement. A manual search of the reference list of selected studies and of review articles was also performed up to November 2013. Randomized Clinical Trials (RCT) that evaluated the systemic administration of antibiotics as adjuvants to scaling and root planning (SRP) at different phases of periodontal treatment were included. Systematic reviews and studies that evaluated subjects with systemic diseases and those that used subantimicrobial doses of antibiotics were excluded. Results The initial search identified 1,039 articles, of which seven were selected, and only one met the inclusion criteria. This study showed that subjects taking metronidazole and amoxicillin at the initial phase of treatment exhibited statistically significantly greater reduction in pocket depth and gain in clinical attachment level in initially deep sites (PD≥7 mm) than subjects taking antibiotics after healing (p<0.05). This comparison was conducted 2 months after antibiotic intake, at the healing phase. Conclusion To date, only one short-term RCT has directly compared different moments of systemic antibiotics administration, as adjuncts to SRP, in the treatment of periodontitis. Although the results of this study suggested some benefits for antibiotics intake during the active phase of therapy, these findings need to be confirmed by larger placebo-controlled randomized clinical trials with longer follow-up periods.

Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas. Methods: The expression of TNF-α, IFN-γ, IL-17A, IL23, IL21, IL-33, IL-10, IL-4, IL-9, IL-22, FOXp3 markers (via RealTimePCR array) was accessed in active/progressive (N=40) versus inactive/stable (N=70) periapical granulomas (as determined by RANKL/OPG expression ratio), and also to compare these samples with a panel of control specimens (N=26). A cluster analysis of 13 cytokine levels was performed to examine possible clustering between the cytokines in a total of 110 granulomas. Results: The expression of all target cytokines was higher in the granulomas than in control samples. TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels were significantly higher in active granulomas, while in inactive lesions the expression levels of IL-4, IL-9, IL-10, IL-22 and FOXp3 were higher than in active granulomas. Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05). Three clusters were identified in active lesions, being the variance in the expression levels of IL-22, IL-10, IFN-γ, IL-17, IL-33, FOXp3, IL-21 and RANKL statistically significant (KW p<0.05). Conclusion: There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions. While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.

Antibiotics are important adjuncts in the treatment of infectious diseases, including periodontitis. The most severe criticisms to the indiscriminate use of these drugs are their side effects and, especially, the development of bacterial resistance. The knowledge of the biological mechanisms involved with the antibiotic usage would help the medical and dental communities to overcome these two problems. Therefore, the aim of this manuscript was to review the mechanisms of action of the antibiotics most commonly used in the periodontal treatment (i.e. penicillin, tetracycline, macrolide and metronidazole) and the main mechanisms of bacterial resistance to these drugs. Antimicrobial resistance can be classified into three groups: intrinsic, mutational and acquired. Penicillin, tetracycline and erythromycin are broad-spectrum drugs, effective against gram-positive and gram-negative microorganisms. Bacterial resistance to penicillin may occur due to diminished permeability of the bacterial cell to the antibiotic; alteration of the penicillin-binding proteins, or production of β-lactamases. However, a very small proportion of the subgingival microbiota is resistant to penicillins. Bacteria become resistant to tetracyclines or macrolides by limiting their access to the cell, by altering the ribosome in order to prevent effective binding of the drug, or by producing tetracycline/macrolide-inactivating enzymes. Periodontal pathogens may become resistant to these drugs. Finally, metronidazole can be considered a prodrug in the sense that it requires metabolic activation by strict anaerobe microorganisms. Acquired resistance to this drug has rarely been reported. Due to these low rates of resistance and to its high activity against the gram-negative anaerobic bacterial species, metronidazole is a promising drug for treating periodontal infections.

The aim of this cross-sectional study was to examine the relationship between the composition of the subgingival microbiota and the vitamin D receptor (VDR) gene polymorphism in Brazilian adults with chronic periodontitis. The clinical parameters of probing depth, clinical attachment level, bleeding on probing, plaque accumulation and suppuration were measured in 60 Caucasian adults who were divided into two groups: 30 healthy individuals (control) and 30 with chronic periodontitis (ChP). Subgingival plaque samples were collected from 6 sites per subject and analyzed for 38 bacterial species using the Checkerboard DNA-DNA Hybridization. DNA was obtained from the subjects' epithelial cells by scraping the buccal mucosa and using a mouthwash containing 3% of glucose. Polymorphism in the VDR gene was analyzed by the polymerase chain reaction (PCR), followed by Taql digestion (RFLP). The healthy subjects presented significantly lower levels (0.3 × 10(7) ± 0.7 × 10(7)) of total microbial counts in comparison with subjects with chronic periodontitis (4.5 × 10(7) ± 2.9 × 10(7)). Regarding the occurrence of VDR polymorphism, it was observed that the Tt genotype was more prevalent in the Periodontitis group (60%) than in the Healthy group (30%), while the prevalences of the TT genotype were 23.3% and 53.3%, respectively (Chi-square test, p < 0.05). No difference was found in the composition of subgingival microbiota among the VDR genotypes evaluated for the Healthy and Periodontitis groups. In conclusion, the Tt genotype was associated with periodontal disease; however, no association with the subgingival microbiota was observed.

Homem de 65 anos com insuficiência cardíaca causada por hipertensão e cardiopatia isquêmica foi internado com dispnéia, escarro sanguinolento e dor pleurítica após uma viagem de ônibus com duração de 52 horas. A avaliação clínica e laboratorial incluiu tomografia helicoidal do tórax, que demonstrou um defeito de enchimento do principal ramo da artéria pulmonar direita e uma opacidade periférica regular de formato triangular no lobo inferior da parte inferior do pulmão. Após o diagnóstico de tromboembolia pulmonar, foi instituída terapia com heparina, seguida por varfarina. O paciente recebeu alta hospitalar. O diagnóstico de embolia pulmonar deve ser levado em consideração em pacientes com sintomas semelhantes aos deste paciente após longa viagem de ônibus.

A 65 year-old man with heart failure due to hypertensive and ischemic heart disease was admitted to the hospital with dyspnea, bloody sputum and pleuritic chest pain after a 52-hour bus trip. Clinical and laboratory evaluation included chest helical tomography that demonstrated a filling defect of the right main branch of the pulmonary artery and a regular peripheral opacity of triangular shape in the inferior lobe of the lower lung. The diagnosis of pulmonary thromboembolism was made and therapy with heparin, followed by warfarin was introduced. The patient was discharged from the hospital. The diagnosis of pulmonary embolism should be considered in patients with complaints like this patient after long-distance bus travel.