Checkerboard DNA-DNA Hybridization Research Articles

Impact of orthodontic movement on clinical microbiological and tomographic parameters in patients with a history of periodontitis a pilot study

Objective The purpose of this study was to evaluate the impact of orthodontic movement on clinical, microbiological, and tomographic parameters in patients with a history of periodontitis. Materials and Methods Twelve individuals (2 males and 10 females, mean age 42.1 years) who had undergone treatment for generalized chronic periodontitis were selected from the Periodontal Clinic of Guarulhos University (Brazil). The patients undergoing orthodontic treatment were submitted to a clinical examination, and cone beam computed tomography scans were performed at baseline and after 12 months of treatment. Subgingival biofilm samples were analyzed by checkerboard DNA-DNA hybridization. Statistical analysis was performed by a Wilcoxon test. Results With regard to the periodontal clinical parameters, when comparing baseline and 12-month evaluations, statistically significant reductions were found in Probing Depth and Clinical Attachment Level (CAL). There was a statistically significant increase in the number of sites that had lost CAL. There were no statistically significant tomographic differences, and no loss in the buccal alveolar bone thickness was detected. Conclusions Considering clinical periodontal parameters, a reduction was observed in the number and percentage of CAL. There was a reduction in A. gerencseriae and an increase in F. nuc. nucleatum, S. constellatus, and T. Forsythia species. Regarding the tomographic parameters, no statistically significant differences were observed.

Microbial contamination profile on esthetic elastomeric ligatures through the checkerboard DNA-DNA hybridization technique : Arandomized split-mouth study.

The aim of this study is to assess the microbial contamination of three different brands of esthetic elastomeric ligatures. Different brands of esthetic ligatures (Unistick Pearl [American Orthodontics, Sheboygan, WI, USA], Power Sticks Pearl [Ortho Technology, Tampa, FL, USA], and Ease [Obscure, 3M Unitek, Monrovia, CA, USA]) were randomly assigned to permanent canines of 25patients (aged 11-18years) undergoing corrective orthodontic treatment. After 30days, the ligatures were removed, processed, and the biofilm composition was analyzed by checkerboard DNA-DNA hybridization for 40bacterial species. The microbiological data were analyzed using anonparametric mixed model. The ligatures presented intense microbial contamination after 30days, but no statistically significant differences were observed among the three groups (p > 0.05). The levels of the evaluated individual species and proportions of the microbial complexes showed no statistically significant differences among the ligature groups (p > 0.05). Esthetic elastomeric ligatures became multicolonized by several bacterial species after 30days of exposure to the oral cavity. However, no relevant differences were observed among the biofilm composition formed on the different ligature brands.

In-vitro polymicrobial oral biofilm model represents clinical microbial profile and disease progression during implant-related infections.

Clinically relevant in-vitro biofilm models are essential and valuable tools for mechanistically dissecting the etiopathogenesis of infectious diseases and test new antimicrobial therapies. Thus, the aim of this study was to develop and test a clinically relevant in-vitro oral polymicrobial biofilm model that mimics implant-related infections in terms of microbial profile. For this purpose, 24-well plate system was used to model oral biofilms, using three different microbial inoculums to grow in-vitro biofilms: (1) human saliva from periodontally healthy patients; (2) saliva as in inoculum 1+Porphyromonas gingivalis strain; and (3) supra and subgingival biofilm collected from peri-implant sites of patients diagnosed with peri-implantitis. Biofilms were grown to represent the dynamic transition from an aerobic to anaerobic community profile. Subsequently, biofilms were collected after each phase and evaluated for microbiological composition, microbial counts, biofilm biomass, structure, and susceptibility to chlorhexidine (CHX). Results showed higher live cell count (P<.05) for biofilms developed from patients' biofilm inoculum, but biomass volume, dry weight, and microbiological composition were similar among groups (P>.05). Interestingly, according to the checkerboard DNA-DNA hybridization results, the biofilm developed from stimulated human saliva exhibited a microbial composition more similar to the clinical subgingival biofilm of patients with peri-implantitis, with proportions of the main pathogens closer to those found in the disease. In addition, biofilm developed using saliva as inoculum was shown to be susceptible to CHX with significant reduction in bacteria compared with biofilms without exposure to CHX (P<.05). The findings suggested that the in-vitro polymicrobial biofilm developed from human saliva as inoculum is a suitable model and clinically relevant tool for mimicking the microbial composition of implant-related infections.

Parkinson’s disease alters the composition of subgingival microbiome

ABSTRACT Aim The current study aimed to test the hypothesis that Parkinson’s disease exacerbates periodontitis by altering its microbiome. Materials and Methods Clinical periodontal parameters were recorded. Subgingival samples from healthy controls, periodontitis patients (PD), and Parkinson’s patients with periodontitis (PA+PD) were analyzed using the checkerboard DNA-DNA hybridization technique for targeting 40 bacterial species typically associated with periodontal disease and health. Next-generation sequencing (NGS) of the 16S ribosomal RNA gene (V1-V3 regions) was performed to analyze the microbiome comprehensively. Results Parkinson’s patients had mild-to-moderate motor dysfunctions. Bleeding on probing was significantly increased in the PA+PD group compared to PD (p < 0.05). With checkerboard analysis, PA was associated with increased Treponema socranskii (p = 0.0062), Peptostreptococcaceae_[G-6] [Eubacterium]_nodatum (p = 0.0439), Parvimona micra (p < 0.0001), Prevotella melaninogenica (p = 0.0002), Lachnoanaerobaculum saburreum (p < 0.0001), and Streptococcus anginosus (p = 0.0020). Streptococcus intermedia (p = 0.0042), P.nodatum (p = 0.0022), P. micra (p = 0.0002), Treponema denticola (p = 0.0045), L.saburreum (p = 0.0267), P.melaninogenica (p = 0.0017), Campylobacter rectus (p = 0.0020), and T.socranskii (p = 0.0002) were higher; Aggregatibacter actinomycetemcomitans (p = 0.0072) was lower in deep pockets in the PA+PD compared to PD. Schaalia odontolytica (p = 0.0351) and A.actinomycetemcomitans (p = 0.002) were lower; C.rectus (p = 0.0002), P. micra (p = 0065), Streptococcus constellatus (p = 0.0151), T.denticola (p = 0.0141), P.melaninogenica (p = 0.0057), and T.socranskii (p = 0.0316) were higher in shallow pockets in the PA+PD. Diversity decreased in PD (p = 0.001) and PA+PD (p = 0.026) compared to control, with minimal differences in alpha and beta diversities among PD and PA+PD based on NGS results. Conclusion These data demonstrated that Parkinson’s disease modifies PD-associated subgingival microbiome.

Assessment of microbial contamination in removable orthodontic appliances with and without the use of antimicrobial agents by checkerboard DNA-DNA hybridization analysis

This clinical, crossover, double-blind trial evaluated the microbial contamination of removable orthodontic appliances used by children and the efficacy of 0.12% chlorhexidine gluconate spray use for disinfection. Twenty children aged 7-11 years were instructed to wear removable orthodontic appliances for 1 week. They were instructed to use a placebo solution (control) or 0.12% chlorhexidine gluconate (experimental) to clean the appliances on days 4 and 7 after installation. After this period, the microbial contamination on the surfaces of the appliance was analyzed using checkerboard DNA-DNA hybridization for 40 bacterial species. Data were analyzed by Fisher exact, t, and Wilcoxon tests (α= 0.05). Removable orthodontic appliances were heavily contaminated by the target microorganisms. Streptococcus sanguinis, Streptococcus oralis, Streptococcus gordonii, and Eikenella corrodens were found in 100% of the appliances. Among cariogenic microorganisms, Streptococcus mutans and Streptococcus sobrinus were more abundant than Lactobacillus acidophilus and Lactobacillus casei. Red complex pathogens were more abundant than orange complex species. Purple complex bacteria were the most prevalent among bacterial complexes not associated with specific pathologies, detected in 34% of the samples. After the use of chlorhexidine, the number of cariogenic microorganisms (S. mutans, S. sobrinus, and L. casei) decreased significantly (P<0.05), and the numbers of periodontal pathogenic species from the orange and red complex also decreased significantly (P<0.05). There was no reduction for Treponema socranskii. Removable orthodontic appliances were densely contaminated by several bacterial species. Twice-a-week application of chlorhexidine spray effectively reduced cariogenic and orange and red complex periodontal pathogens.

Newly identified pathogens in periodontitis: evidence from an association and an elimination study

ABSTRACT We assessed the level of evidence for the presence of new periodontal pathogens by (i) comparing the occurrence of non-classical periodontal taxa between healthy vs. periodontitis patients (Association study); (ii) assessing the modifications in the prevalence and levels of these species after treatments (Elimination study). In the Association study, we compared the prevalence and levels of 39 novel bacterial species between periodontally healthy and periodontitis patients. In the Elimination study, we analyzed samples from periodontitis patients assigned to receive scaling and root planing alone or with metronidazole+ amoxicillin TID/ 14 days. Levels of 79 bacterial species (39 novel and 40 classic) were assessed at baseline, 3 and 12 months post-therapy. All samples were analyzed using Checkerboard DNA-DNA hybridization. Out of the 39 novel species evaluated, eight were categorized as having strong and four as having moderate association with periodontitis. Our findings suggest strong evidence supporting Lancefieldella rimae, Cronobacter sakazakii, Pluralibacter gergoviae, Enterococcus faecalis, Eubacterium limosum, Filifactor alocis, Haemophilus influenzae, and Staphylococcus warneri, and moderate evidence supporting Escherichia coli, Fusobacterium necrophorum, Spiroplasma ixodetis, and Staphylococcus aureus as periodontal pathogens. These findings contribute to a better understanding of the etiology of periodontitis and may guide future diagnostic and interventional studies.

Patients with type 2 diabetes and severe periodontitis harbor a less pathogenic subgingival biofilm than normoglycemic individuals with severe periodontitis.

Whether, and to what extent, diabetes mellitus (DM) can affect the subgingival biofilm composition remains controversial. Thus, the aim of this study was to compare the composition of the subgingival microbiota of non-diabetic and type 2 diabetic patients with periodontitis using 40 "biomarker bacterial species." Biofilm samples of shallow (probing depth [PD] and clinical attachment level [CAL] ≤3mm without bleeding) and deep sites (PD and CAL ≥5mm with bleeding) of patients with or without type 2 DM were evaluated for levels/proportions of 40 bacterial species by checkerboard DNA-DNA hybridization. A total of 828 subgingival biofilm samples from 207 patients with periodontitis (118 normoglycemic and 89 with type 2 DM) were analyzed. The levels of most of the bacterial species evaluated were reduced in the diabetic compared with the normoglycemic group, both in shallow and in deep sites. The shallow and deep sites of patients with type 2 DM presented higher proportions of Actinomyces species, purple and green complexes, and lower proportions of red complex pathogens than those of normoglycemic patients (P<0.05). Patients with type 2 DM have a less dysbiotic subgingival microbial profile than normoglycemic patients, including lower levels/proportions of pathogens and higher levels/proportions of host-compatible species. Thus, type 2 diabetic patients seem to require less remarkable changes in biofilm composition than non-diabetic patients to develop the same pattern of periodontitis.

Bacterial response to Ti-35Nb-7Zr-5Ta alloy incorporated with calcium, phosphate and magnesium

High implant survival rates have been achieved in recent decades due to continual modifications in implant design and surface topography, however there is still an ongoing quest to control peri-implant bone loss. The objective of this work was to develop Ti-35Nb-7Zr-5Ta (TNZT) alloys, perform physicochemical and morphological characterization of their surface modified by electrolytic oxidative plasma technique with ions related to osseointegration and lastly evaluate bacterial colonization in vitro. Three groups were evaluated: C group (polished TNZT), CaP group (sodium β glycerophosphate + calcium acetate) and Mg group (magnesium acetate). Before and after anodizing the surfaces, physicochemical and morphological analyses were performed: scanning electron microscopy with field emission gun (FEG-SEM), energy dispersion spectroscopy (EDS), X-ray diffraction (DRX), wettability (goniometer) and roughness (rugometer). Controlled and treated specimens were contaminated with unstimulated saliva collected from 10 healthy volunteers. Then, biofilm samples were collected and up to 35 microbial species, including commensal and pathogenic microorganisms, were identified and quantified by the Checkerboard DNA-DNA Hybridization method. The CaP group modified the surface morphology in the form of pores, while the Mg group modified it in the form of flakes. The contact angle was significantly smaller in the CaP group. The average roughness was higher in the CaP and Mg groups. A smaller total amount of bacteria was identified in the Mg group and relevant differences were found in the microbial profile associated with different surface treatments. Therefore, considering the microbiological profile and for the prevention of peri-implantitis, the Mg group presented more satisfactory and encouraging results for the manufacture of dental implantsGraphical

Bacterial Leakage Evaluation Through DNA-DNA Checkerboard Hybridization Technique in Morse Taper Implant-Abutment Connections: An In Vitro Study.

Purpose: The objective of this in vitro study was to evaluate the activity of local gel containing metronidazole (MN) in the leakage area, which was analyzed by the DNA-DNA checkerboard hybridization method. Materials and Methods: Thirty-six sets of Morse taper/mini-pillar implants were used in this study. These implants were equally divided into the following three groups: MN gel (test group), no MN gel (negative test group), and no gel (control). The gel was prepared with metronidazole (15%). Unstimulated saliva samples were collected, transferred to a Falcon tube, and stored at 37°C. The sets were partially immersed in microtubes containing 300 μL of saliva and were incubated at 37°C ± 1°C for 7 days. Microbial infiltration was evaluated (37 bacterial species and 5 species of Candida). The results were analyzed with Wald-Type, ANOVA, and multiple comparisons analysis between groups. Results: After comparing the quantity of microorganisms, both gel-treated groups (no MN gel and MN gel) had more significant microorganism presence than the control group (P < .001), and no significant result was found between the no MN gel and MN gel groups (P > .05). Regarding the bacteria found, the most common were Aggregatibacter actinomycetemcomitans, Prevotella melaninogenica, Bacteroides fragilis, and Candida tropicalis. Conclusion: Within the limitations of this study, it was concluded that the gel containing metronidazole used in this study was not effective in preventing the infiltration of microorganisms through the Morse taper implant-abutment interface.

Subgingival Microbiota and Periodontal Clinical Status in Patients with Plaque Psoriasis: A Cross-Sectional Study.

Plaque Psoriasis (PP) and periodontitis are inflammatory disorders with a bidirectional association. They both have a qualitatively similar immune-modulatory cascade, cytokine profile, and a recently described dysbiosis. Different oral bacterial species compositions in the periodontal pocket might play a role in the development of PP. To describe the subgingival microbiota of the Mexican population with PP and the periodontal conditions. Subjects were divided into two groups: periodontal health (PH) (PH-non-PP, PH-PP) and periodontitis (PD) (P-non-PP, PD-PP). Following clinical examination, the patients were classified into three groups according to the degree of psoriasis as measured by the Psoriasis Area Severity Index (PASI) and the periodontal status according to the parameters of the American Academy of Periodontology (AAP). Subgingival microbiota samples of each patient were used to determine 40 species of periodontal bacteria by checkerboard DNA-DNA hybridization. IL-2 and IL-6 were measured by ELISA. Of the forty-eight patients with PP, 21 patients had PH and 27 patients had PD. PD-PP group has a significant increase in the percentage of plaque, gingival redness, pocket probing depth, and clinical attachment loss (P<0.001) compared to PH-PP group. Microbiologically PD-PP exhibited significantly higher mean counts for A. georgiae, A. israelii, A. naeslundii from blue complex (P<0.001) than PD-non-PP. Moreover, the counts of these Actinomyces in PD-PP increased according to the severity of index PASI. The concentration of IL-2 and IL-6 were increased in saliva from PH-PP and PD-PP patients compared to PH non-PP. PP individuals harbored a particular sub-gingival microbiota profile different from non-PP. The severity of psoriasis was related to dysbiosis of microbiota -PASI > 5 related to periodontitis with the predominance of Actinomyces periodontal, irrespective of their periodontal condition. Finally, the severity of psoriasis could be unbalanced in subgingival microbiota and increase the risk to develop periodontitis.

Incorporation of zinc into cetylpyridinium chloride mouthwash affects the composition of multispecies biofilms

This study evaluated the effect of a mouthwash containing 0.075% cetylpyridinium chloride and 0.28% zinc lactate (CPC + Zn) in a multispecies biofilm model. A 7-days 33-species biofilm, formed on Calgary device, was 1-min treated with: 0.12% chlorhexidine (CHX), culture medium (negative control), 0.075% cetylpyridinium chloride (CPC) or CPC + Zn, 2x/day, from day 3 until day 6. The metabolic activity and the microbial composition were evaluated by colorimetric method and checkerboard DNA-DNA hybridization, respectively. The three antimicrobials (CPC, CPC + Zn and CHX) reduced metabolic activity, total biofilm count and several species counts, including Porphyromonas gingivalis, Fusobacterium nucleatum, Parvimonas micra, Campylobacter gracilis and Streptococcus mutans. However, only CPC + Zn reduced counts of the pathogen Prevotella intermedia and did not interfere with the levels of some beneficial species in relation to the negative control. The treatment of multispecies subgingival biofilm with CPC + Zn was effective in controlling periodontal pathogens and favored the colonization of health-associated bacterial species.

Plaque and Dental Caries Risk in Midlife

Dental caries is an endogenous microbial community-based disease resulting from an ecological shift from dynamic stability to metabolic imbalance in a consortium of acidogenic and aciduric bacteria comprising the dental plaque biofilm. Participants were members of the Dunedin Multidisciplinary Health and Development Study, a longitudinal investigation of health and behaviour in a cohort born in Dunedin, New Zealand. Oral biofilm samples (collected at age 32 years) from anterior labial supragingival, posterior lingual supragingival, posterior subgingival, and the dorsum of the tongue habitats for 841 participants were analysed using checkerboard DNA-DNA hybridisation (CKB), focussing on 30 ecologically significant bacteria. Associations of CKB data with dental caries at ages 32 and 45 years were assessed using regression modelling, adjusting for potential confounders including sex, xerostomia, and oral hygiene. The putative periodontitis pathobiont Tannerella forsythia (in the anterior supragingival biofilm) was associated with untreated caries at age 32 years. The percentage of total summed cell number counts for two putative periodontitis-associated species (T. forsythia and Micromonas micros) was associated with greater caries experience at age 32 years and the development of new caries between age 32 and 45 years. Additionally, severe caries (3 + cavities) was associated with putative caries pathobionts (Lactobacillus fermentum, Lactobacillus plantarum), periodontitis-associated species, and commensals (M. micros, Campylobacter rectus, Streptococcus mitis biovar I, Streptococcus mitis biovar II) in the subgingival biofilm. Participants with sustained poor oral hygiene through age 32 years not only had greater experience of caries by that age than those with good oral hygiene (fully adjusted incidence risk ratio = 5.10, 95% CI: 3.30, 7.89) but also experienced greater incidence of new caries from age 32 to 45 years (incidence risk ratio = 3.69, 95% CI: 2.62, 5.20). These findings provide evidence in support of the extended caries ecological plaque hypotheses, the polymicrobial aetiology of caries, and the integrated aetiology of dental caries and periodontal diseases. They also underscore the roles of poor oral self-care (particularly over the life course) and xerostomia in the occurrence and progression of caries.

Impact of N-acetylcysteine (NAC) and calcium hydroxide intracanal medications in primary endodontic infection: a randomized clinical trial.

This RCT investigated the impact of N-acetylcysteine (NAC) and calcium hydroxide [Ca(OH)2] intracanal medications (ICMs) in primary endodontic infection with apical periodontitis (PEIAP). Thirty-six teeth with PEIAP were randomly divided into groups according to the ICM: NAC, Ca(OH)2 + saline solution (SSL), and Ca(OH)2 + 2% chlorhexidine-gel (2% CHX-gel) (all, n = 12). Root canal samples (RCSs) were collected before (s1) and after instrumentation (s2) and after 14days of ICM (s3). Chemomechanical preparation (CMP) was performed with a Reciproc file and 2.5% NaOCl. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species. At s1, bacterial DNA was detected in 100% of RCSs (36/36). All 40 bacterial species were found in PEIAP. The mean number of species per RCS was 17.92 ± 13.18. The most frequent bacteria were S. mitis (65%), E. nodatum (63%), E. faecalis (63%), F. nucl sp vicentii (58%), T. forsythia (58%), and F. periodonticum (56%). CMP reduced the mean number of species per RCS to 6.8 ± 2.36 (p < 0.05). At s3, the intragroup analysis revealed a broader antimicrobial activity for Ca (OH)2 + 2% CHX-gel and NAC than Ca(OH)2 + SSL (p < 0.05). NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs, including P. micra, P. nigrescens, T. denticola, A. israelii, P. endodontalis, P. acnes, C. ochracea, and E. corrodens. Ca (OH)2 + 2% chlorhexidine gel (2% CHX gel) showed a greater bacterial elimination over the number of bacterial species; however, NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs (RBR-3xbnnn). The use of intracanal medication with a broad antimicrobial activity can optimize root canal disinfection. Ca(OH)2 + 2% CHX gel and NAC showed a broader antimicrobial activity than Ca(OH)2 + SSL against endodontic pathogens in primary root canal infection. Brazilian Registry of Clinical Trials (REBEC), No. RBR-3xbnnn.

Replication of gene polymorphisms associated with periodontitis-related traits in an elderly cohort: the Washington Heights/Inwood Community Aging Project Ancillary Study of Oral Health.

We sought to replicate findings from published genome-wide association studies (GWAS), linking specific candidate gene loci with periodontitis-related clinical/microbial traits. In the published GWAS, a total of 2196 single nucleotide polymorphisms associated with periodontitis-related traits at a p≤ 5 × 10-6 and mapped to 136 gene loci. The replication cohort included 1124 individuals, 65-98 years old (67% female, 45% Hispanic, 30% Black, 23% White) with available genome-wide genotypes and full-mouth periodontal status. Microbial profiles using checkerboard DNA-DNA hybridization and 16SrRNA sequencing were available from 912 and 739 participants, respectively. Using gene-specific p-values after linkage disequilibrium pruning, the following gene/phenotype associations replicated successfully: CLEC19A with edentulism and %teeth with pocket depth (PD) ≥4 mm; IL37, HPVC1, TRPS1, ABHD12B, LDLRAD4 (C180rF1), TGM3, and GRK5 with %teeth with PD ≥4 mm; DAB2IP with presence of PD ≥6 mm; KIAA1715(LNPK), ROBO2, RAB28, LINC01017, NELL1, LDLRAD4(C18orF1), and CRYBB2P1 with %teeth with clinical attachment level (CAL) ≥3 mm; RUNX2 and LAMA2 with %teeth with CAL ≥5 mm; and KIAA1715(LNPK) with high colonization by Aggregatibacter actinomycetemcomitans. In addition, CLEC19A, IQSEC1, and EMR1 associated with microbial abundance based on checkerboard data, LBP and NCR2 with abundance based on sequencing data, and NCR2 with microbial diversity based on sequencing data. Several gene loci identified in published GWAS as associated with periodontitis-related phenotypes replicated successfully in an elderly cohort.

Systemic Antibiotics Influence Periodontal Parameters and Oral Microbiota, But Not Serological Markers.

The use of systemic antibiotics may influence the oral microbiota composition. Our aim was to investigate in this retrospective study whether the use of prescribed antibiotics associate with periodontal status, oral microbiota, and antibodies against the periodontal pathogens. The Social Insurance Institution of Finland Data provided the data on the use of systemic antibiotics by record linkage to purchased medications and entitled reimbursements up to 1 year before the oral examination and sampling. Six different classes of antibiotics were considered. The Parogene cohort included 505 subjects undergoing coronary angiography with the mean (SD) age of 63.4 (9.2) years and 65% of males. Subgingival plaque samples were analysed using the checkerboard DNA-DNA hybridisation. Serum and saliva antibody levels to periodontal pathogens were analysed with immunoassays and lipopolysaccharide (LPS) activity with the LAL assay. Systemic antibiotics were prescribed for 261 (51.7%) patients during the preceding year. The mean number of prescriptions among them was 2.13 (range 1–12), and 29.4% of the prescriptions were cephalosporins, 25.7% penicillins, 14.3% quinolones, 12.7% macrolides or lincomycin, 12.0% tetracycline, and 5.8% trimethoprim or sulphonamides. In linear regression models adjusted for age, sex, current smoking, and diabetes, number of antibiotic courses associated significantly with low periodontal inflammation burden index (PIBI, p < 0.001), bleeding on probing (BOP, p = 0.006), and alveolar bone loss (ABL, p = 0.042). Cephalosporins associated with all the parameters. The phyla mainly affected by the antibiotics were Bacteroidetes and Spirochaetes. Their levels were inversely associated with the number of prescriptions (p = 0.010 and p < 0.001) and directly associated with the time since the last prescription (p = 0.019 and p < 0.001). Significant inverse associations were observed between the number of prescriptions and saliva concentrations of Prevotella intermedia, Tannerella forsythia, and Treponema denticola and subgingival bacterial amounts of Porphyromonas gingivalis, P. intermedia, T. forsythia, and T. denticola. Saliva or serum antibody levels did not present an association with the use of antibiotics. Both serum (p = 0.031) and saliva (p = 0.032) LPS activity was lower in patients having any antibiotic course less than 1 month before sampling. Systemic antibiotics have effects on periodontal inflammation and oral microbiota composition, whereas the effects on host immune responses against the periodontal biomarker species seem unchanged.

Efficacy of 6% Sodium Hypochlorite on Infectious Content of Teeth with Symptomatic Irreversible Pulpitis

IntroductionThe objective of this study was to monitor the effects of chemomechanical preparation (CMP) performed with 6% sodium hypochlorite and calcium hydroxide–based intracanal medication (ICM) on the levels and diversity of bacteria, endotoxins (lipopolysaccharides [LPS]), and lipoteichoic acid (LTA) in root canals of teeth with symptomatic irreversible pulpitis. MethodsSamples were collected from 10 teeth with symptomatic irreversible pulpitis before CMP (S1), after CMP (S2), and after ICM (S3). The levels of bacteria, LPS, and LTA were assessed by using checkerboard DNA-DNA hybridization, LAL Pyrogent 5000, and enzyme-linked immunosorbent assay, respectively. Wilcoxon test, repeated-measures analysis of variance, and Tukey post hoc test were used for statistical analysis at a significance level of 5%. ResultsForty species were detected at S1. Two species were eliminated after CMP and 5 after ICM. Resistant and pain-related species were detected in the root canals. Higher levels of culturable bacteria were detected at S1. However, CMP and ICM effectively reduced the microbial load in the root canals. Higher levels of LPS and LTA were detected at S1. CMP was effective in reducing both LPS and LTA (P < .05). ICM produced additional reduction in the levels of LPS (P > .05) and LTA (P < .05). ConclusionsChemomechanical preparation using 6% sodium hypochlorite and calcium hydroxide–based intracanal medication were effective in reducing the levels of bacteria, LPS, and LTA in teeth with vital pulp and irreversibly inflamed pulp.

Associations of sex, oral hygiene and smoking with oral species in distinct habitats at age 32years.

The oral microbiome is ecologically diverse, complex, dynamic, and little understood. We describe the microbiota of four oral habitats in a birth cohort at age 32 and examine differences by sex, oral hygiene, and current smoking status, dental caries, and periodontal health. Oral biofilm samples collected from anterior labial supragingival, posterior lingual supragingival, subgingival, and tongue sites of 841 Dunedin Multidisciplinary Health and Development Study members were analysed using checkerboard DNA-DNA hybridization; focusing on 30 ecologically important bacterial species. The four habitats exhibited distinct microbial profiles that differed by sex. Streptococcus gordonii was more dominant in supragingival and tongue biofilms of males; Porphyromonas gingivalis exhibited higher relative abundance in subgingival biofilm of females. Males had higher scores than females for periodontal pathogens at supragingival sites. The relative abundance of several putative caries and periodontal pathogens differed in smokers and non-smokers. With poor oral hygiene significantly higher proportions of Gram-negative facultative anaerobes were present in subgingival biofilm and there were higher scores for the principal components characterised by putative cariogenic and periodontal pathogens at each site. Distinctive microenvironments shape oral biofilms and systematic differences exist by sex, oral hygiene, and smoking status.

Sports mouthguards: Contamination, roughness, and chlorhexidine for disinfection - A randomized clinical trial.

The aim of the present in vivo study was to evaluate the bacterial contamination of sports mouthguards, surface roughness, and the efficacy of chlorhexidine gluconate spray in the disinfection of these devices. A randomized, blinded cross-over clinical trial was performed with twenty 9 to 13 years old children who practiced martial arts and participated in all phases of the study. They were instructed to wear mouthguards 3 alternated days a week for 1 hour and, after use, to spray sterile tap water or chlorhexidine 0.12%. The mouthguards were analyzed by MTT assay, Checkerboard DNA-DNA hybridization, and confocal laser microscopy prior and after use for 2 weeks. Data were analyzed by Wilcoxon and t-Student, and Pearson correlation tests, with 5% significance level. Were observed that mouthguards of the control group were more contaminated with cariogenic microorganisms than those of the chlorhexidine group (p<0.05). The mouthguards use of spray of chlorhexidine reduced significantly the bacteria contamination compared with control group (p = 0.007). The surface roughness of the mouthguards increased significantly after use, irrespective of application of chlorhexidine spray. A moderate correlation (r=0.59) was observed between surface roughness and the cariogenic microorganism's contamination only for control group. Sports mouthguards had intense microbial contamination and increased surface roughness after its use. The use of chlorhexidine spray was effective for reducing the mouthguards contamination used by children.

The effect of Brazilian propolis type-3 against oral microbiota and volatile sulfur compounds in subjects with morning breath malodor.

To evaluate propolis type-3 mouthrinse effects on the concentration of volatile sulfur compounds (VSCs) and on tongue dorsum microbial profile. A three-step double-blind, crossover, randomized study with 10 individuals divided into three groups: I-placebo (P); II-ethanolic extract of propolis type-3 3% (EEP); and III-chlorhexidine 0.12% (CHX) and instructed to rinse twice daily for 5days. Each experimental period was followed by a 21-day washout interval. Morning mouth breath was assessed by VSC concentrations and microbiological samples were obtained from tongue dorsum at baseline and the end of period of rinses and analyzed using checkerboard DNA-DNA hybridization technique for 39 bacterial species. CHX and EEP presented the lowest VSC concentration when compared with placebo (p < 0.05). Even in the absence of mechanical plaque control, CHX and EEP treatments reduced VSC levels and there were no statistical differences for VSC measurement between CHX and EEP. There was a significant reduction in mean counts of 10 species including some VSC producers (Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia) by EEP. Total counts of organisms, gram-negative and gram-positive bacterial species showed a decrease for EEP and CHX (p < 0.05). In addition, no statistical difference was observed between EEP and CHX (p > 0.05). A positive correlation was observed between decrease of bacterial counts and decrease of VCSs concentration for the EEP and CHX. The use of a 3% propolis type-3 mouthrinse is an effective way to prevent morning bad breath. Thus, propolis may be a promising agent for the treatment of halitosis. Propolis type-3 may be used as adjuvant treatment for morning breath malodor.

Bacterial resistance to minocycline after adjunctive minocycline microspheres during periodontal maintenance: A randomized clinical trial.

Despite widespread use, the impact of minocycline hydrochloride microspheres on the shifts of oral bacterial species resistant to minocycline remains unknown. This study aimed at examining the percentage and taxonomy of minocycline-resistant isolates in saliva and subgingival plaque samples before and after minocycline microspheres application in periodontitis patients during maintenance. Patients received supra- and sub-gingival debridement with (test) or without (control) minocycline microspheres application to sites with probing depth>4mm and were clinically monitored at baseline, 1, 3, and 6 months. Samples were collected at baseline, 1 and 6 months and analyzed via cultivation with or without 4μg/mL minocycline. Percentage of resistant strains was determined by colony counting and taxonomy by checkerboard DNA-DNA hybridization. Significant clinical changes were sought with the Mann-Whitney test and differences in percentage of resistant isolates with the Friedman and Mann-Whitney tests. Groups showed similar clinical improvements. Mean percentage of resistant isolates rose at 1 month and decreased at 6 months in saliva and plaque samples in test group (P<0.05) but remained unchanged in control group. Percentage of resistant isolates of Gemella morbillorum and Eubacterium saburreum increased significantly at 6 months in both groups. Antibiotic resistance by Aggregatibacter actinomycetemcomitans, Tannerella forsythia, and Porphyromonas gingivalis was either absent or infrequent. Minocycline microspheres result in transient selection of minocycline resistant species in saliva and subgingival plaque samples.