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Background Clinical research has shown a relationship between microbial accumulations and oral diseases such as gingivitis and caries. The mouth harbors large densities of bacteria in distinct oral microenvironments, that is, dental plaque on teeth, saliva, and soft tissues such as the tongue, cheek, and gingiva. In this home-use study, the authors compared the effects of brushing with a newly formulated stannous fluoride toothpaste and a sodium monofluorophosphate dentifrice on bacteria of distinct oral microenvironments. Methods Adult participants completed a washout phase before baseline sampling of dental plaque, saliva, and scrapings from tongue, cheek, and gingiva, which were used for microbiological analysis. Treatments were randomly assigned: test (62 participants) and control (67 participants) for twice-daily toothbrushing over 8 weeks. After 4 weeks and 8 weeks, posttreatment samples 12 hours after brushing were collected for analysis. At these posttreatment visits, participants brushed in the dental clinic, and an additional sample was collected 4 hours later for microbiological analysis. Results Relative to the control, 12 hours after brushing, the test toothpaste showed greater reductions of bacteria (ranging from 14% to 27%) at the 4-week evaluation, which increased to 27% to 41% after 8 weeks of brushing. Correspondingly, 4 hours after brushing with the test toothpaste at the 4-week evaluation, there were greater reductions of bacteria in the range of 22% to 59%, which increased to a range of 33% to 61% at the 8-week assessment for participants completing the study. Conclusions The stannous fluoride toothpaste provided bacterial reductions in all oral microenvironments 12 hours after brushing. In addition, the authors observed microbial reductions 4 hours after brushing, which increased after extended use. Practical Implications The results are appropriate for oral hygiene recommendations by dental professionals.

The purpose of this study was to compare color alterations (ΔE) of white-spot lesions (WSLs) bleached before versus after resin infiltration (RI).Using the facial surfaces of bovine maxillary incisors, WSLs were created and the teeth were allocated into 2 groups (n = 45/group): bleach then RI (B-RI group) and RI then bleach (RI-B group). To determine ΔE, Commission Internationale de l'Eclairage L∗ a∗ b∗ (L∗ represents lightness, ranging from black to white [0-100]; a∗ represents green to red chromaticity [-150-+100]; and b∗ represents blue to yellow chromaticity [-100-+150]) measurements were obtained at baseline, after WSL formation, and after RI and bleaching. Representative specimens were evaluated by means of scanning electron microscopy. Statistical analyses included the Mann-Whitney U and Wilcoxon signed rank tests (P ≤ .0016) and repeated measures analysis of variance (P ≤ .05).No differences in ΔE were found comparing B-RI with RI-B groups or when the B-RI group was compared with bleached enamel. A statistically significant difference was found when the RI-B group was compared with bleached enamel (ΔE, 0.81; P.001), but the difference was deemed not clinically significant. Scanning electron microscopy revealed that bleaching after RI increased surface roughness of the resin.There were no clinically significant differences in ΔE of WSLs when bleach was applied before or after RI; however, applying bleaching agent after RI roughened the surface of the resin material.Results indicate that ΔE were not clinically significantly different between WSLs bleached before versus after RI, although it is best to sequence bleaching before RI therapy, as bleaching after RI roughened the restoration's surface.

The purpose of this study was to compare biofilm formation on materials used for the fabrication of implant-supported dental prostheses. Twenty discs (D=15 mm, H=3 mm) were fabricated from one of the following restorative materials: yttria tetragonal zirconia polycrystal (Y-TZP); commercially pure titanium (CP-Ti); or heat-cured polymethyl methacrylate (PMMA). Specimens were polished following standard protocols. A non-contact profilometer (NPFLEX, Bruker, UK) was used to assess the surface roughness of each disk; results were reported as Ra (µm). Five strains of Gram-negative bacteria frequently associated with peri-implantitis, Aggregatibacter actinomycetemcomitans, Candida. albicans, Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia, were cultured on hand-polished discs fabricated from heat-cured PMMA, Y-TZP, or CP-Ti to compare biofilm formation on each type of material. The results were reported as colony-forming units per milliliter (CFU/mL). One-way ANOVA and post hoc tests were used to compare surface roughness and bacterial colonization on the respective materials. Statistical significance was set at a = 0.05. Discs fabricated from Y-TZP had a significantly higher Ra value (350 ± 30 µm) than either PMMA, or CP-Ti discs. Discs fabricated from either Y-TZP and CP-Ti may exhibit less colonization by bacteria associated with peri-mucositis and peri-implantitis. Y-TZP and CP-Ti are suggested materials for fabrication of implant-supported prostheses, considering biofilm formation. Resumo O objetivo deste estudo foi comparar a formação de biofilme em materiais utilizados na confecção de próteses dentárias implantossuportadas. Vinte discos (D = 15 mm, H = 3 mm) foram confeccionados com um dos seguintes materiais restauradores: zircônia tetragonal policristalina estabilizada por ítrio (Y-TZP); titânio comercialmente puro (CP-Ti); ou polimetilmetacrilato (PMMA). As amostras foram polidas seguindo protocolos padrão. Um perfilômetro sem contato (NPFLEX, Bruker, UK) foi usado para avaliar a rugosidade da superfície de cada disco; os resultados foram relatados como Ra (µm). Cinco cepas de bactérias Gram-negativas freqüentemente associadas a peri-implantite, Aggregatibacter actinomycetemcomitans, Candida. albicans, Porphyromonas gingivalis, Prevotella intermedia e Tannerella forsythia, foram cultivadas em discos polidos à mão feitos de PMMA, Y-TZP ou CP-Ti para comparar a formação de biofilme em cada tipo de material. Os resultados foram relatados como unidades formadoras de colônias por mililitro (UFC/mL). Análise de variância a um fator e testes post hoc foram usados para comparar a rugosidade da superfície e a colonização bacteriana nos respectivos materiais. A significância estatística foi estabelecida em a=0,05. Os discos feitos de Y-TZP tiveram um valor Ra significativamente mais alto (350 ± 30 µm) do que os discos de PMMA ou CP-Ti. Os discos fabricados com Y-TZP e CP-Ti podem apresentar menor colonização por bactérias associadas à perimucosite e peri-implantite. Considerando O Y-TZP e CP-Ti são materiais indicados para a confecção de próteses implantossuportadas, considerando a formação de biofilme

The aim of this study was to evaluate the impact of a contemporary preclinical technique skills course on early dental student performance in a preclinical operative course. Ivorine teeth prepared for Class I resin restorations by the Classes of 2020 and 2021 in one U.S. dental school were evaluated in this study (N=184). Ivorine teeth were prepared during the first practical exam of the operative dentistry course. Students in the Class of 2021 had been enrolled in a contemporary technique skills course a semester prior to the operative dentistry course, while students in the Class of 2020 were not. The preparations were randomly evaluated by three calibrated and blinded faculty members using magnification loupes, an explorer, and a periodontal probe to evaluate external outline form, internal form, depth, and margin following an established rubric. The results showed that students in the Class of 2021 (who had taken the technique skills course) performed significantly better than students in the Class of 2020 for all criteria evaluated (p

Recent studies of the dental plaque bacteria associated with the various forms of early-onset periodontitis confirm the importance of target periodontal pathogens such as Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Prevotella intermedia, and Porphyromonas gingivalis in these diseases. A. actinomycetemcomitans strains exhibit a wide range of variability in leukotoxin production. By virtue of a unique promoter for the leukotoxin (ltx) operon, highly leukotoxic A. actinomycetemcomitans strains (e.g., JP2) express 10- to 20-times greater levels of leukotoxin than minimally toxic strains (e.g., 652). In dot blot hybridization and polymerase chain reaction (PCR) assays, the distribution of leukotoxic A. actinomycetemcomitans was examined among 165 fresh isolates and strains from our culture collection obtained from 91 human patients and non-human primates. Highly leukotoxic A. actinomycetemcomitans strains were found in 22% of the subjects and represented 28% of the isolates examined. This is a much higher prevalence than reported in a similar survey of A. actinomycetemcomitans strains from Northern Europe. Patients harboring the highly leukotoxic strains were much younger (mean age 12.7 years) than those harboring minimally toxic A. actinomycetemcomitans (mean age 25.5 years). In addition, patients with localized juvenile periodontitis were shown to have a substantially higher prevalence of highly leukotoxic strains than healthy individuals or those with adult periodontitis. Fifty-seven percent of the localized juvenile periodontitis patients harbored these strains and 64% of the isolates obtained from these patients were highly toxic A. actinomycetemcomitans. No highly toxic strains were identified from healthy individuals or from patients with adult periodontitis. The polymerase chain reaction assay could readily identify and distinguish the ltx promoters from highly toxic and minimally toxic A. actinomycetemcomitans in whole plaque samples. These data point to the importance of specific A. actinomycetemcomitans strains, as characterized by their expression of high levels of leukotoxin, in the pathogenesis of certain types of early-onset periodontitis and, possibly, other forms of rapidly progressing periodontitis. J Periodontol 1996;67:282-290.

Aim: Oral malodor (halitosis) is a widespread condition caused by oral bacteria, particularly sulfur compound-producing species. This study assessed the effect of a triclosan/copolymer-containing dentifrice and a novel toothbrush with attached tongue cleaner on oral malodor and on the bacteria colonizing the dorsal surface of the tongue. Materials and Methods: 14 adult subjects with oral malodor defined as organoleptic scores ≥ 3 (scale 0 - 5) and mouth air sulfur levels ≥ 250 ppb participated in this study. Subjects were examined at baseline and after 28 days use of the triclosan dentifrice and toothbrush/tongue cleaner for: 1) organoleptic assessment; 2) mouth air sulfur levels; 3) tongue coating; and, 4) dorsal tongue surface microorganisms. Total bacterial numbers were assayed by microscopy. Dot-blot hybridization was used to assess a panel of 20 oral bacteria. Results: After 28 days, all subjects had significantly reduced organoleptic scores and mouth air sulfur levels compared to baseline (p 70% reduction in microbial numbers (p Enterococcus faecalis (p Peptostreptococcus micros (p Prevotella melaninogenica (p Porphyromonas gingivalis (p Solobacterium moorei (p

Aim: To test the hypothesis that antimicrobial agents can be incorporated into bis-acryl resin and that the material will inhibit bacterial growth in vitro. Materials and Methods: 0.5%, 1% and 2% triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol) and 0.05% 0.01% and 0.15% cetylpyridinium chloride were incorporated into bis-acryl resins and molded into 1 mm × 10 mm discs. The discs were placed on bacterial culture media inoculated with Aggregatibacter actinomycetemcomitans strain Y4, Porphyromonas gingivalis strain SUNYaB A7A1-28 or Streptococcus gordonii strain 10558, incubated and examined for growth inhibition. Results: Triclosan-containing bisacryl resin discs produced dose-dependent zones of growth inhibition for A. actinomycetemcomitans but not S. gordonii or P. gingivalis. Cetylpyridinium chloride-containing discs produced dose-dependent zones of growth inhibition for Porphyromonas gingivalis and Streptococcus gordonii but not A. actinomycetemcomitans. Conclusion: Triclosan and cetylpyridinium chloride can be incorporated into bis-acryl resin and can inhibit the growth of plaque bacteria in a dose dependent manner. Bis-acryl resin containing antimicrobial agents may inhibit in vivo plaque accumulation and gingival inflammation.

We report the genome sequences of three clinical isolates of Tannerella forsythia from the subgingival plaque of periodontitis patients attending clinics at the School of Dental Medicine, University at Buffalo. The availability of these genome sequences will aid the understanding of the pathogenesis of periodontitis.

This study evaluated the antimicrobial activity of two commercially available 0·05% cetylpyridinium chloride (CPC) mouthrinses with or without alcohol and examined its antimicrobial activity on oral bacterial species including fresh clinical isolates compared to a chlorhexidine mouthrinse and a control fluoride mouthrinse without CPC. Two different approaches were used to evaluate antimicrobial activity. First, the minimum inhibitory concentration (MIC) was determined for each mouthrinse against a panel of 25 micro-organisms including species associated with dental caries, gingivitis and periodontitis. Second, supragingival dental plaque obtained from 15 adults was incubated with the four mouthrinses to evaluate antimicrobial activity on micro-organisms in oral biofilms. Both CPC mouthrinses exhibited lower MIC's, that is, greater antimicrobial activity, against oral Gram-negative bacteria especially periodontal pathogens and species implicated in halitosis such as Aggregatibacter actinomycemcomitans, Campylobacter rectus, Eikenella corrodens, Porphyromonas gingivalis, Prevotella intermedia and Solobacterium moorei than the control mouthrinse. Ex-vivo tests on supragingival plaque micro-organisms demonstrated significantly greater antimicrobial activity by the CPC mouthrinses (>90% killing, P 98% killing, P

To cite this article: Int J Dent Hygiene9, 2011; 136–142 DOI: 10.1111/j.1601-5037.2010.00465.x Sreenivasan PK, Haraszthy VI, Zambon JJ.The effect of a microbead dentifrice on microbial load in oral microenvironments. Abstract: Objectives: The human oral cavity contains several microenvironments or ecologic niches. While mechanical plaque control is well known to reduce the number of supragingival dental plaque bacteria, there is little data on antimicrobial effects in other oral ecologic niches. The present study examined the effects of mechanical plaque control using a microbead dentifrice on bacteria colonizing oral ecologic niches. Methods: Twenty-two adults (aged 18–70 years) including nine generalized moderate chronic periodontitis subjects and 13 periodontally healthy subjects having average gingival indices ≥1 and plaque indices ≥1.5 completed a 1 week washout phase and refrained from oral hygiene the morning of baseline sample collection. Microbial samples from supragingival dental plaque, buccal mucosa, dorsal surface of the tongue and whole mixed saliva were obtained. Subjects brushed with a microbead dentifrice and, after 10 min, sampling was repeated. The number of anaerobic bacteria was determined by culture on non-selective media and transformed to log10 for statistical analyses. Results: Mechanical plaque control using the microbead dentifrice resulted in statistically significant reductions in bacterial numbers in each ecologic niche (P

Background The authors examined the tongue bacteria associated with oral halitosis (bad breath originating from the oral cavity), focusing on noncultivable bacteria—bacteria that cannot be identified by bacterial culture techniques. Methods The authors took samples from the dorsal tongue surface of eight adult subjects with halitosis and five control subjects who did not have halitosis. They identified the bacteria in these samples by using both anaerobic culture and direct amplification of 16S ribosomal DNA, a method that can identify both cultivable and noncultivable microorganisms. They analyzed the resulting microbiological data using χ 2 and correlation coefficient tests. Results Clinical measures of halitosis were correlated highly with each other and with tongue coating scores. Of 4,088 isolates and phylotypes identified from the 13 subjects, 32 species including 13 noncultivable species were found only in subjects with halitosis. Solobacterium moorei was present in all subjects with halitosis but not in any control subjects. Conclusions Subjects with halitosis harbor some bacterial species on their dorsal tongue surfaces that are distinct from bacterial species found in control subjects. This finding is consistent with the hypothesis that halitosis has a microbial etiology. Clinical Implications Like other oral diseases with microbial etiology, halitosis may be amenable to specific and nonspecific antimicrobial therapy targeted toward the bacteria associated with it.

Aims: To determine minimal inhibitory concentrations (MICs) and the percentage of nonsusceptible bacteria – those still cultivable above a threshold concentration – in human supragingival dental plaque and saliva for antiplaque/antimicrobial agents including triclosan (TCS) and trichlorocarbanilide (TCC), and a new potential antimicrobial, 2-t-butyl-5-(4-t-butylphenyl)-phenol (DTBBP). Methods and Results: Broth and agar dilution-based MIC tests were performed using 28 oral and nonoral bacterial strains representing 17 species. MICs for TCS were lowest and more than 100-fold lower than DTBBP (P

Made available in DSpace on 2019-09-12T16:56:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2005 Objectives: Previous studies suggest differences between geographically and racially distinct populations in the prevalence of periodontopathic bacteria as well as greater periodontal destruction associated with infection by highly leucotoxic Actinobacillus actinomycetemcomitans. The present study examined these hypotheses in Brazilians with aggressive or chronic periodontitis. Materials and Methods: Clinical, radiographical, and microbiological assessments were performed on 25 aggressive periodontitis and 178 chronic periodontitis patients including 71 males and 132 females, 15-69 years of age. Results: The prevalence of Porphyromonas gingivalis was similar to that of other South American populations. The prevalence of A. actinomycetemcomitans and its highly leucotoxic subgroup was higher in Brazilians. Highly leucotoxic A. actinomycetemcomitans was more prevalent in aggressive periodontitis (chi(2) = 527.83) and positively associated with deep pockets (46 mm, chi(2) = 18.26) and young age (< 29 years, chi(2) = 18.68). Greater mean attachment loss was found in subjects with highly leucotoxic A. actinomycetemcomitans than in subjects with minimally leucotoxic (p = 0.0029) or subjects not infected (p = 0.0001). Conclusion: These data support the hypothesis of differences between populations in the prevalence of periodontopathic bacteria and of greater attachment loss in sites infected with highly leucotoxic A. actinomycetemcomitans. Detection of highly leucotoxic A. actinomycetemcomitans in children and adolescents may be a useful marker for aggressive periodontitis. SUNY Buffalo, Sch Dent Med, Buffalo, NY 14214 USA; Universidade de Taubaté (Unitau), Sao Paulo, Brazil

In der vorliegenden Studie wurden von 62 Patienten (18 Frauen und 44 Manner) 93 Proben aus verschiedensten Gefasen und Gefasabschnitten wahrend der Gefasoperation entnommen und auf bakterielle 16S rDNS sowie—mittels spezifischen Oligonukleotidprimern—auf die mit Parodontitis assoziierten Keime P. gingivalis, P. intermedia, A. actinomycetemcomitans, C. rectus und B. forsythus untersucht. In 76,34% der 93 untersuchten Gefaswandpraparate waren Bakterien nachzuweisen, in 40,86% einer oder mehrere der parodontalpathogenen Erreger. Diese kamen so gut wie immer (mit einer Ausnahme) zusammen mit anderen Bakterien vor. C. pneumoniae fanden sich in 22,58% der Proben. Ihr Auftreten stand signifikant mit dem Vorkommen von parodontalpathogenen Keimen in Zusammenhang. Herpes-simplex- und Zytomegalieviren kamen in 24,73% bzw. 30,11% der Proben vor, ohne dass ein Zusammenhang mit Parodontalkeimen bestand. Diese Ergebnisse stehen im Einklang mit der Hypothese, dass Mikroorganismen, die mit Parodontitis assoziiert sind, bei der Atiologie und Pathogenese der Arteriosklerose eine wichtige Rolle spielen.

Introduction Recent studies suggest that chronic infections, including those associated with periodontitis, increase the risk for coronary vascular disease. We hypothesize that oral microorganisms including periodontal bacterial pathogens enter the blood stream during transient bacteremias where they may play a role in the development and progression of atherosclerosis. Materials and methods To test this hypothesis, 34 human specimens obtained during carotid endarterectomy or bypass procedures were examined by use of specific oligonucleotide primers for Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans and Bacteroides forsythus in polymerase chain reaction (PCR) assays. Results Twenty (59%) of the 34 specimens tested positive for bacterial 16S rDNA. Subsequent hybridization of the bacterial 16S rDNA positive specimens with species-specific oligonucleotide probes revealed that 32.4% of the 34 atheromas tested positive for at least one of the target periodontal pathogens. Further analysis of the results in the bacterial positive group (n = 20) shows that 55% of the atheromas tested positive for at least one of the target periodontal pathogens. Conclusion These findings indicate that periodontal pathogens are present in atherosclerotic plaques, where they may play a role in the development and progression of atherosclerosis leading to coronary vascular disease and other clinical sequelae.

Recent studies suggest that chronic infections including those associated with periodontitis increase the risk for coronary vascular disease (CVD) and stroke. We hypothesize that oral microorganisms including periodontal bacterial pathogens enter the blood stream during transient bacteremias where they may play a role in the development and progression of atherosclerosis leading to CVD.To test this hypothesis, 50 human specimens obtained during carotid endarterectomy were examined for the presence of Chlamydia pneumoniae, human cytomegalovirus, and bacterial 16S ribosomal RNA using specific oligonucleotide primers in polymerase chain reaction (PCR) assays. Approximately 100 ng of chromosomal DNA was extracted from each specimen and then amplified using standard conditions (30 cycles of 30 seconds at 95 degrees C, 30 seconds at 55 degrees C, and 30 seconds at 72 degrees C). Bacterial 16S rDNA was amplified using 2 synthetic oligonucleotide primers specific for eubacteria. The PCR product generated with the eubacterial primers was transferred to a charged nylon membrane and probed with digoxigenin-labeled synthetic oligonucleotides specific for Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, and Prevotella intermedia.Eighty percent of the 50 endarterectomy specimens were positive in 1 or more of the PCR assays. Thirty-eight percent were positive for HCMV and 18% percent were positive for C. pneumoniae. PCR assays for bacterial 16S rDNA also indicated the presence of bacteria in 72% of the surgical specimens. Subsequent hybridization of the bacterial 16S rDNA positive specimens with species-specific oligonucleotide probes revealed that 44% of the 50 atheromas were positive for at least one of the target periodontal pathogens. Thirty percent of the surgical specimens were positive for B. forsythus, 26% were positive for P. gingivalis, 18% were positive for A. actinomycetemcomitans, and 14% were positive for P. intermedia. In the surgical specimens positive for periodontal pathogens, more than 1 species was most often detected. Thirteen (59%) of the 22 periodontal pathogen-positive surgical specimens were positive for 2 or more of the target species.Periodontal pathogens are present in atherosclerotic plaques where, like other infectious microorganisms such as C. pneumoniae, they may play a role in the development and progression of atherosclerosis leading to coronary vascular disease and other clinical sequelae.

Dental implants and peri-implant tissue are susceptible to disease conditions that may lead to implant loss. The objective of the present study was to describe teeth and implant surroundings as well as clinical health indices and oral microbiologic parameters.A group of 83 adults (42 men and 41 women) were enrolled in the study. Clinical assessments of dental implants and contralateral natural teeth included dental plaque, gingival inflammation, and bleeding on probing. Microbiologic assessments included bacterial culture, light and phase contrast microscopy, and DNA probe hybridization for a panel of 14 target microorganisms. Clinical and microbiologic data were compared by paired t test and ANOVA. P.05 was considered statistically significant.The Plaque Index for the implants was 1.85 ± 0.47, whereas the score for natural teeth was significantly higher, 2.15 ± 0.52. Compared to the samples obtained from the dental implants, the samples from natural teeth demonstrated significantly higher total bacterial cell numbers (P.05). Consistent with the clinical measures of dental plaque, significantly higher numbers of oral bacteria, including aerobic and anaerobic bacteria, were found in dental plaque samples from teeth (aerobic 5.648 ± 0.512, anaerobic 6.243 ± 0.535, P.0001) compared to implants (aerobic 5.430 ± 0.541, anaerobic 5.917 ± 0.523, P.0001). In addition, there were significantly higher numbers of anaerobic (6.243 ± 0.535 and 5.917 ± 0.523, P.0001) than aerobic (5.648 ± 0.512 and 5.430 ± 0.541, P.008) bacteria for samples from teeth and implants, respectively.Clinical and microbiologic analyses provide consistent findings that suggest differences in quantity of plaque and bacterial species between teeth and dental implants. For long-term treatment success, the importance of plaque control and oral hygiene of both periodontal and dental implant therapy is emphasized.

Previous studies suggest that Solobacterium moorei is associated with oral halitosis. In the present study, we examined the prevalence of S. moorei on the dorsal surface of the tongue in 57 adults (21 with and 36 without halitosis) by bacterial culture and direct amplification of nucleic acids. We also examined the S. moorei type strain and four clinical isolates for 16S ribosomal nucleic acid sequence, H(2)S and enzyme production, and antibiotic susceptibility. S. moorei was found on the dorsal surface of the tongue in 100% of the subjects with halitosis and 14% of subjects without halitosis. Infection with S. moorei was correlated with organoleptic measures of halitosis and with volatile sulfur compound levels. Nucleic acid probe detection of S. moorei as a test for halitosis exhibited 100% sensitivity and 86% specificity. The S. moorei type strain and all four clinical isolates showed98% 16S rDNA sequence similarity, produced H(2)S, demonstrated acid phosphatase, beta-galactosidase, alpha-glucosidase, esterase, leucine arylamidase and naphthol phosphohydrolase enzyme activities, and were sensitive to all antibiotics tested except gentamicin, kanamycin, nalidixic acid and rifampin. This study supports the hypothesis that S. moorei is associated with halitosis.

This case-controlled study examined clinical and microbiological parameters in Brazilian children and adolescents receiving orthodontic treatment using fixed orthodontic appliances or removable orthodontic appliances.The plaque index, gingival index, number of decayed, missing and filled teeth, and probing pocket depth was measured on each fully erupted tooth in 30 patients treated with fixed orthodontic appliances and an equal number of age and sex-matched control subjects. The same parameters were also measured in 18 patients treated with removable orthodontic appliances and an equal number of age and sex-matched control subjects. In the patients treated with fixed orthodontic appliances, subgingival plaque samples were collected from four teeth with orthodontic brackets and from four teeth with orthodontic bands. In the patients with removable appliances, subgingival plaque samples were collected from clasped maxillary permanent first molar teeth and from four unclasped permanent teeth. Samples of unstimulated whole saliva and samples from the dorsal surface of the tongue were also obtained from each subject. Each sample was analyzed for the presence of 19 target bacteria by dot blot. A subset of samples was examined by direct amplification of bacterial nucleic acids.Compared to their respective age and sex-matched controls, whole mouth means for plaque index and gingival index were significantly elevated in both the fixed and removable orthodontic groups. There was no difference in the DMFT. Subjects with fixed orthodontic appliances had a higher prevalence of each of the target species except for L. fermentum, Neisseriaceae and S. mutans. The prevalence of A. naeslundii and Streptococcus sp. was significantly higher on teeth with orthodontic brackets alone compared to teeth with both orthodontic bands and brackets. Subjects with removable orthodontic appliances had a higher prevalence of A. actinomycetemcomitans, C. rectus, E. corrodens, L. fermentum, Neisseriaceae, and spirochetes. The prevalence of Neisseriaceae was significantly higher on unclasped teeth compared to clasped teeth. There was no difference between sample sites for the target bacteria except for A. actinomycetemcomitans that was detected less frequently in saliva. Orthodontic patients demonstrated higher proportions of gram negative species by direct amplification of nucleic acids including species frequently associated with periodontal disease as well as rarely cultivable or non-cultivable species such as Abiotrophia defectiva, Gemella haemolysans, Granulicatella adiacens, Lautropia sp., Terrahaemophilus aromaticivorans, and TM7 bacterium.

In vitro testing of antimicrobial agents is an important tool in the testing hierarchy, and may provide interesting insights into their potential clinical efficacy. Agents with demonstrable in vitro antimicrobial activity may be effective against the same microorganisms in vivo, whereas agents without demonstrable in vitro antimicrobial activity are unlikely to exhibit in vivo antimicrobial activity. In addition, these methods may also be useful in screening antimicrobial agents in product formulations because such agents with both in vitro and in vivo activity may have reduced antimicrobial effects when formulated into a dentifrice. Accordingly, this study examined the in vitro and ex vivo antimicrobial activity of three commercial dentifrices: one formulated with 0.243% sodium fluoride (Crest Cavity Protection Toothpaste-Regular); one with 0.454% stannous fluoride, sodium hexametaphosphate, and zinc lactate (Crest Pro-Health), and one with 0.3% triclosan, 2.0% PVM/MA copolymer, and 0.243% sodium fluoride (Colgate Total).The minimum inhibitory concentration (MIC) of each dentifrice was determined for resident oral bacterial species, including bacteria that are associated with dental caries; periodontitis, and oral halitosis. Evaluations were performed on individual laboratory strains, and on oral bacteria from supragingival plaque samples obtained from 10 adults and from oral rinse samples obtained from 18 adults.The lowest MICs against the oral strains and human samples, i.e., greatest antimicrobial activity, were seen for the triclosan/ copolymer dentifrice. There was, in general, a four-fold difference in MICs between the triclosan/copolymer dentifrice and the stannous fluoride/sodium hexametaphosphate/zinc lactate dentifrice. The triclosan/copolymer dentifrice significantly inhibited periodontal pathogens, such as Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum. In ex vivo tests measuring antimicrobial effects, the triclosan/copolymer dentifrice substantially inhibited bacterial growth after 30-, 60-, and 120-second exposures compared to the sodium fluoride or stannous fluoride/sodium hexametaphosphate/zinc lactate dentifrices. Similarly, in ex vivo tests measuring antimicrobial effects on supragingival plaque biofilms, the triclosan/copolymer dentifrice substantially inhibited bacterial growth compared to the other test dentifrices.Different in vitro and ex vivo analyses show that the triclosan/copolymer dentifrice has significant antimicrobial activity on oral bacteria, including species causing dental caries, periodontitis, and oral halitosis, and it provides superior efficacy compared to the stannous fluoride/sodium hexametaphosphate/zinc lactate dentifrice.

Supportive therapy to maintain dental implants is increasingly important. This study examined the effect of a 0.3% triclosan/2% copolymer dentifrice on oral biofilms and gingival inflammation (GI) on dental implants and peri-implant tissues.One hundred and twenty adults with a dental implant and contra-lateral tooth were enrolled in this 6 month, double-blind, two-treatment, parallel group study. Sixty subjects were randomly assigned to a triclosan/copolymer dentifrice test group and 60 subjects to a fluoride dentifrice control group and instructed to brush twice daily for 6 months. At baseline, 3, and 6 months, a calibrated dentist assessed dental plaque, GI and collected supragingival dental plaque for microbiological analysis.Subjects in the triclosan/copolymer group demonstrated significantly lower levels of dental plaque, gingivitis, and bleeding on probing at 3 and 6 months at both the implant and contra-lateral tooth compared with the fluoride group (p0.05). There were significantly fewer Gram-negative anaerobes in the triclosan/copolymer group (p0.05) including90% reductions in Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Eubacterium saburreum, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella melaninogenica, Solobacterium moorei, and Tannerella forsythia.Twice daily use of a triclosan/copolymer dentifrice may enhance dental implant maintenance by reducing dental plaque and GI.

This investigation compared the effects of a fluoride dentifrice and toothpastes formulated with antimicrobial ingredients (stannous fluoride and triclosan/copolymer) on oral micro-organisms, including those found in samples taken from the human oral cavity. Microbiological techniques determined the minimum inhibitory concentrations (MICs) of each dentifrice necessary to inhibit the growth of bacterial strains from the healthy oral cavity, as well as those found in dental caries, periodontal disease, and halitosis. Ex vivo studies utilized oral rinse samples and supragingival plaque from adults to determine antimicrobial effects on the entire microbial diversity of these samples, including biofilm-derived micro-organisms. The triclosan/copolymer dentifrice demonstrated the lowest MICs and significantly inhibited Gram-positive and Gram-negative bacteria (including the periodontal pathogens Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum). In the ex vivo tests, the triclosan/copolymer dentifrice demonstrated substantial inhibition in the oral rinse samples over each treatment period (p0.0005) as compared to either the fluoride or stannous fluoride dentifrices. Similarly, the triclosan/copolymer dentifrice demonstrated the highest inhibition of micro-organisms in the supragingival plaque biofilm (p0.0005). No significant differences were observed between the fluoride and stannous fluoride dentifrices (p0.5).

The aim of the present study is to evaluate the use of anorganic bovine bone (ABB) associated with a collagen membrane (CM) for a sinus graft by means of clinical, histologic, and radiographic parameters in cases with bone availabilityor =7 mm. A preliminary evaluation consisted of a clinical examination, computed tomography (CT), and a panoramic x-ray.Ninety-two patients requiring bilateral sinus grafts and 222 requiring unilateral procedures (total: 406 sinuses) participated in this study. A total of 1,025 implants were placed in the grafted sinuses. A total of 118 implants were placed simultaneously with the sinus graft (one stage), and 907 implants were placed in a subsequent surgery (two stages), 6 to 12 months after the graft was performed. In seven cases, a biopsy was harvested for histomorphometric analysis. Recall appointments were scheduled every 6 months, and panoramic and periapical x-rays were required every year for 3 years.Among 1,025 implants, 19 were lost (survival rate: 98.1%). The difference in survival rates for implants placed in native boneor =3 mm (98.1%),3 toor =5 mm (98.6%), and5 toor =7 mm (97.0%) was not statistically significant (P = 0.3408). The survival rates for implants with rough and machined surfaces (98.6% and 97.0%, respectively) were not statistically significant (P = 0.0840). The histomorphometric analysis showed new bone formation (39.0% +/- 12%), marrow space (52.9% +/- 9.3%), and residual ABB (8% +/- 2.7%).Our results indicated that 1,025 implants placed in sinuses grafted exclusively with ABB combined with CM led to an excellent and predictable survival rate of 98.1%.

Actinobacillus actinomycetemcomitans is an oral pathogen that causes aggressive periodontitis as well as sometimes life-threatening, extra-oral infections. Iron regulation is thought to be important in the pathogenesis of A. actinomycetemcomitans infections and, consistent with this hypothesis, the fur gene has recently been identified and characterized in A. actinomycetemcomitans. In this study, 14 putatively Fur-regulated genes were identified by Fur titration assay (Furta) in A. actinomycetemcomitans, including afuA, dgt, eno, hemA, tbpA, recO and yfe – some of which are known to be Fur regulated in other species. A fur mutant A. actinomycetemcomitans strain was created by selecting for manganese resistance in order to study the Fur regulon. Comparisons between the fur gene sequences revealed that nucleotide 66 changed from C in the wild-type to T in the mutant strain, changing leucine to isoleucine. The fur mutant strain expressed a nonfunctional Fur protein as determined by Escherichia coli-based ferric uptake assays and Western blotting. It was also more sensitive to acid stress and expressed higher levels of minC than the wild-type strain. minC, which inhibits cell division in other bacterial species and whose regulation by iron has not been previously described, was found to be Fur regulated in A. actinomycetemcomitans by Furta, by gel shift assays, and by RT-qPCR assays for gene expression.

Made available in DSpace on 2021-07-14T10:48:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2003-06. Added 1 bitstream(s) on 2021-07-14T11:38:43Z : No. of bitstreams: 1 S1517-74912003000200016.pdf: 129290 bytes, checksum: 46a4a89815dea079ef03f222f2c12885 (MD5) O presente estudo avaliou a prevalência de Actinobacillus actinomycetemcomitans de máxima e mínima leucotoxicidade em indivíduos com doença periodontal. Foram analisadas amostras de placa bacteriana subgengival de 136 indivíduos, entre 14 e 76 anos de idade, com algum tipo de doença periodontal. Os parâmetros clínicos avaliados incluíram profundidade de sondagem (PS), índice de placa bacteriana (IPB) e índice de sangramento gengival (ISG). A presença de amostras de A. actinomycetemcomitans de máxima e mínima leucotoxicidade foi avaliada por reação em cadeia da polimerase (PCR). Os resultados foram analisados empregando-se teste qui-quadrado e análise de regressão logística. Quarenta e sete indivíduos apresentaram gengivite, 70 indivíduos periodontite crônica e 19 indivíduos periodontite agressiva. O teste qui-quadrado não demonstrou correlação significativa entre PS (chi2 = 0,73), IPB (chi2 = 0,35), ISG (chi2 = 0,09) e a presença de amostras de máxima leucotoxicidade. A. actinomycetemcomitans de máxima leucotoxicidade apresentou associação com indivíduos com idade inferior a 28 anos (chi2 = 22,06). Os resultados observados nessa população brasileira confirmam a forte associação existente entre amostras de A. actinomycetemcomitans de máxima leucotoxicidade e periodontite agressiva. This study examined the prevalence of highly and minimally leukotoxic Actinobacillus actinomycetemcomitans in patients with periodontal disease. Pooled subgingival plaque samples from 136 patients with some form of periodontal disease were examined. Subjects were between 14 and 76 years of age. Clinical examinations included periodontal pocket depth (PD), plaque index (PI) and bleeding index (BI). The obtained plaque samples were examined for the presence of highly or minimally leukotoxic A. actinomycetemcomitans strains by the polymerase chain reaction (PCR). Chi-square and logistic regression were performed to evaluate the results. Forty-seven subjects were diagnosed with gingivitis, 70 with chronic periodontitis and 19 with aggressive periodontitis. According to chi-square there was no significant correlation detected between PD (chi2 = 0.73), PI (chi2 = 0.35), BI (chi2 = 0.09) and the presence of the highly leukotoxic A. actinomycetemcomitans. The highly leukotoxic A. actinomycetemcomitans strains were correlated with subjects that were 28 years of age and younger (chi2 = 7.41). There was a significant correlation between highly leukotoxic A. actinomycetemcomitans and aggressive periodontitis (chi2 = 22.06). This study of a Brazilian cohort confirms the strong association between highly leukotoxic A. actinomycetemcomitans strains and the presence of aggressive periodontitis. State University of New York, School of Dental Medicine Universidade Estadual Paulista University of Taubaté Universidade Estadual Paulista

In several bacterial species, iron availability in host tissues is coordinated with the expression of virulence determinants through the fur gene product. Initial experiments showed that a cloned Escherichia coli fur gene probe hybridized to Southern blots of Actinobacillus actinomycetemcomitans strain JP2 (serotype b) chromosomal DNA. The A. actinomycetemcomitans fur gene was then cloned utilizing partial functional complementation of the fur mutant in E. coli strain H1780. Analysis of the cloned DNA sequence revealed a 438-bp open reading frame with a deduced 146-amino-acid sequence exhibiting 80% identity to Haemophilus influenzae Fur and 62% identity to E. coli Fur. The pUC Aafur gene probe (generated from JP2 serotype b) hybridized to representatives from all five A. actinomycetemcomitans serotypes as well as to two strains derived from monkeys, suggesting that fur is widely distributed in A. actinomycetemcomitans. Open reading frames having >70% identity with the E. coli and H. influenzae flavodoxin and gyrase A genes, respectively, were found. Expression of the A. actinomycetemcomitans fur gene product repressed fiu expression and siderophore production in E. coli . A gel shift assay demonstrated that the expressed A. actinomycetemcomitans Fur protein bound the bacterial fur consensus sequence. Further characterization of the fur gene product in A. actinomycetemcomitans may improve our understanding of its role in the pathogenesis of periodontal disease and may lead to specific therapeutic modalities.

Actinobacillus actinomycetemcomitans leukotoxin is thought to be an important virulence factor in the pathogenesis of localized juvenile and other forms of early-onset periodontitis. Some highly leukotoxic A. actinomycetemcomitans strains produce 10 to 20 times more leukotoxin than other minimally leukotoxic strains. The distribution, clonality, and intrafamilial transmission of highly leukotoxic A. actinomycetemcomitans were examined in order to determine the importance of leukotoxin in the pathogenesis of periodontitis.The polymerase chain reaction (PCR) was used to differentiate highly leukotoxic from minimally leukotoxic strains in examining 1,023 fresh A. actinomycetemcomitans isolates and strains from our culture collection. These were obtained from 146 subjects including 71 with localized juvenile periodontitis (LJP), 4 with early-onset periodontitis, 11 with post-localized juvenile periodontitis, 41 with adult periodontitis, and 19 periodontally normal subjects. The arbitrarily primed polymerase chain reaction (AP-PCR) analysis of 30 oral isolates from each of 25 subjects was used to determine the intraoral distribution of A. actinomycetemcomitans clones. AP-PCR was also used to examine the transmission of A. actinomycetemcomitans in 30 members of 6 families. The clonality of 41 highly leukotoxic A. actinomycetemcomitans strains was evaluated by both AP-PCR and ribotyping.Highly leukotoxic A. actinomycetemcomitans was found only in subjects with localized juvenile and early-onset periodontitis. Fifty-five percent of the LJP subjects harbored highly leukotoxic A. actinomycetemcomitans isolates. Seventy-three percent of the A. actinomycetemcomitans isolates in these subjects were highly leukotoxic. Highly leukotoxic A. actinomycetemcomitans infected younger subjects (mean age 13.95 years, range 5 to 28 years) than minimally leukotoxic (mean age 35.47 years, range 6 to 65 years). Most subjects were infected with only one A. actinomycetemcomitans genotype. However, PCR of whole dental plaques and subsequent analysis of up to 130 individual oral isolates suggested a possible shift in A. actinomycetemcomitans over time in that a few subjects harbored both highly leukotoxic and minimally leukotoxic strains. AP-PCR analysis was consistent with intrafamilial A. actinomycetemcomitans transmission. Ribotyping and AP-PCR analysis confirmed a previous report that highly leukotoxic A. actinomycetemcomitans consists of a single clonal type.This study suggests that localized juvenile and other forms of Actinobacillus-associated periodontitis are primarily associated with the highly leukotoxic clone of A. actinomycetemcomitans.

The present study describes a method for amplifying DNA in Actinobacillus actinomycetemcomitans by using short, synthetic oligonucleotides of random sequence as primers in the polymerase chain reaction. Genomic DNA from each of 20 human isolates of A. actinomycetemcomitans was successfully amplified in a thermal cycler with a single synthetic primer (GGGTAACGCC) and reproducibly produced 14 different DNA amplification profiles (amplitypes). A. actinomycetemcomitans isolates from the same subject revealed the same amplitype. The arbitrarily primed polymerase chain reaction appears to be useful in characterizing human isolates of A. actinomycetemcomitans for studies of epidemiology and bacterial transmission.

Actinobacillus actinomycetemcomitans is a key microorganism in the pathogenesis of several different forms of periodontal diseases. Identification of this bacterium from clinical specimens may often be complicated by the fact that the colony morphology on TSBV selective medium closely resembles that of Haemophilus aphrophilus and a key differentiating characteristic, catalase reaction, may be variable. Recent genetic studies have shown that the 23S ribosomal RNA molecule is split into two smaller forms in A. actinomycetemcomitans, but is intact in H. aphrophilus. Based on this finding, we describe a new, rapid method for identifying A. actinomycetemcomitans in which single colonies isolated from culture on TSBV agar in 5% CO2 in air are lysed, electrophoresed on 1.5% submarine agarose gels and visualized by staining with ethidium bromide. Using this assay, A. actinomycetemcomitans can be easily distinguished from morphologically similar colonies such as H. aphrophilus strains by differences in 23S rRNA within 2 h.

Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus which can cause certain severe extra-oral infections as well as forms of human periodontal disease such as localized juvenile periodontitis. In contrast to many prokaryotic and eukaryotic species which exhibit an intact 23S ribosomal RNA (rRNA) molecule, examination of six A. actinomycetemcomitans strains--including three serogroup representative strains and two strains from non-human primates--revealed that this micro-organism does not produce an intact 23S ribosomal RNA (rRNA) molecule but, rather, two smaller forms of 1.8 kb and 1.2 kb designated as 23S alpha and 23S beta fragments. On the other hand, 14 other strains of Actinobacillus, Haemophilus, and Pasteurella species demonstrated intact 23S rRNA. The sequence of the region of the 23S rRNA gene in A. actinomycetemcomitans strain ATCC 43718 containing the cleavage site was determined by dideoxynucleotide sequencing, while the location of the 3' and 5' termini of the 23S alpha and 23S beta fragments was resolved by S1 nuclease mapping and cDNA primer-extension. A deletion of 112 bases was noted in comparisons of base sequences from A. actinomycetemcomitans rRNA and rDNA. The DNA intervening sequence was localized to nucleotide 1180 of the Escherichia coli 23S rRNA map. While the primary structure of the gap region showed little homology with the gap regions described in other organisms, the secondary structure was similar to that previously described in the parasitic helminth Schistosoma japonicum. Restriction enzyme and nucleotide sequence analysis of the gap region in eight other A. actinomycetemcomitans strains showed it to be highly conserved.

Background Triclosan is a broad-spectrum antimicrobial agent used in toothpaste to reduce dental plaque, gingivitis and oral malodor. This community-level assessment evaluated the susceptibility of dental plaque bacteria to triclosan in samples collected over 19 years. Methods A total of 155 dental plaque samples were collected at eleven different times over 19 years from 58 adults using 0.3% triclosan, 2% copolymer, 0.243% sodium fluoride toothpaste and from 97 adults using toothpaste without triclosan. These included samples from 21 subjects who used triclosan toothpaste for at least five years and samples from 20 control subjects. The samples were cultured on media containing 0, 7.5 or 25 μg/ml triclosan. Descriptive statistics and p values were computed and a linear regression model and the runs test were used to examine susceptibility over time. Results Growth inhibition averaged 99.451% (91.209 - 99.830%) on media containing 7.5 μg/ml triclosan and 99.989% (99.670 - 100%) on media containing 25 μg/ml triclosan. There was no change in microbial susceptibility to triclosan over time discernible by regression analysis or the runs test in plaque samples taken over 19 years including samples from subjects using a triclosan-containing dentifrice for at least five years. Conclusions This community-level assessment of microbial susceptibility to triclosan among supragingival plaque bacteria is consistent with the long-term safety of a 0.3% triclosan, 2% copolymer, 0.243% sodium fluoride dentifrice.