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Aim: To compare the subgingival microbiota of patients receiving supportive periodontal care (SPC) with and without subgingival instrumentation, over 2 years., Materials and Methods: This study was a randomized clinical trial that included 62 participants (50.97 ± 9.26 years old; 40 females) who completed non-surgical periodontal therapy. Participants were randomly assigned to receive oral prophylaxis with oral hygiene instructions alone (test) or in combination with subgingival instrumentation (control) during SPC. Pooled subgingival biofilm samples were obtained from four sites per patient at SPC baseline and at 3, 6, 12, 18, and 24 months. Real-time polymerase chain reaction was used for absolute quantification of Eubacteria and the target bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Data were analysed using generalized estimating equations, taking into consideration the clustering of observations within individuals., Results: No significant differences were found between the experimental groups regarding the mean counts of Eubacteria and target bacteria, as well as the periodontal parameters at the sampled sites. Although significant variability in bacterial counts was present during SPC, all counts after 2 years were not statistically different from those at baseline. Bacterial counts were associated with the presence of plaque, bleeding on probing, mean probing depth ≥3 mm, and follow-up period., Conclusions: SPC with or without subgingival instrumentation can result in comparable subgingival microbiological outcomes., Clinical Trial Registration: clinicaltrials.gov: NCT01598155 (https://clinicaltrials.gov/study/NCT01598155?intr=supragingival%20control&rank=4#study-record-dates)., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: This study aims to analyse the association between the baseline microbial load of selected periodontopathogenic bacteria collected from gingival crevicular fluid (GCF) and the primary outcome of steps I and II therapy., Materials and Methods: 222 patients with stage III periodontitis were included into this retrospective analysis that received steps 1 and 2 periodontal therapy without adjunctive systemic antibiotics. Baseline GCF samples were quantitatively analysed using ELISA-based kits for levels of periodontopathogens (Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Treponema denticola (Td), and Tannerella forsythia (Tf)) and associated with the primary therapy outcome using a "treat-to-target" therapy endpoint (TE) defined as ≤ 4 sites with PD ≥ 5 mm six months after therapy., Results: 38.2% of the patients achieved TE. Patients failing to achieve TE revealed significantly increased levels of Pg, Fn, and Tf at baseline (Pg: p = 0.010, Fn: p = 0.008 Tf: p = 0.004). Multivariate binary logistic regression adjusted for sex, mean probing depth, diabetes, and current smoking status showed an independent relationship between Tf and the TE (aOR 2.570, p = 0.023)., Conclusion: Increased microbial load is associated with decreased responsiveness to therapy. The findings suggest that specifically baseline Tf levels are associated with poorer treatment outcomes and might improve the accuracy of periodontal diagnosis., Clinical Relevance: The findings of this study support the concept of a critical biomass that is sufficient to induce and maintain an immune response within the periodontal pocket, which ultimately leads to irreversible tissue destruction. However, calculating this level in advance may serve as an early indicator for intervention., Key Finding: Baseline Tannerella forsythia levels are associated with poorer treatment outcome., (© 2024. The Author(s).)

The most common type of periodontal disease is chronic periodontitis, an inflammatory condition caused by pathogenic bacteria in subgingival plaque. The aim of our study was the development of a real-time PCR test as a diagnostic tool for the detection and differentiation of five periodontopathogenic bacteria, Aggregatibacter actinomycetemcomitans , Porphyromonas gingivalis , Tannerella forsythia , Prevotella intermedia , and Treponema denticola , in patients with periodontitis. We compared the results of our in-house method with the micro-IDent ® semiquantitative commercially available test based on the PCR hybridization method. DNA was isolated from subgingival plaque samples taken from 50 patients and then analyzed by both methods. Comparing the results of the two methods, they show a specificity of 100% for all bacteria. The sensitivity for A. actinomycetemcomitans was 97.5%, for P. gingivalis 96.88%, and for P. intermedia 95.24%. The sensitivity for Tannerella forsythia and T. denticola was 100%. The Spearman correlation factor of two different measurements was 0.976 for A. actinomycetemcomitans , 0.967 for P. gingivalis , 0.949 for P. intermedia , 0.966 for Tannerella forsythia , and 0.917 for T. denticola . In conclusion, the in-house real-time PCR method developed in our laboratory can provide information about relative amount of five bacterial species present in subgingival plaque in patients with periodontitis. It is likely that such a test could be used in dental diagnostics in assessing the efficacy of any treatment to reduce the bacterial burden.

Objectives: Periodontitis is an inflammatory disease caused by bacteria present in the dental biofilm. However, the presence of two oral protozoans, Entamoeba gingivalis and Trichomonas tenax, in patients with the periodontal disease remains largely unknown in Taiwan. Therefore, we investigated the prevalence of oral microbial infections between the sites with mild gingivitis and chronic periodontitis in patients., Materials and Methods: We collected 60 dental biofilm samples from sites with mild gingivitis (probing depth <5 mm) and chronic periodontitis (probing depth ≥5 mm) from 30 patients at the National Cheng Kung University Hospital. The samples were analyzed via polymerase chain reaction and gel electrophoresis., Results: Among oral protozoans, E. gingivalis and T. tenax were detected in 44 (74.07%) and 14 (23.33%) of all samples, respectively. Among oral bacteria, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia were detected in 50 (83.33%), 47 (78.33%), and 48 (80.0%) samples, respectively., Conclusions: This study, which is the first to analyze E. gingivalis and T. tenax presence among patients with periodontitis in Taiwan, revealed an association between periodontitis and oral microbes., (© 2023 Wiley Periodicals LLC.)

Background/aim: Scaling and root planing remain inadequate in periodontitis treatment caused by dysbiotic microbial dental plaque. The aim of this clinical trial is to evaluate the effects of probiotics and kefir consumption in initial periodontal therapy (IPT) on oral microbiota composition and treatment outcomes in patients with periodontitis., Materials and Methods: The study was carried out in the Gazi University Department of Periodontology, including a sample size of 36 individuals and utilizing a randomized controlled design. Thirty-six patients with periodontitis were randomly allocated to three groups: one receiving probiotic treatment, another receiving kefir, and a third serving as the control group. Obtaining subgingival microbial samples, we recorded plaque, gingival index, bleeding on probing, periodontal pocket depth, and clinical attachment level (periodontal clinical indices) and then performed IPT. For 14 days, patients took either probiotics, kefir, or no supplements. Data for the first and third months were collected using periodontal clinical indices. DNA sequencing was performed to detect Tannerella forsythia , Porphyromonas gingivalis , and Treponema denticola in subgingival plaque samples collected at baseline and three months., Results: Significant differences were observed regarding periodontal clinical indices among groups in the intragroup comparisons. Moreover, levels of Tannerella forsythia were significantly decreased in all groups., Conclusion: Kefir can be administered in addition to IPT, providing results similar to those observed with probiotics., (© TÜBİTAK.)

Periodontitis, an inflammatory disease of the oral cavity, was caused by microbes from bacteria to protozoa. In this study, we detected protozoa, Entamoeba gingivalis and other three common pathogenic bacteria, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia by the Polymerase chain reaction (PCR) on patients., Competing Interests: Declaration of Competing Interest All authors declare no conflicts of interest., (Copyright © 2020. Published by Elsevier B.V.)

Background: High levels of periodontopathic bacteria as well as Streptococcus anginosus were detected in cancer tissue from patients with esophageal cancer. An association between oral infectious bacteria and esophageal cancer has been reported., Methods: Characteristics of the oral microbiota and periodontal conditions were studied as clinicopathologic factors in patients with esophageal cancer. The study included 61 patients with esophageal cancer and 62 matched individuals without any cancers. Samples of subgingival dental plaque and unstimulated saliva were collected to evaluate the prevalence and abundance of the following oral bacteria using a real-time polymerase chain reaction assay: Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Treponema denticola, and S. anginosus., Results: In the cancer group, the prevalence of all bacteria, with the exception of F. nucleatum, in dental plaque; the prevalence of A. actinomycetemcomitans in saliva; the abundance of all bacteria, with the exception of F. nucleatum and P. intermedia, in dental plaque; and the abundance of A. actinomycetemcomitans and S. anginosus in saliva were significantly higher. Furthermore, a logistic regression analysis suggested that the prevalence of T. forsythia and S. anginosus in dental plaque and of A. actinomycetemcomitans in saliva, as well as a drinking habit, were associated with a high risk of esophageal cancer, with a high odds ratio., Conclusions: The current findings have potential implications for the early diagnosis of esophageal cancer., (© 2020 American Cancer Society.)

Utilizing the pathology and microbiology found in tissue from patients with documented Alzheimer's disease (AD), the pathogenesis of this fateful disorder has been made clear. Borrelia burgdorferi and Treponema denticola spirochetes enter the brain, mostly via neuronal pathways and the entorhinal circulation. These organisms easily pass through the blood-brain barrier and have an affinity for neural tissue. Once in the brain, the spirochetes make intra- and extracellular biofilms, and it is the biofilms that create the pathology. Specifically, it is the intracellular biofilms that are ultimately responsible for neurofibrillary tangles and dendritic disintegration. The extracellular biofilms are responsible for the inflammation that initially is generated by the first responder, Toll-like receptor 2. The hypothesis that arises from this work is two-pronged: one is related to prevention; the other to treatment. Regarding prevention, it is very likely possible that AD could be prevented by periodic administration of penicillin (PCN), which would kill the spirochetes before they made biofilms; this would prevent the disease and would not allow any of the above deleterious changes generated by the biofilms to occur. As regards treatment, it may be possible to slow or prevent further decline in early AD by administration of PCN together with a biofilm disperser. The disperser would disrupt the biofilm coating and enable the PCN to kill the spirochetes. This protocol could be administered in a trial with the control arm utilizing the current treatment. The progress of the treatment could be evaluated by one of the current blood tests that is semi-quantitative. The specific protocols are listed.

Epigenetic mechanisms, namely DNA and histone modifications, are critical regulators of immunity and inflammation which have emerged as potential targets for immunomodulating therapies. The prevalence and significant morbidity of periodontitis, in combination with accumulating evidence that genetic, environmental and lifestyle factors cannot fully explain the susceptibility of individuals to disease development, have driven interest in epigenetic regulation as an important factor in periodontitis pathogenesis. Aberrant promoter methylation profiles of genes involved in inflammatory activation, including TLR2, PTGS2, IFNG, IL6, IL8, and TNF, have been observed in the gingival tissue, peripheral blood or buccal mucosa from patients with periodontitis, correlating with changes in expression and disease severity. The expression of enzymes that regulate histone acetylation, in particular histone deacetylases (HDACs), is also dysregulated in periodontitis-affected gingival tissue. Infection of gingival epithelial cells, gingival fibroblasts and periodontal ligament cells with the oral pathogens Porphyromonas gingivalis or Treponema denticola induces alterations in expression and activity of chromatin-modifying enzymes, as well as site-specific and global changes in DNA methylation profiles and in histone acetylation and methylation marks. These epigenetic changes are associated with excessive production of inflammatory cytokines, chemokines, and matrix-degrading enzymes that can be suppressed by small molecule inhibitors of HDACs (HDACi) or DNA methyltransferases. HDACi and inhibitors of bromodomain-containing BET proteins ameliorate inflammation, osteoclastogenesis, and alveolar bone resorption in animal models of periodontitis, suggesting their clinical potential as host modulation therapeutic agents. However, broader application of epigenomic methods will be required to create a comprehensive map of epigenetic changes in periodontitis. The integration of functional studies with global analyses of the epigenetic landscape will provide critical information on the therapeutic and diagnostic potential of epigenetics in periodontal disease.

Background: Periodontal bacteria is the major pathogens in the oral cavity and the main cause of adult chronic periodontitis, but their association with incidence and prognosis in cancer is controversial. The aim of this study was to evaluate the effect of periodontal bacteria infection on incidence and prognosis of cancer., Methods: A systematic literature search of PubMed, Embase, Web of Science, and Cochrane Library databases was performed to obtain 39 studies comprising 7184 participants. The incidence of cancer was evaluated as odd ratios (OR) with a 95% confidence interval (95% CI) using Review Manager 5.2 software. Overall survival, cancer-specific survival and disease-free survival, which were measured as hazard ratios (HR) with a 95% CI using Review Manager 5.2 software., Results: Our results indicated that periodontal bacteria infection increased the incidence of cancer (OR = 1.25; 95%CI: 1.03-1.52) and was associated with poor overall survival (HR = 1.75; 95% CI: 1.40-2.20), disease-free survival (HR = 2.18; 95%CI: 1.24-3.84) and cancer-specific survival (HR = 1.85, 95%CI: 1.44-2.39). Subgroup analysis indicted that the risk of cancer was associated with Porphyromonas gingivalis (Pg) infection (OR = 2.16; 95%CI: 1.34-3.47) and Prevotella intermedia (Pi) infection (OR = 1.28; 95%CI: 1.01-1.63) but not Tannerella forsythia (Tf) (OR = 1.06; 95%CI: 0.8-1.41), Treponema denticola (Td) (OR = 1.30; 95%CI: 0.99-1.72), Aggregatibacter actinomycetemcomitans (Aa) (OR = 1.00; 95%CI: 0.48-2.08) and Fusobacterium nucleatum (Fn) (OR = 0.61; 95%CI: 0.32-1.16)., Conclusion: This meta-analysis revealed periodontal bacteria infection increased the incidence of cancer and predicted poor prognosis of cancer.

Periodontal disease, the most prevalent infectious disease in the world, is caused by biofilms formed in periodontal pockets. No specific bacterial species that can cause periodontitis alone has been found in any study to date. Several periodontopathic bacteria are associated with the progress of periodontal disease. Consequently, it is hypothesized that dysbiosis of subgingival microbiota may be a cause of periodontal disease. This study aimed to investigate the relationship between the subgingival microbiota and the clinical status of periodontal pockets in a quantitative and clinically applicable way with the newly developed Oral Care Chip. The Oral Care Chip is a DNA microarray tool with improved quantitative performance, that can be used in combination with competitive PCR to quantitatively detect 17 species of subgingival bacteria. Cluster analysis based on the similarity of each bacterial quantity was performed on 204 subgingival plaque samples collected from periodontitis patients and healthy volunteers. A significant difference in the number of total bacteria, Treponema denticola, Campylobacter rectus, Fusobacterium nucleatum, and Streptococcus intermedia bacteria in any combination of the three clusters indicated that these bacteria gradually increased in number from the stage before the pocket depth deepened. Conversely, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, and Streptococcus constellatus, which had significant differences only in limited clusters, were thought to increase in number as the pocket depth deepened, after periodontal pocket formation. Furthermore, in clusters where healthy or mild periodontal disease sites were classified, there was no statistically significant difference in pocket depth, but the number of bacteria gradually increased from the stage before the pocket depth increased. This means that quantitative changes in these bacteria can be a predictor of the progress of periodontal tissue destruction, and this novel microbiological test using the Oral Care Chip could be effective at detecting dysbiosis., Competing Interests: A research grant was provided by Mitsubishi Chemical Corporation based on the joint research agreement between Mitsubishi Chemical Corporation and Osaka University. Authors AN, HO & NT are affiliated to the funders. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Objectives: To clinically and microbiologically evaluate the effects of hyperbaric oxygen (HBO2) therapy in addition to full-mouth ultrasonic subgingival debridement (FM-UD), in the initial treatment of chronic periodontitis., Methods: Twenty patients presenting moderate to severe generalized forms of chronic periodontitis were included in a three-month randomized, parallel-group, single-blinded, prospective study. At baseline patients were randomly assigned to two treatment groups [Test Group (FM-UD+HBO2) and Control Group (FM-UD)]. Both groups were treated with an FM-UD session. Ten HBO2 sessions (one session per day for 10 days at a pressure of 2.5 ATA) were additionally administered to the Test Group. Soft tissues parameters [probing pocket depth (PPD), bleeding on probing (BOP), clinical attachment level (CAL) and visible plaque index (VPI)] were assessed at baseline (immediately before FM-UD treatment), after two weeks, after six weeks and at three months. For each patient, a site presenting PPD ≥ 6mm and positive BOP was selected as a qualifying site (QS), to be monitored clinically (at T0, T1, T2 and T3) and microbiologically (at T0, T1 and T3)., Results: There were no statistically significant differences between the two groups for any clinical parameter analyzed after three months, except for BOP, which was significantly (p < 0.05) reduced in the Test Group. Reductions in bacterial levels were detected in both groups after therapy. Faster bacterial recolonization occurred after three months in the Control Group., Conclusion: HBO2 therapy in combination with FM-UD may represent an efficacious approach to the treatment of moderate to severe forms of periodontitis., Competing Interests: The authors of this paper declare no conflicts of interest exist with this submission., (Copyright© Undersea and Hyperbaric Medical Society.)

Inflammatory periodontal diseases represent a serious dental and general medical problem due to the high prevalence among the adult population, the presence of clinical forms leading to the destruction of the dentition and tooth loss, insufficient treatment effectiveness and the frequency of relapse, including in connection with the formation of biofilms. A molecular genetic test system has been developed to evaluate the content of periodontopathogenic microorganisms Porphyromonas gingivalis, Treponema denticola, Streptococcus oralis, Streptococcus sanguis and Streptococcus sobrinus in the contents of periodontal pockets. The analytical characteristics of the test system were determined, and testing was carried out on clinical samples of patients with chronic generalized periodontitis of moderate severity. The constructed diagnostic kit allowed us to conduct a comparative analysis of the effectiveness of various types of treatment of inflammatory periodontal diseases based on quantitative data on the content of bacteria in the contents of periodontal pockets., Competing Interests: The authors declare no conflict of interest.

This study aims to investigate the levels of SLIT3 in gingival crevicular fluid (GCF) of healthy and periodontal disease subjects, and their correlations to periodontal disease. A total of 45 periodontal patients and 45 periodontally healthy volunteers were enrolled. The clinical parameters, radiographic bone loss and the levels of SLIT3, receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in GCF were measured. The prevalences of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in subgingival plaque were also analyzed. The expression of SLIT3 and RANKL was detected in the periodontium of experimental periodontitis in rats and lipopolysaccharide (LPS)-induced mouse macrophage. The total amounts and concentrations of SLIT3 and RANKL were significantly higher in periodontitis than those in healthy, while the level of OPG was significantly lower (p < .05). Significant positive correlations were observed between the level of GCF SLIT3 and clinical attachment level and radiographic bone loss (p < .05). There existed a significant positive correlation between SLIT3 and RANKL (p < .05). Increased expression of SLIT3 and RANKL was observed in the periodontium of periodontal rats. SLIT3 expression was induced by LPS stimulation in macrophages. These results suggest that SLIT3 may act as a diagnostic indicator of periodontal disease and should be further investigated., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Aim of this cross-sectional study was to investigate the prevalence of selected potentially periodontal pathogenic bacteria in different sites of patients with tongue piercing (TP) in comparison to a control group (C). Fifty participants in each group were recruited. Samples from the biofilm originating from the piercing surface (TP group), periodontal pocket, tongue as well as cheek surface were examined regarding presence of 11 selected potentially periodontal pathogenic bacteria based on polymerase-chain reaction (PCR). In the periodontal pocket of the participants, the majority of examined bacteria were more frequently detected in TP compared to C group (p i  < 0.05). At tongue and cheek surface, the prevalence of Treponema denticola (P < 0.01) and Prevotella intermedia (P < 0.01) was significantly higher in TP. For the majority of bacteria, a significant correlation between TP surface and periodontal pocket was detected (P < 0.05). In conclusion TP must be considered as potentially important ecological niche and reservoir for periodontal pathogens., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Introduction: The present study aimed to assess the presence of main types of microorganisms involved in the aetiopathogenesis of chronic periodontitis with PCR technique and determinates the presence of composite IL-1 genotype and their associations with founded bacteria., Material and Method: The examined group was consisted from 20 subjects with diagnosed chronic periodontitis and 20 healthy control without periodontitis. Clinical parameters like gingival index (GI), plaque index (PI), bleeding on probing (BOP), periodontal pocket depth (PPD) and clinical attachment lost (CAL) were determinates. Subgingival dental plaque was collected using a sterilized paper point. We used Parodontose Plus test, reverse hybridization kit, for the detection of periodontal marker bacteria, as well as for the detection of composite Interleukin -1 Genotype Results: The most present bacterial species detected from subgingival dental plaque was Treponema denticola and Porfiromonas gingivalis which was present in 65% of examined patients. In relation to the presence of positive genotype in patients, there was no significant difference between the test and control group for p> 0.05 (p = 1.00). For χ2=8,17 (p=0,06, p<0,05) there is an association between Prevotella intermedia, and composite genotype. Between positive genotype and analyzed bacterial species A. actinomycetem comitans for p> 0.05 (p = 1.00), P. gingivalis for p> 0.05 (p = 0.16), T. Forsythia for p> 0.05 (p = 0.20), T. Denticola for p> 0.05 (p = 0.64) no association was found., Conclusion: This investigations confirmed the strong association of these five examined periopathogenes with periodontitis.

Rationale: Halitosis is an unpleasant odor that emanates from the mouth. Studies show halitosis returns in a week, after treatment with PDT. Probably, bacteria living in the periodontal sulcus could recolonize the dorsum of the tongue. Until nowadays, there are no study in adult population that associates halitosis and periodontal treatment with follow-up evaluation. The aim of this randomized, controlled, single-blinded clinical trial is to treat oral halitosis in healthy adults with photodynamic therapy (PDT), associated with periodontal treatment and follow them up for 3 months. PATIENT CONCERNS:: the concerns assessments will be done over the study using anamnesis interviews and specific questionnaire. DIAGNOSES:: halitosis will be evaluated by OralChroma., Interventions: The participants (n = 40) with halitosis will be randomized into 2 groups: G1-treatment with PDT (n = 20) or G2-cleaning of the tongue with a tongue scraper (n = 20)., Outcomes: Halitosis will be evaluated by measuring volatile sulfur compounds using gas chromatography. After the treatments, a second evaluation will be performed, along with a microbiological analysis (RT-PCR) for the identification of the bacteria T. denticola. The assessment of halitosis and the microbiological analysis will be repeated. After that, patients will receive periodontal treatment. The participants will return after 1 week and 3 months for an additional evaluation. Quality of life will be measured by Oral Health Impact Profile questionnaire (OHIP-14)., Lessons: This protocol will determine the effectiveness of phototherapy regarding the reduction of halitosis in adults. clinicaltrials.gov NCT03996915., Ethics and Dissemination: This protocol received approval from the Human Research Ethics Committee of Universidade Nove de Julho (certificate number: 3.257.104). The data will be published in a peer-reviewed periodical.

Current continuous flow polymerase chain reaction (CF-PCR) microfluidic chips require external precision syringe pumps and off-line methods (e.g., electrophoresis and hybridization) to detect PCR products, resulting in complex operations and possible cross-contamination and consequently CF-PCR is still confined to laboratories. Herein, a portable all-in-one microfluidic device is fabricated for rapid diagnosis of pathogens based on an integrated CF-PCR and electrophoresis biochip. A new method was proposed for automatic sample injection into the chip which can substitute the costly external precision syringe pump. It not only achieves rapid DNA amplification and on-site PCR product detection, but also realizes automatic sample injection. As an application, three periodontal pathogens (e.g., Porphyromonas gingivalis, Treponema denticola and Tannerela forsythia) were successfully amplified in the device. Treponema denticola was amplified in as short as 2'31'', and detection of PCR products was completed within 3'43''. The minimum number of bacteria that can be amplified was 125 cfu per μl. The all-in-one device has the potential to be applied in point-of-care nucleic acid testing for diseases.

Background: The periodontal tissues are continuously exposed to specific bacterial components that have the ability to alter many local functions. Normal endogenous infections in healthy mouths cause disease when their numbers increase significantly., Objective: Determine the percentage of different periodontal pathogenic bacteria and their association with periodontal status., Design: Cross-sectional, analytical., Settings: School children of both genders in Saudi Arabia., Patients and Methods: Clinical examination consisted of measurement of the gingival and periodontal supporting tissue including attachment loss, probing pocket depth and furcation involvement following the National Health and Nutrition Examination Survey (NHANES) and taking samples of the subgingival bacterial flora., Main Outcome Measures: The percentage of periodontal pathogenic bacteria and its association with periodontal status in Saudi Arabia., Sample Size: Bacterial samples were collected from 277 subjects., Results: Aggregatibacter actinomycetemcomitans was present in 21.7% of the subjects, Porphyromonas gingivalis in 21.3%; Tannerella forsythia in 10.1%; Treponema denticola in 34.7% and Prevotella inter-media in 12.3%. The red complex bacteria were found in 2.9% of the subjects., Conclusions: The percentages of bacteria varied but only T denticola was significantly associated with periodontal breakdown. In addition, the presence of more than 2 of the 5 species tested were significantly associated with tissue damage., Limitations: Cannot be generalized to all of Saudi Arabia. Larger controlled studies are needed., Conflict of Interest: None.

Objective: To investigate periodontal status of patients with pre-diabetes and evaluate the prevalence of periodontal pathogens in oral cavity. Methods: All the subjects were under regular care in urban area of Beijing, including 88 subjects with normal blood glucose (normal blood glucose group), 27 pre-diabetic patients (pre-diabetic group), 58 well-controlled diabetic patients (glucose well controlled group) and 72 poor-controlled diabetic patients (glucose poor controlled group). Whole unstimulated saliva samples were collected before periodontal examination. Periodontal parameters, including plaque index (PLI), probing depth (PD), bleeding index (BI), bleeding on probing (BOP) and clinical attachment loss (CAL), were examined at mesial-buccal and distal-lingual sites of each tooth. Number of missing teeth was recorded. DNA was extracted from the salivary deposition, Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Campylobacter rectus (Cr), and Prevotella nigrescens (Pn) were detected by using PCR method based on 16SrRNA. Periodontal status and prevalence and quantity of the pathogens under various blood glucose states were compared. Results: The PD scores of four groups had no statistical differences. The CAL [(2.29±1.35) mm] and the number of missing teeth[2.0 (7.0)] in pre-diabetic group were significantly lower than that in glucose poor controlled group [(3.07±1.45) mm, P= 0.04 and 5.0 (10.0), P= 0.04, respectively]. The number of missing teeth in pre-diabetic group [2.0 (7.0)] was significantly lower than that in glucose well controlled group [5.0 (9.0), P= 0.02]. The percent of bleeding on probing [BOP(+)%] in pre-diabetic group [(63.89±20.03)%] was significantly higher than that in normal blood glucose group [(54.51±22.29)%, P= 0.04] and glucose well controlled group [(53.12±21.77)%, P= 0.03]. The prevalence of Pg in pre-diabetic group (81.5%) was significantly higher than that in glucose poor controlled group (54.2%, P= 0.02). The prevalence of Tf in pre-diabetic group (96.3%) was significantly higher than that in glucose poor controlled group (76.4%, P= 0.01). Meanwhile the quantity of Pg [1.58 (4.75)] and Tf [5.46 (7.77)] in pre-diabetic group were significantly higher than that in glucose poor controlled group [0.60 (1.87), P= 0.01 and 1.63 (3.06), P< 0.01, respectively]. The quantity of Pn [0.85 (1.68)] in pre-diabetic group was significantly higher than that in normal blood glucose group [0 (1.02), P= 0.04]. Conclusions: Pre-diabetic patients showed severe periodontal infection and BOP(+)% than other three groups and had high risk-level of periodontitis.

Objective: To evaluate the epidemiological and microbiological aspects of the potential association between bipolar affective disorder (BAPD) and periodontitis., Methodology: The present case-control study comprised 176 individuals with BAPD and 176 controls. All individuals underwent a complete full-mouth periodontal examination and microbiological sampling. Data on bleeding on probing, probing depth, and clinical attachment level in all present teeth were recorded. Quantification of total bacterial load and Aggregatibacter actinomycetemcomitans, Treponema denticola, and Porphyromonas gingivalis counts were performed through qPCR. Data were analyzed using univariate analysis, Spearman correlation and multivariate logistic regression., Results: The prevalence of periodontitis was 39.7% among controls and 58.5% among individuals with BAPD (OR = 2.13; 95% CI 1.39-3.27). A. actinomycetemcomitans and P. gingivalis counts were significantly higher in individuals with BPAD and periodontitis. The final multivariate logistic regression revealed that periodontitis was strongly associated with the total bacterial load (OR = 1.91; 95% IC = 1.0-1.99; P < 0.001) and the depressive phase of BPAD (OR = 28.94; 95% IC = 4.44-177.27; P < 0.001)., Conclusion: BAPD was associated with increased risk for periodontitis. Individuals with BPAD presented higher levels of A. actinomycetemcomitans and P. gingivalis, suggesting that periodontitis could be a co-morbidity frequently found in individuals with BAPD., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Introduction: There is a known connection between periodontitis and atherosclerosis and the presence of periopathogens in blood vessels. However, changes of the oral microflora related to the aging process and its possible effects on atherosclerosis, have yet to be analyzed. The aim of this study was to assess temporal changes in the frequency of periodontal bacteria in the subgingival plaque and in atherosclerotic blood vessels of patients with atherosclerosis., Methodology: The study included 100 patients with atherosclerosis and periodontitis, divided into two groups, below and over 60 years of age. Clinical examinations were performedand subgingival plaque specimens were collected as well as biopsy specimens from the following arteries: coronary (34), carotid (29), abdominal (10), femoral (10), mammary (13) and iliac (4). Subgingival and artery specimens were subjected to PCR detection of 5 major periodontal pathogens: Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), Tannerella forsythensis (Tf) and Treponema denticola (Td)., Results: Tf was the most and Td the least frequent bacteria in both age groups and in both types of samples. The frequencies of bacteria in subgingival versus atherosclerotic samples were: Tf (76%:53%), Pi (71%:31%), Pg (60%:38%), Aa (39%:14%) and Td (21%:6%). Only Aa and Pi showed a significant difference of prevalence between younger and older patients. The most colonized artery was a. coronaria, followed by a. carotis, a. abdominalis, a. mammaria, and a. femoralis., Conclusions: Patient's age and the distance of a given blood vessel from the oral cavity influenced microbiological findings in the atherotic plaque., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2018 Ibrahim Kannosh, Danijela Staletovic, Bosko Toljic, Milena Radunovic, Ana Pucar, Sanja Matic Petrovic, Ivana Grubisa, Milos Lazarevic, Zlata Brkic, Jelena Knezevic Vukcevic, Jelena Milasin.)

Background: Periodontal pathology in children has distinctive characteristics that can be accounted for by the peculiarities of the age, the dynamically changing oral environment, and the processes of formation and stabilization of periodontal structures during teeth eruption., Aim: The aim of this study was to investigate the composition of the subgingival microbiota during permanent dentition formation in puberty., Materials and Methods: The study included 30 children aged between 10 and 15 years without gingivitis (up to 25% PBI), with good oral hygiene, with no systemic diseases or reception of antibiotics for 3 months. The clinical status of all children was assessed using a specially designed chart based on the periodontal examination chart of a child adopted by the Department of Pediatric Dentistry, FDM-Sofia. Gingival sulcus samples were taken with a paper pin from 6 teeth for a PCR - Real Time study of 9 control strains (a comprehensive sample). The samples were sent for investigation in standardized containers., Results: C. gingivalis (green complex) was found in almost all tested children and F. nucleatum (orange complex) - in more than half of them. The rest of the tested samples for microorganisms was isolated in just few of the children (6% - 17%). A. actinomycetemcomitans and P. gingivalis were not isolated in this study., Conclusion: The results of the study suggest that a significant amount of the periodontal pathogens migrate to the subgingival zone relatively early, immediately after the permanent teeth eruption, and become a risk factor for the development of periodontal disease.

Context: Knowledge of the microbial composition of abscessed primary tooth is limited., Aim: The aim was to investigate the presence of 10 oral bacterial species in samples from abscessed primary tooth root canals using microarray technology and to determine their association with clinical findings., Subjects and Methods: The samples were collected from root canals of 20 primary molars with acute primer infection. The bacterial composition of the samples was semi-quantitatively defined using a microarray system (ParoCheck ® ). Clinical parameters included the presence of spontaneous pain, mobility, percussion sensitivity and swelling., Statistical Analysis: Data were statistically analyzed by Student' t-test, Fisher's exact Chi-square test, Freeman-Halton-Fisher's exact test, and Spearman's rho correlation analysis., Results: All the tested species were detected in the samples. Fusobacterium nucleatum was the most frequent bacterium (100%), followed by Parvimonas micra (65%), Provetella intermedia (45%), and Treponema denticola (45%). According to paired bacterial combinations, F. nucleatum was significantly positively correlated with P. intermedia and P. micra (P < 0.05). T. denticola was significantly positively correlated with Porphyromonas gingivalis, Tannerella forsythia, Campylobacter rectus, and P. micra, while it was negatively correlated with Eikenella corrodens (P < 0.05). No statistically significant relationships were found between the presence of any bacteria and clinical findings., Conclusion: Microarray technology used in this study has demonstrated the presence of various bacteria with varying proportions in the root canals of abscessed primary teeth. The results regarding the high rate of certain bacterial combinations suggest the enhanced pathogenicity due to additive or synergistic effects of these microbial combinations., Competing Interests: None

Purpose: The primary prevention of peri-implantitis onset is a key factor in long-term implant success, and the evaluation of the antibacterial efficacy of different implant surfaces is fundamental in this way. The aim of this study was to assess if implants with collars coated with anatase were less subjected to bacterial colonization than implants with noncoated collars, and to investigate how implant bacterial colonization varies over time., Materials and Methods: Eighteen patients in need of implant-supported rehabilitation were selected to have two adjacent implants placed, one with an anatase-coated collar and one with the collar uncoated. Biofilm samples were collected at four sites around each implant at four different time points. Samples were analyzed through polymerase chain reaction (PCR) to detect and calculate the colonization rate of Aggregactibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Prevotella intermedia., Results: Due to one patient dropout and two nonosseointegrated implants, 32 out of 36 placed implants were followed up for 12 months, and 128 samples for each time point were collected: in total, 512 biofilm samples were analyzed. The type and rate of bacterial colonization were not significantly different between the two groups at all the intervals. However, the anatase-coated collar showed no proliferation of T forsythia. A significant difference in marginal bone level could be observed at the 12-month follow-up only. No significant difference in the other clinical and radiographic indexes was observed., Conclusion: In this study, anatase-coated collar implants did not seem to provide significantly different microbiologic outcomes than uncoated collar implants. However, the absence of colonization of the species T forsythia and the slightly smaller peri-implant bone loss at the 12-month follow-up suggest that further investigations on anatase coating are needed. Nevertheless, data on bacterial colonization and crestal bone levels need further investigations to draw meaningful conclusions, due to the statistical power of this pilot study.

This large-scale study cross-sectionally examined the periodontal status and prevalence of "red complex" bacteria (Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia) in Japanese adults. A total of 977 participants were enrolled in the study. Probing depth (PD), bleeding on probing (BOP), and bone crest level (BCL) were recorded, and the presence of red complex bacteria in the saliva was examined using polymerase chain reaction. The mean BCL value and the percentage of sites with a PD ≥4 mm or the presence of BOP were significantly higher in older participants. The detection rates of P. gingivalis, T. denticola, and T. forsythia were 46.3%, 76.4%, and 61.1%, respectively. The P. gingivalis detection rate significantly increased with age, while those of T. denticola and T. forsythia were comparably high for all age groups. A close correlation between P. gingivalis and the percentage of sites with PD ≥4 mm was indicated by nonlinear canonical correlation analysis. Current smokers exhibited a more advanced disease condition and a significantly higher P. gingivalis detection rate than non-smokers. In conclusion, periodontal condition worsens with age, and P. gingivalis appears to be the red complex bacterium most closely associated with periodontitis.

Quantitative analysis of periodontal bacteria is considered useful for clinical diagnosis, evaluation and assessment of the risk of periodontal disease. The purpose of this study was to compare the effectiveness of sampling of saliva, supragingival and subgingival plaque for evaluation of periodontal bacteria. From each of 12 subjects, i) subgingival plaque was collected from the deepest pocket using a sterile paper point, ii) stimulated whole saliva was collected after chewing gum, and iii) supragingival plaque was collected using a tooth brush. These samples were sent to the medical examination laboratory for quantitative analysis of the counts of three periodontal bacterial species: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The proportions of these bacteria in subgingival plaque were higher than those in saliva or supragingival plaque, but lower in subgingival plaque than in saliva or supragingival plaque. In several cases, periodontal bacteria were below the levels of detection in subgingival plaque. We concluded that samples taken from subgingival plaque may be more useful for evaluating the proportion of periodontal bacteria in deep pockets than is the case for other samples. Therefore, for evaluation of periodontal bacteria, clinicians should consider the characteristics of the specimens obtained using different sampling methods.

Objective: Evidence of increased apoptosis is observed in periodontitis and may be associated with destruction of the periodontal tissue caused by the increased cell death, with the release of danger signals and subsequent stimulation of the proinflammatory processes. However, the exact mechanisms associated with these processes remain unclear. This study aimed to investigate the presence of the periodontal pathogen Treponema denticola, apoptosis, high mobility group box 1 as a damage-associated molecular pattern, and several inflammatory markers in periodontitis and gingivitis subjects., Materials and Methods: Soft tissue specimens from gingival tissues of periodontitis and gingivitis patients were used for immunohistochemical and immunofluorescence staining of T. denticola chymotrypsin-like proteinase (CTLP), apoptosis markers, high mobility group box 1, Toll-like receptor 4, inflammatory cell markers, and proinflammatory cytokines., Results: Treponema denticola was detected in all periodontitis-affected tissues. This was associated with a significant increase in the number of apoptotic cells, including macrophages, alterations in the expression of high mobility group box 1 and its receptor, and increased levels of proinflammatory cytokines compared with gingivitis., Conclusions: In summary, the presence of T. denticola (especially its CTLP), apoptosis, high mobility group box 1, and inflammatory markers suggests their potential involvement in the pathogenesis of periodontitis., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

The aim of this study was to determine the relationship between the prevalence of Porphyromonas gingivalis, its fimA genotypes, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, and Treponema denticola and the evolution of periodontal health. In a longitudinal prospective study, samples of subgingival plaque were taken from 114 patients (37 with chronic periodontitis, 17 with gingivitis, and 60 periodontally healthy) in the course of a full periodontal examination. PCR was employed to determine the presence of the periodontopathogenic bacteria. Four years later, a second examination and sample collection were performed in 90 of these patients (20 with chronic periodontitis, 12 with gingivitis, and 58 periodontally healthy). T. forsythia, P. gingivalis, and T. denticola are the most prevalent bacteria in patients with chronic periodontitis (78.4%, 62.2 y 56.8%, respectively). The P. gingivalis bacterium and its fimA genotypes I, II, and IV showed the highest correlation between the baseline and follow-up assessments. P. gingivalis fimA genotype II and T. forsythia were associated to a significant degree with unfavourable periodontal evolution. Of the variables studied, P. gingivalis fimA genotype II and T. forsythia increase the risk of an unfavourable evolution of periodontal status.

Biofilm of the gingival sulcus from 22 patients with type 2 diabetes mellitus and periodontitis, 30 patients with periodontitis not complicated by diabetes mellitus (reference group), and 22 healthy volunteers without signs of gingival disease (control group) was studied by quantitative PCR. Quantitative analysis for the content of P. gingivalis, T. forsythia, A. ctinomycetemcomitans, T. denticola, P. intermedia, F. nucleatum/periodonticum, and P. endodontalis in the dental plaque was performed with a Dentoscreen kit. The presence of other bacterial groups was verified by metagenomic sequencing of the 16S rRNA gene to evaluate some specific features of the etiological factor for periodontitis in type 2 diabetes mellitus. Specimens of the Porphiromonadaceae and Fusobacteriaceae families were characterized by an extremely high incidence in combined pathology. The amount of Sphingobacteriaceae bacteria in the biofilm was shown to decrease significantly during periodontitis. Metagenomic analysis confirmed the pathogenic role of microbiota in combined pathology, as well as the hypothesis on a possible influence of periodontitis on the course and development of type 2 diabetes mellitus.

As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm., (© 2017 The Authors. Molecular Oral Microbiology Published by John Wiley & Sons Ltd.)

Background and Objectives: There has been limited study of the bacterial species associated with aggressive periodontitis (AgP) in high-risk populations in Africa. The aim of this study was to investigate and quantify the presence of four putative periodontal pathogens in the subgingival plaque of Sudanese subjects with AgP. A secondary aim was to investigate the effect of varying the detection threshold on the reported prevalence of the bacterial species investigated., Materials and Methods: Subgingival plaque samples were collected from AgP cases (n=73) and healthy controls (n=71). Bacterial DNA was extracted and analyzed by quantitative polymerase chain reaction for the detection and quantification of four putative periodontal pathogens: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Treponema denticola and Tannerella forsythia., Results: At the lowest detection threshold (>10 1 cells), P. gingivalis (p<0.0001) was more prevalent in AgP cases than controls. T. forsythia and T. denticola had a high prevalence (>70%) in AgP cases at all detection levels. While T. forsythia was significantly more frequently identified in AgP than in controls at all detection thresholds, this was only the case for T. denticola at the intermediate threshold (>10 2 cells). A. actinomycetemcomitans was identified less frequently than the other bacterial species with no difference in its prevalence between AgP cases and controls., Conclusion: The prevalence of the putative periodontal pathogens investigated varied considerably in Sudanese subjects with AgP and in periodontally healthy controls depending on the detection thresholds applied. T. forsythia was identified as having the strongest association with AgP., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Chronic prostatitis (CPr) and benign prostatic hyperplasia (BPH) are complex inflammatory conditions for which etiologic determinants are still poorly defined. Periodontitis is caused by subgingival colonizing bacteria in the oral cavity. The causal effect of periodontal disease on prostatic inflammation has not been established. The purpose of this study is to isolate oral pathogens from expressed prostatic secretions of patients with periodontal disease and CPr or BPH., Methods: Twenty-four men diagnosed with CPr/BPH participated in the study. A complete periodontal examination consisting of probing depth, bleeding on probing, tooth mobility, gingival index, and plaque index was performed on the men, and prostatic secretion was collected for the study. Dental plaque and prostatic secretion samples were used for analysis of bacterial DNA for Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Treponema denticola (Td), and Escherichia coli using reverse transcription-polymerase chain reaction., Results: Six patients were diagnosed with severe, seven with moderate, and four with mild chronic periodontitis. Seventeen of 24 (70.8%) of the prostatic secretion samples showed one or more of the studied oral pathogens. Nine of 10 BPH and eight of 14 patients with CPr had at least one oral pathogen in their prostatic secretions. Pg was found in both prostatic secretion and plaque samples in six of 17 (35.3%) patients, Td was found in both samples in seven of 15 (46.7%) patients, and E. coli was found in both samples in three of 15 (20%) patients. Pi was detected in all dental plaque samples but not in the prostatic secretion., Conclusion: An association between chronic inflammatory prostate and periodontal diseases has been demonstrated by the presence of similar bacterial DNA in both prostatic secretion and subgingival dental plaque from the same individual.

Objectives: The purpose of this study was to compare microbial profiles of saliva, pooled and site-specific subgingival samples in patients with periodontitis. We tested the hypotheses that saliva can be an alternative to pooled subgingival samples, when screening for presence of periopathogens., Design: Site specific subgingival plaque samples (n = 54), pooled subgingival plaque samples (n = 18) and stimulated saliva samples (n = 18) were collected from 18 patients with generalized chronic periodontitis. Subgingival and salivary microbiotas were characterized by means of HOMINGS (Human Oral Microbe Identification using Next Generation Sequencing) and microbial community profiles were compared using Spearman rank correlation coefficient., Results: Pronounced intraindividual differences were recorded in site-specific microbial profiles, and site-specific information was in general not reflected by pooled subgingival samples. Presence of Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Filifactor alocis, Tannerella forsythia and Parvimona micra in site-specific subgingival samples were detected in saliva with an AUC of 0.79 (sensitivity: 0.61, specificity: 0.94), compared to an AUC of 0.76 (sensitivity: 0.56, specificity: 0.94) in pooled subgingival samples., Conclusions: Site-specific presence of periodontal pathogens was detected with comparable accuracy in stimulated saliva samples and pooled subgingival plaque samples. Consequently, saliva may be a reasonable surrogate for pooled subgingival samples when screening for presence of periopathogens. Future large-scale studies are needed to confirm findings from this study.

Background: This study evaluates periodontal conditions and microbiologic findings and their influence on rheumatologic disease parameters in patients with rheumatoid arthritis (RA)., Methods: One hundred and sixty-eight patients with RA were included. A healthy control group (HC, n = 168) was composed according to age, sex, and smoking habits. Rheumatologic data (duration of illness, Disease Activity Score 28, rheumatic factor [RF], anti-cyclic citrullinated peptide [aCCP], medications) were extracted from patients' records. Dental examination included: 1) dental findings (decayed, missing, and/or filled adult teeth [DMF-T] index); 2) gingival inflammation (papillary bleeding index [PBI]); and 3) periodontal status (probing depth [PD], attachment loss [AL]). Periodontal condition was classified as healthy/mild, moderate, or severe periodontitis. Subgingival biofilm was analyzed regarding 11 periodontopathogenic bacteria. Statistical analyses included: 1) Kolmogorov-Smirnov test; 2) Mann-Whitney U test; 3) Pearson χ 2 test; 4) Kruskal-Wallis test; and 5) regression analysis; level of significance α = 5%., Results: Mean DMF-T was significantly higher in patients with RA (19.3 ± 4.8) than in HC group (16.9 ± 5.8), especially owing to number of missing teeth (RA = 6.0 ± 5.4, HC = 3.1 ± 3.3; P <0.01). Patients with RA had a significantly higher proportion of increased PD (P <0.01) and AL compared with HC group (P <0.01). Moderate to severe periodontitis was noted in 98% of patients with RA and 82% of the HC group (P <0.01). RF-positive patients with RA suffered from worse periodontal conditions than RF-negative patients (P = 0.01). Age, PBI, and presence of Treponema denticola (P <0.03) are related to periodontal condition in patients with RA. Although not statistically significant, Porphyromonas gingivalis and Fusobacterium nucleatum occur in higher concentrations more often in aCCP-positive patients with RA (P = 0.06)., Conclusions: Patients with RA had worse periodontal conditions than HC participants. Although a trend for higher F. nucleatum and P. gingivalis concentrations in aCCP-positive patients with RA was found, importance of periodontal pathogenic bacteria and rheumatoid parameters in the interrelationship between periodontitis and RA remains unclear.

Introduction: Apical periodontitis usually results from bacterial accumulation and contamination occurring in the root-canal system, and extending beyond the apical foramen to involve the periapical tissues. Literature has a paucity of the studies that stress on the division and analysis of the pulp canal segments. The reason for this disparity might be the technique used for collecting the samples from the pulp canals. Hence, we carried out the present study to evaluate the microbial flora in the apical part of the roots with necrotic pulp canals., Materials and Methods: The present study included the assessment of 40 freshly extracted teeth that had necrotized pulpal tissue along with the presence of periapical periodontal lesions. Removal of the soft tissue lesions attached to the root portion of the teeth along with apical periodontal lesions was done with the help of scalpel blade, after rinsing them with a sterile solution of saline. Thorough cleaning of the root surfaces was done with hydrogen peroxide followed by rapid disinfection with the help of sodium hypochlorite at varying concentrations. Sectioning of the root portion of all the specimens with the help of a disk was done perpendicular to the long axis of the teeth at a distance of roughly 5 to 6 mm from the teeth's apicalmost point. Cryotubes were used for transferring the specimens of apical portions containing 1 mL of buffer and were subjected to immediate frozen processing at a temperature of -20°C. A 10 K-type file was used for the initial collection of the samples followed by subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al. Paster et al's modification of the reverse-capture checkerboard assay was used in the present study. Semiquantitative data were used for overcoming the difficulties arising due to obtaining the counts of the polymerase chain reaction (PCR)-based analysis of specimens., Results: A positive result for the 16S ribosomal ribonucleic acid (rRNA) gene primer was observed only in two examined specimens of all the samples of the apical portion of the root canals in the present study. Negative result was shown by all the control group specimens, which were sterile samples. Presence of bacteria was confirmed by PCR in 38 out of 40 examined specimens. Amount of bacterial taxa, out of these 24 samples, ranged up to 6. Pseudoramibacter alactolyticus, Porphyromonas endodontalis, Dialister oral species, Bacteroidetes species, Streptococcus species, Olsenella uli, Synergistes species, Fusobacterium nucleatum, Parvimonas micra, Treponema denticola, and Filifactor alocis were the specific species detected. Bacteroidetes species was the only species that were detected at levels at or above 10 5 . Heavy bacterial infections were noticed in more than 45% of the cases at the periradicular part of the root canals., Conclusion: Microbial flora of the apical segment of the root with necrotized pulp tissue comprises a vast variety of pathogenic bacteria., Clinical Significance: For better prognosis of the treatment of such cases, adequate knowledge of the microbial flora of the root, especially the apical portion is necessary.

This study aims to investigate the relation between resistin and periodontopathogenic bacterial levels in the saliva of obese adults compared to healthy control and to examine whether salivary resistin can serve as a biomarker of type 2 diabetes in obese patients. A total of 78 saliva samples were collected from patients attending to the University Dental Hospital, Sharjah, UAE. The patients were divided into three equal groups: obese diabetics, obese nondiabetics, and nonobese nondiabetic control. Salivary resistin was measured using ELISA. The levels of bacterial species associated with periodontitis ( Treponema denticola , Porphyromonas gingivalis , Tannerella forsythia , and Actinobacillus actinomycetemcomitans ) and gingivitis ( Fusobacterium spp.) were measured using real-time PCR. Both salivary resistin and periodontopathogenic bacteria including Fusobacterium spp., P. gingivalis , and T. forsythia were detected in significantly higher quantities in the obese patients (diabetics and nondiabetics) than nonobese nondiabetic control. Resistin concentrations were significantly correlated with BMI; however, its level was not correlated with the blood glucose. In this study, high salivary resistin was associated with obesity, which is a major predisposing factor for type 2 diabetes and also a risk factor for oral diseases. The high levels of salivary periodontopathogenic bacteria could upregulate the local release of salivary resistin in obese people.

Aim: The aim of the present study was to investigate how heavy smoking influences the clinical, microbiological, and host-response characteristics in peri-implant sulcus fluid of patients with healthy dental implants., Methods: A total of 29 individuals with 74 dental implants were included in the present study; 20 implants were in heavy smokers and 54 were in non-smokers. The modified gingival index, modified plaque index, and probing pocket depth were evaluated. Periodontopathogenic bacteria Tannerella forsythia, Treponema denticola, and Porphyromonas gingivalis were evaluated, together with the total bacterial load. Peri-implant sulcus fluid samples were analyzed for the quantification of interleukin-8, interleukin-1β, interleukin-6, interleukin-10, and tumor necrosis factor-α., Results: No significant differences in the clinical parameters evaluated were found between the groups, although smokers had poorer peri-implant parameters. Among the smokers, subgingival microbiota was composed of a greater number of periodontal pathogens; these differences were not statistically significant. Smokers showed a greater expression of interleukin-1β, interleukin-6, interleukin-10, and tumor necrosis factor-α, but interleukin-8 was slightly higher among non-smokers, but not significantly., Conclusions: Although smokers presented deeper probing depths, bleeding on probing, and peri-implant microbiota composed of a greater number of periodontal pathogens than in non-smoking patients, these data did not show significant differences. In the present study, and in relation to the samples analyzed, smoking alone did not influence the immunological and microbiological parameters in dental implants with healthy peri-implant tissues. Further studies with larger samples are required to better evaluate the influence of smoking on dental implants., (© 2015 Wiley Publishing Asia Pty Ltd.)

Aim: The aim of the present study was to evaluate the association between the occurrence of periodontopathogens with cortisol levels in chronic periodontitis patients., Methods: Seventy-five chronic periodontitis patients were invited to participate in the present study. Cortisol levels in serum were measured using an immunoassay method. Porphyromonas gingivalis (P. gingivalis) Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans were detected by polymerase chain reaction using primers designed to target the respective 16S rRNA gene sequences., Results: Severe chronic periodontitis patients showed higher mean levels of cortisol (P < 0.05). Twenty-six patients had hypercortisolemia. High cortisol levels showed a positive significant correlation with P. gingivalis (r = 0.237, P < 0.01). Of the 26 patients with hypercortisolemia, 81% had P. gingivalis, of which 86% had severe chronic periodontitis (P < 0.001). There were higher levels of cortisol with the presence of P. gingivalis (478.65 ± 122.57 vs 402.58 ± 139.60, P = 0.01). The adjusted logistic regression model showed a significant association between high cortisol levels and P. gingivalis (odds ratio = 1.7, 95% confidence interval = 1.6-1.8)., Conclusions: This research offers support for the association between P. gingivalis and higher levels of cortisol in chronic periodontitis patients. These results suggest that high levels of cortisol could increase the occurrence of P. gingivalis in the biofilm., (© 2015 Wiley Publishing Asia Pty Ltd.)

Objective: To assess and compare salivary periodontopathic bacteria between groups of Down syndrome and non-Down syndrome children and adolescents., Materials and Methods: This study included a sample of 30 Down syndrome children and adolescents (G-DS) and 30 age- and sex-matched non-Down syndrome subjects (G-ND). Clinical examination determined the gingival bleeding index (GBI) and plaque index. Unstimulated whole saliva samples were collected from all participants. The fluorescence in situ hybridization (FISH) technique identified the presence and density of eight periodontopathic bacteria in saliva. The statistical analysis included chi-square and Mann-Whitney U tests., Results: In the G-DS group, bleeding on probing was more frequent (p = 0.037) and higher densities of Campylobacter rectus (p = 0.013), Porphyromonas gingivalis (p = 0.025), Treponema denticola (p = 0.026), Fusobacterium nucleatum (p = 0.013), Prevotella intermedia (p = 0.001) and Prevotella nigrescens (p = 0.008) were observed. Besides, in the G-DS, the densities of bacteria from the orange complex were significantly higher in the age group 3-7 years for F. nucleatum (p = 0.029), P. intermedia (p = 0.001) and P. nigrescens (p = 0.006). C. rectus was higher in the age group 8-12 years (p = 0.045)., Conclusion: The results showed that children and adolescents with Down syndrome have higher susceptibility to periodontal disease and number of periodontopathic bacteria., Competing Interests: The authors have declared that no competing interests exist.

Helicobacter pylori (H. pylori), a pathogen inducing peptic disease, is recently found to be binding to the progress of periodontitis. Most previous studies are case-controlled, and they investigate the risk of H. pylori infection in disease the development of while few studies evaluate the correlation between H. pylori and periodontal pathogens. Therefore, we investigated the correlation between H. pylori infection with periodontal parameters, periodontal pathogens and inflammation. The results indicated that patients with H. pylori showed significantly higher probing depth and attachment loss than those without (p < 0.05). Among 28 subgingival plaque samples from 14 patients, the frequencies of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Treponema denticola were significantly higher with H. pylori infection than those without H. pylori infection (p < 0.05). However, the frequency of Aggregatibacter actinomycetemcomitans was lower (p < 0.05). Furthermore, after human acute monocytic leukemia cell line (THP-1) was stimulated with cagA-positive standard strains (cagA+ H. pylori 26695), the expression of periodontitis-related molecules Wnt5a, interleukin 8 (IL-8), interleukin 6 (IL-6) and interferon gamma (IFN-γ) significantly increased (p < 0.05). Conversely, the expression of tumor necrosis factor alpha (TNF-α) was almost stable. Meanwhile, cagA+ H. pylori promoted significantly higher expression of IL-8 and Wnt5a than isogenic cagA mutants strains (cagA- H. pylori 26695) did. Taken together, our data suggested that H. pylori might promote the growth of some periodontal pathogens and aggravate the progress of chronic periodontitis.

Polymorphisms in toll-like receptor (TLR) and β-defensin (DEFB) genes have been recognized as potential genetic factors that can influence susceptibility to and severity of periodontal diseases (PD). However, data regarding associations between these polymorphisms and PD are still scarce in North American populations, and are not available in HIV+ North American populations. In this exploratory study, we analyzed samples from HIV+ adults (n = 115), who received primary HIV care at 3 local outpatient HIV clinics and were monitored for PD status. We genotyped a total of 41 single nucleotide polymorphisms (SNPs) in 8 TLR genes and copy number variation (CNV) in DEFB4/103A. We performed regression analyses for levels of 3 periodontopathogens in subgingival dental plaques (Porphyromonas gingivalis [Pg], Treponema denticola [Td], and Tannerella forsythia [Tf]) and 3 clinical measures of PD (periodontal probing depth [PPD], gingival recession [REC], and bleeding on probing [BOP]). In all subjects combined, 2 SNPs in TLR1 were significantly associated with Td, and one SNP in TLR2 was significantly associated with BOP. One of the 2 SNPs in TLR1 was significantly associated with Td in Caucasians. In addition, another SNP in TLR1 and a SNP in TLR6 were also significantly associated with Td and Pg, respectively, in Caucasians. All 3 periodontopathogen levels were significantly associated with PPD and BOP, but none was associated with REC. Instrumental variable analysis showed that 8 SNPs in 6 TLR genes were significantly associated with the 3 periodontopathogen levels. However, associations between the 3 periodontopathogen levels and PPD or BOP were not driven by associations with these identified SNPs. No association was found between DEFB4/103A CNV and any periodontopathogen level or clinical measure in all samples, Caucasians, or African Americans. Our exploratory study suggests a role of TLR polymorphisms, particularly TLR1 and TLR6 polymorphisms, in PD in HIV+ North Americans., Competing Interests: The authors have declared that no competing interests exist.

Purpose: The purpose of this study was to compare the clinical outcome of and determine the differences in periodonto-pathogenic microbiota around two types of implant collar surfaces: laser-microtextured (test) vs machined (control)., Materials and Methods: Seventeen patients (11 periodontally healthy, and 6 periodontally compromised) were selected to receive the two different implants, placed randomly, in two edentulous sites. Six months following the surgical placement of the dental implants, subgingival plaque samples were collected using paper points from the peri-implant sulcus and from the sulcus of an adjacent tooth. The presence of five putative periodontal pathogens, namely, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Tannerella forsythensis, was assessed using real-time polymerized chain reaction (RT-PCR). Peri-implant parameters and intraoral radiographs were recorded up to 1 year after abutment connection., Results: In the main population, and in the periodontally compromised subgroup, the total number of periodontal pathogens around test implant sites was lower than control implant sites and adjacent tooth sites, with a statistically significant difference (P < .05). In periodontally healthy patients, the mean probing pocket depth for the test implant was 1.31 ± 0.51 mm, compared with 2.66 ± 0.83 mm for the control implant, while in periodontally compromised patients, it was 1.61 ± 0.58 mm for the test implant, compared with a mean value of 2.84 ± 1.0 mm for the control implant., Conclusion: Implants with a laser-microtextured collar surface are not more vulnerable to pathogenic microflora colonization than implants with a machined collar surface. In both of the subgroups of patients (periodontally healthy and periodontally compromised), implants with a laser-microtextured collar surface have a better clinical outcome at 1 year of loading, compared with implants with a machined collar surface.

Objectives: To relate five periodontopathogenic bacteria, including the red complex, to the severity, extent, and inflammation of the periodontal lesion in Caucasian patients with generalized aggressive and chronic periodontitis and to explore whether tobacco use is associated with a specific bacterial profile., Materials and Methods: A cross-sectional and analytic study was conducted in patients with aggressive and chronic periodontitis. Data were gathered on socio-demographic and periodontal variables, and RH-PCR was used to determine subgingival bacterial profile. Linear and logistic regression analyses were performed., Results: The study included 60 patients with aggressive and 123 with chronic periodontitis. Total red complex bacteria count was higher in aggressive periodontitis, mainly due to T. denticola (P = 0.015). In both periodontitis types, models showed an association between T. forsythia count and probing depth (B = 0.157, P = 0.030) and between T. denticola count and higher bleeding scores (B = 2.371, P = 0.027). Smoking did not affect the red complex bacteria count in either disease., Conclusions: The prevalence of red complex bacteria was similar between aggressive and chronic periodontitis, but their count was higher in the former. In both diseases, T. forsythia was associated with greater severity and T. denticola with more severe bleeding. Tobacco smoking was not associated with the presence of red complex bacteria in either disease., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Rheumatoid arthritis (RA) and periodontitis (PD) are chronic inflammatory disorders that cause bone loss. PD tends to be more prevalent and severe in RA patients. Previous experimental studies demonstrated that RA triggers alveolar bone loss similarly to PD. The aim of this study was to investigate if arthritis-induced alveolar bone loss is associated with modification in the oral microbiota. Checkerboard DNA-DNA hybridization was employed to analyze forty oral bacterial species in 3 groups of C57BL/6 mice: control (n = 12; without any challenge); Y4 (n = 8; received oral inoculation of Aggregatibacter Actinomycetemcomitans strain FDC Y4) and AIA group (n = 12; chronic antigen-induced arthritis). The results showed that AIA and Y4 group exhibited similar patterns of bone loss. The AIA group exhibited higher counts of most bacterial species analyzed with predominance of Gram-negative species similarly to infection-induced PD. Prevotella nigrescens and Treponema denticola were detected only in the Y4 group whereas Campylobacter showae, Streptococcus mitis and Streptococcus oralis were only found in the AIA group. Counts of Parvimonas micra, Selenomonas Noxia and Veillonella parvula were greater in the AIA group whereas Actinomyces viscosus and Neisseira mucosa were in large proportion in Y4 group. In conclusion, AIA is associated with changes in the composition of the oral microbiota, which might account for the alveolar bone loss observed in AIA mice., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background and Objectives: Salivary biomarkers may enhance diagnostic sensitivity for peri-implant disease assessment. This study aimed to investigate the association of salivary periodontopathogen count and salivary interleukin-1beta (IL-1β) level with the peri-implant crevicular fluid IL-1β response at peri-implant mucositis (PM) sites among subjects with differing periodontal disease susceptibility., Materials and Methods: Eighty-seven partially edentulous subjects having at least one implant with peri-implant mucositis were included: 40 with history of chronic periodontitis (P) and 47 with no history of periodontitis (NP). Salivary IL-1β, peri-implant crevicular fluid (PICF) IL-1β, and salivary red complex pathogen counts were recorded. Subjects were scored according to a threshold salivary pathogen level of more than 5log (10) counts and assigned a "red complex score." Quartiles of salivary and PICF IL-1β levels were also scored. Area under receiver operating curve (AUC) was computed to predict the highest PICF IL-1β score using salivary biomarker as predictors and age-adjusted logistic regression performed for the significant predictors., Results: In the NP group, red complex score (AUC = 0.758 P = 0.010) (odds ratio = 1.377) and salivary IL-1β (AUC = 0.708 P = 0.038) (odds ratio = 2.506) were significant predictors of highest PICF IL-1β quartile score. In the P group, no significant associations were noted., Conclusions: Salivary biomarkers could distinguish the "high" pro-inflammatory responders at PM sites only in subjects without inherent periodontal disease susceptibility. Periodontal susceptibility may impact the immuno-inflammatory response in sub-peri-implant niches of those with peri-implant mucositis., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Periodontal disease initiated by subgingival pathogens is linked with diminished secretion of saliva, and implies pathogenic bacteria dissemination to or affects secondary sites such as the salivary glands. MicroRNAs activated in response to bacteria may modulate immune responses against pathogens. Therefore, Sprague-Dawley rats were infected by oral lavage consisting of polymicrobial inocula, namely Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, or sham-infected for 12 weeks (n = 6). We quantified inflammatory miRNA expression levels of miRNA-132, miR-146a, and miR-155 at secondary sites to the primary infection of the gingiva, including submandibular salivary glands, lacrimal glands, and pancreas. The presence of bacteria was detected in situ at secondary sites. Infected rat gingiva showed increased relative expression of miR-155. In contrast, miRNA-155 expression was decreased in submandibular salivary glands, along with positive identification of P. gingivalis in 2/6 and T. denticola in 1/6 rat salivary glands. Furthermore, miRNA-132 and miRNA-146a were significantly decreased in the pancreas of infected rats. This study is the first to show primary periodontal infections can alter miRNA profiles in secondary sites such as the salivary gland and pancreas. Whether these alterations contribute to pathologies of salivary glands in Sjögren's syndrome or of pancreas in diabetes warrants further investigation., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Both drinking and periodontal disease are serious health and social problems. Findings on the effect of alcohol consumption on periodontal disease are inconclusive., Objectives: The aim of this study was to evaluate, in patients with alcoholic disease, the composition of the main periopathogens: Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans., Material and Methods: The study was conducted on 25 alcoholics from the Department of Alcohol Addiction Closed Treatment and 25 non-alcoholic patients from the Department of Periodontology, Wroclaw Medical University. Subgingival biofilm samples were obtained from the 4 deepest sites (≥ 4 mm). The presence of 4 bacterial taxa was analysed using the PCR technique., Results: The prevalence of bacterial species was significantly different between groups. Alcoholics showed significantly higher mean DNA counts for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Treponema denticola. In the qualitative analysis, no difference was observed between the groups. The study showed no statistically significant association between the amount of alcohol consumed and the composition of subgingival flora in patients suffering from alcoholism., Conclusions: Alcoholics demonstrated the presence of pathogenic bacteria in similar amounts to people diagnosed with chronic periodontal disease, but showed significantly higher mean DNA counts for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Treponema denticola but there is no correlation between the amount of alcohol consumption and the level of periopathogens.

Real-time quantitative PCR (Dentofl or kit) was used to detect DNA of periodontal pathogens in specimens from 92 patients with chronic periodontitis and from a control sample of 12 normal subjects. A bimodal distribution of patients by periodontium colonization with A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythensis, and T. denticola was demonstrated. A new approach to interpretation of the results of quantitative evaluation of periodontal pathogens, including the notion of pathological colonization level, led to classification of all cases with chronic generalized periodontitis into 3 groups: associated with A. actinomycetemcomitans, with T. forsythensis/T. denticola complex, and cases of uncertain genesis.