Talita Gomes Baêta Lourenço

In the last decades, groundbreaking research on the human microbiome has changed our reductionist conception of the etiology and pathogenesis of several chronic diseases. As a result, we have come to appreciate the significance of a balanced microbiome in maintaining human health. In this context, the upper and lower gastrointestinal tracts (GITs) comprise the most abundant and diverse microbiotas of the human body. In addition to its diversity, functional redundancy, and temporal stability, a healthy GIT microbiome is characterized by its body site specificity. In fact, current evidence has indicated that the translocation of oral species to the gut environment through the oral–gut axis is increased in an array of illnesses, including chronic inflammatory and metabolic diseases, neurological disorders, and cancer. Oral pathogens have also been shown to promote gut dysbiosis and systemic inflammation in animal models. Yet, some level of overlapping between oral and gut microbiomes may occur without disruption of these microbial communities and loss of site specificity. The uniqueness of each host–microbiome entity may hinder our ability to define a “universal” normal GIT microbiome. Despite that, this chapter summarizes the predominant health-related taxa along the human GIT, as well as their role in the physiology and immunity of the digestive system. Some mechanisms that may lead to disturbances and relevant shifts in the oral and gut microbiomes of major inflammatory chronic diseases are also pointed out. Lastly, oral–fecal microbial signatures are presented as potential biomarkers for several oral and systemic disorders. The recognition of such symbiotic/dysbiotic microbial profiles may provide insights into the development of more accurate early diagnosis and therapeutic ecological approaches to restore the balance of the GIT microbiome.

Aim: Evidence suggests that translocation of oral pathogens through the oral–gut axis may induce intestinal dysbiosis. This study aimed to evaluate the impact of a highly leukotoxic Aggregatibacter actinomycetemcomitans (Aa) strain on the gut microbiota, intestinal mucosal integrity and immune system in healthy mice.Methods: Eight‐week‐old male C57BL6 mice were divided into control (n = 16) and JP2 groups (n = 19), which received intragastric gavage with PBS and with a suspension of Aa JP2 (HK921), respectively, twice a week for 4 weeks. Colonic lamina propria, fecal material, serum, gingival tissues, and mandibles were obtained for analyses of leukocyte populations, inflammatory mediators, mucosal integrity, alveolar bone loss, and gut microbiota. Differences between groups for these parameters were examined by non‐parametric tests.Results: The gut microbial richness and the number of colonic macrophages, neutrophils, and monocytes were significantly lower in Aa JP2‐infected mice than in controls (p < .05). In contrast, infected animals showed higher abundance of Clostridiaceae, Lactobacillus taiwanensis, Helicobacter rodentium, higher levels of IL‐6 expression in colonic tissues, and higher splenic MPO activity than controls (p < .05). No differences in tight junction expression, serum endotoxin levels, and colonic inflammatory cytokines were observed between groups. Infected animals presented also slightly more alveolar bone loss and gingival IL‐6 levels than controls (p < .05).Conclusion: Based on this model, intragastric administration of Aa JP2 is associated with changes in the gut ecosystem of healthy hosts, characterized by less live/recruited myeloid cells, enrichment of the gut microbiota with pathobionts and decrease in commensals. Negligible levels of colonic pro‐inflammatory cytokines, and no signs of mucosal barrier disruption were related to these changes.

The dysbiotic biofilm of periodontitis may function as a reservoir for opportunistic human pathogens of clinical relevance. This study explored the virulence and antimicrobial susceptibility of staphylococci isolated from the subgingival biofilm of individuals with different periodontal conditions. Subgingival biofilm was obtained from 142 individuals with periodontal health, 101 with gingivitis and 302 with periodontitis, and cultivated on selective media. Isolated strains were identified by mass spectrometry. Antimicrobial susceptibility was determined by disk diffusion. The mecA and virulence genes were surveyed by PCR. Differences among groups regarding species, virulence and antimicrobial resistance were examined by Chi-square, Kruskal–Wallis or Mann–Whitney tests. The overall prevalence of subgingival staphylococci was 46%, especially in severe periodontitis (> 60%; p < 0.01). S. epidermidis (59%) and S. aureus (22%) were the predominant species across groups. S. condimenti, S. hominis, S. simulans and S. xylosus were identified only in periodontitis. High rates of resistance/reduced sensitivity were found for penicillin (60%), amoxicillin (55%) and azithromycin (37%), but multidrug resistance was observed in 12% of the isolates. Over 70% of the mecA + strains in periodontitis were isolated from severe disease. Higher detection rates of fnB + isolates were observed in periodontitis compared to health and gingivitis, whereas luxF/luxS-pvl + strains were associated with sites with deep pockets and attachment loss (p < 0.05). Penicillin-resistant staphylococci is highly prevalent in the subgingival biofilm regardless of the periodontal status. Strains carrying virulence genes related to tissue adhesion/invasion, inflammation and cytotoxicity support the pathogenic potential of these opportunists in the periodontal microenvironment.

Antarctic camps pose psychophysiological challenges related to isolated, confined, and extreme (ICE) conditions, including meals composed of sealed food. ICE conditions can influence the microbiome and inflammatory responses. Seven expeditioners took part in a 7-week Antarctic summer camp (Nelson Island) and were evaluated at Pre-Camp (i.e., at the beginning of the ship travel), Camp-Initial (i.e., 4th and 5th day in camp), Camp-Middle (i.e., 19th–20th, and 33rd–34th days), Camp-Final (i.e., 45th–46th day), and at the Post-Camp (on the ship). At the Pre-Camp, Camp-Initial, and Camp-Final, we assessed microbiome and inflammatory markers. Catecholamines were accessed Pre- and Post-Camp. Heart rate variability (HRV), leptin, thyroid stimulating hormone (TSH), and thyroxine (T4) were accessed at all time points. Students’ t-tests or repeated-measures analysis of variance (one or two-way ANOVA) followed by Student-Newman-Keuls (post hoc) were used for parametric analysis. Kruskal-Wallis test was applied for non-parametric analysis. Microbiome analysis showed a predominance of Pseudomonadota (34.01%), Bacillota (29.82%), and Bacteroidota (18.54%), followed by Actinomycetota (5.85%), and Fusobacteria (5.74%). Staying in a long-term Antarctic camp resulted in microbiome fluctuations with a reduction in Pseudomonadota—a “microbial signature” of disease. However, the pro-inflammatory marker leptin and IL-8 tended to increase, and the angiogenic factor VEGF was reduced during camp. These results suggest that distinct Antarctic natural environments and behavioral factors modulate oral microbiome and inflammation.

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.

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.

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). To correlate specific consortia of periodontal pathogens with clinical periodontal status and severity of periodontitis. 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. 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). Specific microbial consortia of putative and novel periodontal pathogens, associated with demographic parameters, correlate with severe periodontitis, supporting the multifactorial nature of PD.

Objective: This investigation explored oral‐gut microbial signatures with potential to distinguish among periodontal conditions.Background Data: The interplay between the oral and gut microbiomes may be a critical pathway linking periodontal diseases and systemic inflammatory disorders. The mechanisms by which oral microorganisms translocate to the gut and cause microbial dysbiosis, favoring an inflammatory state, are still unknown. As a first approach, characterization of oral‐gut microbial profiles associated with periodontal health and diseases can provide insights on such mechanisms of etiology and pathogenesis.Methods: Fecal and saliva samples from individuals with periodontal health (PH, 8), gingivitis (GG, 17), and periodontitis (PD, 24) were analyzed for their microbial composition by 16S rRNA gene sequencing. Microbial taxa were compared and correlated to periodontal parameters. Multivariate discriminant analysis (MDA) was carried out to identify profiles related to health and disease.Results: Few significant differences in oral‐gut taxa were detected among clinical groups, although increase in fecal Fusobacterium nucleatum ss vincentii and salivary Aggregatibacter actinomycetemcomitans, Parvimonas micra, and Fretibacterium sp. HMT358 were strongly correlated with deep pockets and inflammation (p < .01). Over 50% of the fecal microbiota comprised microorganisms shared between oral and gut sites, whereas oral taxa were detected in approximately 9%, particularly enriched in GG fecal samples (p = .04). Trends for lower fecal richness and higher salivary diversity in PD compared to PH were observed. MDA was able to classify correctly 82% of the patients into the clinical groups. Main classifiers of periodontitis were high BMI, older age, and enrichment of oral‐fecal Leptotrichia sp. HMT4, Peptostreptococcus stomatis, Dialister invisus, and a novel Lautropia sp. HMTC89‐like organism.Conclusion: Within the limitations of an exploratory investigation, specific profiles of oral‐gut taxa, including known and potential novel organisms, combined with social‐demographic features were able to discriminate individuals with periodontal diseases in this study population.

The oral microbiota under health conditions is in balance between host and microorganisms, but several factors can lead to an imbalance in this relationship, associated with the development and progression of oral diseases, such as periodontal diseases (PD). The mechanical and chemical control of dental biofilm is the main method of prevention and treatment of these diseases, often combined with the use of antimicrobials. Considering the current concern about the indiscriminate use of antimicrobials and the dysbiotic nature of PD, preventive and therapeutic strategies aiming at the ecological balance of the oral microbiota have been explored, among them, the use of probiotics. However, little is known about their real effectiveness in the treatment of oral infections. The aim of this study was to review the literature on the use of probiotics as an adjunct to the treatment of PD. A bibliographic search was carried out in the Scientific Electronic Library Online (SciELO), Virtual Health Library (BVS), US National library of Medicine National Institutes of Health (PubMed) and Google Scholar databases, with the descriptors "Periodontal Disease" and " Probiotics”, from clinical studies conducted between 2011 and 2021. The search resulted in 49 clinical studies and, after a complete reading, 33 were selected, meeting the inclusion criteria. In general, the adjuvant use of probiotics seems to bring some additional clinical benefit to the treatment, and this effect can also be observed in microbiological parameters. However, treatment protocols and outcomes observed in studies are extremely variable, making it difficult to determine the best protocol for administering probiotics.

Background: The oral‐gut axis may be a route linking periodontal and systemic diseases. Probiotics could be an alternative for the treatment of microbial dysbiotic conditions, including periodontitis. This randomized placebo‐controlled clinical trial evaluated the short‐term efficacy of systemic probiotics adjunctive to subgingival instrumentation (SI) in promoting a better restoration of the oral‐gut microbiotas and greater periodontal clinical outcome.Methods: Systemically healthy adults with untreated periodontitis were recruited from a Dental School setting and allocated to receive SI plus placebo (n = 24) or probiotics (n = 24), one capsule/day for 30 days. Subgingival biofilm and stool were obtained at baseline and 2‐months post‐therapy for microbiological analyses by checkerboard and 16S rRNA gene sequencing. Differences in all parameters between placebo (n = 23) and probiotics (n = 19) groups were assessed by non‐parametric tests.Results: Most subgingival species and α‐diversity decreased after therapies (P P P = 0.07). Strong correlations between oral and fecal species were found (P P P <0.05).Conclusion: Systemic probiotics combined with SI did not provide short‐term additional clinical or microbiological benefits in the treatment of periodontitis; however, response to therapies seemed to correlate with distinct oral‐gut microbial profiles.