Differential analysis of culturable and unculturable subgingival target microorganisms according to the stages of periodontitis.

OBJECTIVES: Culturable and unculturable microorganisms have been associated with periodontitis. Their differential proportions and composition have not been evaluated by their severity and complexity defined by stages in the 2018 AAP-EEP classification. METHODS: One hundred eighty subgingival biofilm samples were collected in Spain and Colombia from subjects categorized as health/gingivitis: periodontitis stages I/II periodontitis stages III/IV. Target culturable microorganisms (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Treponema denticola, and Eubacterium nodatum) and target unculturable microorganisms (Filifactor alocis, Eubacterium saphenum, Eubacterium brachy, Desulfobulbus oralis) were evaluated by quantitative PCR analysis. In addition, their differences and association with periodontal status were analyzed by ANCOVA and logistic regression models once adjusted to age, current smoking, and country. RESULTS: P. gingivalis was significantly associated with periodontitis stages I/II, OR 2.44 (CI 95% 1.08-5.47) and stages III/V, OR 6.43 (CI 95% 2.43-16.9). T forsythia, OR 7.53 (CI 95% 2.07-27.4); D. oralis, OR 5.99 (CI 95% 2.71-13.23); F. alocis, OR 10.9 (CI 95% 4.56-23.2); E. brachy, 3.57 (CI 95% 1.40-9.11); and E. saphenum, 4.85 (CI 95% 1.99-11.7) were significantly associated only with stages III/IV periodontitis. P. gingivalis evidenced significant differences with the increase in the severity of the periodontal lesion: 2.97 colony forming unit (CFU)/µL (CI 95% 2.32-3.54) health/gingivitis, and 4.66 CFU/µL (CI 95% 4.03-5.30) and 5.90 CFU/µL (CI 95% 5.20-6.48) in stages I/II and III/IV respectively (p < 0.0001). Unculturable microorganisms only evidenced differences in concentration in stages III/IV compared with health-gingivitis (p ≤ 0.001). CONCLUSION: Culturable and unculturable are strongly associated with stages III/IV periodontitis. Classic culturable microorganisms are more sensitive to differentiate between stages of periodontitis in the quantitative analysis. CLINICAL RELEVANCE: Future interventional studies of periodontal disease should include Filifactor alocis, Eubacterium saphenum, Eubacterium brachy, and Desulfobulbus oralis as possible markers of therapy response and as indicators of progressive disease.

Porphyromonas gingivalis Placental Atopobiosis and Inflammatory Responses in Women With Adverse Pregnancy Outcomes.

The microbiome modulates inflammation at the fetal maternal interface on both term and preterm labor. Inflammophilic oral bacteria, such as Porphyromonas gingivalis, as well as urogenital microorganisms (UGM) could translocate to the placenta and activate immune mechanisms in decidual tissue that is associated with adverse pregnancy outcomes (APO). This study establishes the associations between the presence of microbes in the placenta and placental cytokine patterns in women who presented APO, e.g., low birth weight (LBW), preterm premature rupture of membranes (PPROM), preterm birth (PTB) and other clinical signs related to Chorioamnionitis (CA). A total of 40 pregnant women were included in the study and divided into five groups according to placental infection (PI) and APO, as follows: (1) women without PI and without APO (n = 17), (2) women with P. gingivalis-related PI and APO (n = 5), (3) women with P. gingivalis-related PI and without APO (n = 4), (4) women with PI related to UGM and APO (n = 5) and (5) women without PI with APO (n = 9). Obstetric, clinical periodontal status evaluation, and subgingival plaque sampling were performed at the time of delivery. Placental levels of interleukin IL-1ß, IL-6, IL-10, IL-15, IL-17A, IL-17F, IL-21, IL-12p70, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 α (MCP-1α), granzyme B, and interferon-γ (IFN-γ) were determined using a multiplex flow cytometry assay. All patients showed a predominant Th-1 cytokine profile related to labor, characterized by IFN-γ overexpression. The analysis by groups suggests that Th-1 profile was trending to maintain cytotoxic cell activity by the expression of IL-15 and granzyme B, except for the group with P. gingivalis-related PI and APO, which exhibited a reduction of IL-10 and IL-17F cytokines (p < 0.05) and a Th-1 profile favoring macrophage activation by MCP-1 production (p < 0.05). This study confirms a pro-inflammatory pattern associated with labor, characterized by a Th-1 profile and the activity of cytotoxic cells, which is enhanced by PI with UGM. However, PI associated with P. gingivalis suggests a switch where the Th-1 profile favors an inflammatory response mediated by MCP-1 and macrophage activity as a mechanistic explanation of its possible relationship with adverse outcomes in pregnancy.

Periodontal condition is associated with adverse perinatal outcomes and premature rupture of membranes in low-income pregnant women in Bogota, Colombia: a case-control study.

Objectives: To assess the periodontal condition as a factor associated with adverse perinatal outcomes, premature rupture of membranes (PRM), and preeclampsia in low-income pregnant women treated at public hospitals in Bogotá, Colombia.Methods: Pregnant women with preterm birth (PTB) and low birth weight (LBW) or both conditions (n = 107/428), or only PTB (n = 73/292) or LBW (n = 74/296) or with PRM (n = 98/392) or preeclampsia (n = 76/304) in a ratio of four controls for each case, coming from three hospitals of the public Northern Network of Bogotá, Colombia were studied. Sociodemographic, perinatal adverse outcome history, antenatal care, chronic infections, periodontal condition, threatened abortion, bleeding in the second half of pregnancy, oligohydramnios, diabetes, gestational diabetes, alcohol consumption, hypertension, smoking, alcohol during pregnancy were determined. Logistic regression was conducted to establish factors associated to perinatal adverse outcomes. Multiple correspondence analysis was conducted as secondary analysis.Results: Threatened abortion, absence of antenatal care, hypertension, chronic infections, and periodontal condition were the most important factors associated with perinatal adverse outcomes. The presence of periodontal pockets was associated with LBW OR 2.52 (IC95% 1.36-4.70), PTB OR 2.04 (IC95% 1.10-3.64), PTB-LBW or both OR 2.08 (IC95% 1.18-3.31), PRM OR 2.04 (IC95% 1.17-3.56). Periodontal pockets presence was not associated with preeclampsia. Multiple correspondence analyses showed high correlation between PRM with chronic infection and presence of periodontal pockets.Conclusions: Periodontal condition is a factor independent of other important risk factors for a perinatal adverse outcome and PRM. Prevention of periodontal disease should be included in preconception and prenatal care programs.

Microbiome and Microbial Biofilm Profiles of Peri-Implantitis: A Systematic Review.

BACKGROUND: This systematic review assesses microbiologic profiles of peri-implantitis, periodontitis, and healthy implants based on studies that evaluated microbial biofilms and entire microbiomes to establish their similarities and differences. METHODS: The Medical Literature Analysis and Retrieval System Online via PubMed, Excerpta Medica Database, and Cochrane Central Register of Controlled Trials, were searched without language restrictions through July 30, 2016. Observational studies that evaluated microbial profiles or entire microbiomes of peri-implantitis compared with healthy implants or periodontitis were considered eligible for inclusion. A descriptive summary was created to determine quantity of data and interstudy variations. RESULTS: Of 126 potentially eligible articles, 26 were included in this study. Twenty-one of these articles evaluated the microbiologic profile of peri-implantitis versus healthy implants or periodontitis using conventional microbiologic techniques. Five articles evaluated the entire microbiome using genomic sequencing. Teeth with periodontitis, healthy implants, or implants with peri-implantitis were colonized by periodontal microorganisms. Porphyromonas gingivalis and especially Prevotella intermedius/nigrescens were often identified at peri-implantitis sites. Peri-implantitis sites were also colonized by uncultivable asaccharolytic anaerobic Gram-positive rods and anaerobic Gram-negative rods, which were not frequently identified in teeth with periodontitis or healthy implants. Opportunistic microorganisms were not found very frequently in peri-implantitis sites. CONCLUSIONS: Peri-implantitis represents a heterogeneous mixed infection that includes periodontopathic microorganisms, uncultivable asaccharolytic anaerobic Gram-positive rods and other uncultivable Gram-negative rods, and, rarely, opportunistic microorganisms such as enteric rods and Staphylococcus aureus. Sequencing methods that evaluate the entire microbiome improve identification of microorganisms associated with peri-implantitis.