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PubMed2005

[Detection of Porphyromonas gingivalis in the subgingival plaque of puberty gingivitis].

PURPOSE: To detect the presence of Porphyromonas gingivalis (Pg) in the subgingival plaque of puberty gingivitis and to investigate its relationship with gingival health. METHODS: The subjects of 14- to 17-year-old children in this study were divided into two groups, puberty gingivitis group and gingival healthy group, with 36 in each group. Periodontal parameters were recorded. A 16S rRNA-based polymerase chain reaction (PCR) detection method was used to determine the prevalence of Pg in the subgingival plaque samples. The AVG of positive band in the agarose gel electrophoresis was analyzed using the automatic gel documentation and image analyzer software. The relative quantity of Pg was determined. The data were statistically analyzed using SPSS11.5 software. Chi-square test was used to compare the prevalence of Pg in the two groups. Wilcoxon rank sum test was used to analyze the relative quantity of Pg and the severity between the Pg-positive and the Pg-negative subjects. Spearman correlation analysis was used to test the relationship between the relative quantity of Pg and periodontal parameters. RESULTS: The prevalence of Pg in the puberty gingivitis group and gingival healthy group was 47.22% and 25.00%, respectively (P<0.05). The relative quantity of Pg in the two groups was 48.02% and 21.46%, respectively(P<0.01). In all the Pg-positive subjects of puberty gingivitis group, there was positive correlation between the relative quantity of Pg and the gingival index (GI),sulcus bleeding index (SBI) and the probe depth (PD). There were statistically differences between them. The GI and SBI index of the Pg-positive subjects were statistically higher than that of the Pg-negative subjects in the puberty gingivitis group (P<0.01). CONCLUSION: Periodontal pathogens can colonize in the subgingival plaque in puberty subjects and colonization of Pg is relevant to the status of gingival health.

Adolescent↗
PubMed1976

Subgingival plaque and loss of attachment in periodontosis as observed in autopsy material.

Histologic sections from six autopsy cases which from a clinical point of view fitted into the criteria of periodontosis were examined with the aim at evaluating the role of subgingival plaque in the etiology of the loss of attachment in this condition. The following pertinent observations were made: 1. The subgingival plaque in most instances was not calcified to form calculus. 2. The thickness of the subgingival plaque varied between 20 and 200 mu (0.02-0.2 mm). 3. Where loss of attachment had taken place, the distance from the most apical part of the subgingival plaque to the most apical point of the epithelial cuff varied between 0.2 and 1.1 mm; this distance was never found to be more than 1.1 mm. This would indicate a cause and effect relationship, the plaque being the obvious cause. 4. There was very severe chronic inflammation in the soft tissue bordering upon the plaque with resulting collagenolysis. 5. The cellular infiltration and the collagenolysis may be limited to a zone of 1 to 2 mm in the immediate vicinity of the plaque. Between the inflamed area and the surface of the gingivae buccally and lingually there may be a fairly wide zone of healthy tissue which hides the symptoms of inflammation from being observed on a clinical examination. In turn this could leave the clinician with the impression that attachment has been lost and bone resorbed because of degenerative changes. The following conclusions can be made: In these six cases of "alveolar bone loss vastly out of proportion to what one would expect from the local etiologic factors in the patient at that age" there was no morphologic evidence that degenerative changes were responsible for the loss of attachment. On the contrary inflammatory changes induced by the subgingval plaque dominated the histopathologic picture.

Adult↗
PubMed1999

Identification by PCR of Helicobacter pylori in subgingival plaque of adult periodontitis patients.

The PCR was used to detect the presence of Helicobacter pylori in subgingival plaque samples from patients with adult periodontitis. Primers based upon the 16S ribosomal RNA (rRNA) gene sequence of H. pylori were used in a single round of PCR to amplify a 295-bp DNA fragment and the identity of the amplified products was confirmed by Southern blot hybridisation to a digoxigenin-labelled H. pylori probe. Further confirmation of product identity was obtained by DNA sequencing of a proportion of the amplified products. The assay was demonstrated to be specific for H. pylori with a lower limit of detection of 100 fg of bacterial genomic DNA. In all, 73 samples from 29 patients were analysed, of which 24 (33%) were H. pylori-positive by PCR; the proportion of patients carrying H. pylori in at least one sampled site was 38% (11 of 29). This is the first study to demonstrate the presence of H. pylori in the subgingival plaque of patients with adult periodontitis and indicates that, in this patient group at least, subgingival plaque may be a reservoir for H. pylori infection.

Adult↗
PubMed2003

Comparison of real-time PCR and culture for detection of Porphyromonas gingivalis in subgingival plaque samples.

Porphyromonas gingivalis is a major pathogen in destructive periodontal disease in humans. Detection and quantification of this microorganism are relevant for diagnosis and treatment planning. The prevalence and quantity of P. gingivalis in subgingival plaque samples of periodontitis patients were determined by anaerobic culture and real-time PCR amplification of the 16S small-subunit rRNA gene. The PCR was performed with primers and a fluorescently labeled probe specific for the P. gingivalis 16S rRNA gene. By the real-time PCR assay, as few as 1 CFU of P. gingivalis could be detected. Subgingival plaque samples from 259 adult patients with severe periodontitis were analyzed. P. gingivalis was detected in 111 (43%) of the 259 subgingival plaque samples by culture and in 138 (53%) samples by PCR. The sensitivity, specificity, and positive and negative predictive values of the real-time PCR were 100, 94, 94, and 100%, respectively. We conclude that real-time PCR confirms the results of quantitative culture of P. gingivalis and offers significant advantages with respect to the rapidity and sensitivity of detection of P. gingivalis in subgingival plaque samples.

Bacteroidaceae Infections↗
PubMed1992

A sensitive enzymatic method (SK-013) for detection of Treponema denticola, Porphyromonas gingivals and Bacteroides forsythus in subgingival plaque samples.

An enzymatic method, SK-013, was developed for rapid detection of the peptidase activity in subgingival plaque samples. This method was found to have specificity for Porphyromonas gingivalis, Treponema denticola, Bacteroides forsythus, and some Capnocytophaga strains. The purpose of this study was to determine whether SK-013 could indicate the presence of periodontopathic bacteria, including T. denticola, P. gingivalis and B. forsythus, which produce trypsin-like enzymes. Subgingival plaque samples were taken from 10 clinically healthy sites and 30 periodontally diseased sites with 3 paper points. SK-013 activity of plaque samples was assayed, and the numbers of T. denticola, P. gingivalis and B. forsythus in the sample were counted by immunofluorescence technique. In diseased sites, the SK-013 activity was significantly correlated with clinical parameters such as Gingival Index, Plaque Index, probing depth and bleeding on probing. A significant correlation was found between the presence of these organisms and SK-013 activity. Correlation coefficients between the presence of T. denticola and SK-013 activity were higher than those with other organisms. These findings indicate that the SK-013 is useful as an indicator of cell population of T. denticola, P. gingivalis and B. forsythus in subgingival plaque.

Bacteroides↗
PubMed1990

Darkground microscopy of subgingival plaque from the top to the bottom of the periodontal pocket.

Part of the results of a study of subgingival plaque by darkground microscopy was used to investigate morphotype distribution at different levels within the periodontal pocket. Subgingival plaque was sampled at coronal, middle and apical levels from each of 28 approximal surfaces on 28 teeth extracted from 28 adults with untreated moderate to advanced periodontitis. Sample preparation and darkground microscopy were as described previously. The data recorded only the subgingival level at which the maximum count occurred for a given morphotype and subject. It was found that the maximum count of spirochaetes occurred most frequently at the apical level (p less than 0.05) and the maximum count of 'others' at the coronal level (p less than 0.05). Also, coccal counts showed a trend to increase the more apical the sample. Standardised darkground microscopy was able to show that the sampled subgingival level had a significant influence on plaque bacterial morphotype distribution, indicating the importance of accurate sampling of subgingival plaque at the most apical level.

Adult↗
PubMed1999

Identical clonal types of Porphyromonas gingivalis or Prevotella nigrescens recovered from infected root canals and subgingival plaque.

Clinical samples from 10 infected root canals and from subgingival plaque in 10 patients were screened by anaerobic culture for black-pigmented anaerobes. A total of 17 Porphyromonas gingivalis and 9 Prevotella nigrescens were obtained from four patients and were identified by species-specific polymerase chain reaction (PCR) amplification. The arbitrarily primed PCR reaction used to examine the genetic diversity of the isolates revealed that the P. gingivalis or P. nigrescens simultaneously present in the root canal system and in subgingival plaque of all four patients were genotypically indistinguishable. These data indicate that the endodontium and the periodontium can be colonized by the same clonal types of black-pigmented anaerobes.

Clone Cells↗
PubMed1989

The in vitro effects of chlorhexidine on subgingival plaque bacteria.

The purpose of this study was to determine the susceptibility to chlorhexidine of a range of bacteria which may be isolated from subgingival plaque. In addition, the effect of chlorhexidine on the survival of bacteria in subgingival plaque samples from patients with chronic inflammatory periodontal disease was investigated. The minimum inhibitory concentration (MIC) of chlorhexidine for 52 strains of bacteria ranged from 8 to 500 micrograms/ml. The modal value of the MIC was found to be 62 micrograms/ml, 64% of the strains tested being inhibited at this concentration. A concentration of 250 micrograms/ml of chlorhexidine inhibited the growth of all bacteria in the 25 subgingival plaque samples investigated. The MIC of chlorhexidine for the samples ranged from 31 to 250 micrograms/ml, the modal value being 125 micrograms/ml.

Adult↗
PubMed2002

Frequency of reactivity for Porphyromonas gingivalis and Prevotella spp. in supra- and subgingival plaques, and periodontal clinical parameters according to subject age.

BACKGROUND: The present study was conducted to assess the association between selected clinical parameters and the distribution of Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Prevotella nigrescens (Pn), and Prevotella melaninogenica (Pm) in supra- and subgingival plaque samples measured by an immunoslot blot assay (IBA) using their monoclonal antibodies. METHODS: Plaque samples from 299 patients aged 6 to 69 randomly chosen from a group of dental outpatients were examined. Plaque index, gingival index, and probing depths were evaluated according to the criteria of positive (cell number > or = 10(6)) or negative (<106) reactivity to the 4 different monoclonal antibodies. RESULTS: An increase in probing depth in subjects exhibiting either a positive or negative reaction for the 4 test bacteria was associated with increasing age. Comparing bacteria-positive subgingival plaque samples to their corresponding bacteria-negative counterparts, we found an increased plaque index in children positive for any of the 4 bacteria; in addition, that for Pg and Pi was increased in subjects 40 to 49 years old. The gingival index increased with increasing amount of Pi and Pn, but not with Pg and Pm in those 20 to 29 years of age. The frequency of Pg reactivity in subgingival plaque was markedly enhanced in subjects older than 30 to 39 years of age, and was significantly higher than that in supragingival plaque. The frequency of Pi and Pn reactivity was significantly increased in adults aged 20 to 29 and plateaued at older ages. The frequency of Pm reactivity was relatively low and independent of subject age. CONCLUSIONS: The increase in probing depth with increasing age was not affected by the occurrence of periodontopathic bacteria. The high rate of occurrence of Pg, together with Pi and Pn, in subgingival plaque of the adult age groups was suggested to be associated with the high frequency of periodontal disease in the older age groups (above 30 to 49 years of age). The IBA appears to be useful for the efficient and rapid detection of periodontopathic bacteria.

Adolescent↗
PubMed1998

Microbial complexes in subgingival plaque.

It has been recognized for some time that bacterial species exist in complexes in subgingival plaque. The purpose of the present investigation was to attempt to define such communities using data from large numbers of plaque samples and different clustering and ordination techniques. Subgingival plaque samples were taken from the mesial aspect of each tooth in 185 subjects (mean age 51 +/- 16 years) with (n = 160) or without (n = 25) periodontitis. The presence and levels of 40 subgingival taxa were determined in 13,261 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments were made at 6 sites per tooth at each visit. Similarities between pairs of species were computed using phi coefficients and species clustered using an averaged unweighted linkage sort. Community ordination was performed using principal components analysis and correspondence analysis. 5 major complexes were consistently observed using any of the analytical methods. One complex consisted of the tightly related group: Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. The 2nd complex consisted of a tightly related core group including members of the Fusobacterium nucleatum/periodonticum subspecies, Prevotella intermedia, Prevotella nigrescens and Peptostreptococcus micros. Species associated with this group included: Eubacterium nodatum, Campylobacter rectus, Campylobacter showae, Streptococcus constellatus and Campylobacter gracilis. The 3rd complex consisted of Streptococcus sanguis, S. oralis, S. mitis, S. gordonii and S. intermedius. The 4th complex was comprised of 3 Capnocytophaga species, Campylobacter concisus, Eikenella corrodens and Actinobacillus actinomycetemcomitans serotype a. The 5th complex consisted of Veillonella parvula and Actinomyces odontolyticus. A. actinomycetemcomitans serotype b, Selenomonas noxia and Actinomyces naeslundii genospecies 2 (A. viscosus) were outliers with little relation to each other and the 5 major complexes. The 1st complex related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.

Adult↗
PubMed1986

A new method for assessing the % of spirochaetes in subgingival plaque.

A new method for counting the numbers of spirochaetes in subgingival plaque is described. The technique involves the combined use of a negative stained preparation, dark field microscopy, and an image analysing system. Advantages of the method include: smears need not be made until 2 days after sampling, and can be examined microscopically for up to 1 year after preparation; reproducibility of counts are more than 90% and the data can be computerised. The new method should prove useful in the routine assessment of subgingival plaque from patients with various forms of periodontitis, and in epidemiological surveys.

Aniline Compounds↗
PubMed1998

A statistical approach to the ecology of Actinobacillus actinomycetemcomitans in subgingival plaque.

Structured complexity may be a key feature of the microbiota residing in periodontal ecosystems. Recently, several attempts have been made to analyse the mutual interdependence of certain organisms in subgingival plaque and/or to identify clinical conditions which might influence presence or absence of these bacteria in subgingival plaque. The aim of the present analysis was to compare the results of different models derived from logistic regression of several clinical factors on the presence of Actinobacillus actinomycetemcomitans in subgingival plaque of adult periodontitis patients all harbouring this organism. Models assuming independence of observations or assuming independent observations with the level of response depending on the individual subject were compared to models considering the correlated structure of the acquired data by employing Generalized Estimating Equation (GEE) methods. Significant associations found when neglecting the correlated observations within a given subject as, e.g., between A. actinomycetemcomitans and amount of supragingival plaque, bleeding on probing, or attachment loss, became generally spurious in the GEE approach. Thus, presence of A. actinomycetemcomitans only depended on periodontal probing depth. It was concluded that the correlated structure of observations in an oral cavity should be carefully considered when associations between clinical factors are calculated.

Adult↗
PubMed1990

Microbial patterns in pooled subgingival plaque samples from young adults with advanced marginal periodontitis.

Different bacterial profiles of subgingival plaque have been described for different clinical categories of marginal periodontitis. In the present investigation the subgingival microflora was studied in young adults with advanced marginal periodontitis. From 12 patients pooled subgingival plaque samples from 5 advanced stages of diseased sites were examined by direct differential phase-contrast microscopy and by cultivation on enriched and selective media. The proportions of the following genera and species were calculated: black-pigmented Bacteroides sp., B. gingivalis, Fusobacterium nucleatum, Veillonella sp., Actinobacillus actinomycetemcomitans, Capnocytophaga sp., Eikenella corrodens, Campylobacter sp., and Candida sp. Spirochete identification was carried out ultrastructurally. Calculated percent minimum similarity levels between the individuals revealed that each patient harbored its characteristic cultivable subgingival microflora different from the other individuals. The spirochetes seemed to constitute the subgingival bacterial group with the smallest variation as certain morphotypes were dominating in almost all individuals.

Adult↗
PubMed1990

The prevalence of beta-lactamase producing bacteria in subgingival plaque and their sensitivity to Augmentin.

Subgingival plaque samples from 20 patients with chronic periodontitis who had received no antibiotics for at least 3 months were screened for the presence of beta-lactamase-producing bacteria. Thirteen of the patients harboured beta-lactamase producing bacteria, most of which were members of the genus Bacteroides. The most frequently isolated species were Bacteroides melaninogenicus and Bacteroides capillosus which are often implicated in acute oral infections. All of the beta-lactamase-producing bacteroides strains were sensitive to a combination of amoxycillin with clavulanic acid (Augmentin).

Adult↗
PubMed2003

Subgingival plaque removal in buccal and lingual sites using a novel low abrasive air-polishing powder.

AIM OF THE STUDY: This study was aimed at assessing the efficacy of subgingival plaque removal in buccal and lingual sites during supportive periodontal therapy (SPT) using a novel low abrasive air-polishing powder. MATERIAL AND METHODS: In 27 SPT patients, subgingival debridement was performed using the novel air-polishing powder (test) and hand instruments (positive control) in a randomized split mouth design. Before and immediately after treatment, subgingival plaque samples were taken from two teeth with pockets of 3-5 mm depth in both groups. To assess the influence of plaque sampling on the microflora, samples were also taken twice from two untreated teeth (negative control). The mean reduction in total colony forming units (CFU) was assessed by anaerobic culture. The patients' perception of treatment was assessed by a visual analog score (VAS). Therapy and plaque sampling were repeated after a 3-month interval. RESULTS: Test treatment resulted in a significantly greater reduction in mean CFU than positive control treatment (log 1.7+/-0.98 and log 0.61+/-0.79 respectively; p<0.05). Following both treatments, the CFU reduction was significantly greater compared to negative control treatment (log 0.06+/-0.49; p<0.05). In addition, test treatment was perceived as significantly more pleasant than hand instrumentation (p<0.05). CONCLUSION: The novel low abrasive air-polishing powder is superior to curettes in removing subgingival plaque from pockets of 3-5 mm depth in supportive periodontal therapy and offers greater patient comfort.

Adolescent↗
PubMed2006

Biofilm structure and cell vitality in a laboratory model of subgingival plaque.

The accumulation of dental plaque below the gingival margin (i.e. subgingival plaque) is responsible for the most prevalent microbe-induced diseases of humans--the periodontal diseases. Access to this plaque is difficult, making studies of its structure in vivo very difficult. We have, therefore, used a constant-depth film fermenter to grow microcosm subgingival dental plaques under conditions similar to those existing in vivo to enable us to study certain aspects of its structure. Confocal laser scanning microscopy revealed that the biofilms consisted of pillar-like stacks of bacteria separated by water channels. In terms of their structure, these microcosm dental plaques reached a pseudo steady-state after 4 days. Individual optical sections generally showed the outer layers of the biofilm stacks to contain a high proportion of nonviable cells surrounding an inner core of predominantly viable cells with "veins" of nonviable bacteria penetrating from the outer layers through into the inner core. Such a structure differs from that classically described for biofilms growing in an aerobic atmosphere where the arrangement of viable and nonviable bacteria is usually reversed. The reasons for the preponderance of nonviable bacteria in the outer layer of the bacterial stacks remain to be established.

Biofilms↗
PubMed1987

The presence of methanobacteria in human subgingival plaque.

Through a procedure of setting up a strictly anaerobic chain from sampling subgingival plaque to incubation in an anaerobic cabinet (O2 less than 10 ppm), it was possible to, for the first time, isolate methanobacteria from human subgingival plaque.

Adult↗
PubMed1989

Relationship of subgingival plaque flora to lysosomal and cytoplasmic enzyme activity in gingival crevicular fluid.

Examining the relationships among indicators of the acute inflammatory response in gingival crevicular fluid (GCF) and specific bacterial species in subgingival plaque may provide indications of which bacterial species or groups of species may be associated with potentially destructive host-derived processes. Here we report on the relationship of the subgingival plaque flora to the activity of mammalian forms of the enzymes beta-glucuronidase (beta G), lactate dehydrogenase (LDH), and arylsulfatase (AS) in GCF from a total of 54 4-6 mm periodontal sites from 13 periodontitis patients. Sites were scored for probing depth (PD) and bleeding on probing, and GCF was collected using filter paper strips inserted into the sulcus for 30 s, eluted in buffer and assayed for enzyme activity. 1 week later, the patients were again evaluated for PD and bleeding, and subgingival plaque was removed with a curette oriented toward the pocket epithelium. Plaque samples were examined by darkfield microscopy and cultured anaerobically on selective and non-selective media. Various groups of bacteria, including species of black pigmenting Bacteroides (BPB), Fusobacterium sp., Capnocytophaga sp, Streptococcus sanguis, and total facultative organisms were enumerated. Relationships among the enzymes and bacterial groups expressed as colony-forming unit (CFU) counts or as a % of the total cultivable flora were assessed by Spearman correlation analysis. beta G levels were significantly correlated with populations of spirochetes, B. intermedius, B. gingivalis, and total lactose negative BPB's. Correlation between beta G and F. nucleatum sp. or Capnocytophaga sp. approached but did not reach statistically significant levels. In contrast, LDH activity showed a significant positive correlation with levels of B. gingivalis and total lactose negative BPB's. AS levels were significantly correlated only with B. gingivalis. beta G and LDH showed a significant negative correlation with levels of coccoid forms. Thus, beta G, an acid hydrolase which can serve as a marker for primary granule release from polymorphonuclear leukocytes, was most closely correlated with the micro-organisms found in other studies to be associated with chronic adult periodontitis.

Arylsulfatases↗