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Effect of phenoxyethanol, chlorhexidine and their combination on subgingival plaque bacteria.

The effects of phenoxyethanol, chlorhexidine and a mixture of both on subgingival plaque samples from 44 patients with chronic periodontitis were investigated in vitro. At a concentration of 0.5 mg/ml, chlorhexidine inhibited the growth of all cultivable bacteria in all 44 samples while a comparable effect was achieved with phenoxyethanol only at a concentration of 20 mg/ml. All cultivable bacteria in the samples were inhibited by a mixture of the two agents containing 5 mg/ml of phenoxyethanol and 0.125 mg/ml of chlorhexidine i.e. considerably lower concentrations than when the agents were used separately. Kill times for 99.9% of bacteria in 19 of the plaque samples were less than 15 min using a chlorhexidine concentration of 0.125 mg/ml, but were from 24 to greater than 240 min with 10 mg/ml phenoxyethanol. A mixture of the two agents at these concentrations was more effective than either agent alone with 99.9% kill times of less than 10 min. This investigation has shown that the addition of phenoxyethanol to chlorhexidine results in a mixture which is effective against bacteria found in subgingival plaque samples from patients with chronic periodontitis. It also implies that formulations with lower concentrations of chlorhexidine than those currently in use may be effective as adjuncts in the treatment of chronic periodontitis.

Adult↗

Proportion of antibiotic resistance in subgingival plaque samples from Mexican subjects.

AIM: To determine the proportion of bacteria resistant to amoxicillin and doxycycline in subgingival plaque samples from Mexican subjects. MATERIALS AND METHODS: Two subgingival plaque samples were taken from 20 Mexican subjects. Samples were dispersed, diluted and plated on non-antibiotic agar plates and on plates containing 0.5, 1, 2, 4, 8 and 16 microg/ml of either amoxicillin or doxycycline. The proportion of resistant bacteria was calculated based on the total number of colony-forming units present in the non-antibiotic containing plates. RESULTS: On average, 0.4-13.4% and 0.9-20.4% of the total cultivable subgingival microbiota was resistant to the concentrations tested of amoxicillin and doxycycline, respectively. The differences between antibiotics were statistically significant for the 0.5, 2 and 4 mug/ml concentrations (p < 0.05, Wilcoxon's test). CONCLUSIONS: Our findings revealed that a relatively small proportion of the total cultivable subgingival microbiota from Mexican subjects was resistant to amoxicillin and doxycycline.

Adult↗

Dark-field microscopy of subgingival plaque microflora in Indian and English subjects.

This present investigation reports on the microbial pattern of subgingival plaque from English and Indian subjects living in the West Midlands of England. Subgingival plaque from healthy and diseased sites was studied using darkfield microscopy. The results indicate that significant differences exist in microbial flora of clinically normal and diseased sites of English as well as Indian subjects. Coccoid cells predominated in healthy sites, with an increase of 21% in the spirochaetes in diseased sites. The ratio of non-motiles to motiles was 1:0.6 in healthy sites, whereas in diseased sites the ratio observed was 1:1.8 in English subjects and 1:3.6 in Indian subjects. Comparison of healthy sites in Indians with healthy sites in English subjects revealed significant differences between numbers of rods (p less than 0.05). Comparing diseased sites of Indians with diseased sites in English subjects revealed significant difference between both cocci (p less than 0.05) and motile rods (p less than 0.01). A positive correlation between chronic inflammatory periodontal disease and spirochaete burden, and a negative one between the disease and coccal burden, was found.

Adult↗

Effect of metronidazole on development of subgingival plaque and experimental periodontitis.

Active tissue destruction in experimental periodontitis has been positively correlated with subgingival spirochetes and total number of organisms. The present study was designed to inhibit spirochete populations and evaluate the effect upon periodontal destruction. Metronidazole was administered orally to four squirrel monkeys (100 mg/kg/bwt) for 17 days. After 3 days, marginal periodontitis was induced around bicuspids and molars by tying silk ligatures at the gingival margins. Subgingival plaque samples were taken baseline and 14 days after ligature placement. Dark-field microscopy quantitated motile forms, spirochetes, straight and curved rods, filament, cocci and fusiforms. Periodontal destruction was evaluated at 2 weeks by histometric analysis of connective tissue attachment, crestal alveolar bone and infiltrated connective tissue. Bacterial and histometric comparisons were made with experimental periodontitis data from four animals (control) which had not received metronidazole. Subgingival plaque prior to periodontitis induction was dominated by cocci, but fusiforms and straight rods were also present. Straight rods formed a greater proportion of the plaque which developed in metronidazole-receiving animals. The absence of spirochetes and motile rods contrasted with the control group where spirochetes were the predominant type. The total number of bacteria was also larger in the control animals. Histometric analysis showed that areas of infiltrated supracrestal connective tissue were similar in both groups. Loss of connective tissue attachment and alveolar bone was significantly less in experimental animals, and the latter values did not differ significantly from baseline dimensions. The results indicated that the subgingival bacterial populations which developed during metronidazole administration did not result in an experimental periodontitis.

Animals↗

Subgingival plaque microbiota in Saudi Arabians after use of miswak chewing stick and toothbrush.

BACKGROUND: The chewing stick, the miswak, is used in many developing countries as the traditional means for oral hygiene. It is prepared from the roots, twigs and stem of Salvadora persica or other alternative local plants. OBJECTIVES: To compare the effects of the chewing stick miswak (from S. persica) and toothbrush on subgingival plaque microflora among Saudi Arabian individuals. Further, to investigate whether components extracted from S. persica may interfere with the subgingival plaque micro-organisms. MATERIAL AND METHODS: Fifteen healthy Saudi Arabian male volunteers aged 21-36 years were included in a single-blind, randomized cross-over study. The participants were taught how to use each device properly. Plaque sampling for DNA test was performed at the baseline, 1 week after professional tooth cleaning, and after 3 weeks of either miswak or toothbrush use. Identification and quantification of microbial species were performed by the checkerboard method, using whole genomic, digoxigenin-labelled DNA probes. Inhibition zones around miswak were examined on agar plates with Actinobacillus actinomycetemcomitans and the leukotoxicity of this bacterium was analyzed in a bioassay with macrophages+/-extracts of miswak. RESULTS: Miswak and toothbrushing had a similar influence on the levels of the subgingival microbiota. However, A. actinomycetemcomitans was significantly more reduced by miswak (p<0.05) than by toothbrushing. These results were supported by our in vitro results which, indicated that extracts from S. persica might interfere with the growth and leukotoxicity of A. actinomycetemcomitans. CONCLUSIONS: In contrast to toothbrush use, miswak use significantly reduced the amount of A. actinomycetemcomitans in the subgingival plaque.

Adult↗

Comparison of polymerase chain reaction and culture methods for detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in subgingival plaque samples.

In this study, the major periodontal pathogens Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis were detected in subgingival plaque samples from patients with periodontal disease by polymerase chain reaction (PCR) and conventional culture methods. 170 plaque samples from 43 patients were analysed; A. actinomycetemcomitans and P. gingivalis were each detected in 40 (24%) of samples by PCR, whereas conventional culture methods detected A. actinomycetemcomitans and P. gingivalis in 25 (15%) and 18 (11%) of samples, respectively. The proportion of patients carrying A. actinomycetemcomitans in at least 1 sampled periodontal site was 17/43 (40%) by PCR and 13/43 (30%) by culture; for P. gingivalis this was 12/43 (28%) by PCR and 9/43 (21%) by culture. Only 5 samples, from 3 patients, harboured both A. actinomycetemcomitans and P. gingivalis. It is concluded that PCR is more accurate than conventional culture methods for identification of these periodontal pathogens in subgingival plaque samples and has a higher frequency of detection.

Aggregatibacter actinomycetemcomitans↗

[Subgingival plaque due to gingivitis and inactive periodontitis sites in the adult periodontitis patient].

There are several discrepancies with respect to the composition of subgingival plaque in gingivitis and in inactive adult periodontitis (AP). In this study we compared subgingival plaque samples taken from gingivitis sites to those from inactive periodontitis sites of the same patients. Of 44 patients plaque samples from 86 gingivitis sites and 92 periodontitis sites were analysed. Darkfield microscopy showed a higher proportion of spirochetes and mobile rods in the periodontitis sites. Analysis of cultures revealed a higher and statistically significant number of anaerobes in the periodontitis sites (1.7 x 10(7) vs 3 x 10(6), p = 0.006). The following bacterial species were isolated more frequently from periodontitis sites than from gingivitis sites: Actinobacillus actinomycetemcomitans (18% vs 10%), as well as the black-pigmented Prevotella intermedia (68% vs 48%) and Porphyromonas gingivalis (48% vs 28%). On the one hand, these small differences in the bacteriological parameters can be explained by the fact that both gingivitis and periodontitis plaque samples were taken from the same periodontitis patients. An infection of the gingivitis sites from the parodontitis sites within the same patient could not be excluded. On the other hand, the mean probing depth of the gingivitis sites was relatively high, 3.6 mm (measuring point interdental plus pseudo-pocket) which may favor the growth of anaerobic bacteria.

Adult↗

Distribution of 3-hydroxy iC17:0 in subgingival plaque and gingival tissue samples: relationship to adult periodontitis.

Gram-negative organisms incorporate hydroxy fatty acids into the lipid A moiety of lipopolysaccharide (LPS), and in the case of some members of the family Enterobacteriaceae, hydroxy fatty acids are incorporated exclusively into lipid A. However, a limited number of Bacteroides species have been shown to incorporate several classes of 3-hydroxy fatty acids, particularly 3-hydroxy iC17:0, into constitutive lipids as well as LPS. The present study examined the distribution of hydroxy fatty acids in two periodontal pathogens, Prevotella intermedia and Porphyromonas gingivalis, by employing a phospholipid extraction procedure (E. G. Bligh and W. J. Dyer, Can. J. Biochem. Physiol. 37:911-917, 1959) which partitioned constitutive lipids into the organic solvent phase and LPS into the aqueous phase. The distribution of hydroxy fatty acids within organic solvent and aqueous extracts of these bacterial species was then compared with the distribution in subgingival plaque samples isolated from either gingivitis or severe periodontitis sites as well as the distribution in gingival tissue samples. The organic solvent and aqueous extracts were hydrolyzed under strong alkaline conditions, and the free fatty acids were treated to form pentafluorobenzyl-ester, trimethylsilyl-ether derivatives. Hydroxy fatty acid levels were quantified by using gas chromatography-negative-ion chemical ionization-mass spectrometry. By using this approach, the mean values of the 3-hydroxy iC17:0 recovered within organic solvent extracts of P. gingivalis strains ranged from 56 to 63% of total 3-hydroxy iC17:0. Substantially less 3-hydroxy iC17:0 (< 5%) was recovered in organic solvent extracts of P. intermedia. By comparison, 75% of the 3-hydroxy iC17:0 in periodontitis subgingival plaque samples was recovered in organic solvent extracts, while only 43% of the 3-hydroxy iC17:0 in gingivitis plaque samples from the same patients was recovered in organic solvent extracts. However, 3-hydroxy iC17:0 was recovered essentially only in organic solvent extracts of both healthy or mildly inflamed and periodontitis gingival tissue samples. The preferential recovery of 3-hydroxy iC17:0 in tissue lipids indicates that gingival tissues do not harbor significant levels of subgingival plaque organisms which contain 3-hydroxy iC17:0. Furthermore, these results indicate that LPS from these organisms is not prevalent in gingival tissues. Finally, these results indicate either selective penetration of certain bacterial lipids into gingival tissues or that 3-hydroxy iC17:0 is metabolically transferred from bacterial lipids into gingival tissue lipids.

Bacteroides↗

A feasibility study on the use of direct light silver stain compared with dark field microscopy for differential counting of subgingival plaque.

The purpose of the present study was to compare direct light microscopy using a silver stain, with dark field microscopy for differential counting of subgingival plaque samples from patients with periodontitis. The feasibility of using the staining method was assessed on subgingival plaque samples which were collected at 34 sites from patients with untreated adult periodontitis. Differential counts of plaque morphotypes assessed by both methods showed close agreement. The proportions of spirochaetes assessed by both methods were significantly associated with probing depth. The silver stain proved a simple, rapid and inexpensive method for differential counting of plaque composition. This method is suitable for possible use in general dental practice where smears could be made at the chair-side and allowed to dry ready for staining and counting at the dentist's convenience using a simple light microscope. A permanent record is also produced which could allow for comparison of samples from before and after treatment and could be used as an aid to patient motivation in suitable cases.

Adult↗

[The study of major anaerobic bacteria from subgingival plaques of juvenile periodontitis].

The studies of the subgingival plaques from juvenile periodontitis (JP) have shown that JP is associated with Haemophilus actinomycetemcomitans (H. a), Capnocytophaga (Capno.) and other species. This study was designed to study these species with Chinese JP patients using selective cultivable technique. The media used include TSBV to support H. a, TBBP to support Capno. and selective media for Bacteroides gingivalis. A total of 303 subgingival samples were collected from 43 JP, 31 gingivitis and 13 normal juvenile. It was found that the recovery rates of H. a and Capnocytophaga in JP group were higher than that in two other groups. The Black-pigmented Bacteroides had a similar recovery rate in JP and gingivitis groups, but higher than that in periodontal healthy group. The bacterial counts and the correlation analysis between bacteria findings and clinical indices were consistent with the above results.

Adolescent↗

Effect of gingival fluid collection on subgingival plaque sampling.

The purpose of this study was to determine whether gingival crevicular fluid (GCF) sampling by paper strip removes sufficient bacteria to affect subsequent subgingival plaque sampling using a curette. In 25 subjects, one healthy, gingivitis and periodontitis site was sampled for GCF using a strip followed by subgingival plaque sampling with a curette. Bacterial assays indicated that GCF strips removed significant numbers of bacteria when placed intracrevicularly for 5 s. A greater proportion of total bacteria was removed with strip sampling at healthy rather than gingivitis or periodontitis sites. Qualitative assessment of presence or absence of spirochaetes and dark-pigmented species indicated potential for significant interference of curette sampling by the strip at gingivitis and healthy sites. We concluded that paper strip GCF sampling may significantly affect curette sampling at the same sites. The magnitude of this impact depended on the clinical classification of specific sites and the assay performed.

Bacteria↗

[A study on sex hormones in gingival crevicular fluid and black pigmented bacteria in subgingival plaque of pregnant women].

Gingivitis is one of the most common oral diseases. It is caused by dental plaque and by the factors produced/released from it. The black pigmented bacteria in subgingival dental plaque are thought to be the periodontopathogens. Prevotella intermedia and Porphyromonas gingivalis have been shown to be closely associated with human gingivitis. Prevetella intermedia can use female sex hormones such as progesterone or estradiol as a source of nutrients. In pregnant women, the concentrations of progesterone and estradiol are markedly increased in serum and both are accumulated and found in the gingival tissue. The purpose of this study was to test levels of female sex hormones in gingival crevicular fluid and to observe the relationship between hormones and black pigmented bacteria in subgingival plaque. The results showed that the amount of progesterone found in the gingival crevicular fluid and percentage of black pigmented bacteria in subgingival plaque of pregnant women were markedly higher than in the postpartum stage. The percentage of black pigmented bacteria was positively correlated with the progesterone level, pregnancy and the severity of the gingivitis. Severity of the gingivitis was positively correlated with both the plaque index and the percentage of black pigmented bacteria in subgingival plaque.

Adult↗

[Distribution of 3 kinds of periodontal pathogens in subgingival plaques of patients with chronic periodontitis].

OBJECTIVE: To observe the prevalence of Porphyromonas gingivalis(Pg), Treponema denticola(Td) and Bacterides forsythus(Bf) subgingivally in diseased sites of chronic periodontitis (CP) patients. METHODS: Samples of subgingival plaque were detected by 3 kinds of oligonucleotide DNA probe from 60 sites of CP patients and 10 healthy sites of 10 healthy people. RESULTS: The positive rates of Pg, Bf and Td in patients were 91.67%, 90.00% and 95.67% respectively; Pg, Bf and Td were detected simultaneously in 83.33% patients and Pg, Bf and Td were found to be related with each other(P < 0.01). CONCLUSION: Pg, Bf and Td were prominent periodontopathic bacteria and related each other, might exist in complexes in subgingival plaque and coaggretate together.

Adult↗

Bacterial profiles of subgingival plaques in periodontitis.

In this report over 400 subgingival plaque samples taken from over 110 patients were examined microscopically and culturally for 30 bacterial parameters. The patients could be placed into six disease categories based upon clinical criteria. The bacterial profile of each clinical category was generally distinctive of that category. Periodontal patients who had been successfully treated and maintained had plaques that were populated by significantly higher proportions of Streptococcus sanguis, Actinomyces viscosus, A. odontolyticus and S. mutans and significantly lower proportions of B. gingivalis and spirochetes compared to the five untreated disease categories. The spirochetes were the overwhelming microbial type in the plaques of adult periodontitis (AP) patients, averaging about 45% of the microscopic count. The bacteriological results could not distinguish between ADA Type III and IV periodontitis, suggesting that the same type of infection was occurring in an active site in any AP patient. The patients designated as early onset periodontitis (EOP) differed from the other patients by their relative youth and by their significantly higher proportions of Bacteroides gingivalis and/or B. intermedius. Two types of EOP were recognized in which the most diseased variant was characterized by having an average of 49% spirochetes in the plaque. Four localized juvenile periodontitis (LJP) patients were notable in not having detectable A. actinomycetemcomitans. The data indicate that the various types of periodontitis, with the possible exception of LJP are specific anaerobic infections involving spirochetes and to a lesser extent B. gingivalis and B. intermedius.

Actinomyces↗

Eikenella corrodens in subgingival plaque: relationship to age and periodontal condition.

BACKGROUND: The purpose of this study was to determine the prevalence and distribution of Eikenella corrodens (E. corrodens) in subgingival plaque in different age and periodontitis groups and to examine whether its presence is related to periodontal diseases. METHODS: A total of 273 subgingival plaque samples from 213 periodontitis patients and 60 healthy subjects were assessed. Smears from each plaque sample were made and E. corrodens was detected by means of indirect immunofluorescent technique. Mean percentage of E. corrodens per total bacteria (distribution) was calculated in each sample. The prevalence (% of positive samples) and distribution of E. corrodens were statistically analyzed based on age or diagnosis by means of Fisher's exact test and analysis of variance (ANOVA). RESULTS: Prevalence of E. corrodens decreased by age in the healthy control group; however, prevalence did not change in periodontitis groups. Distribution of E. corrodens was highest in juvenile periodontitis (JP) (2.3 +/- 1.5%) followed by post-JP (1.7 +/- 2.1%), prepubertal periodontitis (1.4 +/- 1.1%), rapidly progressive periodontitis (0.8 +/- 0.7%), adult periodontitis (0.7 +/- 0.6%), and healthy subjects (0.3 +/- 0.3%) (ANOVA, P<0.0001). The <20-year-old age group with periodontitis showed the highest distribution of E. corrodens (2.2 +/- 1.6%) compared to the older age groups who were either healthy or had periodontitis (ANOVA, P<0.0001). CONCLUSIONS: Since the distribution of E. corrodens is significantly higher in JP, post-JP, and PP, E. corrodens might play an important role in the occurrence or progression of periodontitis in young patients.

Adult↗

Periodontal diseases, caries, and microbial composition of the subgingival plaque in children: a longitudinal study.

The present study compares periodontal parameters, caries, and levels of colony forming units (CFU) of bacteria from subgingival plaque of permanent teeth, to those of primary teeth examined 4 years previously. Six children who had periodontitis and 5 who had no periodontitis in primary teeth (groups A and B respectively) were examined. The microbial examination included the number of CFU of the total anaerobic count, Actinobacillus actinomycetemcomitans, Streptococcus mutans and Porphyromonas gingivalis. The differences in CFU values for the permanent teeth between groups A and B were not significant. Group A had significantly higher gingival inflammation values in the permanent teeth than group B. Permanent teeth had significantly higher CFU values of P. gingivalis than the primary teeth. Based on the present limited sample, the number of CFU from bacteria of the subgingival plaque of primary teeth are not an adequate predictor of periodontal disease or caries in the permanent teeth.

Adolescent↗

[The prognostic significance of the presence of Actinobacillus actinomycetemcomitans in the subgingival plaque of young orthodontic patients].

The aim of present study was to evaluate the prognostic significance of the presence of Actinobacillus actinomycetemcomitans in subgingival plaque from young patients wearing fixed orthodontic appliances. Fifteen orthodontic patients, 3 males and 12 females, aged 10 to 27, systemically healthy, participated in the study. First examination took place in the spring of 1992 (T1) and consisted in the collection of clinical (pocket depth, PD; Gingival Index, GI; Plaque Index, P1I; Gingival Bleeding Index, GBI) and microbiological (Actinobacillus actinomycetemcomitans presence) parameters in 4-8 periodontal sites in each patient. The same clinical and microbiological parameters were collected again in the spring of 1994 (T2). Results showed that the presence of Actinobacillus actinomycetemcomitans in the examined periodontal sites underwent fluctuation, 25% of sites being positive for the sought after bacterium in T2 whereas the percentage of Actinobacillus actinomycetemcomitans positivity in T1 was 47.5% with a statistically significant difference (p < 0.005). On the contrary clinical parameters did not show significant modifications through the study period T1-T2. Differential analysis of clinical parameter modifications in relation to Actinobacillus actinomycetemcomitans positivity in T1 or Actinobacillus actinomycetemcomitans-positivity fluctuations in T1-T2 period did not evidence any significant difference among various microbiological conditions. The authors conclude that the mere presence of Actinobacillus actinomycetemcomitans in subgingival plaque from specific periodontal sites in young orthodontic patients does not seem sufficient to consider those sites at risk for periodontal destruction in the medium-short term.

Adolescent↗

A rapid DNA probe test compared to culture methods for identification of subgingival plaque bacteria.

There has been a significant amount of interest in developing a more rapid and cost-effective test to identify bacterial pathogens in plaque. DNA probe technology may meet both these objectives, it is more rapid and cost-effective when compared to culture methods. The purpose of this study was to compare an automated DNA probe test with classical culture methods for identifying Bacteroides forsythus and Porphyromonas gingivalis in subgingival plaque of patients with adult periodontitis. Subgingival plaque samples were collected from sites with moderate to severe periodontitis and divided into two aliquots for analysis by either DNA probe or culture methods. When the DNA probe method was compared with the culture method (gold standard), the sensitivity and specificity for B. forsythus were 92.0% (SE = 3.4%) and 50.5% (SE = 7.8%), respectively; for P. gingivalis they were 52.2% (SE = 8.7%) and 74.7% (SE = 5.9%), respectively. Detection of B. forsythus and P. gingivalis by DNA probe correlated with probing depth (P = 0.01 for B. forsythus and P = 0.03 for P. gingivalis). It was concluded the DNA probe test was comparable to culture methods in detecting B. forsythus. In addition, when compared to the culture method, a better correlation was obtained with DNA probe detection of B. forsythus or P. gingivalis and clinical parameters.

Adult↗