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Background: This study determined the prevalence of aggressive (molar-incisor pattern) (Ag/MI) periodontitis and assessed the associated subgingival bacterial-herpesvirus microbiota in Pueblo Indian adolescents in the southwestern United States., Methods: The study included 240 Pueblo Indian adolescents, aged 13-20 years old, residing in three Rio Grande River villages in New Mexico and the Hopi Pueblo reservation in Arizona. Adolescents with Ag/MI periodontitis or periodontal health provided subgingival samples for culture of bacterial pathogens and for polymerase chain reaction detection of periodontal herpesviruses., Results: Ag/MI periodontitis was detected in 22 (9.2%) Pueblo Indian adolescents, with 21 exhibiting a localized molar-incisor breakdown pattern. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and other red/orange complex bacterial pathogens predominated in Ag/MI periodontitis, whereas periodontal health yielded mainly viridans streptococci and Actinomyces species. Periodontal herpesviruses demonstrated a 3.5 odds ratio relationship with Ag/MI periodontitis. The only adolescent with generalized Ag/MI periodontitis harbored viral co-infection by cytomegalovirus plus Epstein-Barr virus Type 1, in addition to A. actinomycetemcomitans, P. gingivalis, and several other periodontopathic bacteria., Conclusions: Pueblo Indian adolescents showed an unusually high prevalence of early-age Ag/MI periodontitis predominated by periodontopathic bacteria and herpesviruses suspected to be major etiologic agents of the disease., (© 2023 American Academy of Periodontology.)

Localized juvenile (aggressive) periodontitis starts at puberty in otherwise healthy individuals and involves the proximal surfaces of permanent incisors and first molars. The disease destroys a sizeable amount of periodontal bone within a few months despite minimal dental plaque and gingival tissue inflammation. Cytomegalovirus and Epstein-Barr virus, as well as the two main periodontopathic bacteria Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, are linked to juvenile periodontitis. Juvenile periodontitis-affected teeth show cementum hypoplasia. We hypothesize that an active herpesvirus infection, at the time of root formation, hampers cementum formation and, at puberty, herpesvirus reactivation triggers an upgrowth of bacterial pathogens which produce rapid periodontal destruction on teeth with a defective periodontium. A pathogenic interaction between active herpesviruses and bacterial pathogens can potentially explain the etiology and incisor-first molar destructive pattern of juvenile periodontitis. Effective treatment of juvenile periodontitis may target the herpesvirus-bacteria co-infection., (© 2023 The Authors. Periodontology 2000 published by John Wiley & Sons Ltd.)

The prevalence of periodontitis among Thai schoolchildren is unknown. In a cross-sectional study, the prevalence and severity of periodontal diseases, in a group of Thai schoolchildren, along with the presence and numbers of bacterial species commonly associated with periodontitis were investigated. A consent form was sent out to 192 schoolchildren in one school (Chanachanupathom School) in Chana, Southern Thailand (in the age range of 12-18 years) and 119 attended for a clinical and microbiological examination. Clinical recordings included number of teeth present, DMFT, plaque index, bleeding index, clinical attachment loss (CAL), and probing pocket depth (PPD). Pooled plaque samples were analyzed with culture and qPCR against bacteria associated with periodontitis. The children had low caries experience (DMFT = 3.2 ± 2.3), poor oral hygiene, high bleeding scores, and 67 (56.3%) had at least one interproximal site with CAL ≥ 1 mm. Thirty-seven (31.1%) of the children were diagnosed with periodontitis stage I, and sixteen (13.4%) were classified as periodontitis Stage II. Aggregatibacter actinomycetemcomitans was sparsely found in all but the healthy clinical groups (gingivitis, periodontitis Stage I and II), while the groups showed a high prevalence of Fusobacterium spp., Prevotella intermedia/nigrescens, and Campylobacter species as well as of the periodontitis-associated species Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. Thai schoolchildren have poor oral hygiene with abundant amounts of plaque and high presence of bleeding. Early onset periodontitis is common but mostly in its mild form and is not associated with the presence of A. actinomycetemcomitans., (© 2023. The Author(s).)

Porphyromonas gingivalis and Tannerella forsythia have been thought to be associated with periodontitis; however comprehensive histopathological localization of bacteria in affected human periodontal tissues is not well documented. In the present study, we examined formalin-fixed paraffin-embedded gingival and subgingival granulation tissues from 71 patients with chronic periodontitis and 11 patients with aggressive periodontitis, using immunohistochemistry with novel monoclonal antibodies specific to P. gingivalis or T. forsythia, together with quantitative real-time polymerase chain reaction for each bacterial DNA. Immunohistochemisty revealed both bacterial species extracellularly, as aggregates or within bacterial plaque, and intracellularly in stromal inflammatory cells, squamous epithelium, and capillary endothelium of granulation tissue. Combined analysis with the results from polymerase chain reaction suggested that localization and density of T. forsythia is closely associated with those of P. gingivalis, and that bacterial density is a factor responsible for the cell-invasiveness and tissue-invasiveness of these periodontal bacteria. Detection of these bacteria in the capillary endothelium in some samples suggested possible bacterial translocation into the systemic circulation from inflamed gingival and subgingival granulation tissues. Immunohistochemistry with the novel antibodies showed high specificity and sensitivity, and can be used to locate these periodontal bacteria in routinely-used formalin-fixed paraffin-embedded human tissue sections from systemic locations.

In vitro studies revealed that Porphyromonas gingivalis (Pg), a pathogen intimately associated with the onset and progression of periodontitis, is able to activate platelets, thus linking periodontal inflammation with the endangerment of vascular health. As wild-type Pg strains are characterized by major genetic heterogeneity, the commonness of platelet-activating Pg strains in periodontitis patients is unknown as of yet. Therefore, this study evaluated the platelet activation capacity of wild-type Pg isolates sampled from patients with aggressive periodontitis., Methods: Extent and velocity of platelet aggregation were determined by light transmission aggregometry. Platelet surface expression of P-selectin was measured by flow cytometry, activation of p38 MAP kinase, and protein kinase C by Western blot using phospho-specific antibodies., Results: Pg isolates displayed high variability regarding extent and velocity of platelet activation, as well as the involved activating pathways. Corresponding results were observed for platelet P-selectin expression, activation of p38 MAP kinase, or protein kinase C. Inhibitors of platelet immune receptor FcγRIIA and protease-activated receptors revealed several, diverging pathways of activation. Some isolates induced platelet aggregation even in the presence of potent therapeutical platelet inhibitors., Conclusions: Chronic bacteremia involving specific, platelet-activating Pg strains may constitute a substantial hazard for the integrity of cardiovascular health., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Periodontal disease (PD) is generated by microorganisms. These microbes can enter the general circulation causing a bacteraemia. The result can be adverse systemic effects, which could promote conditions such as cardiovascular disease. Level A evidence supports that PD is independently associated with arterial disease. PD is a common chronic condition affecting the majority of Americans 30 years of age and older. Atherosclerosis remains the largest cause of death and disability. Studies indicate that the adverse cardiovascular effects from PD are due to a few putative or high-risk bacteria: Aggregatibacter actinomycetemcomitans , Porphyromonas gingivalis , Tannerella forsythia , Treponema denticola or Fusobacterium nucleatum There are three accepted essential elements in the pathogenesis of atherosclerosis: lipoprotein serum concentration, endothelial permeability and binding of lipoproteins in the arterial intima. There is scientific evidence that PD caused by the high-risk pathogens can influence the pathogenesis triad in an adverse manner. With this appreciation, it is reasonable to state PD, due to high-risk pathogens, is a contributory cause of atherosclerosis. Distinguishing this type of PD as causal provides a significant opportunity to reduce arterial disease., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.)

Background: Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) have been associated with aggressive (AgP) and chronic periodontitis., Objective: The aim of this study was to evaluate the levels of Aa and Pg in gingival crevicular fluid (GCF) of patients with AgP and its relation with clinical parameters., Design: Sixteen females and fourteen males with clinical diagnosis of AgP aged 17-23 years and their match's controls, were included in this study. Clinical recording concerning probing pocket depth, clinical attachment level, plaque index and gingival bleeding index were performed at baseline, 30 and 60 days after baseline. After clinical examination GCF samples were analyzed for Aa and Pg with a real-time polymerase chain reaction technique. Patients group was treated with a combined of mechanical and oral antibiotic therapy (doxycycline 100 mg/day, during 21 days). A multivariate analysis was used to determine the relationship between Aa and Pg counts with clinical parameters., Results: GCF from all subjects was positive for Aa and PG. In controls Pg concentration was higher than Aa (Pg: 42,420 ± 3,034 copies/ml; Aa: 66.6 ± 5.4 copies/ml p < 0.001) while in patients both microbes showed the same concentration (Aa: 559,878 ± 39,698 Pg: 572,321 ± 58,752). A significant and positive correlation was observed between counts of Aa and Pg (R square: 0.7965, p < 0.0001). Female showed more counts/ml. Aa might be closely associated with clinical parameters while Pg did not. At 30 and 60 days Aa counts in patients were similar to controls while Pg counts were equal to baseline. However, in spite of Pg presence a clinical improvement was observed in all patients., Conclusions: In our population the presence of Aa may be associated with AgP while Pg may be in GCF as an opportunistic pathogen which might caused disease when the ecological balance was favorable., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Cigarette smokers are more susceptible to periodontal diseases and are more likely to be infected with Porphyromonas gingivalis than non-smokers. Furthermore, smoking is known to alter the expression of P. gingivalis surface components and compromise immunoglobulin (Ig)G generation. The aim of this study is to evaluate whether the overall IgG response to P. gingivalis is suppressed in smokers in vivo and whether previously established in vitro tobacco-induced phenotypic P. gingivalis changes would be reflected in vivo., Methods: The authors examined the humoral response to several P. gingivalis strains as well as specific tobacco-regulated outer membrane proteins (FimA and RagB) by enzyme-linked immunosorbent assay in biochemically validated (salivary cotinine) smokers and non-smokers with chronic periodontitis (CP: n = 13) or aggressive periodontitis (AgP: n = 20). The local and systemic presence of P. gingivalis DNA was also monitored by polymerase chain reaction., Results: Smoking was associated with decreased total IgG responses against clinical (10512, 5607, and 10208C; all P <0.05) but not laboratory (ATCC 33277, W83) P. gingivalis strains. Smoking did not influence IgG produced against specific cell-surface proteins, although a non-significant pattern toward increased total FimA-specific IgG in patients with CP, but not AgP, was observed. Seropositive smokers were more likely to be infected orally and systemically with P. gingivalis (P <0.001), as determined by 16S RNA analysis., Conclusion: Smoking alters the humoral response against P. gingivalis and may increase P. gingivalis infectivity, strengthening the evidence that mechanisms of periodontal disease progression in smokers may differ from those of non-smokers with the same disease classification.

Background: The microbiologic feature of aggressive periodontitis (AgP) in Chinese patients has not yet been determined. This study aims to investigate the prevalence of eight periodontal microorganisms and the distribution of the Porphyromonas gingivalis fimA genotype in a cohort of Chinese patients with AgP., Methods: Saliva and pooled subgingival plaque samples were collected from 81 patients with AgP (25 with incisor-first molar type and 56 with generalized type [GAgP]) and 34 periodontally healthy controls. Eight periodontal microorganisms, including Aggregatibacter actinomycetemcomitans, P. gingivalis, Tannerella forsythia, Treponema denticola, Campylobacter rectus, Prevotella intermedia, Prevotella nigrescens, and Fusobacterium nucleatum were detected in these samples by the polymerase chain reaction (PCR). In addition, the distribution of fimA genotypes was assessed in P. gingivalis-positive individuals by PCR., Results: The prevalence of P. gingivalis, T. forsythia, T. denticola, C. rectus, P. intermedia, F. nucleatum, and A. actinomycetemcomitans in patients with AgP was significantly higher than that in healthy controls. The prevalence of A. actinomycetemcomitans in patients with GAgP was relatively low (30.4%) compared with other pathogens. Results of logistic regression analysis showed that younger patients were more likely to harbor A. actinomycetemcomitans (odds ratio = 2.85). Type II was the most prevalent fimA genotype of P. gingivalis in patients with AgP., Conclusions: P. gingivalis, T. forsythia, T. denticola, C. rectus, P. intermedia, and F. nucleatum were the predominant periodontal pathogens of patients with GAgP in China. Type II of fimA was the most prevalent genotype of P. gingivalis in patients with AgP. The prevalence of A. actinomycetemcomitans in patients with GAgP was relatively low.

In the eighties of the former century it was observed that individuals had different modes of development of periodontitis, largely for unknown reasons. It was assumed that periodontal disease developed as a result of the presence of virulent micro-organisms in combination with an inadequate host response. The concept of this (im)balance in host parasite interactions developed further, in part because of new research techniques. With respect to oral bacteria it was shown that periodontal pathogens like Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia are risk factors for periodontitis. Furthermore, genetic polymorphisms (SNP's) have been shown to modulate the host response against the periopathogens. Medical and lifestyle factors, e.g. diabetes mellitus, smoking, psychological stress and, to a minor extent, also hormonal changes can have an impact on the progression of periodontitis. All this information contributes to determining the individual risk profile of a patient for developing periodontitis and for estimating the prognosis of periodontal treatment and maintenance care on an individual basis.

Objective: The microbial eatiology of aggressive perodontitis is different worldwidly. The purpose of this study was to determine the presence or absence of two anaerobic, highly pathogenic bacteria, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis (P.g) in aggressive periodontitis in a Moroccan population., Material and Methods: Our study involved a sample of 32 subjects, 20 with aggressive periodontitis and 12 with a healthy periodontium. A pool of bacterial samples was performed in each patient looking for Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis by the method of bacterial culture., Results and Conclusion: Microbiological analysis showed an association of the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis with aggressive periodontitis in Morocco. Porphyromonas gingivalis would be less present than Aggregatibacter actinomycetemcomitans in aggressive periodontitis in Morocco (30% vs 83%)(p < or = 0.05%). However, given the small sample size other studies are necessary.

This study aimed to investigate the prevalence and levels of major periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in subgingival plaque samples of a group of Japanese patients with aggressive periodontitis (AgP) and chronic periodontitis (CP). A total of 40 patients with clinical diagnosis of AgP or CP and 10 periodontally healthy volunteers were subjected to clinical and microbiological analysis. Subgingival plaque samples were analyzed for A. actinomycetemcomitans, P. gingivalis and T. forsythia with a real-time polymerase chain reaction (PCR) technique. The prevalence of P. gingivalis and T. forsythia was relatively high in patients with periodontitis: over 60% of AgP or CP patients harbored these pathogens whereas they were not detected in the subgingival plaque samples from periodontally healthy individuals. P. gingivalis and T. forsythia were relatively frequently detected together in AgP and CP patients. No significant differences in the prevalence or level of the 3 pathogens were found between periodontitis groups. The proportion of T. forsythia was approximately 4-fold higher in CP group than in AgP group (P = 0.02). In periodontitis patients, a significant positive correlation was found between periodontal parameters (probing depth and clinical attachment level) and the numbers of total bacteria, P. gingivalis and T. forsythia. No distinct pattern of the subgingival profile of these pathogens was discerned between the two disease entities, except for the difference in the proportion of T. forsythia. The red complex bacteria, P. gingivalis and T. forsythia were highly prevalent in this population of Japanese AgP and CP patients, collaborating their roles in periodontitis., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background and Objectives: Immunoglobulin (Ig) G1 plays an important role in the adaptive immune response. Kgp, a lysine-specific cysteine protease from Porphyromonas gingivalis, specifically hydrolyses IgG1 heavy chains. The purpose of this study was to examine whether cleavage of IgG1 occurs in gingival crevicular fluid (GCF) in vivo, and whether there is any association with the presence of Porphyromonas gingivalis and other periodontopathogens., Material and Methods: GCF was obtained from nine patients with aggressive periodontitis, nine with chronic periodontitis and five periodontally healthy individuals. The bacterial loads of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Treponema denticola, Prevotella intermedia and Tannerella forsythia were analysed by real-time polymerase chain reaction, and the presence and cleavage of IgG1 and IgG2 were determined using Western blotting. Kgp levels were measured by ELISA., Results: Cleaved IgG1 was identified in the GCF from 67% of patients with aggressive periodontitis and in 44% of patients with chronic periodontitis. By contrast, no cleaved IgG1 was detectable in healthy controls. No degradation of IgG2 was detected in any of the samples, regardless of health status. Porphyromonas gingivalis was found in high numbers in all samples in which cleavage of IgG1 was detected (P < 0.001 compared with samples with no IgG cleavage). Furthermore, high numbers of Tannerella forsythia and Prevotella intermedia were also present in these samples. The level of Kgp in the GCF correlated with the load of Porphyromonas gingivalis (r = 0.425, P < 0.01). The presence of Kgp (range 0.07-10.98 ng/mL) was associated with proteolytic fragments of IgG1 (P < 0.001). However, cleaved IgG1 was also detected in samples with no detectable Kgp., Conclusion: In patients with periodontitis, cleavage of IgG1 occurs in vivo and may suppress antibody-dependent antibacterial activity in subgingival biofilms especially those colonized by Porphyromonas gingivalis., (© 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: Short chain fatty acids (SCFAs), such as succinic acid, acetic acid, propionic acid, butyric acid, etc. are metabolic product of putative periodontal pathogens, which play significant roles in periodontitis. The aim of this study was to analyze the relationship between Porphyromonas gingivalis (P. gingivalis), Treponema denticola (T. denticola), and the concentration of SCFAs in gingival crevicular fluid (GCF) of patients with aggressive periodontitis (AgP)., Methods: GCF was sampled from 4 sites per individual in 20 patients with AgP and 14 healthy controls. Concentrations of SCFAs, including succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid in the supernant of GCF were analyzed by high performance capillary electrophoresis (HPCE), P. gingivalis and T. denticola in the deposit of the same GCF were detected by PCR with their electrophoretic band quantified., Results: The concentrations of succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid, the prevalence and PCR band quantity of P. gingivalis and T. denticola in GCF were all significantly higher in patients with AgP than that of healthy controls. In patients with AgP, butyric acid concentration was significantly higher in P. gingivalis positive sites than negative sites [2.87 (0.99, 4.36) mmol/L vs. 0.33 (0.00, 1.44) mmol/L, P<0.05], the concentrations of succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid were positively correlated with PCR band quantity of P. gingivalis (r value was 0.334, 0.548, 0.411, 0.493, 0.273, respectively, P<0.05); the concentrations of SCFAs were significantly higher in T. denticola positive sites than negative sites: succinic acid, 1.67 (1.15, 2.11) mmol/L vs. 0.80 (0.48, 1.06) mmol/L; acetic acid, 31.95 (23.77, 43.13) mmol/L vs.12.51 (7.57, 15.69) mmol/L; propionic acid, 11.86 (6.55, 14.98) mmol/L vs. 2.82 (1.71, 7.03) mmol/L; butyric acid, 3.45 (2.41, 4.78) mmol/L vs. 0.54 (0.00, 1.56) mmol/L; isovaleric acid, 2.23 (1.05, 3.85) mmol/L vs. 0.62 (0.00, 2.33) mmol/L. The concentrations of succinic acid, acetic acid, propionic acid, butyric acid were positively correlated with PCR band quantity of T. denticola (r value was 0.443, 0.702, 0.625, 0.557, respectively, P<0.05)., Conclusion: SCFAs concentrations reflect the quantity of P. gingivalis and T. denticola in patients with AgP, and may be an indicator to the disease progression in patients with AgP.

Objective: To evaluate the changes in the clinical parameters, and the prevalence and quantities of three major periodontopathic bacteria, namely Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia, in subgingival plaque collected from patients with generalized chronic periodontitis (GCP) or generalized aggressive periodontitis (GAgP) in response to nonsurgical periodontal therapy., Method and Materials: 73 GCP patients and 57 GAgP patients were enrolled in this study. Clinical parameters, including probing depth (PD), clinical attachment loss (CAL), and Sulcus Bleeding Index (SBI) were measured. The prevalence and quantities of the three bacteria collected from the subgingival plaque were detected by real-time PCR. Both clinical and microbiologic parameters were evaluated at baseline, 4, and 12 weeks after the nonsurgical periodontal treatment., Results: PD, CAL, and SBI were significantly improved in GCP and GAgP groups at 4 and 12 weeks after nonsurgical periodontal therapy, compared to the baseline levels. The prevalence and quantities of P gingivalis in GCP at baseline (82.19% and 3.35E+5) were statistically higher than those found in GAgP (66.67% and 1.08E+5; P < .05). After therapy, the prevalence and quantities of the three bacteria were sufficiently reduced in both groups at 4 and 12 weeks. There was no significant difference in improvement of clinical and microbiologic parameters between the GCP and GAgP patients after treatment., Conclusion: There was a difference in P gingivalis prevalence and quantity between the GCP and GAgP patients at baseline. In addition, nonsurgical periodontal therapy was effective in the treatment of clinical symptoms and the major periodontopathic bacterial control between GCP and GAgP patients.

Objectives: Only a few studies have attempted to detect differences in microbiologic profiles of patients with chronic periodontitis (CP) and aggressive periodontitis (AgP). The aim of this analysis was to assess if clinical diagnosis or other subject factors showed association with the presence of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in a cohort of periodontitis patients., Method and Materials: Statistical analysis for association between bacterial detection and clinical diagnosis was performed on a total of 267 consecutive periodontitis cases diagnosed with either CP (n = 183) or AgP (n = 84). All subjects had microbiologic samples collected from the four deepest pockets and analyzed by nested polymerase chain reaction., Results: A actinomycetemcomitans was detected in 54% and 48% of CP and AgP subjects, respectively. A slightly higher detection of P gingivalis was found in CP (67% ) compared with AgP (52%) cases. The detection of P gingivalis was associated with older age (P = .002), less disease severity (P = .015), and IL6-1480 genotypes (P = .026), while A actinomycetemcomitans was associated with IL6-1480 genotypes (P = .001)., Conclusion: Detection of known periodontopathogenic bacteria is not able to discriminate different forms of periodontitis.

In periodontitis, an effective host-response is primarily related to neutrophils loaded with serine proteases, including elastase (NE) and protease 3 (PR3), the extracellular activity of which is tightly controlled by endogenous inhibitors. In vitro these inhibitors are degraded by gingipains, cysteine proteases produced by Porphyromonas gingivalis. The purpose of this study was to determine the level of selected protease inhibitors in gingival crevicular fluid (GCF) in relation to periodontal infection. The GCF collected from 31 subjects (nine healthy controls, seven with gingivitis, five with aggressive periodontitis and 10 with chronic periodontitis) was analyzed for the levels of elafin and secretory leukocyte protease inhibitor (SLPI), two main tissue-derived inhibitors of neutrophil serine proteases. In parallel, activity of NE, PR3 and arginine-specific gingipains (Rgps) in GCF was measured. Finally loads of P. gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia and Treponema denticola were determined. The highest values of elafin were found in aggressive periodontitis and the lowest in controls. The quantity of elafin correlated positively with the load of P. gingivalis, Ta. forsythia and Tr. denticola, as well as with Rgps activity. In addition, NE activity was positively associated with the counts of those bacterial species, but not with the amount of elafin. In contrast, the highest concentrations of SLPI were found in periodontally healthy subjects whereas amounts of this inhibitor were significantly decreased in patients infected with P. gingivalis. Periodontopathogenic bacteria stimulate the release of NE and PR3, which activities escape the control through degradation of locally produced inhibitors (SLPI and elafin) by host-derived and bacteria-derived proteases., (© 2011 John Wiley & Sons A/S.)

Background and Objective: Aggressive periodontitis pathogenesis still is not completely understood in the literature regarding the relationship between microbial and inflammatory aspects. So this study aimed to compare microbial and inflammatory patterns in the gingival crevicular fluid of generalized aggressive and chronic periodontitis patients., Material and Methods: Forty aggressive and 28 chronic periodontitis patients were selected. Biofilm and gingival crevicular fluid were collected from a deep pocket (periodontal probing depth >7 mm) and a moderate pocket (periodontal probing depth = 5 mm) of each patient, and microbiological and immunoenzymatic assays were performed. Real-time PCR was used to determine quantities of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Enzyme-linked immunosorbent assay (ELISA) was employed to determine gingival crevicular fluid levels of interleukin-1beta, interferon-gamma, prostaglandin E(2) and interleukin-10. In addition, immunoglobulin G (IgG) levels against A. actinomycetemcomitans and P. gingivalis lipopolysaccharide were also determined by ELISA. Analysis of variance/Tukey test, Mann-Whitney U-test and the Pearson correlation test were used to determine differences and correlations between variables analysed (alpha = 5%)., Results: Patients suffering from generalized aggressive periodontitis had their mouth colonized by higher amounts of A. actinomycetemcomitans and P. gingivalis than chronic periodontitis patients. Conversely, the gingival crevicular fluid levels of IgG against both pathogens were statistically inferior in aggressive periodontitis patients (p < 0.05). With regard to gingival crevicular fluid levels of cytokines, aggressive periodontitis patients presented reduced levels of interleukin-10 (p < 0.05)., Conclusion: In comparison to chronic periodontitis, generalized aggressive periodontitis patients have an imbalance in the host response, with reduced levels of interleukin-10 and IgG, and increased periodontal pathogens., ((c) 2010 John Wiley & Sons A/S.)

Generalized aggressive periodontitis (GAgP) is an inflammatory condition resulting in destruction of tooth-supporting tissues. We examined the production of IL-1beta, IL-6, tumour necrosis factor (TNF)-alpha, IL-12 and IL-10 in cultures of peripheral mononuclear cells (MNC) from 10 patients with GAgP and 10 controls stimulated with periodontal pathogens or a control antigen, tetanus toxoid (TT) in the presence of autologous serum. The pathogens used were Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, either as type strains or bacteria isolated from the participants' inherent oral flora. The P. gingivalis -induced production of IL-6 was approximately 2.5-fold higher in patients with GAgP than in healthy controls (P < 0.05), while the corresponding TNF-alpha production was non-significantly elevated. IL-1beta production induced by P. gingivalis, as all cytokine responses induced by Pr. intermedia, F. nucleatum and TT was similar in the two groups. A reduced IL-12p70 response to Pr. intermedia and F. nucleatum was observed in smokers compared to non-smoking patients (P < 0.02). To assess the role of serum factors in the elevated IL-6 response to P. gingivalis, MNC from two donors free of disease were stimulated with this bacterium in the presence of the various patient and control sera. An elevated IL-6 and TNF-alpha response was observed in the presence of patient sera (P < 0.01 and P < 0.04, respectively). The data suggest that an exaggerated production of IL-6 occurs in GAgP, and that pro-inflammatory serum factors play an essential role in the response.

Background and Objective: As a pro-inflammatory cytokine, interleukin-2 mediates the activation, growth and differentiation of T and B lymphocytes and natural killer cells. Promoter polymorphisms of the interleukin-2 gene have been associated with altered interleukin-2 production or identified as prognostic markers for various infectious diseases. Therefore, the aim of this study was to evaluate two polymorphisms at positions -330 T/G and 166 G/T in patients with generalized chronic periodontitis (n = 58) or generalized aggressive periodontitis (n = 73) in comparison with periodontitis-free controls (n = 69)., Material and Methods: Both interleukin-2 polymorphisms were analyzed using the polymerase chain reaction with sequence-specific primers. Distributions of single alleles, genotypes and haplotypes were calculated using the chi-square test. Risk factor analyses were carried out by logistic regression with respect to established cofactors for periodontitis. The presence of subgingival bacteria in an individual were analyzed using a molecular biological method (the micro-Ident test)., Results: The interleukin-2 genotype -330 TG occurred less frequently in patients with chronic periodontitis (25.9% vs. 49.3%). Moreover, this genotype decreased the adjusted odds ratio for chronic periodontitis (odds ratio = 0.394), whereas the interleukin-2 genotype 166 TT and the haplotype combination interleukin-2 -330,166 TT : TT were associated with an increased adjusted odds ratio (odds ratio = 2.82 or 2.97). For the latter interleukin-2 combination, a positive association for the subgingival presence of Porphyromonas gingivalis (81.3% vs. 59.5%) and bacteria of the 'red complex' (78.1% vs. 56.0%) was shown., Conclusion: The interleukin-2 genotypes -330 TG and 166 TT, as well as the combination genotype interleukin-2 TT : TT, could be putative prognostic factors for chronic periodontitis.

Background and Objective: This study analyzed the interaction of Porphyromonas gingivalis ATCC 33277 and Aggregatibacter actinomycetemcomitans Y4 with peripheral blood polymorphonuclear neutrophils taken from patients with aggressive periodontitis and chronic periodontitis., Material and Methods: Peripheral blood polymorphonuclear neutrophils obtained from 12 patients with chronic periodontitis, six patients with aggressive periodontitis and 12 healthy controls were exposed to P. gingivalis and A. actinomycetemcomitans following opsonization of the bacteria using the patient's own serum. Serum immunoglobulin G (IgG) levels against both periodontopathogens were measured. Phagocytosis and killing of the bacteria, as well as the extracellular human neutrophil elastase activity, were quantified. The total amount and the extracellular release of reactive oxygen species were measured using luminol-dependent and isoluminol-dependent chemiluminescence., Results: Polymorphonuclear neutrophils from patients with chronic (62.16 +/- 19.39%) and aggressive (43.26 +/- 26.63%) periodontitis phagocytosed more P. gingivalis than the healthy controls (24.43 +/- 19.87%) at the 30-min time point after exposure to the bacteria (p < 0.05). High serum IgG levels against P. gingivalis and A. actinomycetemcomitans were detected in subjects with periodontitis. Polymorphonuclear neutrophils from subjects with chronic and aggressive periodontitis released significantly more reactive oxygen species and demonstrated greater human neutrophil elastase activity in the absence of any stimulus than polymorphonuclear neutrophils from healthy controls (p < 0.05). Polymorphonuclear neutrophils in chronic periodontitis released significantly more reactive oxygen species when exposed to P. gingivalis and A. actinomycetemcomitans than polymorphonuclear neutrophils in aggressive periodontitis., Conclusion: High serum IgG levels against P. gingivalis and A. actinomycetemcomitans promote phagocytosis in periodontitis. The extracellular release of reactive oxygen species and neutrophil elastase by polymorphonuclear neutrophils may also contribute to damage of the surrounding periodontal tissues.

Objective: To measure the anti-Pg (porphyromonas gingivalis, Pg) IgG antibody titers in serum in patients with aggressive periodontitis (AgP); to study the association of IgG titers with periodontal status (alveolar bone loss); to analyze the age, gender or smoking status on IgG levels in patients with aggressive periodontitis., Methods: (1) Ninety-one AgP subjects with an average age of 27.6 years (females 56, males 35, and 14 male-smokers) were collected. Thirty-five healthy subjects with an average age of 29.4 years (males 10 and females 25) were also collected as control group. (2) Questionnaire, clinical evaluations and assessments of alveolar bone loss were made. (3) Patients were categorized into two groups: severe AgP group (58 subjects, with >14 teeth of moderate-severe alveolar bone loss); light-moderate group (33 subjects). (4) To collect serum sample of patients and determine the anti-Pg IgG antibody titers in serum using ELISA., Results: (1) The mean IgG antibody titers to Pg in AgP group (8.55 +/-3.24) were significantly higher than that in control group (4.78+/-2.58, P=0.0004). (2) No significant difference [(8.38+/-3.73) and (8.67+/-2.90), respectively] of mean anti-Pg IgG titers was found between male and female. (3) The mean of anti-Pg IgG titers (9.26+/-2.58) in elder group (aged>25) was significantly higher than that in younger patients (6.96+/-3.98, P=0.0014). (4) Smokers had significantly lower anti-Pg IgG titers (7.33+/- 3.72) than non-smokers (8.78+/-3.12, P=0.016). (5) Anti-Pg IgG titers in severe AgP group was significantly higher than that in light-moderate AgP group. (6)The results of multi-factor regression analysis showed that smoking, age and status of alveolar bone loss were the factors affecting anti-Pg IgG titers., Conclusion: The titers of IgG antibody to Pg in serum were related to age, smoking and periodontal status in AgP patients.

Objective: To examine Porphyromonas gingivalis (Pg) in subgingival plaque of the patients with periodontitis and to find out the rules of Pg colonization after periodontal initial treatment., Methods: A total of 1620 subgingival plaque samples were collected from 180 subjects including chronic periodontitis (CP) patients (n = 90), and aggressive periodontitis (AgP) patients (n = 90) in different periods of periodontal initial therapy-the baseline, 6 weeks, and 12 weeks after treatment. The following periodontal clinical parameters were recorded with Florida probe at sampled sites: probing depth (PD), clinical attachment loss (CAL), and bleeding on probing (BOP). Quantities of Pg were examined by AmpliFluor endpoint quantitative polymerase chain reaction., Results: At the 6th week of periodontal initial therapy, there were 61 (22.6%) and 66 (24.4%) Pg increased sites respectively, in which no significant difference was detected (P > 0.05). At baseline of periodontal initial therapy, more severe periodontal clinical parameters of Pg increased sites were observed than those of Pg stationary sites. At the 12th week, however, there were 96 (35.6%) and 18 (6.7%) Pg increased sites respectively, significant difference detected (P < 0.05). At 6th week of periodontal initial therapy, more severe periodontal clinical parameters of Pg increased sites were observed than those of Pg stationary sites., Conclusions: Pg colonization in AgP and CP patients started 6 weeks after periodontal initial therapy, but the recolonization pattern was different between these two groups of patients. Severe periodontitis sites in baseline seemed to place them at risk of Pg colonization after periodontal initial therapy.

Objective: The aim of this study was to investigate effects of immunodeficiency on the periodontal infection characters of the specific pathogens of juvenile periodontitis., Methods: A total of 36 immunodeficient guinea pigs produced by twice whole-body irradiation with 60Co were divided randomly into four groups, in which Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and A. actinomycetemcomitans with P. gingivalis were inoculated into the gingival sulcus of two mandibular incisors respectively. The pigs in the control group did not receive any inoculation. At 2, 3 and 6 weeks after inoculation, three animals in each group were sacrificed successively. Clinical and histological examinations were used to examine the changes in the periodontal tissues. The other 36 normal guinea pigs were divided into four groups and treated in a similar way described above., Results: Significant periodontal damages were noted in immunodeficient pigs inoculated with A. actinomycetemcomitans, P. gingivalis or A. actinomycetemcomitans and P. gingivalis in 2 and 3 weeks after bacterial inoculation. The damages were more severe than in the normal groups. The immunodeficient groups demonstrated larger numbers of osteoclasts than the normal groups (P < 0.05)., Conclusion: The loss of periodontal tissue in immunodeficient hosts is much serious than those with normal defence system, after they are infected with A. actinomycetemcomitans and P. gingivalis. Abnormal defence system in hosts may play an important role in onset and development of juvenile periodontitis.

The terms 'early onset periodontitis' (EOP) and 'juvenile periodontitis' (JP) were replaced by that of 'aggressive periodontitis' in a recent international workshop for the classification of periodontal diseases and conditions. The chief etiologic agent for aggressive periodontitis is considered to be Actinobacillus actinomycetemcomitans in localized juvenile periodontitis. Porphyromonas gingivalis is also mentioned as the etiologic agent of the aggressive periodontitis, although to date its role remains questionable. This communication describes three cases of aggressive periodontitis found to be associated with P. gingivalis but not A. actinomycetemcomitans by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Our findings clarify the role of P. gingivalis as an etiologic agent in this type of periodontitis and confirm its inclusion in the current definition of aggressive periodontitis.

Microorganisms typically adapt to environmental cues by turning on and off the expression of virulence genes which, in turn, allows for optimal growth and survival within different environmental niches. This adaptation strategy includes sensing and responding to changes in nutrients, pH, temperature, oxygen tension, redox potential, microbial flora, and osmolarity. For a bacterium to adhere to, penetrate, replicate in, and colonize host cells, it is critical that virulence genes are expressed during certain periods of the infection process. Thus, throughout the different stages of an infection, different sets of virulence factors are turned on and off in response to different environmental signals, allowing the bacterium to effectively adapt to its varying niche. In this review, we focus on the regulation of virulence gene expression in two pathogens which have been implicated as major etiological agents in adult and juvenile periodontal diseases: Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. Understanding the mechanisms of virulence gene expression in response to the local environment of the host will provide crucial information in the development of effective treatments targeted at eradication of these periodontal disease pathogens.

Background, Aims: The aim of this study was to evaluate antibody responses against Porphyromonas gingivalis (P. gingivalis) infection in early-onset periodontitis (EOP) patients to elucidate further the host-parasite interactions in the pathogenesis of EOP., Method: 16 P. gingivalis-infected EOP and 20 adult periodontitis (AP) patients, and 18 periodontally healthy subjects (HS) participated in this study. Serum immunoglobulin G (IgG) antibody levels and avidities against extracted P. gingivalis whole cells were measured. The components of P. gingivalis outer membrane antigens (OMA) reacting to patients' sera were analysed from the molecular weights by Western blotting. Serum antibody levels against P. gingivalis lipopolysaccharide (LPS) were also measured. The ability of the patients' sera to block interleukin-1beta (IL-1beta) production by human mononuclear cells in response to P. gingivalis LPS was examined., Results: Antibody levels were positively correlated with antibody avidities in both EOP and AP patients (r=0.91, r=0.72, p<0.0005, respectively), while not significantly so in HS (r=0.09). There was variability in the antigen recognition of P. gingivalis OMA in EOP and AP patients. Smear and 53-kDa protein were more frequently recognized by sera of EOP and AP patients rather than that of HS (p<0.05). The smear was partly diminished by absorption with P. gingivalis LPS, indicating the smear antigen was partly composed of LPS. There was high correlation between antibody levels against P. gingivalis whole-cell extracts and LPS in EOP and AP patients (r=0.81, p=0.0002, r=0.87, p<0.0001, respectively), while not significant in HS (r=0.22). The sera of EOP and AP patients with high IgG titre to P. gingivalis LPS blocked IL-1beta production more effectively than that of the patients with low IgG titre to P. gingivalis LPS., Conclusions: These results indicate that EOP patients' antibody response against P. gingivalis infection does not differ significantly from that of AP patients. The person-to-person heterogeneous antibody production against P. gingivalis LPS could contribute to our understanding of the relationship between the defensive ability of EOP patients and their chronic infection with this pathogen.

Background: The aim of this study was to examine crevicular polymorphonuclear neutrophils (PMN) of patients with rapidly progressive periodontitis (RPP) for their in vitro phagocytic activity and intracellular killing of Porphyromonas gingivalis ATCC 33277 and two strains of Actinobacillus actinomycetemcomitans (NCTC 9710 - type strain and Tanner FDC 44 - leukotoxin producing strain)., Patients and Methods: 18 patients with RPP and nine healthy controls were included in the study. Phagocytosis and intracellular killing were assessed by fluorescence microscopy after staining with acridine orange. The percentage of phagocytosing PMN was determined. The phagocytic cells were then separated into two groups; those containing < 10 phagocytosed bacteria and those containing > 10 bacteria. The percentage of PMN containing viable bacteria was also determined., Results: The leukotoxic A. actinomycetemcomitans strain was phagocytosed to a lesser degree than the corresponding type strain. The number of phagocytosing cells obtained from the RPP patients did not differ from the controls. However, in healthy subjects there were more phagocytes with more than ten ingested P. gingivalis than in RPP patients. The intracellular killing was diminished in the periodontitis group for P. gingivalis and for both A. actinomycetemcomitans strains., Conclusion: The PMN of patients with RPP show deficiencies in phagotcytic activity and in the intracellular killing or peridontopathogenic bacteria.

Background, Aims: This study investigated the prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans among various periodontitis patients and healthy individuals in Japan, and correlated it with clinical parameters., Method: Subgingival plaque samples were collected from 21 patients with adult periodontitis (AP), 8 with rapidly progressive periodontitis (RPP) and 15 healthy individuals., Results: The frequency detected in culture was as follows: B. forsythus was found in 47.6% of AP sites and in 37.5% of RPP sites. P. gingivalis was identified in 64.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in 4.8% of AP and 3.1% of RPP sites. The 3 species were detected in only 2 of the healthy individuals. The proportion of B. forsythus in the total microflora in culture was 0.07% in the healthy group, 4.1% in AP and 2.4% in RPP. The proportions of P. gingivalis were 0% in the healthy group, 18.8% in AP and 16.2% in RPP. The proportion of A. actinomycetemcomitans was very low in all 3 groups. A DNA probe detected B. forsythus in 78.6% of AP and 65.6% of RPP sites, as well as P.gingivalis in 58.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in only 1.2% of AP sites. The 3 species were undetectable in the healthy group., Conclusions: The prevalence and the proportion of B. forsythus and P. gingivalis were significantly correlated with clinical parameters, suggesting that B. forsythus and P. gingivalis are closely related to AP and RPP in the Japanese population.

Background: Although herpesviruses have been associated with adult periodontitis, their relationship with juvenile periodontitis (JP) has not been established. This case-control study examined possible associations between JP and pathogenic bacteria, the human cytomegalovirus (HCMV), and the Epstein-Barr type 1 virus (EBV-1)., Methods: Subjects were participants in a larger survey of schoolchildren in North-Central Jamaica. Subgingival plaque samples from 15 subjects with JP, 20 with incipient periodontitis (IP), and 65 randomly-selected healthy controls were assayed for Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans using a 16S rRNA polymerase chain reaction (PCR) identification method, and for HCMV and EBV-1 using nested PCR identification., Results: Strong bivariate associations were found between JP and P. gingivalis (odds ratio [OR] = 12.7; 95% CI = 2.6, 61.4), HCMV (OR = 10.0; 95% CI = 2.7, 36.3), and A. actinomycetemcomitans (OR = 8.0; 95% CI = 2.3, 27.5), but not EBV-1. In multivariate analyses, P. gingivalis remained a significant explanatory variable (OR = 7.8; 95% CI = 1.5, 40.9); however, the associations were marginal for HCMV (OR = 4.6; 95% CI = 0.9, 22.7), and non-significant for A. actinomycetemcomitans (OR = 2.0; 95% CI = 0.4, 9.7). The associations with JP and the extent of attachment loss were even stronger when both P. gingivalis and HCMV were detected together. P. gingivalis (OR = 3.9; 95% CI = 1.3, 12.0) and EBV-1 (OR = 3.3; 95% CI = 1.0, 10.3) were the only significant explanatory variables in the multivariate analysis of IP., Conclusions: P. gingivalis is the strongest and most stable indicator of periodontitis in Jamaican adolescents. Co-infection with P. gingivalis and HCMV appears to be particularly deleterious to periodontal health.

The potential involvement of the inducible cyclooxygenase isoform (COX-2) and the role of novel lipid mediators were investigated in the pathogenesis of periodontal disease. Crevicular fluids from localized juvenile periodontitis (LJP) patients contained prostaglandin (PG)E(2) and 5-lipoxygenase-derived products, leukotriene B(4), and the biosynthesis interaction product, lipoxin (LX)A(4). Neutrophils from peripheral blood of LJP patients, but not from asymptomatic donors, also generated LXA(4), suggesting a role for this immunomodulatory molecule in periodontal disease. To characterize host responses of interest to periodontal pathogens, Porphyromonas gingivalis was introduced within murine dorsal air pouches. In the air pouch cavity, P. gingivalis elicited leukocyte infiltration, concomitant with elevated PGE(2) levels in the cellular exudates, and upregulated COX-2 expression in infiltrated leukocytes. In addition, human neutrophils exposed to P. gingivalis also upregulated COX-2 expression. Blood borne P. gingivalis gave significant increases in the murine tissue levels of COX-2 mRNA associated with both heart and lungs, supporting a potential role for this oral pathogen in the evolution of systemic events. The administration of metabolically stable analogues of LX and of aspirin-triggered LX potently blocked neutrophil traffic into the dorsal pouch cavity and lowered PGE(2) levels within exudates. Together, these results identify PMN as an additional and potentially important source of PGE(2) in periodontal tissues. Moreover, they provide evidence for a novel protective role for LX in periodontitis, limiting further PMN recruitment and PMN-mediated tissue injury that can lead to loss of inflammatory barriers that prevent systemic tissue invasion of oral microbial pathogens.

Background: Six serotypes of Porphyromonas gingivalis have recently been described. We sought to test the hypothesis that serotype specific carbohydrates from these strains are important antigens that elicit potent immune responses., Methods: Serum concentrations of IgG reactive with P. gingivalis serotypes K1-K6 were determined for 28 adult (AP) and 28 generalized early-onset (G-EOP) periodontitis patients previously determined to be seropositive for a broken cell preparation of P. gingivalis. To confirm relationships suggested for K1, K2, and K6 in the analysis of initial data, the study population was increased to 133., Results: Frequency of seropositivity for the 6 serotypes ranged from 26 to 54% of subjects. IgG concentrations ranged from 0 to 453 microg/ml with many subjects seropositive to more than one serotype. Concentrations for the subset of patients who was seropositive were high (mean responses ranged from 20 to 105 microg/ml for the 6 serotypes). Significant correlations between seropositivity to serotypes K1 and K5 as well as between K5 and K6 were found., Conclusions: We examined the relationship of diagnosis, race, gender, smoking, probing depth, attachment loss, and antibody reaction with the P. gingivalis serotypes by analysis of variance. Initial findings suggested potential relationships between diagnosis, smoking, race, gender, and antibody reactive with serotypes K1, K2, and K6. A significant relationship did exist between smoking and decreased antibody reactive with P. gingivalis serotype K2. No other relationships were substantiated. We also examined the IgG subclass distribution and found that responses were almost exclusively IgG2. These data support the concept that antibody responses to all 6 serotypes are common in both AP and G-EOP and that these K serotype carbohydrates elicit potent IgG2 responses.

Recent work has indicated that Bacteroides forsythus and Porphyromonas gingivalis are significant local risk factors for periodontitis. Several reports find that both organisms are frequently associated with periodontitis lesions and often are present together. We have previously shown that early-onset periodontitis patients seropositive for P. gingivalis have less attachment loss than seronegative patients. In this study, we determined serum IgG antibody concentrations reactive with B. forsythus in adult and early-onset periodontitis patients using an ELISA and used P. gingivalis in the same populations as a positive control. The results for P. gingivalis were consistent with previous work and indicated that 47%, 36%, and 33% of adult, generalized early-onset, and localized juvenile patients were seropositive, respectively. Mean serum IgG concentrations for the three groups were 5.36 microg/ml, 5.65 microg/ml, and 5.44 microg/ml for adult, generalized early-onset, and localized juvenile patients, respectively. In contrast, for B. forsythus only 11%, 14%, and 10% of adult, generalized early-onset, and localized juvenile patients were seropositive, with mean serum IgG concentrations of 0.46 microg/ml, 0.46 microg/ml, and 0.47 microg/ml, respectively. This suggests that B. forsythus is either poorly immunogenic or less invasive than P. gingivalis. If most patients fail to mount an immune response to B. forsythus and it is invasive, it may explain why this organism is a risk factor for disease.

Porphyromonas gingivalis is considered to be an important pathogen in periodontitis. The present study investigates the relationship between serum anti-P. gingivalis IgG antibody levels and the subgingival distribution of P. gingivalis in patients with periodontitis. We examined subgingival plaque samples from 15 patients with adult periodontitis (AP), 8 patients with early-onset periodontitis (EOP), and 6 clinically healthy individuals. The samples were collected from periodontal pockets or gingival crevices of all remaining teeth in each subject. The total number of samples was 3,024, ranging from 76 to 120 per subject. Probing depth and bleeding at each sample site were recorded. P. gingivalis was detected using a non-radioactive whole genomic DNA probe. Serum samples were taken from the subjects, and the serum anti-P. gingivalis IgG antibody titer was determined by enzyme-linked immunosorbent assay (ELISA). P. gingivalis was recovered from all AP and EOP patients, and from 3 of the 6 healthy subjects. Two significant positive correlations were observed among the subjects. The serum anti-P. gingivalis IgG antibody titer correlated with detection frequency of P. gingivalis, and the antibody titer correlated with the amount of P. gingivalis detected. Higher levels of P. gingivalis were detected in the EOP group than in the AP group. However, no significant difference was found in the serum IgG titer levels between EOP and AP patients. These findings suggest a direct relationship between the serum anti-P. gingivalis IgG levels and subgingival P. gingivalis colonization; however, the functional capabilities of IgG antibodies may vary among the various types of periodontitis patients.

The aim of the study was to compare the occurrence and levels of A. actinomycetemcomitans, P. gingivalis, and P. intermedia in the subgingival plaque from sites with and without early periodontitis in adolescents using an ELISA. 47, 15- to 16-year-old adolescents (39 Indo-Pakistani, 8 white Caucasian) were examined for clinical attachment level, probing depth, supragingival plaque, subgingival calculus and bleeding on probing on the mesio-buccal and disto-buccal aspects of the 1st molars and the incisors. Based on the clinical data, 2 sites per subject were selected for subgingival plaque sampling 3 weeks later: in 32 subjects with loss of attachment > or = 1 mm, a diseased site (D) and a healthy comparison control site (C) were sampled; in 15 subjects in whom loss of attachment had not yet developed, 1 of the upper molar sites was selected, called the at-risk site (R), together with a C site. The presence and levels of A. actinomycetemcomitans, P. gingivalis, and P. intermedia were determined using an ELISA. The loss of attachment subgroup had significantly more pockets > or = 4 mm, subgingival calculus and bleeding on probing (p < 0.05). Significantly more of the D than C sites had P. gingivalis both at detectable and at measurable levels (p < 0.05). In subjects who had no loss in clinical attachment levels, fewer sampled sites harboured any of the suspected periodontopathogens investigated, and no significant differences were found between the R or C sites (p > 0.05). Although there was a significantly higher prevalence and extent of loss of attachment > or = 1 mm in the Indo-Pakistani subjects compared with the Caucasians (p < 0.05), no differences could be identified in the distribution of the bacteria. It is concluded that monitoring of the subgingival plaque may be useful in studies of early periodontitis in adolescents, and the role of P. gingivalis needs to be elucidated in prospective longitudinal investigations.

The occurrence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque in 24 juvenile periodontitis patients was determined using DNA probe. 36 samples of subgingival plaque from 36 pockets having > or = 6 mm depth, > or = 3 mm of loss of attachment, and bleeding on probing and/or suppuration were taken from 18 patients with localized juvenile periodontitis (LJP, age range 12-24 years); and 12 samples from 6 patients with generalized juvenile periodontitis (GJP, age range 23-26 years). As control, an equal numbers of samples from healthy sites in the same patients were studied. P. gingivalis was found in 17 of 18 LJP patients, and in 31 of 36 diseased sites in those patients. P. intermedia was found in 15 out of the 18 LJP patients and in 28 of the 36 diseased sites. A. actinomycetemcomitans was present in 7 of the 18 LJP patients, and in 9 of the 36 diseased sites, and was not found in any GJP patients. All GJP patients had P. gingivalis (11 out of 12 diseased sites) and P. intermedia (all of the diseased sites). None of the three bacterial species was detected in healthy sites of GJP patients, and were found in healthy sites in only 2 of 18 LJP patients. The high prevalence and high levels of P. gingivalis and P. intermedia found in the LJP and GJP patients studied, suggest that there are populations affected by juvenile periodontitis in which this type of periodontitis is more associated with these species than with A. actinomycetemcomitans.

A population of 33 subjects were selected on the basis that all had tested positive for A, actinomycetemcomitans at some time during the prior 7 years. Most subjects (31/33) belonged to families with a proband with confirmed localized juvenile periodontitis (JP); however, most subjects had no evidence of the typical lesions associated with JP. Two additional subjects with rapidly progressive periodontitis, known to be positive for A. actinomycetemcomitans, were also recruited. The patients with a history of JP had been treated, but were no longer enrolled in a regular maintenance program. With 3 exceptions, the subjects had not received any dental treatment or antibiotics in the past 3 months. One aim of the study was to determine the prevalence of A. actinomycetemcomitans, P. gingivalis, and B. forsythus in this population. The main purpose was to compare the relative sensitivity of various methods for detecting these periodontal pathogens. Pooled subgingival plaque samples were collected from all the mesial surfaces and aliquots of the suspension processed for the detection of A. actinomycetemcomitans by culture and indirect immunofluorescence (IF) to serotypes a, b, and c. P. gingivalis and B. forsythus were monitored with a DNA probe and IF. With culture, A. actinomycetemcomitans was detected in 39.4% of the samples, at a mean level of 0.64% of the cultivable counts. With IF, A. actinomycetemcomitans was detected in 81.8% of the samples, at levels of 0.40, 0.79, and 0.17% of the total counts for serotypes a, b and c respectively. Overall, IF was more likely to detect A. actinomycetemcomitans, P. gingivalis, and B. forsythus than any of the other methods.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunological studies examining the homogeneity of the major antigenic components of P. gingivalis have suggested 3 serotypes and have indicated a limited distribution of the serotypes in an individual patient. These studies prompted us to define the immunodominant antigens and distribution of immune responses to P. gingivalis serotypes. Serum IgG antibody levels in periodontitis patients in the present study were most frequently elevated above the normal subjects when tested against P. gingivalis serotype A (i.e., 33277). Nearly 1/3 of the patients showed significantly elevated antibody to multiple serotypes of the P. gingivalis apparently resulting from cross-reacting antigens. We determined distinctive differences among outer envelope protein and antigen patterns obtained from the three serotypes. Moreover, the results identified considerable similarities in the qualitative and quantitative antigen response patterns among patients to a particular serotype. There was a strong positive correlation between IgG antibody levels (ELISA) and the total level of reactivity determined in the immunoblots, as well as a positive correlation to the proportion of antibody to particular antigens. These findings suggest that responses to these antigens comprised a major portion of the response to the intact microorganism. Additionally, the detection of antibody to particular antigen bands was indicative of early responses to each of the P. gingivalis serotypes. The results of our study indicate that a subpopulation of periodontitis patients develop an extensive serum antibody response often to multiple serotypes of P. gingivalis and may define a patient population with a P. gingivalis disease. Finally, our results indicate a more consistent antigenic composition for P. gingivalis which may enhance the potential for strategies to immunologically interfere with disease caused by this microorganism.

The treatment of children with prepubertal periodontitis (PP), may be complicated by the extent of the lesions and the possibility of tetracycline stain of the developing permanent dentition. Therefore, with the purpose of preventing the infection of permanent teeth during the mixed dentition, it has been recommended that the treatment of children with PP, should include the early extraction of the primary teeth affected with alveolar bone loss (ABL). Still, there is little evidence which confirms that extraction of the affected primary teeth do in fact reduce the periodonto-pathogens load of the subgingival plaque. The present study reports values of colony forming units (CFU) of total anaerobic bacteria, Actinobacillus actynomicetemcomitans (Aa) and Porphyromonas gingivalis (Pg) from the subgingival plaque from a child with PP, collected immediately before and 1 year after extractions of the primary teeth affected with ABL. CFU of Aa and Pg developed only from the subgingival plaque collected before the extraction of the primary teeth affected with ABL. These findings suggest that in cases of PP, extraction of the affected primary teeth may reduce the possibility of infection of the periodontum of the permanent teeth during the mixed dentition period.

A panel of 15 monoclonal antibodies (MAbs) was raised to Porphyromonas gingivalis and used to characterise the antigens which they recognise by ELISA, immunofluorescence (IF), Western blotting and patient serum inhibition studies. All the MAbs were specific for P. gingivalis and did not recognise 24 other species. Eight MAbs gave bright ring-like staining patterns against the majority of serotypes of P. gingivalis tested by IF. The antibodies could be grouped into 5 different antigen recognition profiles on Western blotting, and ELISA indicated that 8 of them bound to a capsular extract. Five antibodies bound to antigenic determinants recognised by sera from patients with periodonitis and 4 of these (1A1, 2B/H9, 3B1, 7D5) bound to a P. gingivalis 47kDa protease preparation on Western blotting. These antibodies are potentially useful for the purification and characterisation of biologically active components of P. gingivalis that are recognised by sera from patients with periodontitis.

The distribution and transmission of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in 4 families were studied. The families were included, based on the isolation of P. gingivalis from a young child or adolescent. The probands of these 4 families were: a 5-year old periodontally healthy boy; a 17-year old girl with severe generalized juvenile periodontitis; an 11-year old girl with prepubertal periodontitis; 2 sisters, 5 and 17-years old, with untreated severe periodontitis as a component of the Papillon-Lefèvre syndrome. All members of the 4 families were examined clinically and microbiologically for the presence of P. gingivalis and A. actinomycetemcomitans. Most of the parents appeared to be adult periodontitis patients; the parents of one proband were edentulous. Results showed that in all cases at least one of the parents was positive for P. gingivalis. On the basis of indistinguishable restriction endonuclease patterns (REPs) of P. gingivalis and A. actinomycetemcomitans isolates from parents and their children, and distinct REPs from unrelated individuals, the present study indicates that P. gingivalis and A. actinomycetemcomitans were transmitted between parents and their children.

The adherence of Porphyromonas gingivalis (P. gingivalis) to the periodontal tissue may be an initial step in the pathogenesis of periodontal disease, and fimbriae is believed to play an important role in such processes. However, the heterogeneity of the size, sequence and antigenic reactivity of the fimbriae have been reported. The aim of the present study was to investigate the heterogeneity of P. gingivalis strains on the fimbrillin gene locus by RFLPs analysis. Fourty-seven P. gingivalis strains including 5 reference strains were used in this study. The plasmid (pUC13Bg 12.1) with the insert of fimA381 was modified with bisulfite and used as the probe. The genomic DNAs from P. gingivalis were digested with the restriction endonuclease SacI and PstI, electrophoresed, transferred and hybridized with the DNA probe. All Sac I digests generated one major band and the band size was almost the same (ca 2.5-kb), except strain W50. The Pst I digests showed one or two major bands and could be divided into 9 groups based on the band patterns. Moreover, isolates from one patient showed different band patterns. By RFLPs analysis, genetic heterogeneity seems to exist within the fimbrillin gene locus of P. gingivalis strains. Such genetic heterogeneity may reflect the previously reported difference of P. gingivalis fimbriae and moreover a single patient could be infected with more than one genotype of P. gingivalis.

Solubilized surface-associated material (SAM) from a number of periodontopathogenic bacteria have been shown to be potent stimulators of bone resorption in vitro in the murine calvarial bone culture assay. Antibodies to the constituents of SAM are also found in patients with periodontal diseases. Serum from patients with severe generalized periodontitis (SGP) containing high titers of antibodies to the SAM of Porphyromonas gingivalis completely inhibited the bone resorption induced by SAM from this organism. In contrast, serum from patients with low titers of antibodies to SAM from P. gingivalis failed to inhibit bone resorption. High-titer sera (containing antibodies to SAM from Actinobacillus actinomycetemcomitans) from patients with localized juvenile periodontitis (LJP) were added to calvarial cultures stimulated with SAM from A. actinomycetemcomitans. Of 6 high-titer sera tested, only 4 inhibited bone breakdown, the other 2 sera having no effect on resorption. Low-titer sera were also ineffective at blocking bone resorption. This suggests that the antibody response to SAM may have a protective effect in patients with periodontal disease.

Several forms of periodontal disease are considered to be infectious diseases with associated specific bacteria. This study examined the humoral antibody levels as assayed by ELISA to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in adult periodontitis (AP), localized juvenile periodontitis (LJP), rapidly progressive periodontitis (RPP), and in periodontally healthy subjects (HS). Sixty-two of the 64 (96.9%) patients had significantly elevated antibody levels to at least one of the three organisms. Elevated levels of antibodies to P. gingivalis occurred in 82.8% of the RPP, LJP, and AP patients with all 3 disease groups showing greater responses than HS controls. Antibodies to A. actinomycetemcomitans were found in 59.4% of the RPP, LJP, and AP patients and were significantly higher in both LJP and RPP patients. Only 21.9% of the RPP, LJP, and AP patients showed elevated levels to P. intermedia with only significantly higher levels in the RPP and LJP groups. Antibodies to A. actinomycetemcomitans and P. intermedia were rarely found alone (only 5.1% and 2.6% of the patients respectively) but were usually accompanied by P. gingivalis. These results suggest that one or more combinations of these 3 bacteria may play a role in the pathogenesis of these forms of periodontal disease.