Subgingival microbiome in periodontal health, gingivitis and different stages of periodontitis.

AIM: To characterize the subgingival microbiome in subjects with different periodontal health statuses. MATERIALS AND METHODS: In this cross-sectional observational study, subgingival samples were harvested from Spanish subjects with different periodontal health statuses, based on the 2018 Classification of Periodontal and Peri-Implant Diseases and Conditions. Samples were processed using high-throughput sequencing technologies (Illumina MiSeq). Taxa differentially abundant were identified using Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC). α- and ß-diversity metrics were calculated using q2-diversity in QIIME2. The analyses were adjusted for age, gender and smoking status. RESULTS: The identified subgingival microbiome showed statistically significant differences among subjects, categorized into periodontal health, gingivitis and stages I-II and III-IV periodontitis (p < .05). In patients with severe (stages III-IV) periodontitis, the genera Filifactor and Fretibacterium were detected 24 times more frequently than in periodontally healthy subjects. Similarly, the genera Porphyromonas, Prevotella and Tannerella were detected four times more frequently (p < .05). The genera Granulicatella, Streptococcus, Paracoccus, Pseudomonas, Haemophilus, Actinobacteria, Bergeyella and Capnocytophaga were significantly associated with healthier periodontal status (p < .05). CONCLUSIONS: Significant differences were detected in the subgingival microbiome among periodontal health, gingivitis and stages I-II or III-IV periodontitis, suggesting overlapping, yet distinguishable microbial profiles.

Neuroinflammation related to the blood-brain barrier and sphingosine-1-phosphate in a pre-clinical model of periodontal diseases and depression in rats.

AIM: To explore the potential mechanisms of neuroinflammation (microglia, blood-brain barrier [BBB] permeability, and the sphingosine-1-phosphate [S1P] pathways) resulting from the association between periodontitis and depression in rats. MATERIALS AND METHODS: This pre-clinical in vivo experimental study used Wistar rats, in which experimental periodontitis (P) was induced by using oral gavages with Porphyromonas gingivalis and Fusobacterium nucleatum. Then, a chronic mild stress (CMS) model was implemented to induce a depressive-like behaviour, resulting in four groups: P with CMS (P+CMS+), P without CMS (P+CMS-), CMS without P (P-CMS+), and control (P-CMS-). After harvesting brain samples, protein/mRNA expression analyses and fluorescence immunohistochemistry were performed in the frontal cortex (FC). Results were analysed by ANOVA. RESULTS: CMS exposure increased the number of microglia (an indicator of neuroinflammation) in the FC. In the combined model (P+CMS+), there was a decrease in the expression of tight junction proteins (zonula occludens-1 [ZO-1], occludin) and an increase in intercellular and vascular cell adhesion molecules (ICAM-1, VCAM-1) and matrix metalloproteinase 9 (MMP9), suggesting a more severe disruption of the BBB. The enzymes and receptors of S1P were also differentially regulated. CONCLUSIONS: Microglia, BBB permeability, and S1P pathways could be relevant mechanisms explaining the association between periodontitis and depression.

Adjunctive efficacy of systemic metronidazole in the surgical treatment of periodontitis: a double-blind parallel randomized clinical trial.

OBJECTIVES: The purpose of this study was to evaluate the clinical and microbiological impact of adjunctive metronidazole to periodontal surgery. MATERIALS AND METHODS: Systemically healthy patients, with stages III-IV, grades B-C periodontitis, were randomly assigned to receive metronidazole or placebo adjunctive to periodontal surgery, after subgingival instrumentation. Clinical variables were recorded at the initial visit, 6 weeks after subgingival instrumentation, and 3, 6, and 12 months after surgery. Microbiological samples were taken at initial and final visits and analyzed by quantitative polymerase chain reaction. RESULTS: Our results showed no statistically significant differences in the reduction of probing depth between the initial and final (1 year) visits in the two treatment groups. Additionally, no statistically significant differences were observed between study groups when comparing the post-subgingival instrumentation and final visits. However, 3 months after surgery, probing depth (mean difference, MD = 0.31 mm, 95% confidence interval, CI [0.13; 0.49]; p = 0.001) and clinical attachment level (MD = 0.64 mm, 95% CI [0.02; 1.27]; p = 0.044) were significantly lower in the test group. CONCLUSIONS: The adjunctive use of systemic metronidazole to periodontal surgery has a limited clinical and microbiological impact in the present study, and therefore, its use is not recommended. CLINICAL RELEVANCE: There are no studies that have evaluated the clinical and microbiological impact of the adjunctive use of systemic metronidazole to periodontal surgery (step 3 of periodontal therapy). The results of the present study do not support the adjunctive use of systemic metronidazole to periodontal surgery.

Periodontal diseases and depression: A pre-clinical in vivo study.

AIM: To analyse, through a pre-clinical in vivo model, the possible mechanisms linking depression and periodontitis at behavioural, microbiological and molecular levels. MATERIALS AND METHODS: Periodontitis (P) was induced in Wistar:Han rats (oral gavages with Porphyromonas gingivalis and Fusobacterium nucleatum) during 12 weeks, followed by a 3-week period of Chronic Mild Stress (CMS) induction. Four groups (n = 12 rats/group) were obtained: periodontitis and CMS (P+CMS+); periodontitis without CMS; CMS without periodontitis; and control. Periodontal clinical variables, alveolar bone levels (ABL), depressive-like behaviour, microbial counts and expression of inflammatory mediators in plasma and brain frontal cortex (FC), were measured. ANOVA tests were applied. RESULTS: The highest values for ABL occurred in the P+CMS+ group, which also presented the highest expression of pro-inflammatory mediators (TNF-α, IL-1ß and NF-kB) in frontal cortex, related to the lipoprotein APOA1-mediated transport of bacterial lipopolysaccharide to the brain and the detection of F. nucleatum in the brain parenchyma. A dysregulation of the hypothalamic-pituitary-adrenal stress axis, reflected by the increase in plasma corticosterone and glucocorticoid receptor levels in FC, was also found in this group. CONCLUSIONS: Neuroinflammation induced by F. nucleatum (through a leaky mouth) might act as the linking mechanism between periodontal diseases and depression.

Clinical and microbiological effects of the adjunctive use of probiotics in the treatment of gingivitis: A randomized controlled clinical trial.

AIM: To evaluate the efficacy of a probiotic combination in the treatment of gingivitis and to assess its impact on the subgingival microbiota. MATERIALS AND METHODS: A placebo-controlled clinical trial was conducted in gingivitis subjects during 6 weeks. Test treatment consisted of the administration of two oral tablets per day containing the probiotic strains Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici; the control group received the same tablets but without live bacteria. The main outcome variable was the changes in gingival index (GI). Subgingival samples were collected and analysed by quantitative polymerase chain reaction (qPCR) for five putative periodontal pathogens. Outcome variables were compared between and within groups, and multiple regression analysis was performed. RESULTS: A total of 59 patients (29 tests, 30 placebos) were included in the analysis. Both treatment groups experienced a statistically significant improvement in mean GI (p < .0001), but no differences between treatment groups were found for any clinical index. A significantly higher reduction in the number of sites with higher GI scores (GI = 3 at baseline) was observed in the test group. In subgingival samples, a significant reduction in T. forsythia was significant only in the test group (p < .008). CONCLUSIONS: The use of probiotic tablets containing L. plantarum, L. brevis and P. acidilactici did not lead to significant changes in mean GI; although a significant reduction occurred in the number of sites with severe inflammation. Furthermore, the adjunctive use of this probiotic promoted a significant microbiological impact.

Validation of ATP bioluminescence as a tool to assess antimicrobial effects of mouthrinses in an in vitro subgingival-biofilm model.

OBJECTIVES: The aim of this investigation was to evaluate whether the adenosine triphosphate (ATP) bioluminescence method is an appropriate tool to assess the efficacy of antiseptic mouthrinses in terms of quantitative reductions of total viable microbial counts in mixed biofilm populations in vitro. STUDY DESIGN: Three mouthrinses, containing respectively, chlorhexidine and cetylpyridinium chloride (CHX/CPC), essential oils (EO) and amine fluoride/stannous fluoride (AFSF), as well as Phosphate Buffered Saline (PBS) used as control, were tested in an in vitro static biofilm model by ATP bioluminescence and compared to culture method. Biofilms were grown on saliva-coated hydroxyapatite disks for 72 hours and then exposed for 1 minute to the mouthrinse or control by immersion. The antibacterial effect of the rinses was tested by analysis of variance. The reliability of the ATP bioluminescence method was assessed by calculating the Pearson correlation coefficients when compared to the viable cell counts obtained by culture. RESULTS: Using ATP bioluminescence, the antimicrobial activity of the tested mouthrinses was demonstrated when compared to the PBS control. The ATP bioluminescence values were significantly correlated (0.769, p<0.001) to the viable cell counts. CHX/CPC and AFSF showed similar antimicrobial activity, although AFSF had a less homogeneous effect, being both more effective than the EO rinse. CONCLUSION: ATP bioluminescence viability testing may be considered a useful tool to assess the in vitro efficacy of antibacterial compounds. In the proposed model, CHX/CPC and AFSF containing mouthrinses demonstrated superior antimicrobial activity, as compared to EO rinses, in a multispecies biofilm model.

Detection and quantification of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in bacteremia induced by interdental brushing in periodontally healthy and periodontitis patients.

OBJECTIVE: This study was aimed to compare the presence and amounts of bacteremia induced by interdental brushing in periodontally healthy (H) and periodontitis (P) individuals using culture based (direct culture [DC]) and molecular based techniques (real-time quantitative polymerase chain reaction [qPCR]) in a cross-sectional study model. MATERIALS AND METHODS: After a full mouth periodontal evaluation, blood samples were taken before and 1 min after professionally-administered interdental brushing. These samples were analyzed by DC and qPCR, targeting Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. Subgingival samples were also collected and analyzed. Student t-test, chi-squar tests and correlations were used for analyzing the data. RESULTS: Thirty individuals per group were included. P. gingivalis and A. actinomycetemcomitans were detected with qPCR methods, but not with DC. At baseline, bacteremia was observed in 5 P patients (16.7%) and in 2 H individuals (6.6%) (p = 0.421). After interdental brushing, bacteremia was only observed in 2 P patients (6.6%) (p = 0.901). A positive correlation between subgingival and blood levels of A. actinomycetemcomitans was observed (r = 0.3; p = 0.013). CONCLUSION: Bacteremia related to A. actinomycetemcomitans and P. gingivalis did not significantly increase after a single session of use of interdental brushes.

Detection and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis in blood samples with different microbiological identification methods: An in vitro study.

BACKGROUND: Culture-based methods (culture broth bottles or lysis methods) have been the standard for detecting bacteremia. More recently, quantitative polymerase chain reaction (qPCR) was proposed as a more sensitive and specific test although none of them has been validated for the identification of periodontal pathogens (fastidious growing bacteria) in blood samples. OBJECTIVE: To compare the ability to detect and quantify Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis (alone or in combination) in blood samples with three culture techniques [direct anaerobic culturing (DAC), haemo-culture (BACTEC), and lysis-centrifugation (LC)] and a non-culture dependent approach (qPCR) in an in vitro study. MATERIAL AND METHODS: Blood samples from 12 periodontally healthy volunteers were contaminated with three concentrations [104,102 and 101 colony forming units (CFU)/mL] of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination. Samples were analysed by DAC, BACTEC, LC and qPCR. Sensitivity, specificity, predictive values, kappa index and Lins correlation coefficients were calculated. RESULTS: DAC, LC and qPCR were able to detect the three target species at all concentrations. An excellent concordance (correlation coefficient r: 0.92-1) was observed between DAC and the reference standard (sensitivity raging 93.33-100% and specificity 88.89-100%) values. BACTEC was not able to identify P. gingivalis in any of the performed experiments. qPCR provided false negative results for S.oralis. CONCLUSIONS: DAC showed the best results for the proper identification and quantification of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination, in blood samples.

Quantification of periodontal pathogens in vascular, blood, and subgingival samples from patients with peripheral arterial disease or abdominal aortic aneurysms.

BACKGROUND: The aim of this investigation is to quantify periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, and Tannerella forsythia) in vascular, blood, and subgingival samples. As a secondary objective, two molecular bacterial identification methods (nested polymerase chain reaction [PCR] and quantitative PCR [qPCR]) are compared. METHODS: Seventy consecutive patients provided a vascular lesion, a blood sample, and 36 subgingival samples. Bacterial DNA was extracted, and qPCR was used to determine the prevalence and amounts of the target pathogens in each sample. Nested PCR was performed only in the samples from vascular lesions. Periodontal examination was performed in 42 patients. Mann-Whitney U or χ(2) tests were used to compare microbiologic results according to periodontal diagnosis. RESULTS: All targeted periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, T. forsythia, or C. rectus) were detected in subgingival samples, with a prevalence rate of 72.2%, 47.2%, 74.3%, and 82.9%, respectively. In 7.1% and 11.4% of vascular and blood samples, bacterial DNA was detected. One patient was positive for A. actinomycetemcomitans in the three types of samples. No differences were found in the levels of targeted bacteria when comparing patients with and without periodontitis. Prevalence rates obtained with nested PCR were significantly higher than those obtained with qPCR. CONCLUSIONS: The presence of A. actinomycetemcomitans was demonstrated in vascular, blood, and subgingival samples in one of 36 patients. These results, although with a very low frequency, may support the hypothesis of a translocation of periodontal pathogens from subgingival microbiota to the bloodstream and then to atheromatous plaques in carotid or other peripheral arteries. Nested PCR is not an adequate method for identifying DNA of periodontal pathogens in low quantities because of the high number of false-negative results.

Formação na Modalidade a Distância pela Universidade Aberta do SUS: Estudo Qualitativo sobre o Impacto do Curso na Prática Profissional / Education in Distance Mode Conducted by Open University of Brazilian Unique Health System (SUS): Qualitative Study about the Impact of the Course on Professional Practice

RESUMO Introdução As propostas da atual Política Nacional de Saúde impõem a necessidade de transformação nas formas de cuidado em saúde e, consequentemente, nas relações de trabalho da equipe para se obter uma visão ampliada sobre o processo saúde-doença. Existe, portanto, necessidade de instrumentalizar os profissionais da saúde nos aspectos técnicos, éticos e políticos para a transformação de seus processos de trabalho, sendo que para isso, vem sendo proposto pelo Ministério da Saúde cursos na modalidade a distância. Objetivo Compreender o impacto do Curso de Especialização em Saúde da Família desenvolvido na modalidade a distância na prática profissional de egressos. Método Estudo qualitativo realizado por meio de 24 entrevistas com egressos (11dentistas, oito enfermeiros e cinco médicos) de um curso de especialização em Saúde da Família na modalidade a distância realizado pela Universidade Aberta do Sistema Único de Saúde UNA-SUS em parceria com a Universidade Federal de São Paulo (Unifesp). A análise teve como direção os pressupostos do pensamento hermenêutico-dialético, que se insere na perspectiva interpretativa das metodologias qualitativas. Resultados Para os egressos, foi possível incorporar uma visão ampliada das necessidades de saúde da população e do estabelecimento de vínculo, o que é essencial ao processo de cuidado ampliado e inovador; reconhecer o papel e a importância dos diferentes profissionais da equipe, levando a novas formas de fazer o cuidado e a uma visão diferenciada sobre o sentido da Estratégia Saúde da Família; melhorar o processo de gestão por meio de decisões compartilhadas, organizar o processo de trabalho e considerar os dados epidemiológicos no planejamento das ações, o que envolve a gestão de pessoas, dos processos de trabalho e da informação. Conclusões Movimentos de Educação Permanente como este são essenciais à implantação efetiva dos princípios e diretrizes do SUS, uma vez que aproxima os profissionais de conceitos e práticas necessárias a um novo modo de agir e pensar em saúde.

ABSTRACT Introduction The proposals of the current National Health Policy imposes the need for transformation in the forms of health care, consequently, in team work behaviors to achieve a larger view on the health-disease process. Therefore, there is a need to train health professionals in the technical, ethical and political aspects to the transformation of work processes. Then it's has been proposed by the Ministry of health for courses in distance mode. Objective To understand the impact of The Specialization Course in Family Health Care developed in the distance mode (DE) in professional practice of graduates. Method qualitative study accomplished through 24 interviews with graduates (11 dentists, 08 nurses, 05 doctors) from a Specialization Course in Family Health Care developed in DE mode conducted by Open University of Brazilian Unique Health System UNA-SUS in collaboration with the Federal University of São Paulo (Unifesp). The analysis was directed by assumptions of hermeneutical thinking-dialectic, which is included in interpretative perspective of qualitative methodologies. Results For the graduates, it was possible to incorporate a broader view of the health needs of the population and the bond establishment, what is essential to the process and innovative care; recognition of the role and the importance of the different team members, leading to new ways to make care and a different vision about the meaning of the family health strategy; to improve the managing process through shared decisions, to organize the work process and to considerate the epidemiological data for the action planning, what involves managements of people, process and information. Conclusions Permanent Education Movements like this are essential for the effective implementation of the elements and guideline of SUS, this brings the necessary concepts and practices to a new way of acting and thinking about health.

A atenção à saúde do idoso: ações e perspectivas dos profissionais / Health attention to the elderly: actions and perspectives of professionals

O crescente aumento da população idosa e os complexos problemas de saúde que a envolvem tornam evidente a necessidade de atenção adequada à sua saúde, e espera-se que a Estratégia de Saúde da Família (ESF) possa trazer contribuições. O presente estudo propõe-se a analisar a percepção de profissionais que atuam na ESF quanto à saúde do idoso, as ações que vêm sendo desenvolvidas e as perspectivas dos profissionais visando à promoção, prevenção, cura e reabilitação das condições de saúde deles. Trata-se de um estudo qualitativo, realizado por meio de entrevistas com dentistas, enfermeiros e médicos que atuam na ESF. A análise dos resultados possibilitou a identificação de três categorias temáticas: 1. Falta de adesão ao cuidado e de apoio dos familiares; 2. Assistência centrada no aspecto curativo e no atendimento a demanda; 3. Vislumbre de mudança no modelo de atenção. Os dados revelam consonância entre as respostas dos entrevistados com o momento histórico e social das transformações que perpassam o modelo de atenção, levando a acreditar na ESF como um caminho de consolidação do Sistema Único de Saúde, desde que haja investimentos visando a instrumentalizar as equipes para atuar frente à nova lógica