Antimicrobial activity of violacein against oral bacteria associated with halitosis: an in vitro study

Introduction: violacein is a natural purple pigment produced by environmental bacteria that presents antimicrobial activity, particularly against Gram-positive bacteria. Intraoral halitosis (IOH) is a condition defined by the unpleasant odor emanating from the mouth, whose main source are volatile sulfur compounds, produced by Gram-negative oral bacteria on the tongue coating. In IOH treatment, antimicrobials have been indicated as chemical adjuncts, including natural products. Objective: thus, this study tested the antimicrobial activity of a violacein extract on key IOH-related bacteria (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, Prevotella intermedia, Solobacterium moorei). Materials and Methods: bacteria were cultured in fastidious anaerobe blood agar in anaerobiosis, and 109 cells/ml suspensions were plated. Crude extract of violacein obtained from Chromobacterium violaceum was diluted in a 25% ethanol aqueous solution to 8, 4, 2, 1, 0.5 and 0.25 mg/ml. Using the disk agar diffusion method, 10 µl aliquots of each dilution were deposited on the seeded plates. Chlorohexidine (0.1%) and 25% ethanol solution were used as controls. Plates were incubated in anaerobiosis at 37°C for 72h, and the inhibition halos were recorded. Results: although chlorhexidine showed higher inhibition halos than the violacein extract, most species were inhibited at 4 and 8 mg/ml concentrations (p<0.05). P. gingivalis followed by F. nucleatum were the most affected species in relation to the other bacteria, although statistical significance was only reached for P. gingivalis (p<0.05). Conclusion: crude violacein extract from C. violaceum demonstrated antimicrobial activity against IOH-associated oral bacteria, being a potential antimicrobial to be studied as an adjunct in the control of IOH.

Introdução: a violaceína é um pigmento roxo natural produzido por bactérias ambientais que apresenta ação antimicrobiana, particularmente contra bactérias Gram-positivas. A halitose intraoral (HIO) é uma condição definida pelo odor desagradável que emana da boca, cuja principal fonte são os compostos sulfurados voláteis produzidos por bactérias Gram-negativas da saburra lingual. No tratamento da HIO, antimicrobianos têm sido indicados como adjuvantes, incluindo produtos naturais. Objetivo: assim, este estudo avaliou o potencial antimicrobiano de um extrato de violaceína em patógenos-chave da HIO (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, Prevotella intermedia, Solobacterium moorei). Materiais e Métodos: bactérias foram cultivadas em meio ágar sangue para fastidiosos, em anaerobiose, e suspensões de 109 células/ml foram semeadas. O extrato bruto de violaceína obtido de Chromobacterium violaceum foi diluído em solução aquosa com 25% de etanol nas concentrações de 8, 4, 2, 1, 0,5 e 0,25 mg/ml. Através do método de disco difusão, 10 µl de cada diluição foram depositados nas placas semeadas. A clorexidina (0,1%) e a solução etanólica a 25% foram usadas como controles. As placas foram incubadas em anaerobiose a 37°C por 72h, e os halos de inibição foram registrados. Resultados: embora a clorexidina tenha apresentado os maiores halos de inibição do do que o extrato, a maioria das espécies foi inibida nas concentrações de 4 e 8 mg/ml (p<0,05). P. gingivalis e F. nucleatum foram as espécies mais afetadas em relação às outras bactérias, porém só foi observada significância estatística para P. gingivalis (p<0,05). Conclusão: o extrato bruto de violaceína de C. violaceum demonstrou atividade antimicrobiana contra bactérias orais associadas a HIO, sendo um potencial antimicrobiano a ser estudado como adjuvante no controle da HIO.

Colonization dynamics of subgingival microbiota in recently installed dental implants compared to healthy teeth in the same individual: a 6-month prospective observational study

Abstract Objectives To evaluate the colonization dynamics of subgingival microbiota established over six months around newly installed dental implants in periodontally healthy individuals, compared with their corresponding teeth. Methodology Seventeen healthy individuals assigned to receive single dental implants participated in the study. Subgingival biofilm was sampled from all implant sites and contralateral/ antagonist teeth on days 7, 30, 90, and 180 after implant installation. Microbiological analysis was performed using the Checkerboard DNA-DNA hybridization technique for detection of classical oral taxa and non-oral microorganisms. Significant differences were estimated by Mann-Whitney and Friedman tests, while associations between implants/teeth and target species levels were assessed by linear regression analysis (LRA). Significance level was set at 5%. Results Levels of some species were significantly higher in teeth compared to implants, respectively, at day 7 ( V.parvula , 6 × 10 5 vs 3 × 105 ; Milleri streptococci , 2 × 10 6 vs 6 × 10 5 ; Capnocytophaga spp., 2 × 10 6 vs 9 × 10 5 ; E.corrodens , 2 × 10 6 vs 5 × 10 5 ; N. mucosa , 2 × 10 6 vs 5 × 10 5 ; S.noxia , 2 × 10 6 vs 3 × 10 5 ; T.socranskii , 2 × 10 6 vs 5 × 10 5 ; H.alvei , 4 × 10 5 vs 2 × 10 5 ; and Neisseria spp., 6 × 10 5 vs 4 × 10 4 ), day 30 ( V.parvula , 5 × 10 5 vs 10 5 ; Capnocytophaga spp., 1.3 × 10 6 vs 6.8 × 10 4 ; F.periodonticum , 2 × 10 6 vs 10 6 ; S.noxia , 6 × 10 5 vs 2 × 10 5 ; H.alvei , 8 × 10 5 vs 9 × 10 4 ; and Neisseria spp., 2 × 10 5 vs 10 6 ), day 120 ( V.parvula , 8 × 10 5 vs 3 × 10 5 ; S.noxia , 2 × 10 6 vs 0; and T.socranskii , 3 × 10 5 vs 8 × 10 4 ), and day 180 ( S.enterica subsp. enterica serovar Typhi, 8 × 10 6 vs 2 × 10 6 ) (p<0.05). Implants showed significant increases over time in the levels of F.nucleatum , Gemella spp., H.pylori , P.micra , S.aureus , S.liquefaciens , and T.forsythia (p<0.05). LRA found that dental implants were negatively correlated with high levels of S. noxia and V. parvula (β=-0.5 to -0.3; p<0.05). Conclusions Early submucosal microbiota is diverse and only a few species differ between teeth and implants in the same individual. Only 7 days after implant installation, a rich microbiota can be found in the peri-implant site. After six months of evaluation, teeth and implants show similar prevalence and levels of the target species, including known and new periodontopathic species.

Subgingival microbiota and immune response modulation of osseointegrable implants / Modulação da microbiota subgengival e resposta imune dos implantes osseointegráveis

ABSTRACT Osseointegrable dental implants are biomaterials made of titanium or other alloys mixed with titanium, which have high biocompatibility and allow osseointegration. However, this process can be modulated by changes in the complex mechanisms between microbiota, immune response and host. The present study aims to present how the immune system-microbiota-host interaction influences the osseointegration process of titanium dental implants and its alloys. A literature review was performed through electronic and manual searches in several databases, including PubMed, LILACS, Google Scholar, SciELO and Web of Science for articles published in the last 20 years in English and Portuguese. The formation of a temporary fibrin matrix on the implants surface after implantation implies the recruitment, adhesion and activity of immune cells at this site, with the release of pro-inflammatory molecules and recruitment of neutrophils. In the second moment, monocytes and macrophages (M1) are recruited, producing, in this step, reactive oxygen species. In the later stage of inflammation, macrophages (M2) help in tissue regeneration with expression of anti-inflammatory cytokines. In addition, the implants surface provides a site for microbial colonization mediated by salivary pellicle and topographical features. Thus, in symbiosis the modulation of the immune response will be favorable to osseointegration. However, the dysbiotic process exacerbates the inflammatory progression modulating the immune response influencing abnormal tissue healing or scar and fibrosis formation, compromising osseointegration. Different conditions of the subgingival microbiota will influence different immunological cascades, generating different cellular responses and positive or negative modulation of the osseointegration process.

RESUMO Os implantes dentários osseointegráveis são biomateriais constituídos de titânio ou outras ligas misturadas com titânio, as quais possuem alta biocompatibilidade e permitem a osseointegração. Esse processo pode ser modulado por alterações nos mecanismos complexos entre microbiota, resposta-imune e hospedeiro. O presente estudo busca apresentar como a interação sistema imune-microbiota-hospedeiro influenciam no processo de osseointegração proveniente de implantes dentários de titânio. Foi realizada uma revisão de literatura através de busca eletrônica e manual em diversas bases de dados, incluindo PubMed, LILACS, Google Acadêmico, SciELO e Web of Science para artigos publicados nos últimos 20 anos em inglês e português. A formação de uma matriz provisória de fibrina na superfície dos implantes após a implantação implica no recrutamento, adesão e atividade das células imunes, com a liberação de moléculas pró-inflamatórias e recrutamento de neutrófilos. No segundo momento, monócitos e macrófagos (M1) são recrutados, produzindo espécies reativas a oxigênio. Já no estágio posterior da inflamação, macrófagos (M2) ajudam na regeneração do tecido com expressão de citocinas anti-inflamatórias. Além disso, a superfície dos implantes oferece um local para colonização microbiana mediada pela película salivar e características topográficas. Assim, em simbiose a modulação da resposta imune vai ser favorável à osseointegração. Contudo, em estado de doença periodontal, o processo disbiótico exacerba a progressão inflamatória modulando a resposta imune influindo em um processo cicatricial comprometendo a osseointegração. Diferentes condições da microbiota subgengival vão influenciar em cascatas imunológicas diferentes gerando respostas celulares diferentes e modulação positiva ou negativa do processo de osseointegração.

Periodontal disease severity is associated to pathogenic consortia comprising putative and candidate periodontal pathogens

Abstract Based on a holistic concept of polymicrobial etiology, we have hypothesized that putative and candidate periodontal pathogens are more frequently detected in consortia than alone in advanced forms of periodontal diseases (PD). Objective To correlate specific consortia of periodontal pathogens with clinical periodontal status and severity of periodontitis. Methodology Subgingival biofilm was obtained from individuals with periodontal health (113, PH), gingivitis (91, G), and periodontitis (209, P). Genomic DNA was purified and the species Aggregatibacter actinomycetemcomitans (Aa), Aa JP2-like strain, Porphyromonas gingivalis (Pg), Dialister pneumosintes (Dp), and Filifactor alocis (Fa) were detected by PCR. Configural frequency and logistic regression analyses were performed to correlate microbial consortia and PD. Results Aa + Pg in the presence of Dp (phi=0.240; χ2=11.9, p<0.01), as well as Aa JP2 + Dp + Fa (phi=0.186, χ2=4.6, p<0.05) were significantly more associated in advanced stages of P. The consortium Aa + Fa + Dp was strongly associated with deep pocketing and inflammation (p<0.001). The best predictors of disease severity (80% accuracy) included older age (OR 1.11 [95% CI 1.07 - 1.15], p<0.001), Black/African-American ancestry (OR 1.89 [95% CI 1.19 - 2.99], p=0.007), and high frequency of Aa + Pg + Dp (OR 3.04 [95% CI 1.49 - 6.22], p=0.002). Conclusion Specific microbial consortia of putative and novel periodontal pathogens, associated with demographic parameters, correlate with severe periodontitis, supporting the multifactorial nature of PD.

The role of dentistry in the hospital environment: the oral microbiota control as secondary infections prevention

Objective: The purpose of this review is to highlight the oral health care importance on patients in the hospital Intensive Care Unit. Sources of data: The review was performed within articles published in English and Portuguese in the past 11 years, and the search was performed on the following platforms: Google Scholar, PubMed (MEDLINE), Scielo, and Bireme (LILACS). Synthesis of data: When it comes to the hospital environment, especially intensive care units, the lack of dental care could compromise the well evolution of the hospitalized patient. The gap between what would be best proposed to the patient and what, indeed, is done by the current professionals in the intensive care units, highlights the necessity to study this subject more carefully so that there is a resumption of concepts about the expected management pattern, as well as how the multiprofessionalism develops in practice. If the structure cracks in any level, it's often seen the rise of diseases commonly associated with intensive care, such as ventilator-associated pneumonia. The importance of a dentist in the intensive care units could avoid these diseases or, at least, reduce them. Conclusion: From this same perspective, the presence of a dentist working in an integrated way with physicians, nurses, and nursing technicians is valuable, especially concerning oral hygiene for patients using a dental prosthesis.

Objetivo: Esta revisão busca destacar a importância dos cuidados com a saúde bucal em pacientes internados em Unidade de Terapia Intensiva hospitalar. Fonte dos dados: A revisão foi feita com artigos em inglês e português, nos últimos 11 anos, nas plataformas: Google Scholar, Pubmed (MEDLINE), Scielo e Bireme (LILACS). Síntese dos dados: No que se refere ao ambiente hospitalar, principalmente unidades de terapia intensiva, a falta de atendimento odontológico pode comprometer a boa evolução do paciente internado. A lacuna entre o que seria melhor proposto ao paciente e o que de fato é realizado pelos atuais profissionais das unidades de terapia intensiva evidencia a necessidade de estudar criteriosamente esse assunto, a fim de que haja uma retomada dos conceitos sobre o padrão de gestão esperado, bem como a forma como o multiprofissionalismo se desenvolve na prática. Se a estrutura sofrer ruptura em qualquer nível, geralmente ocorre o aumento de doenças comumente associadas à terapia intensiva, como a pneumonia associada ao ventilador. A importância do dentista nas unidades de terapia intensiva pode evitar essas doenças ou, pelo menos, reduzi-las. Conclusão: Sob essa mesma perspectiva, a presença do cirurgião-dentista atuando de forma integrada com médicos, enfermeiros e técnicos de enfermagem é valiosa, sobretudo no tocante a higiene bucal a portadores de prótese oral.

Nutritional influences on oral infections: the oral microbiota modulation

Introduction: The oral microbiota has approximately 700 symbiotic microorganisms responsible for oral health homeostasis. However, changes in oral microbiota can generate dysbiotic processes that favor the worsening of infections such as caries and periodontal disease. These oral infections, in addition to systemic involvement, can compromise the teeth integrity as well as oral health. Thus, inadequate nutrition proves to be a risk factor influencing the prevention and treatment of these oral infections. Objective: This study aims to evidence the nutritional influence on the oral microbiota modulation affecting, in the longterm, the gut microbiota, highlighting the use of probiotics and prebiotics in the treatment of oral infections by a literature review. Synthesis of data : Supplements of certain nutrients and the intake of an adequate diet in macronutrients and micronutrients directly influence nutritional status and consequently in the maintenance of oral-systemic homeostasis. In addition, due to microbial multidrug resistance, therapies using probiotics and prebiotics have been adopted as aids to the treatment of oral infections. Conclusion: Personalized Dentistry must integrate multidisciplinary knowledge of attention for health care. This in addition to knowingwhen to refer and work together with a Nutritionist.

Introdução: A microbiota oral possui aproximadamente 700 microrganismos simbiontes responsáveis pela homeostase da saúde bucal. Contudo, alterações na microbiota oral podem gerar processos disbióticos que favorecem o agravamento de infecções como a cárie e a doença periodontal. Essas infecções orais, além do acometimento sistêmico, podem comprometer a integridade dos dentes e também da saúde bucal. Dessa forma, a alimentação inadequada mostra-se um fator de risco que influência na prevenção e no tratamento dessas infecções orais. Objetivo: Este estudo visa evidenciar a influência nutricional na modulação da microbiota oral afetando a longo prazo a microbiota intestinal, destacando o uso de probióticos e prebióticos no tratamento de infecções orais através de uma revisão de literatura. Síntese de dados: A suplementação de determinados nutrientes e a ingestão de uma dieta adequada em macronutrientes e micronutrientes influenciam diretamente no estado nutricional e consequentemente na manutenção da homeostase oral-sistêmica. Além disso, devido à multirresistência microbiana, terapias com probióticos e prebióticos têm sido adotadas como auxiliares no tratamento de infecções orais. Conclusão: A Odontologia Personalizada deve integrar conhecimentos multidisciplinares de atenção à saúde. Isso além de saber quando encaminhar e trabalhar junto com o Nutricionista.

Long-term evaluation of the antimicrobial susceptibility and microbial profile of subgingival biofilms in individuals with aggressive periodontitis

This study evaluates the antimicrobial susceptibility and composition of subgingival biofilms in generalized aggressive periodontitis (GAP) patients treated using mechanical/antimicrobial therapies, including chlorhexidine (CHX), amoxicillin (AMX) and metronidazole (MET). GAP patients allocated to the placebo (C, n = 15) or test group (T, n = 16) received full-mouth disinfection with CHX, scaling and root planning, and systemic AMX (500 mg)/MET (250 mg) or placebos. Subgingival plaque samples were obtained at baseline, 3, 6, 9 and 12 months post-therapy from 3–4 periodontal pockets, and the samples were pooled and cultivated under anaerobic conditions. The minimum inhibitory concentrations (MICs) of AMX, MET and CHX were assessed using the microdilution method. Bacterial species present in the cultivated biofilm were identified by checkerboard DNA-DNA hybridization. At baseline, no differences in the MICs between groups were observed for the 3 antimicrobials. In the T group, significant increases in the MICs of CHX (p < 0.05) and AMX (p < 0.01) were detected during the first 3 months; however, the MIC of MET decreased at 12 months (p < 0.05). For several species, the MICs significantly changed over time in both groups, i.e., Streptococci MICs tended to increase, while for several periodontal pathogens, the MICs diminished. A transitory increase in the MIC of the subgingival biofilm to AMX and CHX was observed in GAP patients treated using enhanced mechanical therapy with topical CHX and systemic AMX/MET. Both protocols presented limited effects on the cultivable subgingival microbiota.