• Journal of Periodontal and Implant Science
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• v.46 no.1
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• pp.2-9
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• 2016

Purpose: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. Methods: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. Results: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. Conclusions: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.265-272
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• 2006

Dental plaque, a biofilm consisting of more than 500 different bacterial species, is an etiological agent of human periodontal disease, It is therefore important to characterize interactions among periodontopathic microorganisms in order to understand the microbial pathogenesis of periodontal disease. Previous data have suggested a synergistic effect of tow major periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia in the periodontal lesion. In the present study, to better understand interaction between P. gingivalis and T. forsythia, the coaggregation activity between these bacteria was characterized. The coaggregation activity was observed by a direct visual assay by mixing equal amount (1 ${\times}$ $10^9$)of T. forsythia and P. gingivaJis cells. It was found that the first aggregates began to appear after 5-10 min, and that the large aggregates completely settled within 1 h. Electron and epifluorescence microscopic studies confirmed cell-cell contact between two bacteria. The heat treatment of P. gingivalis completely blocked the activity, suggesting an involvement of a heat-labile component of P. gingivalis in the interaction. On the other hand, heat treatment of T. forsythia significantly increased the coaggregation activity; the aggregates began to appear immediately. The coaggregation activity was inhibited by addition of protease, however carbohydrates did not inhibit the activity, suggesting that coaggregation is a protein-protein interaction. The results of this study suggest that coaggregation between P. gingivalis and T. forsythia is a result of cell-cell physical contact, and that coaggregation is mediated by a heat-labile component of P. gingivalis and T. forsythia component that can be activated on heat treatment.

• Journal of Periodontal and Implant Science
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• v.38 no.3
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• pp.543-550
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• 2008

Purpose: The purpose of this study was to investigate induction of cytokine expression in human gingival fibroblasts (HGFs) by whole cell and the components of T. forsythia. Material and Methods: After HGFs were treated with lipopolysaccharide (LPS), membrane protein isolated from T. forsythia or culture media of T. forsythia, the induction of interleukin (IL)-1, IL-6 and IL-8 was examined with real-time PCR and ELISA. Their induction ability of cytokines was compared with whole bacteria. Result: The expression of IL-6 and IL-8 was significantly induced in HGFs by whole bacteria and membrane protein. The expression of IL-$1{\beta}$ was induced by membrane protein of T. forsythia, not by whole bacteria. LPS and condition media of T. forsythia slightly activated HGFs. Conclusion: The membrane protein of T. forsythia could be one of virulence factors.

• Journal of Periodontal and Implant Science
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• v.38 no.4
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• pp.691-698
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• 2008

Purpose: Periodontal pathogens can invade the host tissue. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The objective of this study was to visualize and evaluate presence of Porphyromonas gingivalis and Tannerella forsythia in crevicular epithelial cells of periodontally healthy subjects and chronic periodontitis patients. Materials and Methods: A total of 666 crevicular epithelial cells in the samples obtained from 27 chronic periodontitis patients and 9 healthy volunteers were examined. Specific probes for P. gingivalis and T. forsythia and a universal probe for detection of all eubacteria targeting 168 rRNA for fluorescence in situ hybridization was used in conjunction with confocal laser scanning microscopy. Results: 98.99% of sulcular epithelial cells from healthy volunteers and 84.40% of pocket epithelial cells from periodontitis patients were found to harbor bacteria. P. gingivalis and T. forsythia were discovered more often in crevicular epithelial cells from periodontitis patients. Conclusion: P. gingivalis and T. forsythia can invade crevicular epithelial cells and intracellular bacteria may act as a source of bacteria for persistent infection.

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.237-243
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• 2004

The aim of this study is to investigate the detection rate of putative periodontopathogens, Porphyromonas gingivalis, Tannerella forsythia, and Actiobacillus actinomycetemcomitans, related to cardiovascular diseases(CVD). Plaques were sampled from 15 subjects (4 sites of denture base and/or tooth) with sterilized explorers and were transported in IX PBS. The detection of periodontopathogens was performed by polymerase chain reaction with species-specific primers based on 16S rDNA. The PCR products were cloned into pGEM-T easy vector and its nucleotides were sequenced in order to confirm the specificity. Our data showed that the detection rate of P. gingivalis and T forsythia in denture base of edentulous patients was 25％ and 75％, respectively. And the detection rate of P. gingivalis and T.forsythia in denture base of patient having one more tooth was 91％. The results indicate that plaque of denture base may serve as reservoirs of oral bacteria related to CVD.

• Journal of Periodontal and Implant Science
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• v.51 no.6
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• pp.409-421
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• 2021

Purpose: The aim of this study was to compare the prevalence and bacterial load of 6 main periodontal pathogens between pairs of periodontal patients with and without type 2 diabetes mellitus. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans genotypes were also investigated. Methods: Twenty patients affected by chronic periodontitis and type 2 diabetes were retrospectively selected and matched to 20 patients without diabetes on the basis of the degree and severity of periodontal disease. Microbiological data of subgingival biofilms were analysed and compared for the examined pathogens: A. actinomycetemcomitans, P. gingivalis, Prevotella intermedia, Treponema denticola, Fusobacterium nucleatum, and Tannerella forsythia. Results: The pairs were balanced in terms of demographic and clinical parameters, except for bleeding on probing and suppuration. In the microbiological test sites (4 for each patient), the mean probing pocket depth was 6.34±1.63 mm in patients with diabetes and 6.41±1.78 mm in patients without diabetes. No significant difference between pairs in the prevalence of P. gingivalis or the distribution of its genotypes was recorded. Patients with diabetes had a significantly greater amount of total bacterial load, P. gingivalis, T. denticola, T. forsythia, and F. nucleatum (P<0.05). Moreover, patients with diabetes had a higher number of sites with a greater cell count than patients without diabetes. When compared to the total bacterial load, only T. forsythia maintained its relative load in patients with diabetes (P=0.001). Conclusions: This retrospective matched study supports the hypothesis that microbiological differences exist among periodontal patients with and without diabetes mellitus.

• Journal of Periodontal and Implant Science
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• v.44 no.6
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• pp.274-279
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• 2014

Purpose: To evaluate whether the levels of immunoglobulin G (IgG) antibody to Tanerella forsythia are associated with periodontal status. Methods: Patients with a diagnosis of chronic periodontitis were considered candidates for the study; thus 80 chronic periodontitis patients and 28 healthy persons (control group) were invited to participate in this investigation. The presence of T. forsythia was detected by polymerase chain reaction (PCR) analysis using primers designed to target the respective 16S rRNA gene sequences. Peripheral blood was collected from each subject to identify the IgG1 and IgG2 serum antibodies against T. forsythia. All microbiological and immunological laboratory processes were completed blindly, without awareness of the clinical status of the study patients or of the periodontal sites tested. Results: The bivariate analysis showed that lower mean levels of clinical attachment level (CAL) and probing depth were found in the presence of the IgG1 antibody titers against whole-cell T. forsythia; however, only the difference in CAL was statistically significant. In the presence of the IgG2 antibody titers against whole-cell T. forsythia, the periodontal parameters evaluated were higher but they did not show statistical differences, except for plaque. The unadjusted linear regression model showed that the IgG1 antibody against whole-cell T. forsythia in periodontitis patients was associated with a lower mean CAL (${\beta}=-0.654$; 95% confidence interval [CI], -1.27 to -0.28; P<0.05). This statistically significant association remained after adjusting for possible confounders (${\beta}=-0.655$; 95% CI, -1.28 to -0.29; P<0.05). On the other hand, smoking was a statistically significant risk factor in the model (${\beta}=0.704$; 95% CI, 0.24 to 1.38; P<0.05). Conclusions: Significantly lower mean levels of CAL were shown in the presence of the IgG1 antibody titers against whole-cell T. forsythia in periodontitis patients. Thus, the results of this study suggest that IgG1 antibody to T. forsythia may have been a protective factor from periodontitis in this sample.

• International Journal of Clinical Preventive Dentistry
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• v.14 no.4
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• pp.264-270
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• 2018

Objective: This study performed a quantitative analysis using the real-time polymerase chain reaction technique to examine the oral microbial prevalence in adults and intended to examine the correlations between risk factors of periodontal disease and oral bacteria and correlation between oral test scores and oral microorganisms. Methods: We examined papillary marginal attached (PMA) index, modified patient hygiene performance (M-PHP) index, probing depth (PD), modified gingival index, and oral bacteria counts and surveyed 117, 20 years or older adult males and females who visited dental clinics in the Daejeon region to analyze the prevalence and oral health. Results: The prevalence was 100% for Fusobacterium nucleatum, meaning it was observed in all examined subject, 85.5% for Parvimonas micra, 76.1% for Prevotella intermedia, and 72.6% for Tannerella forsythia. The averages of P. gingivalis and T. forsythia increased as the examined subjects were older, and there was a statistically significant difference between T. forsythia and E. nodatum in relation to medical history, between P. intermedia and P. micra in relation to gender, and between P. intermedia and E. corrodens in relation to smoking (p<0.05). For a correlation between the oral test scores and oral microorganisms, P. gingivalis and F. nucleatum was highly correlated with PD (correlation coefficient of 0.51 and 0.41) (p<0.01) while P. gingivalis, P. micra, C. rectus, and E. nodatum were significantly correlated with M-PHP index, gingival index, PD, and PMA index (p<0.01, p<0.05). Conclusion: For oral health management of adults, the age, systemic disease, and smoking are closely related to oral bacteria, and P. gingivalis, T. forsythia, F. nucleatum, P. intermedia, P. micra, C. rectus, E. corrodens, and E. nodatum are considered to be the oral microorganisms that indicate periodontal health.

• International Journal of Oral Biology
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• v.38 no.2
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• pp.81-85
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• 2013

There are estimated to be about 700 species of bacteria in the oral cavity. Based on epidemiological investigations, some of these strains have been proposed as the pathogens responsible for oral diseases such as dental caries, gingivitis and periodontitis. Since electrolyzed hydrogen-rich water has been shown to have beneficial effects on human immunity, its use has increased. In our study, the antibacterial activity of hydrogen-rich water for oralagainst bacteria associated with oral disease was evaluated. The bacterial strains Streptococcus mutans, Fusobacterium nucleatum, Porphyromonas gingivalis and Tannerella forsythia were cultured in specific growth medium. S. mutans, F. nucleatum and P. gingivalis were soaked to thein both hydrogen water and tap water for 30 sec and then inoculated onto mitis-salivarius agar and brain heart infusion agar including supplemented withvitamin K and hemin, respectively. The numbers of bacterial colonies were then measured after cultivation for 48 hours. In the case of T. forsythia, which does not grow well on agar plates, inoculations into modified new oral spirochete (NOS) broth were performed and growth curve analysis was undertaken every day with a spectrophotometer. Hydrogen water showed antibacterial activity against all four bacterial strains in comparison with tap-water. We conclude from this that hydrogen water may have a positive impact on oral hygiene by helping to remove cariogenic bacteria and periodontopathogens.

• International Journal of Oral Biology
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• v.48 no.3
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• pp.19-31
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• 2023

Peri-implantitis is a disease affecting the tissue surrounding dental implants, destroying both soft and hard tissues. A total of 2,015 studies were collected by searching items in the National Library of Medicine, including keywords, such as "peri-implantitis," "microbiota," and "microbiome." Of them, 62 studies were screened and considered eligible for analysis. Only 16 studies qualified all criteria mentioned here: "Using PCR methods for microorganism detection," "Suggesting quantified results," "Stating obvious clinical diagnosis criteria ("Bleeding on probing," "Probing pocket depth," "Suppuration," and "Radiographic bone loss")." Only 8 studies were included in the meta-analysis because the others had special issues. Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, and Epstein-Barr virus were the microbiological subjects of analysis. The odds ratio (OR) between the healthy implants and peri-implantitis were calculated for each microorganism to compare two groups, and the forest plots were suggested as the visual materials. P. gingivalis (1.392 < OR < 2.841), T. forsythia (1.345 < OR < 3.221), T. denticola (2.180 < OR < 5.150), A. actinomycetemcomitans (1.975 < OR < 6.456), P. intermedia (1.245 < OR < 3.612), and Epstein-Barr virus (1.995 < OR < 9.383). The species showed that their 95% confidence interval of odds ratio was higher than 1, indicating that they were detected more frequently in periimplantitis than in healthy implants. Meanwhile, other species, such as Fusobacterium nucleatum and Staphylococcus aureus, were not included in the meta-analysis because the number of studies was insufficient.