Biomedical subjects

A D Haffajee

Publications and source records attributed to A D Haffajee.

At least 19 recordsLinked to original sources

PubMed2006

Association of Eubacterium nodatum and Treponema denticola with human periodontitis lesions.

BACKGROUND: The purpose of the present investigation was to compare the levels, proportions and percentage of sites colonized by 40 bacterial species in subgingival plaque samples from periodontally healthy subjects and patients with chronic periodontitis to seek possible pathogens other than the consensus pathogens Porphyromonas gingivalis and Tannerella forsythia. METHOD: Subgingival plaque samples were taken from the mesial aspect of each tooth in 635 subjects with chronic periodontitis and 189 periodontally healthy subjects. The samples were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples = 21,832). Mean counts, % DNA probe counts and percentage of sites colonized at >10(5) were determined for each species in each subject and then averaged in each clinical group. Significance of difference between groups was determined using the Mann-Whitney test. Association between combinations of species and periodontal status was examined by stepwise logistic regression analysis. Analyses were repeated using a subset of subjects from both clinical groups who had proportions of P. gingivalis plus T. forsythia less than the median (4.42%) found in periodontally healthy subjects. All analyses were adjusted for multiple comparisons. RESULTS: For the 824 subjects the consensus pathogens P. gingivalis and T. forsythia as well as Eubacterium nodatum and Treponema denticola had significantly higher mean counts, proportions and percentage of sites colonized in samples from subjects with periodontitis than from periodontally healthy subjects. There were significantly more Capnocytophaga gingivalis, Streptococcus gordonii and Veillonella parvula in periodontally healthy subjects. E. nodatum, T. denticola, Streptococcus oralis, Streptococcus intermedius, Fusobacterium nucleatum ssp. vincentii all had higher counts and proportions in diseased than healthy subjects who had low proportions of P. gingivalis and T. forsythia. Logistic regression analysis indicated that the same species groups were associated with disease status after adjusting for the proportions of the other species. CONCLUSIONS: This investigation confirmed the strong association of P. gingivalis and T. forsythia with chronic periodontitis and emphasized a strong association of E. nodatum and T. denticola with periodontitis whether in the presence or absence of high levels of the consensus pathogens. Other species, including S. oralis, Eikenella corrodens, S. intermedius and F. nucleatum ssp. vincentii, were associated with disease when P. gingivalis and T. forsythia were present in low proportions.

Adult↗

PubMed2005

The salivary microbiota as a diagnostic indicator of oral cancer: a descriptive, non-randomized study of cancer-free and oral squamous cell carcinoma subjects.

BACKGROUND: The purpose of the present investigation was to determine if the salivary counts of 40 common oral bacteria in subjects with an oral squamous cell carcinoma (OSCC) lesion would differ from those found in cancer-free (OSCC-free) controls. METHODS: Unstimulated saliva samples were collected from 229 OSCC-free and 45 OSCC subjects and evaluated for their content of 40 common oral bacteria using checkerboard DNA-DNA hybridization. DNA counts per ml saliva were determined for each species, averaged across subjects in the 2 subject groups, and significance of differences between groups determined using the Mann-Whitney test and adjusted for multiple comparisons. Diagnostic sensitivity and specificity in detection of OSCC by levels of salivary organisms were computed and comparisons made separately between a non-matched group of 45 OSCC subjects and 229 controls and a group of 45 OSCC subjects and 45 controls matched by age, gender and smoking history. RESULTS: Counts of 3 of the 40 species tested, Capnocytophaga gingivalis, Prevotella melaninogenica and Streptococcus mitis, were elevated in the saliva of individuals with OSCC (p < 0.001). When tested as diagnostic markers the 3 species were found to predict 80% of cancer cases (sensitivity) while excluding 83% of controls (specificity) in the non-matched group. Diagnostic sensitivity and specificity in the matched group were 80% and 82% respectively. CONCLUSION: High salivary counts of C. gingivalis, P. melaninogenica and S. mitis may be diagnostic indicators of OSCC.

Adolescent↗

PubMed2005

Differences in the subgingival microbiota of Swedish and USA subjects who were periodontally healthy or exhibited minimal periodontal disease.

BACKGROUND: Previous studies have shown differences in the mean proportions of subgingival species in samples from periodontitis subjects in different countries, which may relate to differences in diet, genetics, disease susceptibility and manifestation. The purpose of the present investigation was to determine whether there were differences in the subgingival microbiotas of Swedish and American subjects who exhibited periodontal health or minimal periodontal disease. METHOD: One hundred and fifty eight periodontally healthy or minimally diseased subjects (N Sweden=79; USA=79) were recruited. Subjects were measured at baseline for plaque, gingivitis, BOP, suppuration, pocket depth and attachment level at 6 sites per tooth. Subgingival plaque samples taken from the mesial aspect of each tooth at baseline were individually analyzed, in one laboratory, for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples=4345). % DNA probe counts comprised by each species was determined for each site and averaged across sites in each subject. Significance of differences in proportions of each species between countries was determined using ancova adjusting for age, mean pocket depth, gender and smoking status. p values were adjusted for multiple comparisons. Cluster analysis was performed to group subjects based on their subgingival microbial profiles using a chord coefficient and an average unweighted linkage sort. RESULTS: On average, all species were detected in samples from subjects in both countries. After adjusting for multiple comparisons, 5 species were in significantly higher adjusted mean percentages in Swedish than American subjects: Actinomyces naeslundii genospecies 1 (9.7, 3.3); Streptococcus sanguis (2.5, 1.2); Eikenella corrodens (1.7, 1.0); Tannerella forsythensis (3.5, 2.3) and Prevotella melaninogenica (6.3, 1.8). Leptotrichia buccalis was in significantly higher adjusted mean percentages in American (5.5) than Swedish subjects (3.0). Cluster analysis grouped 121 subjects into 8 microbial profiles. Twenty four of the 40 test species examined differed significantly among cluster groups. Five clusters were dominated by American subjects and 2 clusters by Swedish subjects. Fifty eight of 79 (73%) of the Swedish subjects fell into 1 cluster group dominated by high proportions of A. naeslundii genospecies 1, Prevotella nigrescens, T. forsythensis and P. melaninogenica. Other clusters were characterized by high proportions of Actinomyces gerencseriae, Veillonella parvula, Capnocytophaga gingivalis, Prevotella intermedia, Eubacterium saburreum, L. buccalis and Neisseria mucosa. CONCLUSIONS: The microbial profiles of subgingival plaque samples from Swedish and American subjects who exhibited periodontal health or minimal disease differed. The heterogeneity in subgingival microbial profiles was more pronounced in the American subjects, possibly because of greater genetic and microbiologic diversity in the American subjects sampled.

Adult↗

PubMed2005

Scaling and root planing, systemic metronidazole and professional plaque removal in the treatment of chronic periodontitis in a Brazilian population II--microbiological results.

OBJECTIVE: The current investigation evaluated changes in levels and proportions of 39 bacterial species in subgingival plaque samples after scaling and root planing (SRP) alone or in combination with systemic metronidazole and/or professional cleaning in subjects with chronic periodontitis. METHODS: Forty-four adult subjects (mean age 45+/-6 years) with periodontitis were randomly assigned in four treatment groups, a control (C, n=10) that received SRP and placebo and three test groups treated as follows: T1 (n=12): SRP and metronidazole (M, 400 mg tid) for 10 days; T2 (n=12): SRP, weekly professional supragingival plaque removal for 3 months (PC) and placebo; and T3 (n=10): SRP, M and PC. Subgingival plaque samples were taken from seven sites per subject at baseline and 90 days post-therapy. Counts of 39 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Wilcoxon signed ranks test and among groups using ancova. RESULTS: The mean counts of the majority of the species were reduced post-therapy in the 4 treatment groups. Counts (x 10(5)+/-SEM) of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola were significantly reduced in groups T2 and T3. Levels of beneficial species, such as some Actinomyces species, Veillonella parvula, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii were minimally affected in levels when the combined therapy was applied (T3). Mean proportions of red complex species decreased from 18.4% at baseline to 3% at 90 days post-therapy in group T3 (p<0.01), from 25.8% to 2.3% in group T2 (p<0.01), from 17.7% to 5.6% in group T1 (p<0.05) and from 19.4% to 8.8% in group C (NS). Proportions of the suspected periodontal pathogens from the orange complex were also markedly reduced in groups T2 and T3. CONCLUSIONS: All treatments reduced counts and proportions of red complex species. Adjunctive therapy appeared to have a greater effect and also affected members of the orange complex.

Administration, Oral↗

PubMed2005

Initial subgingival colonization of 'pristine' pockets.

The treatment of periodontitis/peri-implantitis involves the reduction/eradication of periopathogens. After therapy, beneficial and pathogenic species recolonize the subgingival area. The dynamics of recolonization and especially the role of the supragingival environment in this process are still not well-understood. This prospective, split-mouth study followed the early colonization of 'pristine' pockets created during implant surgery (16 partially edentulous patients), to record the time needed before a complex subgingival flora could be established with the supragingival area as the single source. Four subgingival plaque samples were taken from shallow and medium pockets around implants (test), and neighboring teeth (undisturbed microbiota as reference) 1, 2, and 4 wks after abutment connection. Checkerboard DNA-DNA hybridization and culture data revealed a complex microbiota (including several pathogenic species) in the pristine pockets within a wk, with a minimal increase in counts up to 4 wks. Analysis of these data demonstrated that, even with the supragingival environment as the single source for colonizing bacteria, a complex subgingival microbiota can develop within 1 wk.

Adult↗

PubMed2004

Identification of early microbial colonizers in human dental biofilm.

AIMS: To elucidate the first colonizers within in vivo dental biofilm and to establish potential population shifts that occur during the early phases of biofilm formation. METHODS AND RESULTS: A 'checkerboard' DNA-DNA hybridization assay was employed to identify 40 different bacterial strains. Dental biofilm samples were collected from 15 healthy subjects, 0, 2, 4 and 6 h after tooth cleaning and the composition of these samples was compared with that of whole saliva collected from the same individuals. The bacterial distribution in biofilm samples was distinct from that in saliva, confirming the selectivity of the adhesion process. In the very early stages, the predominant tooth colonizers were found to be Actinomyces species. The relative proportion of streptococci, in particular Streptococcus mitis and S. oralis, increased at the expense of Actinomyces species between 2 and 6 h while the absolute level of Actinomyces remained unaltered. Periodontal pathogens such as Tannerella forsythensis(Bacteroides forsythus), Porphyromonas gingivalis and Treponema denticola as well as Actinobacillus actinomycetemcomitans were present in extremely low levels at all the examined time intervals in this healthy group of subjects. CONCLUSION: The data provide a detailed insight into the bacterial population shifts occurring within the first few hours of biofilm formation and show that the early colonizers of the tooth surface predominantly consist of beneficial micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The early colonizers of dental plaque are of great importance in the succession stages of biofilm formation and its overall effect on the oral health of the host.

Actinomyces↗

PubMed2004

Use of checkerboard DNA-DNA hybridization to study complex microbial ecosystems.

It has been difficult to conduct large scale studies of microbiologically complex ecosystems using conventional microbiological techniques. Molecular identification techniques in new probe-target formats, such as checkerboard DNA-DNA hybridization, permit enumeration of large numbers of species in very large numbers of samples. Digoxigenin-labeled whole genomic probes to 40 common subgingival species were tested in a checkerboard hydridization format. Chemifluorescent signals resulting from the hybridization reactions were quantified using a Fluorimager and used to evaluate sensitivity and specificity of the probes. Sensitivity of the DNA probes was adjusted to detect 10(4) cells. In all, 93.5% of potential cross-reactions to 80 cultivable species exhibited signals <5% of that detected for the homologous probe signal. Competitive hybridization and probes prepared by subtraction hybridization and polymerase chain reaction were effective in minimizing cross-reactions for closely related taxa. To demonstrate utility, the technique was used to evaluate 8887 subgingival plaque samples from 79 periodontally healthy and 272 chronic periodontitis subjects and 8126 samples from 166 subjects taken prior to and after periodontal therapy. Significant differences were detected for many taxa for mean counts, proportion of total sample, and percentage of sites colonized between samples from periodontally healthy and periodontitis subjects. Further, significant reductions were observed post therapy for many subgingival species including periodontal pathogens. DNA probes used in the checkerboard DNA-DNA format provide a useful tool for the enumeration of bacterial species in microbiologically complex systems.

Bacterial Typing Techniques↗

PubMed2004

Clinical and microbiological changes associated with the use of combined antimicrobial therapies to treat "refractory" periodontitis.

BACKGROUND: The present investigation examined clinical and microbial changes after a combined aggressive antimicrobial therapy in subjects identified as "refractory" to conventional periodontal therapy. METHOD: Fourteen subjects were identified as "refractory" based on full-mouth mean attachment loss and/or >3 sites with attachment loss > or =3 mm following scaling and root planing (SRP), periodontal surgery and systemic antibiotics. After baseline monitoring, subjects received SRP, locally delivered tetracycline at pockets > or =4 mm, systemically administered amoxicillin (500 mg, t.i.d. for 14 days)+metronidazole (250 mg, t.i.d. for 14 days) and professional removal of supragingival plaque weekly for 3 months. Subjects were monitored clinically every 3 months post-therapy for 2 years. Subgingival plaque samples were taken at the same time points from the mesial aspect of each tooth and the levels of 40 subgingival taxa were determined using checkerboard DNA-DNA hybridization. Mean levels of each species were averaged within a subject at each visit. Significance of changes in clinical and microbiological parameters over time were evaluated using the Friedman or Wilcoxon signed ranks test. RESULTS: On average, subjects showed significant improvements in all clinical parameters after therapy. Mean (+/-SEM) full-mouth pocket depth reduction was 0.83+/-0.13 mm and mean attachment level "gain" was 0.44+/-0.12 at 24 months. Clinical improvement was accompanied by major reductions in multiple subgingival species during the first 3 months of active therapy that were maintained for most species to the last monitoring visit. Reductions occurred for three Actinomyces species, "orange complex" species including Campylobacter showae, Eubacterium nodatum, three Fusobacterium nucleatum subspecies, Peptostreptococcus micros, Prevotella intermedia as well as the "Streptococcus milleri" group, Streptococcus anginosus, Streptococcus constellatus and Streptococcus intermedus. Subjects differed in their response to therapy; six modest response subjects exhibited less attachment level gain and were characterized by reductions in the microbiota from baseline to 3 months, but re-growth of many species thereafter. CONCLUSIONS: The combined antibacterial therapy was successful in controlling disease progression in 14 "refractory" periodontitis subjects for 2 years.

Adult↗

PubMed2004

Subgingival microbiota of chronic periodontitis subjects from different geographic locations.

BACKGROUND: Most clinical studies assume that the subgingival microbiota is similar from one geographic location to another. The purpose of the present investigation was to examine the composition of the subgingival microbiota in chronic periodontitis subjects from four countries. METHOD: Subjects with chronic periodontitis (N, Sweden=101; USA=115; Brazil=58; Chile=26) were recruited. Subjects were measured at baseline for plaque, gingivitis, bleeding on probing (BOP), suppuration, pocket depth (PD) and attachment level (AL) at six sites per tooth. Subgingival plaque samples taken from the mesial aspect of each tooth at baseline were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples=6036). % DNA probe counts comprised by each species was determined for each site and averaged across sites in each subject. Significance of differences in proportions of each species among countries was determined using ancova adjusting for age, mean pocket depth, gender and smoking status. p-Values were adjusted for multiple comparisons. RESULTS: On average, all species were detected in samples from subjects in the four countries. Thirteen species differed significantly in adjusted mean proportions among countries even after adjusting for multiple comparisons. Porphyromonas gingivalis, one species that differed in proportions among countries, comprised adjusted means of 7.5, 11.9, 1.6 and 6.6% of the microbiota in subjects from Brazil, Chile, Sweden and USA (p<0.001), while mean proportions of Treponema denticola were 6.7, 4.2, 0.8 and 2.3, respectively (p<0.001). In contrast, a key periodontal pathogen, Tannerella forsythensis, exhibited mean proportions ranging from 6.2-8.5% and did not differ significantly among countries. Besides these species, prominent species in Brazil were Actinomyces naeslundii genospecies 1 and 2 (8.4%, 7.2%) and Prevotella intermedia (6.5%); in Chile, Prevotella melaninogenica (6.4%) and Neisseria mucosa (5.3%); in Sweden A. naeslundii genospecies 2 (8.4%), Capnocytophaga gingivalis (7.1%) and Peptostreptococcus micros (5.0%); in USA A. naeslundii genospecies 2 (7.5%), P. intermedia (6.8%) and C. gingivalis (6.1%). CONCLUSIONS: The microbial profiles of subgingival plaque samples from chronic periodontitis subjects in four countries showed surprisingly marked differences. These differences persisted after adjusting for age, mean pocket depth, gender and smoking status.

Adult↗

PubMed2004

Scaling and root planing, systemic metronidazole and professional plaque removal in the treatment of chronic periodontitis in a Brazilian population. I. clinical results.

OBJECTIVE: The current investigation evaluated the clinical effects of scaling and root planing (SRP) alone or in combination with systemic metronidazole and/or repeated professional removal of supragingival plaque in subjects with chronic periodontitis. METHODS: Fourty-four adult subjects (mean age: 45+/-6 years) with periodontitis were randomly assigned to four treatment groups; a control (C, n=10) that received SRP and placebo and three test groups treated as follows: Test 1 (T1) (n=12) received SRP and metronidazole (400 mg t.i.d., M) for 10 days; Test 2 (T2) (n=12) received SRP, weekly professional supragingival plaque removal for three months (professional cleaning (PC)) and placebo; and Test 3 (T3) (n=10) received SRP, M and PC. Pocket depth (PD), attachment level (AL), bleeding on probing (BOP) and presence of visible plaque and suppuration were measured at six sites per tooth at baseline and at 90 days post-therapy. Significance of differences over time was determined using the Wilcoxon test, and among groups using ancova. RESULTS: A reduction in full-mouth mean clinical parameters was observed at 90 days after all therapies. Sites with baseline PD<4 mm showed an increase in mean PD in the control group and in mean AL in all treatment groups. Sites with baseline PD of 4-6 mm in subjects who received PC as part of therapy (T2, T3) showed a marked reduction in PD, AL and in the % of sites with BOP. Subjects who received metronidazole (T1 and T3) showed the best clinical response at sites with an initial PD of >6 mm. The major clinical benefit occurred when the combination of SRP, M and PC was used. Group T3 showed the least attachment loss in initially shallow pockets. This group also exhibited the greatest reduction in the % of sites with BOP and suppuration as well as in mean PD and AL at sites with baseline PD>4 mm. CONCLUSION: The data suggest a significant clinical benefit in combining SRP, systemic metronidazole and weekly professional supragingival plaque removal for the treatment of chronic periodontitis.

Adult↗

PubMed2003

Accumulation of methylglyoxal in the gingival crevicular fluid of chronic periodontitis patients.

BACKGROUND, AIMS: Methylglyoxal (MG), a toxic product of cellular metabolism, is elevated in tissues and fluids in a number of human diseases. A cross-sectional study was undertaken to determine whether MG accumulates in the gingival crevicular fluid (GCF) of chronic periodontitis patients. METHODS: GCF samples were collected for 30 s each from three teeth with pocket depths greater than 3 mm (DD sites), from 14 chronic periodontitis patients. Control samples were taken from three healthy sites (DH sites) in the same patients, as well as from seven subjects who were periodontally healthy (HH sites). Fluid volumes were determined and the strips were placed in 0.5 N perchloric acid. Subsequently, samples were derivatized with o-phenylenediamine and the resulting methylquinoxaline was assayed by high-performance liquid chromatography on Lichrospher(R)-100 RP-18, with UV detection. RESULTS: Mean pocket depths were 5.7+/-0.7, 2.7+/-0.6 and 2.7+/-0.5 mm (mean+/-SD) for the DD, DH and HH sites, respectively. Mean MG levels were found to be 208.7+/-241.7 and 142.9+/-235.7 pmol/site in the GCF from DD and DH sites, respectively (p=0.0023), but only 11.5+/-4.4 pmol/site for the HH sites. Bacteroides forsythus has been found to accumulate high levels of MG in culture (unpublished data) and, consistent with this, the sampled diseased sites contained higher levels of B. forsythus than the corresponding healthy sites (2.7+/-4.2 x 10(5) versus 0.7+/-1.1 x 10(5), respectively; p=0.022). Total "red complex" microorganisms were significantly elevated in the DD sites. CONCLUSIONS: In view of the known protein- and DNA-modifying effects of MG, the finding of elevated levels of MG in the GCF from chronic periodontitis patients supports the hypothesis that MG may contribute to destructive tissue damage in this disease.

Bacteroides↗

PubMed2003

Relationship between periodontal pocket sulfide levels and subgingival species.

BACKGROUND: Many species implicated in the pathogenesis of periodontal disease produce volatile sulfur compounds (VSC). This investigation examined the relationship between levels of sulfide and subgingival bacterial species in the same periodontal pockets. MATERIAL AND METHODS: Twenty chronic periodontitis subjects were measured clinically at six sites per tooth for plaque, gingivitis, bleeding on probing, suppuration, pocket depth and attachment level. Subgingival plaque samples, taken from the mesial aspect of each tooth, were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. Sulfide levels were measured at the same sites using a Diamond Probe/Perio 2000 system. Clinical and microbiological data were averaged for sulfide-positive and -negative sites separately in each subject and then averaged across subjects. Significance differences in clinical and microbial parameters between sulfide-positive and -negative sites were sought using the Wilcoxon signed ranks test. RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) at sulfide-negative and -positive sites were 44.0 +/- 9.9 and 65.0 +/- 13.3, respectively (p < 0.01). Seventeen species were found at significantly higher levels in sulfide-positive than -negative sites. These included abundant producers of VSC such as members of the genera Fusobacterium, Campylobacter, Prevotella, Treponema and Eubacterium, and Bacteriodes forsythus, Selenomonas noxia and Propionibacterium acnes. Prevotella intermedia, Bacteriodes forsythus, Prevotella nigrescens, Fusobacterium nucleatum ss vincentii and Treponema denticola exhibited the greatest difference in mean counts between sulfide-negative and -positive sites. Orange and red complex species were at higher counts at shallow (< 4 mm) sulfide-positive than shallow sulfide-negative sites. Although not statistically significant, mean clinical parameters were somewhat higher at sulfide-positive than sulfide-negative sites. CONCLUSIONS: Intra-pocket sulfide levels reflect the levels of sulfide-producing species and may provide useful diagnostic information.

Adult↗

PubMed2003

Controlling the plaque biofilm.

AIM: To examine the effect of supragingival plaque removal in conjunction with different periodontal therapies on subgingival plaque composition in different subject populations. METHOD: Four different studies are presented which examined the effect of repeated removal of supragingival plaque performed by professionals or by tooth brushing on subgingival plaque composition. The studies were performed in different populations including chronic periodontitis, periodontal maintenance and refractory subjects. For all studies, each subject was examined for clinical parameters at up to 168 sites and subgingival plaque samples were taken from the mesial aspect of each tooth and examined for their content of specific bacterial species using checkerboard DNA-DNA hybridisation techniques. RESULTS: Repeated supragingival plaque removal used in conjunction with scaling and root planing only or combined with other periodontal therapies resulted in improvements in clinical parameters as well as significant decreases in the counts of subgingival species, including those associated with periodontal disease aetiology. Meticulous tooth brushing provided similar clinical and microbial improvements. CONCLUSIONS: Meticulous removal of supragingival plaque has beneficial effects on clinical parameters of periodontal disease and on the nature of the microbiota that colonises both above and below the gingival margin and appropriately has been a major focus in the prevention and control of dental diseases, particularly periodontal disease.

Anti-Bacterial Agents↗

PubMed2002

Antibiotic resistance of subgingival species during and after antibiotic therapy.

AIM: The purpose of the present investigation was to determine the percentage and identity of antibiotic-resistant species in subgingival plaque and saliva samples from chronic periodontitis patients treated by scaling and root planing followed by orally administered amoxicillin or metronidazole. METHOD: In all, 20 chronic periodontitis patients were selected for study. After clinical and microbiological monitoring, subjects were randomly assigned to receive either orally administered amoxicillin at the dosage of 500 mg, 3 times daily for 14 days or orally administered metronidazole at the dosage of 250 mg, 3 times daily for 14 days. For the antibiotic resistance determinations, subgingival plaque samples were taken from six posterior teeth at baseline, and 90 days; and from two randomly selected teeth at 3, 7 and 14 days during and after antibiotic administration. Samples were plated on enriched blood agar plates with or without either 2 micro g/mL metronidazole or 2 micro g/mL amoxicillin. Colonies were counted at 7 days. Significant differences in percentage of resistant organisms over time were determined by the Quade test. Microbial growth was washed from antibiotic-containing media and the identity of species determined using checkerboard DNA-DNA hybridization. Data were compared with those obtained in a previous study from subjects receiving SRP only or SRP followed by 14 days of orally administered doxycycline. The level of doxycycline used to determine antibiotic resistance in that study was 4 micro g/mL. RESULTS: The mean percentage of resistant isolates increased during antibiotic administration and returned to baseline levels by 90 days post therapy. The mean percentages (+/- SEM) of isolates resistant to 2 micro g/mL metronidazole were 53 +/- 9, 65 +/- 9, 79 +/- 4 and 69 +/- 7 at baseline, 3, 7 and 14 days during antibiotic administration, and 57 +/- 4, 64 +/- 5, 62 +/- 7 and 47 +/- 6 at 3, 7, 14 and 90 days after antibiotic administration. At the same time points, the percentage of resistant isolates to amoxicillin was 0.5 +/- 0.2, 22 +/- 12, 14 +/- 5 and 37 +/- 11 during, and 31 +/- 11, 8 +/- 3, 3 +/- 2 and 3 +/- 0.6 after, administration. Antibiotic-resistant isolates of resistant species detected during or after therapy were also detected prior to therapy. The most prevalent resistant species in the metronidazole-treated group were: A. naeslundii 1, S. constellatus, A. naeslundii 2, S. mitis, S. oralis, A. odontolyticus, S. sanguis, and in the amoxicillin-treated group: S. constellatus, P. nigrescens, E. saburreum, A. naeslundii 1, S. oralis, P. melaninogenica and P. intermedia. CONCLUSIONS: Systemic antibiotic administration transiently increased the percentage of resistant subgingival species, but a major component of subgingival plaque remained sensitive to the agents during their administration. Antibiotic-resistant isolates of resistant species could be detected in samples both prior to and after therapy. However, % antibiotic-resistant isolates returned to baseline levels 90 days after antibiotic administration.

Adult↗

PubMed2001

Relationship of cigarette smoking to attachment level profiles.

OBJECTIVES: The present investigation examined clinical features of periodontal disease and patterns of attachment loss in adult periodontitis subjects who were current, past or never smokers. MATERIAL AND METHODS: 289 adult periodontitis subjects ranging in age from 20-86 years with at least 20 teeth and at least 4 sites with pocket depth and/or attachment level >4 mm were recruited. Smoking history was obtained using a questionnaire. Measures of plaque accumulation, overt gingivitis, bleeding on probing, suppuration, probing pocket depth and probing attachment level were taken at 6 sites per tooth at all teeth excluding 3rd molars at a baseline visit. Subjects were subset according to smoking history into never, past and current smokers and for certain analyses into age categories <41, 41-49, >49. Uni- and multi-variate analyses examined associations between smoking category, age and clinical parameters. RESULTS: Current smokers had significantly more attachment loss, missing teeth, deeper pockets and fewer sites exhibiting bleeding on probing than past or never smokers. Current smokers had greater attachment loss than past or never smokers whether the subjects had mild, moderate or severe initial attachment loss. Increasing age and smoking status were independently significantly related to mean attachment level and the effect of these parameters was additive. Mean attachment level in non smokers <41 years and current smokers >49 years was 2.49 and 4.10 mm respectively. Stepwise multiple linear regression indicated that age, pack years and being a current smoker were strongly associated with mean attachment level. Full mouth attachment level profiles indicated that smokers had more attachment loss than never smokers particularly at maxillary lingual sites and at lower anterior teeth. CONCLUSIONS: In accord with other studies, smokers had evidence of more severe periodontal disease than past or never smokers. At all levels of mean attachment loss, smokers exhibited more disease than never smokers. Difference in mean attachment level between smokers and never smokers at individual sites was not uniform. Significantly more loss was observed at maxillary lingual sites and lower anterior teeth suggesting the possibility of a local effect of cigarette smoking.

Adult↗

PubMed2001

Relationship of cigarette smoking to the subgingival microbiota.

BACKGROUND: The relationship of cigarette smoking to the composition of the subgingival microbiota is not clear. Some studies indicated higher levels of certain species in smokers, while other studies failed to detect differences in the microbiota between subjects with different smoking histories. Thus, the purpose of the present investigation was to examine the prevalence, proportions and levels of the subgingival species in adult subjects who were current, past or never smokers. METHOD: 272 adult subjects ranging in age from 20-86 years with at least 20 teeth were recruited for study. Smoking history was obtained using a questionnaire. Clinical measures were taken at 6 sites per tooth at all teeth excluding third molars at a baseline visit. Subgingival plaque samples were taken from the mesial surface of all teeth excluding third molars in each subject at baseline and assayed individually for counts of 29 subgingival species using checkerboard DNA-DNA hybridization. Subjects were subset according to smoking history into never (n=124), past (n=98) and current smokers (n=50). Uni-variate and multi-variate analyses were used to seek associations between smoking category and the counts, proportions and prevalence of subgingival species. RESULTS: Greater differences were observed for the prevalence (% of sites colonized) of the test species in the 3 smoking groups than were observed for counts or proportions of total counts. Members of the orange and red complexes including E. nodatum, F. nucleatum ss vincentii, P. intermedia, P. micros, P. nigrescens, B. forsythus, P. gingivalis and T. denticola were significantly more prevalent in current smokers than in the other 2 groups. The difference in prevalence between smokers and non-smokers was due to greater colonization at sites with pocket depth <4 mm. Stepwise multiple linear regression analysis indicated that combinations of the prevalence of 5 microbial species and pack years accounted for 44% of the variance for mean pocket depth (p<0.000001), while the prevalence of 3 microbial taxa along with age, pack years, current smoking and gender accounted for 31% of the variance in mean attachment level (p<0.000001). The difference in prevalence between current and never smokers of all members of the red complex and 8 of 12 members of the orange complex was significantly greater in the maxilla than in the mandible. CONCLUSIONS: The major difference between the subgingival microbiota in subjects with different smoking history was in the prevalence of species rather than counts or proportions. The greater extent of colonization in smokers appeared to be due to greater colonization at pocket depths <4 mm. Differences in colonization patterns between current and never smokers were greater in the maxilla than in the mandible.

Adult↗

PubMed2001

Prevalence and risk indicators for destructive periodontal diseases in 3 urban American minority populations.

BACKGROUND, AIMS: Destructive periodontal diseases have been reported disproportionately more prevalent and severe in African-Americans relative to other American populations. Differences in subgingival microbiota and host immune response have also been reported for African-Americans, implying that risk factors for disease progression may also differ for these populations. Since it is not clear whether these differences are truly genetic or due to confounding variables such as social economic status, we examined a series of clinical, environmental, demographic, and microbiologic features associated with periodontal disease status in a group of 185 urban minority subjects resident within the greater New York metropolitan area. METHODS: The study population consisted of 56 Asian-American, 71 African-American and 58 Hispanic subjects. Clinical data recorded included pocket depth, attachment level, gingival erythema, bleeding upon probing, suppuration, and the presence of supragingival plaque. Environmental and demographic data recorded included smoking history, years resident in the United States, whether the subject reported a private dentist and occupational status. Subgingival plaque was sampled from the mesial aspect of all teeth exclusive of third molars and the levels of 40 subgingival species enumerated using checkerboard DNA-DNA hybridization. RESULTS: The African-American group had more missing teeth, deeper periodontal pocket depth and more attachment loss than the Asian-American or Hispanic groups. However, the African-American group were less likely to report having a private dentist, had a greater proportion of smokers and a greater proportion of unskilled individuals. The profile of subgingival species differed among the three ethnic/racial groups with A. actinomycetemcomitans, N. mucosa, S. noxia and T. socranskii significantly elevated in the Asian-American group and P. micros significantly elevated in the African-American group. When subset by occupational status, numbers of missing teeth, pocket depth, attachment level and prior disease activity were all found increased in the unskilled relative to the professional group. Local factors including the mean % of sites with plaque, marginal gingival erythema, bleeding upon probing and suppuration were also elevated in the unskilled group. The microbial profile differed among the 3 occupational groups with the unskilled group having elevated numbers of species associated with destructive periodontal diseases. CONCLUSIONS: Although greater destructive periodontal disease prevalence and severity were found in the African-American group, these results suggest that environmental and demographic variables, such as occupational status, may have a greater influence on risk indicators associated with disease prevalence and progression in these populations.

Adult↗

PubMed2001

Change in subgingival microbial profiles in adult periodontitis subjects receiving either systemically-administered amoxicillin or metronidazole.

AIM: The current investigation evaluated changes in levels and proportions of 40 bacterial species in subgingival plaque samples during, immediately after and up to 1 year after metronidazole or amoxicillin therapy combined with SRP. METHOD: After baseline clinical and microbiological monitoring, 17 adult periodontitis subjects received full mouth SRP and 14 days systemic administration of either metronidazole (250 mg, TID, n=8) or amoxicillin (500 mg, TID, n=9). Clinical measurements including % of sites with plaque, gingival redness, bleeding on probing and suppuration, pocket depth (PD) and attachment level (AL) were made at baseline, 90, 180 and 360 days. Subgingival plaque samples were taken from the mesial surface of all teeth in each subject at baseline, 90, 180 and 360 days and from 2 randomly selected posterior teeth at 3, 7, and 14 days during and after antibiotic administration. Counts of 40 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Quade test and between groups using ANCOVA. RESULTS: Mean PD was reduced from 3.22+/-0.12 at baseline to 2.81+/-0.16 (p<0.01) at 360 days and from 3.38+/-0.23 mm to 2.80+/-0.14 mm (p<0.01) in the amoxicillin and metronidazole treated subjects respectively. Corresponding values for mean AL were 3.21+/-0.30 to 2.76+/-0.32 (p<0.05) and 3.23+/-0.28 mm to 2.94+/-0.23 mm (p<0.01). Levels and proportions of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were markedly reduced during antibiotic administration and were lower than baseline levels at 360 days. Counts (x10(5), +/-SEM) of B. forsythus fell from baseline levels of 0.66+/-0.16 to 0.04+/-0.02, 0.13+/-0.04, 0.10+/-0.03 and 0.42+/-0.19 in the amoxicillin group at 14, 90, 180 and 360 days respectively (p<0.001). Corresponding values for metronidazole treated subjects were: 1.69+/-0.28 to 0.02+/-0.01, 0.20+/-0.08, 0.22+/-0.06 and 0.22+/-0.08 (p<0.001). Counts of Campylobacter species, Eubacterium nodatum, Fusobacterium nucleatum subspecies, F. periodonticum and Prevotella nigrescens were also detected at lower mean levels during and immediately after therapy, but gradually increased after withdrawal of the antibiotics. Members of the genera Actinomyces, Streptococcus and Capnocytophaga were minimally affected by metronidazole. However, amoxicillin decreased the counts and proportions of Actinomyces species during and after therapy. CONCLUSIONS: The data suggest that metronidazole and amoxicillin are useful in rapidly lowering counts of putative periodontal pathogens, but must be accompanied by other procedures to bring about periodontal stability.

Actinomyces↗