Resumo O objetivo do presente estudo in vivo foi avaliar a contaminação bacteriana de protetores bucais esportivos, a rugosidade da superfície e a eficácia do spray de gluconato de clorexidina na desinfecção desses dispositivos. Um ensaio clínico randomizado, cego, cruzado foi realizado com vinte crianças de 9 a 13 anos, que praticavam artes marciais, participaram de todas as fases do estudo. As crianças foram orientadas a usar o protetor bucal por 3 dias alternados durante 1 hora e, após o uso, borrifar água de torneira estéril ou clorexidina 0,12%. Os protetores foram analisados por ensaio MTT, Hibridização DNA-DNA e microscopia confocal a laser antes e após o uso por 2 semanas. Os dados foram analisados pelos teste de Wilcoxon, teste t de Student, e correlação de Pearson, com nível de significância de 5%. Observou-se que os protetores bucais do grupo controle estavam mais contaminados com microrganismos cariogênicos do que os do grupo experimental (clorexidina) (p <0,05). O uso de protetores bucais com spray de clorexidina reduziu significativamente a contaminação bacteriana em relação ao grupo controle (p = 0,007). A rugosidade da superfície dos protetores bucais aumentou significativamente após o uso, independentemente da aplicação de spray de clorexidina. Uma correlação moderada (r = 0,59) foi observada entre a rugosidade da superfície e a contaminação do micro-organismo apenas para o grupo controle. Os protetores bucais esportivos apresentam intensa contaminação microbiana e aumento da rugosidade superficial após o uso. O uso de spray de clorexidina foi eficaz para reduzir a contaminação dos protetores bucais usados por crianças.

Abstract The aim of the present in vivo study was to evaluate the bacterial contamination of sports mouthguards, surface roughness, and the efficacy of chlorhexidine gluconate spray in the disinfection of these devices. A randomized, blinded cross-over clinical trial was performed with twenty 9 to 13 years old children who practiced martial arts and participated in all phases of the study. They were instructed to wear mouthguards 3 alternated days a week for 1 hour and, after use, to spray sterile tap water or chlorhexidine 0.12%. The mouthguards were analyzed by MTT assay, Checkerboard DNA-DNA hybridization, and confocal laser microscopy prior and after use for 2 weeks. Data were analyzed by Wilcoxon and t-Student, and Pearson correlation tests, with 5% significance level. Were observed that mouthguards of the control group were more contaminated with cariogenic microorganisms than those of the chlorhexidine group (p<0.05). The mouthguards use of spray of chlorhexidine reduced significantly the bacteria contamination compared with control group (p = 0.007). The surface roughness of the mouthguards increased significantly after use, irrespective of application of chlorhexidine spray. A moderate correlation (r=0.59) was observed between surface roughness and the cariogenic microorganism’s contamination only for control group. Sports mouthguards had intense microbial contamination and increased surface roughness after its use. The use of chlorhexidine spray was effective for reducing the mouthguards contamination used by children.

Abstract Randomized clinical trials (RCTs) are human studies carried out to compare different treatments or interventions, and their results are used to support clinical decision-making and improve patient care. Herein, the aim of this study was to review the selection process of study outcomes in periodontology. Primary outcomes should draw the main conclusions of the study, whereas secondary outcomes should only be used to help explain the main findings and generate future research hypothesis. Outcomes are classified as clinically relevant (CROs) or surrogate outcomes. CROs – the first option for primary outcome variables - should convey not only substantial health benefits, but also be deemed important by patients. In periodontology, tooth loss/retention and oral health–related quality of life (OHRQoL) are examples of CROs. While tooth loss has main limitations as a primary outcome, emerging evidence suggest that patient-reported outcome measures (PROMs) can accurately detect OHRQoL following periodontal therapy. When CROs cannot be assessed, validated surrogate outcomes can be used as proxies. Primary outcome variables should reflect a treatment endpoint at the patient level that can be easily used to inform decision-making in daily practice. These outcomes should allow the implementation of a treat-to-target concept in which the intervention can be clearly judged against a prespecified treatment target. Recently, the presence of at most 4 sites with periodontal probing depth ≥5 mm post-treatment was suggested as an effective endpoint for periodontal trials. In perspective, a combination of validated clinical parameters and PROMs will provide a more comprehensive assessment of periodontal treatments.

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: Periodontal diseases are considered a worldwide public health problem, owing to their high prevalence in developed and developing countries. Periodontitis may lead to tooth loss, which can impact oral health-related quality of life. Gingivitis and periodontitis have been extensively studied regarding their etiopathogenesis, epidemiology, prevention and treatment outcomes. However, most of these aspects are studied and discussed globally, which may hamper a clear interpretation of the findings and the design of effective plans of action for specific regions or populations. For example, in Latin America, epidemiological data about the distribution of periodontal diseases is still scarce, mainly when it comes to nationwide representative samples. This Consensus aimed to address the following topics related to periodontal diseases in Latin America: a) The impact of the global burden of periodontal diseases on health: a global reality; b) Periodontal diseases in Latin America; c) Strategies for the prevention of periodontal diseases in Latin America; d) Problems associated with diagnosis of periodontal conditions and possible solutions for Latin America; e) Treatment of Periodontitis. This consensus will help to increase awareness about diagnosis, prevention and treatment of periodontal diseases, in the context of Latin American countries.

Abstract: Gingivitis and periodontitis are associated with a negative impact on Oral Health Related Quality of Life (OHRQoL), exerting a significant influence on aspects related to the patients' function and esthetics. Periodontitis has been associated with several systemic conditions, including adverse pregnancy outcomes, cardiovascular diseases, type 2 diabetes mellitus (DM), respiratory disorders, fatal pneumonia in hemodialysis patients, chronic renal disease and metabolic syndrome. The aim of this paper was to review the results of different periodontal treatments and their impacts on patients' OHRQoL and systemic health. Non-surgical and surgical periodontal treatments are predictable procedures in terms of controlling infection, reducing probing pocket depth and gaining clinical attachment. In addition, the treatment of periodontitis may significantly improve OHRQoL and promote a reduction in the levels of systemic markers of inflammation, including some cytokines associated with cardiovascular diseases. Studies have also suggested that periodontal treatment may improve glycemic control in patients with DM. Strategies and actions for preventing the onset and recurrence of periodontitis, and the challenges facing the field of periodontology in the XXI century are presented in this review.

Abstract Objective: The microbial composition of pericoronitis (Pc) is still controversial; it is not yet clear if the microbial profile of these lesions is similar to the profile observed in periodontitis (Pd). Therefore, the aim of the present study was to describe the microbial profile of Pc lesions and compare it directly with that of subjects with Pd. Methodology: Subjects with Pc and Pd were selected, and subgingival biofilm samples were collected from (i) third molars with symptomatic Pc (Pc-T), (ii) contralateral third molars without Pc (Pc-C) and (iii) teeth with a probing depth >3 mm from subjects with Pd. Counts and proportions of 40 bacterial species were evaluated using a checkerboard DNA-DNA hybridization technique. Results: Twenty-six patients with Pc and 18 with Pd were included in the study. In general, higher levels of microorganisms were observed in Pd. Only Actinomyces oris and Eubacterium nodatum were present in higher mean counts in the Pc-T group in comparison with the Pc-C and Pd-C groups (p<0.05). The microbiota associated with Pc-T was similar to that found in Pc-C. Sites with Pc lesions had lower proportions of red complex in comparison with the Pd sites. Conclusion: The microbiota of Pc is very diverse, but these lesions harbour lower levels of periodontal pathogens than Pd.

Abstract Additive manufacturing (AM) is an emerging process for biomaterials and medical devices. Direct Laser Metal Sintering (DLMS) is an AM technique used to fabricate Ti–6Al–4V implant materials with enhanced surface-related properties compared with wrought samples; thus, this technique could influence microbial adsorption and colonization. Therefore, this in vitro study was conducted to evaluate the impact of different implant production processes on microbial adhesion of periodontal pathogens. Titanium discs produced using two different processes—conventional and AM—were divided into three groups: conventional titanium discs with machined surface (G1), AM titanium discs with chemical treatment (G2) and AM titanium discs without chemical treatment (G3). Subgingival biofilm composed of 32 species was formed on the titanium discs, and positioned vertically in 96-well plates, for 7 days. The proportions of microbial complexes and the microbial profiles were analyzed using a DNA–DNA hybridization technique, and data were evaluated using Kruskal–Wallis and Dunnett tests (p < 0.05). Lower proportions of the red complex species were observed in the biofilm formed in G2 compared with that in G1 (p < 0.05). Moreover, the proportions of the microbial complexes were similar between G2 and G3 (p > 0.05). Compared with G1, G2 showed reduced levels of Porphyromonas gingvalis , Actinomyces gerencseriae, and Streptococcus intermedius , and increased levels of Parvimonas micra , Actinomyces odontolyticus, and Eikenella corrodens (p < 0.05). The microbial profile of G3 did not differ from G1 and G2 (p > 0.05). The results of this in vitro study showed that titanium discs produced via AM could alter the microbial profile of the biofilm formed around them. Further clinical studies should be conducted to confirm these findings.

Abstract The aim was of this study was to determine the current weight of evidence for the existence of specific differences between the microbiota of healthy teeth and healthy implants, or of teeth with periodontitis and implants with peri-implantitis. A systematic review was conducted according to the PRISMA statement. The MEDLINE, EMBASE and Cochrane databases were searched up to February 2018 for studies comparing microbiological data of biofilm samples collected from healthy teeth and implants or from teeth with periodontitis and implants with peri-implantitis. The weight of evidence was defined in three categories (strong, moderate and mild/some), according to the difference in number of studies showing statistically significantly higher counts and/or proportions and/or abundance and/or prevalence of microorganisms in health or in disease. Of the 132 articles identified, 8 were included. A wide range of microorganisms were present in different conditions but no microorganisms showed strong, moderate or mild/some evidence for a specific association with either teeth or implants. The results of this systematic review indicated that there is insufficient evidence in the literature to support specific differences between microorganisms colonizing teeth and implants, either in health or in disease.

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 Objectives Using two groups of mini-implants (successful and failed) the objectives of this in vivo study were: to evaluate the microbial contamination by the checkerboard DNA-DNA hybridization technique and to quantify the bacterial endotoxin by the limulus amebocyte lysate assay. Material and Methods The 15 successful and 10 failed mini-implants (1.6 mm diameter × 7.0 or 9.0 mm long), placed in the maxilla and/or mandible, were obtained from 15 patients undergoing orthodontic treatment. Data were analyzed statistically by the Wilcoxon rank-sum test using the SAS software (a=0.05). Results All 40 microbial species were detected in both groups of mini-implants, with different frequencies. No differences were observed between the groups with respect to microbial complexes (blue, purple, yellow, green, orange, red and other species) and endotoxin quantification (p>0.05). Conclusion Neither microbial contamination nor endotoxin quantification was determinant for the early loss of stability of the mini-implants.

RESUMO Objetivo: Identificar e quantificar os níveis de três espécies bacterianas que foram recentemente apontadas como possíveis “novos” patógenos periodontais (Enterococcus faecalis, Staphylococcus warneri e Staphylococcus aureus), em indivíduos periodontalmente saudáveis e com periodontite crônica generalizada. Métodos: A amostra foi composta por 30 indivíduos adultos com periodontite crônica generalizada e 10 periodontalmente saudáveis. Nove amostras de biofilme subgengival foram coletadas por indivíduo e analisadas individualmente pela técnica de checkerboard DNA-DNA hybridization. Resultados: Os níveis médios de E. faecalis e S. warneri foram maiores no grupo com periodontite crônica do que no de saúde periodontal (p<0,05). Além disso, um maior percentual de indivíduos com periodontite estavam colonizados pelas três espécies avaliadas em comparação com os indivíduos saudáveis (p<0,05). Essa diferença entre os dois grupos foi de 40 pontos percentuais para E. faecalis (presente em 90% dos indivíduos com periodontite e 50% dos saudáveis) e S. warneri (100% e 60%, respectivamente), e de 26 pontos percentuais para S. aureus (86% e 60%, respectivamente). Conclusão: E. faecalis e S. warneri tem potencial para serem patógenos periodontais. O papel do S. aureus foi menos evidente, uma vez que esta espécie estava mais prevalente e em níveis relativamente mais altos do que as outras duas espécies em saúde. Estes dados podem guiar futuros estudos sobre o papel dessas espécies na etiologia das periodontites e ajudar a estabelecer tratamentos mais eficazes para essas infecções.

ABSTRACT Objective: To identify and quantify the levels of three bacterial species that have recently been identified as potential “new” periodontal pathogens (Enterococcus faecalis, Staphylococcus aureus and Staphylococcus warneri) in subjects with periodontal health and generalized chronic periodontitis. Methods: Thirty adults with generalized chronic periodontitis and 10 periodontally healthy were included in this study. Nine subgingival biofilm samples were collected per subject and individually analyzed by checkerboard DNA-DNA hybridization technique. Results: The mean levels of E. faecalis and S. warneri were higher in chronic periodontitis than in periodontal health (p<0.05). Furthermore, a higher percentage of subjects with periodontitis were colonized by the three species evaluated in comparison with healthy subjects (p<0.05). This represented a difference of 40 percentage points between the two groups, for E. faecalis (present in 90% of individuals with periodontitis and 50% of the healthy individuals) and S. warneri (100% and 60%, respectively), and 26 percentage points for S. aureus (86% and 60%, respectively). Conclusion: E. faecalis and S. warneri have the potential to be periodontal pathogens. The role of S. aureus was less evident, since this species was more prevalent and at relatively higher levels in health than the other two species. These data might guide future studies on the role of these microorganisms in the etiology of periodontitis and help to establish more effective treatments for these infections.

Abstract Objective This study evaluated the influence of glycemic control on the levels and frequency of subgingival periodontal pathogens in patients with type 2 diabetes mellitus (DM) and generalized chronic periodontitis (ChP). Material and Methods Fifty-six patients with generalized ChP and type 2 DM were assigned according to the levels of glycated hemoglobin (HbA1c) into one of the following groups: HbA1c<8% (n=28) or HbA1c≥8% (n=28). Three subgingival biofilm samples from sites with probing depth (PD)<5 mm and three samples from sites with PD≥5 mm were analyzed by quantitative Polymerase Chain Reaction (PCR) for the presence and levels of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Eubacterium nodatum, Parvimona micra, Fusobacterium nucleatum ssp. and Prevotella intermedia. Results The mean counts of F. nucleatum ssp. were statistically significantly higher in the sites with PD≥5 mm of the HbA1c≥8% group (p<0.05). Frequencies of detection of T. forsythia, E. nodatum, P. micra and F. nucleatum ssp. were all higher in the sites with PD≥5 mm of the patients with HbA1c≥8%, compared with those of patients with HbA1c<8% (p<0.05). Frequency of detection of P. intermedia was higher in the sites with PD<5 mm of the patients with HbA1c≥8% than those of the patients with HbA1c<8% (p<0.05). Conclusions Poor glycemic control, as indicated by HbA1c≥8%, is associated with increased levels and frequencies of periodontal pathogens in the subgingival biofilm of subjects with type 2 DM and ChP.

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.