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Background: The objective was to analyze clinical, microbiological, and immunological periodontal parameters in patients in corrective orthodontic treatment., Materials and Methods: Twenty-eight patients were selected. Plaque index (PI), bleeding on probing (BOP), width of keratinized gingiva, levels of 40 bacterial species, and of 3 cytokines (IL-1β, MMP-8, and TNF-α) in gingival crevicular fluid (GCF) were evaluated at T0, before orthodontic treatment; T1, 6 months; and T2, 12 months post-treatment. Non-parametric, Friedman, Wilcoxon, ANOVA, and Spearman correlation coefficient tests were used for statistical analyses, with the significance level of 5%., Results: No significant difference was found for the width of keratinized gingiva, but PI presented a significant increase at T1 and T2 (p < 0.05) when compared with T0. The percentage of sites with BOP increased significantly from T0 to T1 (p < 0.05); however, at T2, the values decreased and did not differ anymore from T0 (p > 0.05). In the microbiological analysis, red complex pathogens were in significantly greater proportions in T2 compared with T0 (p < 0.05). There was no statistically significant difference in the cytokine levels between the periods but there was a positive correlation between BOP and IL-1β (r = 0.49 p = .01) and TNF-α (r = 0.39 and p = .05)., Conclusion: In conclusion, corrective orthodontic treatment caused clinical periodontal alterations regarding biofilm accumulation and gingival bleeding, with alteration of periodontopathogens.

Objectives: This study evaluated the effects of type 2 diabetes mellitus (DM), smoking, and these two factors combined on gingival crevicular fluid levels and ratios of pro-/anti-inflammatory cytokines. Associations between cytokines with each other and with key periodontal pathogens in periodontal sites under the challenge of one or both of these risk factors were also assessed., Methods: A total of 102 subjects with periodontitis were included in this cross-sectional study and assigned to one of the following groups: non-diabetic non-smokers (control group, n = 25), non-smokers with DM (DM group, n = 30), non-diabetic smokers (S group, n = 26), and smokers with DM (S + DM group, n = 21). The levels of 13 pro-inflammatory (IFN-γ, TNF-α, MIP-1α, GM-CSF, IL-1β, IL-2, IL-6, IL-7, IL-8, IL-12, IL-17, IL-21, and IL-23) and 5 anti-inflammatory (IL-4, IL-5, IL-10, IL-13, and TGF-β) cytokines were assessed in healthy and diseased sites, using multiplex immunoassay. Ratios of pro-/anti-inflammatory cytokines were obtained in all possible permutations. The levels of 7 key periodontal pathogens were evaluated by qPCR., Results: Overall, the ratios of pro-/anti-inflammatory cytokines were higher in healthy and diseased sites of the DM group and in healthy sites of the S + DM group, and lower in diseased sites of the S group, compared with the control (p < .05). The proportion of the pro-inflammatory cytokines in relation to the 18 cytokines studied was higher in the DM group and lower in the S group, whereas the proportion of the anti-inflammatory cytokines was lower in both diabetic groups and higher in the S group, compared to the control (p < .05). A cluster of six common cytokines (IL-4, IL-5, IL-12, IL-13, IL-21, and IL-23) was observed in the diseased sites of all groups studied. Eight common cytokines (IL-4, IL-5, IL-12, IL-13, IL-17, IL-21, IL-23, and IFN-γ) grouped closely in the healthy sites of both diabetic groups. Significant associations between pathogens and cytokines occurred mainly in the diseased sites of the S + DM group (p < .05)., Conclusion: Diabetes mellitus induced an overall pro-inflammatory state, while smoking mainly stimulated immunosuppression in periodontal sites. When the two risk factors overlapped, smoking seemed to partially assuage the hyperinflammatory effect of DM., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

Objective: The microbial composition of pericoronitis (Pc) is still controversial; it is not yet clear if the microbial profile of these lesions is similar to the profile observed in periodontitis (Pd). Therefore, the aim of the present study was to describe the microbial profile of Pc lesions and compare it directly with that of subjects with Pd., Methodology: Subjects with Pc and Pd were selected, and subgingival biofilm samples were collected from (i) third molars with symptomatic Pc (Pc-T), (ii) contralateral third molars without Pc (Pc-C) and (iii) teeth with a probing depth >3 mm from subjects with Pd. Counts and proportions of 40 bacterial species were evaluated using a checkerboard DNA-DNA hybridization technique., Results: Twenty-six patients with Pc and 18 with Pd were included in the study. In general, higher levels of microorganisms were observed in Pd. Only Actinomyces oris and Eubacterium nodatum were present in higher mean counts in the Pc-T group in comparison with the Pc-C and Pd-C groups (p<0.05). The microbiota associated with Pc-T was similar to that found in Pc-C. Sites with Pc lesions had lower proportions of red complex in comparison with the Pd sites., Conclusion: The microbiota of Pc is very diverse, but these lesions harbour lower levels of periodontal pathogens than Pd.

The aim was of this study was to determine the current weight of evidence for the existence of specific differences between the microbiota of healthy teeth and healthy implants, or of teeth with periodontitis and implants with peri-implantitis. A systematic review was conducted according to the PRISMA statement. The MEDLINE, EMBASE and Cochrane databases were searched up to February 2018 for studies comparing microbiological data of biofilm samples collected from healthy teeth and implants or from teeth with periodontitis and implants with peri-implantitis. The weight of evidence was defined in three categories (strong, moderate and mild/some), according to the difference in number of studies showing statistically significantly higher counts and/or proportions and/or abundance and/or prevalence of microorganisms in health or in disease. Of the 132 articles identified, 8 were included. A wide range of microorganisms were present in different conditions but no microorganisms showed strong, moderate or mild/some evidence for a specific association with either teeth or implants. The results of this systematic review indicated that there is insufficient evidence in the literature to support specific differences between microorganisms colonizing teeth and implants, either in health or in disease.

The aim of this study was to evaluate the effects of adjunct systemic antibiotic treatment with metronidazole (MTZ) and amoxicillin (AMX) in patients receiving non-surgical subgingival debridement (NSD) for peri-implantitis. Forty subjects presenting with at least one implant with severe peri-implantitis were randomized into an experimental group [treated with NSD plus MTZ (400 mg) and AMX (500 mg) three times a day for 14 days] and a control group treated with NSD plus placebo. Clinical parameters and submucosal biofilm profiles were evaluated up to 1 year post-treatment. Overall, both treatments improved clinical parameters over time. At 1 year, mean probing depth (PD), mean clinical attachment (CA) level and proportions of red complex pathogens did not differ significantly between the two groups. In addition, mean PD and CA changes to 1-year posttreatment did not differ significantly between the two groups between baseline and 1-year post-treatment. These results suggest that the addition of MTZ and AMX to the treatment protocol of patients undergoing NSD for with severe peri-implantitis does not improve the clinical and microbiological outcomes of NSD. The fact that half of the implants in both groups did not achieve clinical success (PD < 5 mm, no BoP, no bone loss) suggest that neither of the tested protocols were effective for treating severe peri-implantitis.

Additive manufacturing (AM) is an emerging process for biomaterials and medical devices. Direct Laser Metal Sintering (DLMS) is an AM technique used to fabricate Ti-6Al-4V implant materials with enhanced surface-related properties compared with wrought samples; thus, this technique could influence microbial adsorption and colonization. Therefore, this in vitro study was conducted to evaluate the impact of different implant production processes on microbial adhesion of periodontal pathogens. Titanium discs produced using two different processes-conventional and AM-were divided into three groups: conventional titanium discs with machined surface (G1), AM titanium discs with chemical treatment (G2) and AM titanium discs without chemical treatment (G3). Subgingival biofilm composed of 32 species was formed on the titanium discs, and positioned vertically in 96-well plates, for 7 days. The proportions of microbial complexes and the microbial profiles were analyzed using a DNA-DNA hybridization technique, and data were evaluated using Kruskal-Wallis and Dunnett tests (p < 0.05). Lower proportions of the red complex species were observed in the biofilm formed in G2 compared with that in G1 (p < 0.05). Moreover, the proportions of the microbial complexes were similar between G2 and G3 (p > 0.05). Compared with G1, G2 showed reduced levels of Porphyromonas gingvalis , Actinomyces gerencseriae, and Streptococcus intermedius , and increased levels of Parvimonas micra , Actinomyces odontolyticus, and Eikenella corrodens (p < 0.05). The microbial profile of G3 did not differ from G1 and G2 (p > 0.05). The results of this in vitro study showed that titanium discs produced via AM could alter the microbial profile of the biofilm formed around them. Further clinical studies should be conducted to confirm these findings.

Background and Objective: Little is known about the correspondence of cytokines detected among the different biological matrices used in periodontal research and whether locally-detected cytokine mRNA can also be identified at the translated protein level. The aims of this study were to compare: (a) the detection of classic cytokines at protein level in corresponding samples of serum, gingival tissue and gingival crevicular fluid (GCF) in subjects with periodontitis and (b) the detection of cytokines at protein level in GCF and tissue with their detection at mRNA level in tissue., Material and Methods: Protein concentrations of IL-1β, IL-6 and TNF-α were measured in corresponding samples of GCF, gingival tissue and serum from 125 subjects with chronic periodontitis by multiplex immunoassay. The mRNA levels of these cytokines were evaluated in the same gingival tissue by qPCR., Results: The mean protein concentrations of IL-1ß and TNF-α were higher in tissue and GCF than in serum (P < 0.05). The mean protein concentration of IL-6 was higher in tissue, followed by serum and GCF, respectively (P < 0.05). Using serum as the gold standard, detection of IL-1ß in GCF and IL-6 in tissue had the highest sensitivity (100%), while detection of IL-6 in the GCF had the lowest sensitivity (77.9%). Using tissue as the gold standard, the detection sensitivities of IL-1ß, IL-6 and TNF-α were 93.6%, 78.4% and 94.2%, respectively, in GCF. Using mRNA as the gold standard, the detection of IL-1ß and IL-6 at the protein level in tissue (100.0%) had the highest sensitivity, while the detection of IL-6 in GCF (77.9%) had the lowest sensitivity., Conclusion: Tissue and GCF exhibited overall higher protein concentrations of cytokines than serum. GCF presented good reliability for identifying the target cytokines at protein level, when compared with detecting tissue cytokines at protein level. GCF and tissue presented high sensitivities for detecting cytokines at the protein level, when compared with detecting tissue cytokines at the mRNA level. The sensitivity for the identification of cytokines in the GCF depended on the target cytokine, where IL-6 was the least likely cytokine to be detected in this matrix., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This study investigated the antimicrobial effects of the ethanolic extract of Brazilian red propolis (BRP) on multispecies biofilms. A seven-day-old subgingival biofilm with 32 species was grown in a Calgary device. Biofilms were treated with BRP (1,600, 800, 400 and 200 μg ml -1 ) twice a day for 1 min, starting from day 3. Chlorhexidine (0.12%) and dilution-vehicle were used as positive and negative controls, respectively. On day 7, metabolic activity and the microbial composition of the biofilms by DNA-DNA hybridization were determined. The viability data were analyzed by one-way ANOVA followed by Tukey's post hoc , whereas the microbial composition data were transformed via BOX-COX and analyzed using Dunnett's post hoc . BRP (1,600 μg ml -1 ) decreased biofilm metabolic activity by 45%, with no significant difference from chlorhexidine-treated samples. BRP (1,600 μg ml -1 ) and chlorhexidine significantly reduced levels of 14 bacterial species compared to the vehicle control. Taken together, BRP showed promising antimicrobial properties which may be useful in periodontal disease control.

Background: Because sucrose may change the composition of biofilms formed on dental surfaces, the aim of this study was to evaluate in situ the effect of this dietary sugar on biofilm formation on titanium surface., Methods: In this blind, crossover, in situ study, 10 volunteers wore, in 3 phases of 7 days each, a palatal appliance containing titanium specimens. In each phase, the specimens were treated extraorally with 20% sucrose solution at a frequency of 4 or 8 times per day. As control, no treatment was rendered (0×). At the end of each phase, the biofilms were collected for biochemical analysis of biofilm wet weight (biomass), protein concentration, soluble (S-EPS), and insoluble (I-EPS) extracellular polysaccharides and intracellular polysaccharides (IPS), and for microbiologic analysis by checkerboard DNA-DNA hybridization (for levels and proportions of 40 bacterial species). Biochemical data were analyzed by linear regression and microbiological findings by Friedman and Dunn tests (α = .05)., Results: A positive significant linear relationship was found among sucrose exposure (0×, 4×, and 8×) and biomass, S-EPS, I-EPS and IPS (p < 0.05). The biofilms treated with sucrose (4× and/or 8×) presented higher mean total levels of the 40 bacterial species evaluated, higher proportions of red complex species and lower proportions of the host-compatible green complex species, in comparison with the control group (p < 0.05)., Conclusion: The findings of the present study suggest that daily sucrose exposure has a harmful effect on the composition of biofilms formed on titanium surfaces., (© 2018 American Academy of Periodontology.)

Objective: This study assessed the impact of chronic periodontitis (CP) and CP associated with type 2 diabetes mellitus (DM) and/or smoking on the serum ratios of pro- to anti-inflammatory cytokines., Materials and Methods: Subjects were assigned into one of the following groups: control (n = 25, non-diabetic non-smokers with no history of periodontitis), CP (n = 26, non-diabetic non-smokers with CP), DMCP (n = 30, non-smokers with DM and CP), SCP (n = 27, non-diabetic smokers with CP), and SDMCP (n = 22, smokers with type 2 DM and CP). Serum levels of 18 cytokines were measured using multiplex immunoassays., Results: Six ratios of pro-inflammatory to anti-inflammatory cytokines were significantly higher in the CP group than in the control group (p < 0.05). Eleven, seventeen and nine ratios of pro-inflammatory to anti-inflammatory cytokines were significantly higher in the DMCP, SCP and SDMCP groups than in the control group, respectively (p < 0.05). The SCP group presented higher serum ratios of tumor necrosis factor (TNF)-α/interleukin (IL)-4, TNF-α/IL-5, IL-17/IL-13 and IL-6/IL-13 (p < 0.05) than the CP group. Cluster analysis revealed a relevant cluster composed of ten cytokines (IL-17, IL-23, interferon-γ, IL-12, IL-1β, IL-2, IL-21, IL-6, IL-4 and granulocyte-macrophage colony-stimulating factor [GM-CSF]) in the serum of subjects from the DMCP group., Conclusions: The ratios of pro- to anti-inflammatory cytokines shift to favor a pro-inflammatory status in the serum of patients with CP and even more when CP is associated with one or both risk factors., Clinical Relevance: CP and CP associated with hyperglycemia and/or smoking might contribute to a systemic inflammatory burden and increased risk of systemic complications.

This study aimed to test the dose-response effect of chlorhexidine on multispecies biofilms formed on commercially pure titanium (cpTi) and titanium-zirconium (TiZr) alloy. Biofilms were formed on cpTi and TiZr discs and treated two times per day with five different chlorhexidine concentrations (0.12, 0.20, 0.50, 1, 2%). The biofilms were collected for microbiological, biochemical and microscopic analyses. The significance of differences among groups was evaluated by linear regression, ANOVA, Bonferroni and Tukey tests. The mean number of colony-forming units decreased as the chlorhexidine concentration increased for both cpTi and TiZr (p < 0.05). The maximum effect was observed with the 0.5% concentration. Confocal microscopy images suggested an increase in the number of dead bacterial cells with increased chlorhexidine concentration. The biofilm pH increased after chlorhexidine exposure (p < 0.05). Chlorhexidine showed an antimicrobial dose-response effect in controlling biofilm on cpTi and TiZr. 0.5% chlorhexidine can be used to achieve the maximum antimicrobial effect on both materials.

Purpose: The All-on-4 treatment concept has been shown to be an effective clinical procedure; however, to date, no studies have analyzed the subgingival microbiota present in these restorations. The purpose of this study was to evaluate the microbial profile of the subgingival biofilm around dental implants placed in the All-on-4 protocol and compare the microbial profile around axial and tilted implants., Materials and Methods: Fourteen subjects treated by the All-on-4 concept were evaluated clinically and microbiologically. Subgingival biofilm was collected from each patient, and the amount of 40 species of bacteria was assessed using the checkerboard DNA-DNA hybridization technique., Results: The results for the indices of probing depth (PD), bleeding on probing, marginal bleeding, and visible plaque were 2.32 mm, 46%, 60%, and 57%, respectively. Tilted implants presented a significantly higher mean PD and Plaque Index compared with axial implants (P < .05). Fusobacterium nucleatum ssp vincentii, Veillonella parvula, and Fusobacterium nucleatum ssp polymorphum were found in higher levels; however, no difference in the microbial composition was observed between tilted and axial implants (P > .05). Tilted implants presented statistically higher mean levels for the orange complex in relation to the axial implants (P < .05)., Conclusion: Despite the clinical success rate of the All-on-4 protocol, the subgingival biofilm of tilted implants presented a higher proportion for the orange complex pathogens in comparison to axial implants. These data could suggest that subjects with this modality of implant-supported restoration must be aware that they need a more rigorous maintenance protocol.

Aim: To evaluate if non-diabetic subjects with periodontitis respond better than subjects with type 2 diabetes to the treatment protocol of scaling and root planing (SRP), metronidazole (MTZ) and amoxicillin (AMX)., Material and Methods: Diabetic and nondiabetic subjects with severe periodontitis received SRP + MTZ (400 mg/thrice a day [TID]) + AMX (500 mg/TID) for 14 days. Subgingival biofilm samples were analyzed by checkerboard DNA-DNA hybridization for 40 bacterial species. Subjects were monitored at baseline, 3, 6 and 12 months post-therapy., Results: Twenty-nine type 2 diabetics and 29 non-diabetic subjects participated of this study. Of the non-diabetics and diabetics, 68.9% and 75.9%, respectively, reached the clinical endpoint for treatment (≤4 sites with probing depth [PD] ≥5 mm) at 1 year post-therapy (p > 0.05). The diabetic group presented lower mean clinical attachment gain from baseline to 1 year post-therapy and higher mean proportions of the red and orange complexes than the non-diabetic group (p < 0.05)., Conclusions: Non-diabetic subjects with severe periodontitis did not respond better than type 2 diabetic subjects to the treatment protocol of SRP + MTZ + AMX, both in terms of achieving the clinical endpoint for treatment and of PD improvement. Diabetic subjects exhibited a slightly worse microbiological response and showed a healing process more associated with gingival recession than the non-diabetics., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To evaluate the effects of a toothpaste containing 0.3% triclosan in volunteers that have been treated for peri-implantitis and were enrolled in a maintenance program., Material and Methods: Subjects presenting at least one implant with peri-implantitis were selected. They received anti-infective surgical therapy, and sixty days post-surgery (baseline) were randomized into two groups, brushing twice/day for 2 years (a) with a toothpaste containing 0.3% triclosan+2.0% PVM/MA copolymer (Gantrez TM )+1,450 ppm fluoride (test) or (b) with a toothpaste containing 1,450 ppm fluoride (control). They received clinical and microbiological monitoring at baseline, 3, 6, 12, 18, and 24 months, and professional maintenance every 3 months., Results: A total of 102 subjects were enrolled (test: 48; control: 54). The control group showed loss of clinical attachment (CA) around treated implants over the course of the study (p < 0.05), while the test group was stable for this parameter. The difference between groups (0.55 mm) for CA change between baseline and 24 months (primary outcome variable) was statistically significant (p < 0.05). Red complex pathogens were only reduced in the test group at 24 months. The implants with no history of peri-implantitis in the test group had a significant reduction in the percentage of sites with bleeding on probing and in mean probing depth, throughout the study (p < 0.05). This improvement was not observed in the control group., Conclusion: A toothpaste containing 0.3% triclosan was more effective than a toothpaste without triclosan in maintaining a healthy peri-implant environment around treated implants and implants with no history of peri-implantitis during a 2-year maintenance program (ClinicalTrials.govNCT03191721)., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: Using two groups of mini-implants (successful and failed) the objectives of this in vivo study were: to evaluate the microbial contamination by the checkerboard DNA-DNA hybridization technique and to quantify the bacterial endotoxin by the limulus amebocyte lysate assay., Material and Methods: The 15 successful and 10 failed mini-implants (1.6 mm diameter × 7.0 or 9.0 mm long), placed in the maxilla and/or mandible, were obtained from 15 patients undergoing orthodontic treatment. Data were analyzed statistically by the Wilcoxon rank-sum test using the SAS software (a=0.05)., Results: All 40 microbial species were detected in both groups of mini-implants, with different frequencies. No differences were observed between the groups with respect to microbial complexes (blue, purple, yellow, green, orange, red and other species) and endotoxin quantification (p>0.05)., Conclusion: Neither microbial contamination nor endotoxin quantification was determinant for the early loss of stability of the mini-implants.

Objective: To critically evaluate the existing literature on acute lesions occurring in the periodontium (periodontal abscesses [PA], necrotizing periodontal diseases [NPD], and endo-periodontal lesions [EPL]) to determine the weight of evidence for the existence of specific clinical conditions that may be grouped together according to common features. The ultimate goal is to support an objective classification system., Importance: Although PA, NPD, and EPL occur with relatively low frequency, these lesions are of clinical relevance, because they require immediate management and might severely compromise the prognosis of the tooth., Findings: In general, the evidence available to define these three conditions was considered limited. PA and EPL are normally associated with deep periodontal pockets, bleeding on probing, suppuration, and almost invariably, with pain. EPL are also associated with endodontic pathology. NPDs have three typical features: pain, bleeding, and ulceration of the gingival interdental papilla. The available data suggested that the prognosis of PA and EPL are worse in periodontitis than in nonperiodontitis patients. Lesions associated with root damage, such as fractures and perforations, had the worst prognosis. NPD progression, extent and severity mainly depended on host-related factors predisposing to these diseases., Conclusions: PA should be classified according to the etiological factors involved, with the most frequent being those occurring in pre-existing periodontal pockets. NPD are clearly associated with the host immune response, which should be considered in the classification system for these lesions. EPLs should be classified according to signs and symptoms that have direct impact on their prognosis and treatment, such as presence or absence of fractures and perforations, and presence or absence of periodontitis., (© 2018 American Academy of Periodontology and European Federation of Periodontology.)

A new periodontitis classification scheme has been adopted, in which forms of the disease previously recognized as "chronic" or "aggressive" are now grouped under a single category ("periodontitis") and are further characterized based on a multi-dimensional staging and grading system. Staging is largely dependent upon the severity of disease at presentation as well as on the complexity of disease management, while grading provides supplemental information about biological features of the disease including a history-based analysis of the rate of periodontitis progression; assessment of the risk for further progression; analysis of possible poor outcomes of treatment; and assessment of the risk that the disease or its treatment may negatively affect the general health of the patient. Necrotizing periodontal diseases, whose characteristic clinical phenotype includes typical features (papilla necrosis, bleeding, and pain) and are associated with host immune response impairments, remain a distinct periodontitis category. Endodontic-periodontal lesions, defined by a pathological communication between the pulpal and periodontal tissues at a given tooth, occur in either an acute or a chronic form, and are classified according to signs and symptoms that have direct impact on their prognosis and treatment. Periodontal abscesses are defined as acute lesions characterized by localized accumulation of pus within the gingival wall of the periodontal pocket/sulcus, rapid tissue destruction and are associated with risk for systemic dissemination., (© 2018 American Academy of Periodontology and European Federation of Periodontology.)

Chronic periodontitis is the most prevalent form of inflammatory destructive bone disease and has been affecting humans since antiquity. Evidence suggest that genetic factors can highly influence periodontitis risk, modulating disease elements such as the susceptibility to microbial colonization and the nature of subsequent host-microbe interaction. Several single-nucleotide polymorphisms (SNPs) have been associated with the occurrence of periodontitis, but the full range of genetic influence in periodontitis outcomes remains to be determined. In this context, this study comprises an analysis of possible correlation between periodontitis-related genetic variants with changes in the subgingival microbiological pattern performed in a Brazilian population ( n = 167, comprising 76 chronic periodontitis patients and 91 healthy subjects). For the genetic characterization, 19 candidate SNPs were selected based on the top hits of previous large genome wide association studies (GWAS), while the subgingival microbiota was characterized for the presence and relative quantity of 40 bacterial species by DNA-DNA checkerboard. The case/control association test did not demonstrate a significant effect of the target SNPs with the disease phenotype. The polymorphism rs2521634 proved significantly associated with Tannerella. forsythia , Actinomyces gerencseriae , Fusobacterium periodonticum, and Prevotella nigrescens ; rs10010758 and rs6667202 were associated with increased counts of Porphyromonas gingivalis ; and rs10043775 proved significantly associated with decreased counts of Prevotella intermedia . In conclusion, we present strong evidence supporting a direct connection between the host's genetic profile, specifically rs2521634, rs10010758, rs6667202, and rs10043775 polymorphisms, and the occurrence of chronic periodontitis-associated bacteria., Competing Interests: The authors declare no conflict of interest.

Background: The combination of systemic metronidazole (MTZ) and amoxicillin (AMX) with scaling and root planing (SRP) has shown to be an effective periodontal treatment. However, some essential issues associated with the use of these antibiotics remain unanswered, such as the ideal time of administration during the course of periodontal treatment. Although these agents are often prescribed after the healing phase of the SRP procedure, there is biological plausibility to support its use in conjunction with the mechanical treatment. However, to date, no placebo controlled randomized clinical trial (RCT) has directly compared these two protocols. Therefore, the aim of this RCT is to compare the clinical, microbiological and immunological effects of the adjunctive systemic MTZ + AMX administered in different phases of the treatment of severe periodontitis., Methods: Subjects with severe periodontitis (n = 180) are being randomly assigned into three groups (n = 60/group): (i) SRP-only (control group), SRP in combination with 400 mg MTZ + 500 mg AMX, starting (ii) at the first SRP session (active phase group), or (iii) after 3 months of its completion (healing phase group). All volunteers are receiving clinical and microbiological evaluation at baseline, 3, 6 and 12 months, and immunological assessment at baseline and 12 months post-therapy. Nine subgingival biofilm samples are being collected per subject and analyzed for counts and proportions of 40 bacterial species by checkerboard DNA-DNA hybridization, and six gingival crevicular fluid samples are being collected and analyzed for the levels of 20 chemokines by multiplex immunoassay. The primary outcome variable is the number of volunteers reaching the clinical endpoint for treatment (≤ 4 sites with probing depth ≥5 mm) at 1 year post-therapy. Differences in clinical, microbiological and immunological parameters among groups and over time will be evaluated using analysis of variance, analysis of covariance and the Chi-square and Tukey tests. Microbiological and immunological analyses will be performed using adjustments for multiple comparisons. Statistical significance will be set at 5%., Trial Registration: ClinicalTrials.gov , NCT02954393 . Registered on 3 November 2016.

Aim: This study evaluated the levels of sclerostin (SOST) and Dickkopf (DKK)-1 in the chronic periodontitis (CP) associated with type 2 diabetes (DM) and/or smoking. Relationships between SOST, DDK1, RANKL, OPG, IL-1ß, IL-6 and TNF-α, and pathogens were assessed., Material and Methods: The study population included non-diabetic non-smokers (control), non-smokers with DM (DM group), non-diabetic smokers (S group) and smokers with DM (SDM group), all with CP. Serum and gingival levels of SOST, DKK1, RANKL, OPG, IL-1ß, IL-6 and TNF-α were evaluated by multiplex immunoassay. Gene expressions of these biomarkers and subgingival levels of pathogens were assessed by qPCR., Results: Gingival protein and/or mRNA levels of DKK1 and SOST were higher in subjects with DM and/or smoking than in controls (p < .05). Serum levels of SOST were higher in the DM group than in controls (p < .05). DKK1 positively correlated with SOST in the DM, SDM and control groups (p < .05) at mRNA levels. DKK-1 and SOST correlated with pathogens, especially in both groups with DM., Conclusions: SOST and DKK1 were upregulated in patients with CP presenting DM and/or smoking. DM, alone or with smoking, particularly influenced the correlations of SOST and DKK1 with each other and with the other biomarkers mostly at mRNA levels, as well as with periodontal pathogens., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The existence of specific microbial profiles for different periodontal conditions is still a matter of debate. The aim of this study was to test the hypothesis that 40 bacterial species could be used to classify patients, utilising machine learning, into generalised chronic periodontitis (ChP), generalised aggressive periodontitis (AgP) and periodontal health (PH)., Method: Subgingival biofilm samples were collected from patients with AgP, ChP and PH and analysed for their content of 40 bacterial species using checkerboard DNA-DNA hybridisation. Two stages of machine learning were then performed. First of all, we tested whether there was a difference between the composition of bacterial communities in PH and in disease, and then we tested whether a difference existed in the composition of bacterial communities between ChP and AgP. The data were split in each analysis to 70% train and 30% test. A support vector machine (SVM) classifier was used with a linear kernel and a Box constraint of 1. The analysis was divided into two parts., Results: Overall, 435 patients (3,915 samples) were included in the analysis (PH = 53; ChP = 308; AgP = 74). The variance of the healthy samples in all principal component analysis (PCA) directions was smaller than that of the periodontally diseased samples, suggesting that PH is characterised by a uniform bacterial composition and that the bacterial composition of periodontally diseased samples is much more diverse. The relative bacterial load could distinguish between AgP and ChP., Conclusion: An SVC classifier using a panel of 40 bacterial species was able to distinguish between PH, AgP in young individuals and ChP., (© 2017 FDI World Dental Federation.)

Aim: To perform a thorough characterization of the subgingival microbiota of shallow, moderate and deep sites in subjects with chronic periodontitis (ChP)., Material and Methods: Subgingival samples were collected from subjects with ChP (n = 3/category of probing depth: ≤3, 4-6 and ≥7 mm) and periodontal health (PH). Individual samples were submitted to 16S rDNA high- throughput sequencing and the analysis was made using mothur and R packages., Results: Nine subjects with ChP and seven with PH were included and 101 samples were evaluated. Thirteen phyla, 118 genera and 211 OTUs were detected. Taxa from Chloroflexi and Spirochaetes phyla were associated with initial stages of disease. Fretibacterium, Eubacterium[XI][G-6], Desulfobulbus, Peptostreptococcaceae[XI][G-1] and [G-3], Bacteroidetes[G-3], Bacteroidaceae[G-1] genera and Filifactor alocis, Fretibacterium fastidiosum, Johnsonella spHOT166, Peptostreptococcaceae[XIII][G-1]HOT113, Porphyromonas endodontalis and Treponema sp. HOT258, which are not conventionally associated with disease, increased with the deepening of the pockets and/or were elevated in ChP; while Streptococcus, Corynebacterium and Bergeyella genera were associated with PH (p < .05)., Conclusion: Striking differences were observed between the microbiota of shallow and moderate/deep sites, but not between moderate and deep sites in ChP subjects. Differences between shallow sites in PH and ChP were also observed. The characterized microbiota included known oral microorganisms and newly identified periodontal taxa, some of them not-yet-cultured., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: To evaluate the clinical effects of different dosages of metronidazole (MTZ) and durations of MTZ + amoxicillin (AMX) in the treatment of generalized chronic periodontitis (GChP)., Material and Methods: Subjects with severe GChP were randomly assigned to receive scaling and root planing (SRP)-only, or combined with 250 or 400 mg of MTZ + AMX (500 mg) thrice a day (TID), for 7 or 14 days. Subjects were monitored for 1 year., Results: One hundred and nine subjects were enrolled. At 1 year, 61.9% and 63.6% of the subjects receiving AMX + 250 or 400 mg of MTZ for 14 days, respectively, reached the clinical endpoint for treatment (≤4 sites with probing depth ≥5 mm), against 31.8% of those taking 250 or 400 mg of MTZ for 7 days (p < .05) and 13.6% of those receiving SRP-only (p < .05). Fourteen days of MTZ + AMX was the only significant predictor of subjects reaching the clinical endpoint at 1 year (OR, 5.26; 95% CI, 2.3-12.1, p = .0000). The frequency of adverse events did not differ among treatment groups (p > .05)., Conclusion: The adjunctive use of 400 or 250 mg of MTZ plus 500 mg of AMX/TID/14 days offers statistically significant and clinically relevant benefits over those achieved with SRP alone in the treatment of severe GChP. The added benefits of the 7-days regimen in this population were less evident. (ClinicalTrials.gov NCT02735395)., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The aim of this randomized, single blinded clinical trial was to evaluate the effect of a pre-procedural mouthwash containing cetylpyridinium chloride (CPC), zinc lactate (Zn) and sodium fluoride (F) in the reduction of viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler. Sixty systemically healthy volunteers receiving dental prophylaxis were randomly assigned to one of the following experimental groups (15 per group): (i) rinsing with 0.075% CPC, 0.28% Zn and 0.05% F (CPC+Zn+F), (ii) water or (iii) 0.12% chlorhexidine digluconate (CHX), and (iv) no rinsing. Viable bacteria were collected from different locations in the dental office on enriched TSA plates and anaerobically incubated for 72 hours. The colonies were counted and species were then identified by Checkerboard DNA-DNA Hybridization. The total number of colony-forming units (CFUs) detected in the aerosols from volunteers who rinsed with CPC+Zn+F or CHX was statistically significantly (p<0.05) lower than of those subjects who did not rinse or who rinsed with water. When all locations were considered together, the aerosols from the CPC+Zn+F and CHX groups showed, respectively, 70% and 77% fewer CFUs than those from the No Rinsing group and 61% and 70% than those from the Water group. The mean proportions of bacterial species from the orange complex were statistically significantly (p<0.05) lower in aerosols from the CPC+Zn+F and CHX groups compared with the others two groups. In conclusion, the mouthwash containing CPC+Zn+F, is effective in reducing viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler.

Objective: This study evaluated the influence of glycemic control on the levels and frequency of subgingival periodontal pathogens in patients with type 2 diabetes mellitus (DM) and generalized chronic periodontitis (ChP)., Material and Methods: Fifty-six patients with generalized ChP and type 2 DM were assigned according to the levels of glycated hemoglobin (HbA1c) into one of the following groups: HbA1c<8% (n=28) or HbA1c≥8% (n=28). Three subgingival biofilm samples from sites with probing depth (PD)<5 mm and three samples from sites with PD≥5 mm were analyzed by quantitative Polymerase Chain Reaction (PCR) for the presence and levels of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Eubacterium nodatum, Parvimona micra, Fusobacterium nucleatum ssp. and Prevotella intermedia., Results: The mean counts of F. nucleatum ssp. were statistically significantly higher in the sites with PD≥5 mm of the HbA1c≥8% group (p<0.05). Frequencies of detection of T. forsythia, E. nodatum, P. micra and F. nucleatum ssp. were all higher in the sites with PD≥5 mm of the patients with HbA1c≥8%, compared with those of patients with HbA1c<8% (p<0.05). Frequency of detection of P. intermedia was higher in the sites with PD<5 mm of the patients with HbA1c≥8% than those of the patients with HbA1c<8% (p<0.05)., Conclusions: Poor glycemic control, as indicated by HbA1c≥8%, is associated with increased levels and frequencies of periodontal pathogens in the subgingival biofilm of subjects with type 2 DM and ChP.

Aim: To evaluate whether obesity affects the subgingival microbial composition of patients with periodontal health or chronic periodontitis (CP)., Materials and Methods: Based on periodontal parameters, body mass index and waist-hip ratio, 166 patients were allocated into one of the following groups: Normal weight (NW) patients with periodontal health (n = 44), NW patients with CP (n = 40), obese patients with periodontal health (n = 40) and obese patients with CP (n = 42). Six subgingival biofilm samples per patient were analysed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization., Results: Obese patients with CP harboured higher levels and/or higher proportions of several periodontal pathogens than those with NW and CP, including Aggregatibacter actinomycetemcomitans, Eubacterium nodatum, Fusobacterium nucleatum ss vincentii, Parvimonas micra, Prevotella intermedia, Tannerella forsythia, Prevotella melaninogenica and Treponema socranskii. The proportions of most of these pathogens, as well Campylobacter rectus and Eikenella corrodens, were more increased in the diseased sites of the obese patients than in those with NW. Furthermore, the healthy sites of the obese patients, presenting or not CP, also exhibited higher proportions of some of the pathogens than patients with NW., Conclusions: Obesity is associated with increased levels and proportions of periodontal pathogens, especially in patients with CP., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: There is currently no consensus regarding microorganisms that may be considered true peri-implant pathogens. Therefore, the aim of this systematic review is to determine the weight of evidence for microorganisms related to peri-implantitis based on results of association studies., Methods: This review was performed following the Preferred Reporting Items for Systematic Reviews and MetaAnalyses (PRISMA). Two independent researchers searched PubMed/Medline, Embase, and Cochrane Library databases up to August 4, 2015, for studies comparing microbiologic outcomes of subgingival biofilm samples from healthy implants and implants with peri-implantitis., Results: A total of 799 titles was identified and 11 studies were included in this review. All data were extracted using a predefined form. Microorganisms found in increased count/abundance/frequency in peri-implantitis belonged to Bacteria domain and viruses, and included a total of six bacterial phyla, 17 bacterial genera, 23 bacterial species, and two genera of viruses. The main bacterial species associated with peri-implantitis are recognized as periodontal pathogens., Conclusion: Results of this systematic review suggest moderate evidence supporting association of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia and some evidence supporting association of Prevotella intermedia and Campylobacter rectus with the etiology of peri-implantitis.

Aim: To evaluate the clinical and microbiological effects of irrigation with 10% povidone-iodine (PVP-I) as an adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis (ChP)., Methods: Thirty-six subjects with ChP were randomly assigned to receive SRP + irrigation with saline solution (control group) or SRP + irrigation with 10% PVP-I (test group). Subjects received clinical and microbiological evaluation at baseline, 30 and 90 days post-treatment. Six subgingival plaque samples/subject were collected at each time point and individually analyzed for 40 bacterial species by checkerboard DNA-DNA hybridization., Results: No statistically significant differences were observed between the two groups at 90 days post-treatment. At 30 days, the test group presented with lower levels of Tannerella forsythia and lower proportions of red complex pathogens in comparison to the control group. However, these benefits were not maintained at 90 days., Conclusion: The results of this study were unable to show a significant clinical benefit from the adjunctive use of 10% PVP-I in the treatment of chronic periodontitis. However, the microbiological benefit observed with the use of this substance at 30 days might indicate a role for PVP-I in periodontal care, possibly through its repeated application during supportive therapy., (Copyright© by the International Academy of Periodontology.)

Different mechanisms have been hypothesized to explain the increase in prevalence and severity of periodontitis in older adults, including shifts in the periodontal microbiota. However, the actual impact of aging on the composition of subgingival biofilms remains unclear. In the present article, we provide an overview of the composition of the subgingival biofilm in older adults and the potential effects of age on the oral microbiome. In particular, this review covers the following topics: (i) the oral microbiota of an aging mouth; (ii) the effects of age and time on the human oral microbiome; (iii) the potential impact of inflammaging and immunosenescence in the host-oral microbiota interactions; and (iv) the relationship of the aging oral microbiota and Alzheimer's disease. Finally, we present analyses of data compiled from large clinical studies that evaluated the subgingival microbiota of periodontally healthy subjects and patients with periodontitis from a wide age spectrum (20-83 years of age)., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: The objective of this cross-sectional study was to examine the potential of peri-implant crevicular fluid (PICF) analytes to discriminate between peri-implant health and disease using a multi-biomarker approach., Methods: We collected PICF samples from the mesio-buccal site of every implant (n = 145) from 52 subjects with peri-implantitis and measured the levels of 20 biomarkers using Luminex. We grouped implants and subjects based on the clinical characteristic of the sampled sites and implants into: healthy sites from healthy implants (HH), diseased sites from diseased implants (DD) and healthy sites from diseased implants (HD). The significance of the differences between the HH and DD groups was determined using general linear models controlling for false discovery rate. We used logistic regression to determine the best multi-biomarker models that could distinguish HH from DD subjects and HH from HD subjects., Results: There were statistically significant differences between HH and DD groups for 12/20 biomarkers. Logistic regression resulted in a 6-biomarker model (Flt-3L, GM-CSF, IL-10, sCD40L, IL-17 and TNFα) that discriminated HH from DD subjects (AUC = 0.93) and a 3-biomarker model (IL-17, IL-1ra and vascular endothelial growth factor) that distinguished HH from DD subjects (AUC = 0.90)., Conclusion: PICF biomarkers might help discriminate peri-implant health from disease., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The aim of this study was to assess the changes occurring in subgingival biofilm composition and in the periodontal clinical parameters of subjects with periodontitis and type 2 diabetes mellitus (DM) treated by means of scaling and root planing (SRP) only or combined with systemic metronidazole (MTZ) and amoxicillin (AMX). Fifty-eight subjects were randomly assigned to receive SRP only (n = 29) or with MTZ (400 mg/thrice a day [TID]) and AMX (500 mg/TID) (n = 29) for 14 d. Six subgingival plaque samples/subject were analyzed by checkerboard DNA-DNA hybridization for 40 bacterial species at baseline and 3 mo, 1 y, and 2 y posttherapy. At 2 y posttherapy, the antibiotic-treated group harbored lower mean proportions (5.5%) of red complex pathogens than the control group (12.1%) (P < 0.05). The proportions of the Actinomyces species remained stable in the antibiotic group but showed a statistically significant reduction in the control group from 1 to 2 y in subjects achieving a low risk clinical profile for future disease progression (i.e., ≤4 sites with probing depth [PD] ≥5 mm). The test group also had a lower mean number of sites with PD ≥5 mm (3.5 ± 3.4) and a higher percentage of subjects reaching the low risk clinical profile (76%) than the control group (14.7 ± 13.1 and 22%, respectively) (P < 0.05) at 2 y posttreatment. MTZ + AMX intake was the only significant predictor of subjects achieving the low risk at 2 y (odds ratio, 20.9; P = 0.0000). In conclusion, the results of this study showed that the adjunctive use of MTZ + AMX improves the microbiological and clinical outcomes of SRP in the treatment of subjects with generalized chronic periodontitis and type 2 DM up to 2 y (ClinicalTrials.gov NCT02135952)., (© International & American Associations for Dental Research 2016.)

In recent years, several new periodontal taxa have been associated with the etiology of periodontitis. A recent systematic review provides further support for the pathogenic role of 17 species/phylotypes. Thus, the aim of this study was to assess the prevalence and levels of these species in subjects with generalized chronic periodontitis (GChP; n = 30), generalized aggressive periodontitis (GAgP; n = 30), and periodontal health (PH; n = 30). All subjects underwent clinical and microbiological assessment. Nine subgingival plaque samples were collected from each subject and analyzed for their content of 20 bacterial species/phylotypes through the RNA-oligonucleotide quantification technique. Subjects from the GChP and GAgP groups presented the highest mean values for all clinical parameters in comparison with the PH group (P < 0.05). Subjects with GChP and GAgP showed significantly higher mean levels of Bacteroidetes sp. human oral taxon (HOT) 274, Fretibacterium sp. HOT 360, and TM7 sp. HOT 356 phylotypes, as well as higher mean levels of Filifactor alocis, Fretibacterium fastidiosum, Porphyromonas gingivalis, Tannerella forsythia, and Selenomonas sputigena species than PH subjects (P < 0.05). GAgP subjects presented higher mean levels of TM7 sp. HOT 356 and F. alocis than GChP subjects (P < 0.05). A significantly higher mean prevalence of Bacteroidales sp. HOT 274, Desulfobulbus sp. HOT 041, Fretibacterium sp. HOT 360, and Fretibacterium sp. HOT 362 was found in subjects with GChP and GAgP than in PH subjects. Mean levels of P. gingivalis (r = 0.68), T. forsythia (r = 0.62), F. alocis (r = 0.51, P = 0.001), and Fretibacterium sp. HOT 360 (r = 0.41) were correlated with pocket depth (P < 0.001). In conclusion, Bacteroidales sp. HOT 274, Desulfobulbus sp. HOT 041, Fretibacterium sp. HOT 360, Fretibacterium sp. HOT 362, and TM7 sp. HOT 356 phylotypes, in addition to F. alocis, F. fastidiosum, and S. sputigena, seem to be associated with periodontitis, and their role in periodontal pathogenesis should be further investigated., (© International & American Associations for Dental Research 2016.)

Objective The aim of this study was to evaluate the association of Porphyromonas endodontalis, Filifactor alocis and Dialister pneumosintes with the occurrence of periodontitis. Material and Methods Thirty subjects with chronic periodontitis (ChP) and 10 with periodontal health (PH) were included in the study. Nine subgingival biofilm samples were collected as follows: i) PH group - from the mesial/buccal aspect of each tooth in two randomly chosen contralateral quadrants; ii) ChP group - from three sites in each of the following probing depth (PD) categories: shallow (≤3 mm), moderate (4-6 mm) and deep (≥7 mm). Checkerboard DNA-DNA hybridization was used to analyze the samples. Results We found the three species evaluated in a higher percentage of sites and at higher levels in the group with ChP than in the PH group (p<0.05, Mann-Whitney test). We also observed these differences when the samples from sites with PD≤4 mm or ≥5 mm of subjects with ChP were compared with those from subjects with PH (p<0.05, Mann-Whitney test). In addition, the prevalence and levels of D. pneumosintes, and especially of F. alocis were very low in healthy subjects (0.12x105 and 0.01x105, respectively). Conclusion F. alocis and D. pneumosintes might be associated with the etiology of ChP, and their role in the onset and progression of this infection should be further investigated. The role of P. endodontalis was less evident, since this species was found in relatively high levels and prevalence in the PH group.

Introduction: Oral-derived bacteremia may occur after several dental procedures and routine daily activities. Some conditions of the oral cavity may favor episodes of bacteremia. This would be the case of patients with diabetes mellitus and periodontitis, who exhibit exacerbated gingival inflammation and may be more prone to developing oral-derived bacteremia. , Objective: To compare the effectiveness of an independent culture method (quantitative real-time PCR- qCR) and the most commonly used method (BacT-ALERT 3D®) for the diagnosis of bacteremia. , Materials and Methods: Blood samples were drawn from subjects with type 2 diabetes mellitus and chronic periodontitis before and after apple chewing. Samples were processed by an automated blood culture system (BacT-ALERT 3D®) monitored for 15 days with suitable subculture of positive cultures. In parallel, whole DNA from blood samples was purified using a commercial kit and screened by qPCR using a universal primer set of16S rDNA for bacteria detection. , Results: Blood cultures taken before apple chewing were shown to be negative by the two diagnostic methods. After chewing, two samples (11%) showed bacterial growth by BacT-ALERT 3D® whereas qPCR did not detect the presence of bacteria in any sample. , Conclusions: qPCR did not show greater effectiveness than the BacT-ALERT 3D® in the detection of bacteremia of oral origin.

Background and Objective: To compare the subgingival microbial diversity between non-HIV-infected and HIV-infected individuals with chronic periodontitis using denaturing gradient gel electrophoresis (DGGE)., Material and Methods: Thirty-two patients were selected: 11 were HIV-infected and 21 were non-HIV-infected, and all had chronic periodontitis. Periodontal measurements included probing depth, clinical attachment level, visible supragingival biofilm and bleeding on probing. Subgingival biofilm samples were collected from periodontal sites (50% with probing depth ≤ 4 mm and 50% with probing depth ≥ 5 mm) and whole-genomic-amplified DNA was obtained. The DNA samples were subjected to amplification of a 16S rRNA gene fragment using universal bacterial primers, followed by DGGE analysis of the amplified gene sequences., Results: The non-HIV-infected group presented higher mean full-mouth visible supragingival biofilm (p = 0.004), bleeding on probing (p = 0.006), probing depth (p < 0.001) and clinical attachment level (p = 0.001) in comparison with the HIV-infected group. DGGE analysis revealed 81 distinct bands from all 33 individuals. Banding profiles revealed a higher diversity of the bacterial communities in the subgingival biofilm of HIV-infected patients with chronic periodontitis. Moreover, cluster and principal component analyses demonstrated that the bacterial community profiles differed between these two conditions. High interindividual and intra-individual variability in banding profiles were observed for both groups., Conclusion: HIV-infected patients with chronic periodontitis present greater subgingival microbial diversity. In addition, the bacterial communities associated with HIV-infected and non-HIV-infected individuals are different in structure., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: This study compared the efficacy of two oral hygiene regimens in reducing oral malodour and the proportions of bacterial species involved in the production of volatile sulphur compounds., Material and Methods: Seventy subjects who participated in a halitosis-induction phase and achieved an organoleptic score of ≥ 3.0 [time point 0 (T0)] randomised into two groups: brushing with regular fluoride toothpaste alone (control group) or brushing with regular fluoride toothpaste followed by rinsing with a 0.075% cetylpyridinium chloride (CPC) mouthwash (CPC group). Subjects followed their assigned oral hygiene regimen for 21 days. Then, they underwent an organoleptic examination and measurement of volatile sulphur compounds (VSCs) using a portable gas chromatograph, 12 hours after their last oral hygiene procedure (T1) and 4 hours after an on-site oral hygiene (T2). Microbiological samples (supragingival biofilm, tongue coating and saliva) were analysed using checkerboard DNA-DNA hybridisation., Results: Both therapies statistically significantly improved the organoleptic scores (P < 0.05), but the VSC levels and/or concentrations were reduced only in the CPC group (P < 0.05). In subjects rinsing with CPC, oral malodour scores were reduced by 49% at the 4-hour assessment (T2) compared with those not rinsing (P < 0.05). Red-complex pathogens were reduced more effectively in the CPC group than in the control group., Conclusions: Brushing followed by rinsing with a 0.075% CPC mouthwash provided statistically significantly greater reductions in oral malodour, measured organoleptically and instrumentally, and in the proportions of red-complex species when compared with brushing alone., (© 2015 FDI World Dental Federation.)

Purpose: The aim of this prospective controlled study was to evaluate the influence of osteopenia on the levels of osteoclastogenesis-related factors in the peri-implant crevicular fluid (PICF) and on the clinical parameters of immediately loaded implants., Materials and Methods: This study included 24 patients who received at least two implants in the mandible, with restorations delivered 48 hours after implant placement. Patients were divided into control (n = 11) and osteopenia (n = 13) groups. Seven days after implant placement (baseline) and 4 months after implant placement, PICF samples were obtained, and clinical parameters (Plaque Index, Gingival Index, bleeding on probing, suppuration, probing depths, clinical attachment levels) were measured. A commercially available enzyme-linked immunosorbent assay was used to analyze PICF samples for levels of soluble receptor activator of nuclear factor of κB ligand (sRANKL) and osteoprotegerin (OPG). At the 4-month follow-up visit, the implant-supported restorations were removed and periapical radiographs were acquired to evaluate bone loss around the implants., Results: Eighty-eight immediately loaded implants were included in this study (38 in the control group, 50 in the osteopenia group). The RANKL and OPG levels, the RANKL/OPG ratio, and the clinical parameters were similar between the groups at both time points. However, the levels of these factors in PICF differed significantly between baseline and 4 months after surgery., Conclusion: Within the limitations of this short-term study, it can be concluded that osteopenia does not influence the PICF levels of osteoclastogenesis-related factors in immediately loaded implants after 4 months of loading.

Aim: The aim of this study was to assess the effect of systemic antibiotic therapy on the treatment of aggressive periodontitis (AgP)., Methods: This study was conducted and reported in accordance with the PRISMA statement. The MEDLINE, EMBASE and CENTRAL databases were searched up to June 2014 for randomized clinical trials comparing the treatment of subjects with AgP with either scaling and root planing (SRP) alone or associated with systemic antibiotics. Bayesian network meta-analysis was prepared using the Bayesian random-effects hierarchical models and the outcomes reported at 6-month post-treatment., Results: Out of 350 papers identified, 14 studies were eligible. Greater gain in clinical attachment (CA) (mean difference [MD]: 1.08 mm; p < 0.0001) and reduction in probing depth (PD) (MD: 1.05 mm; p < 0.00001) were observed for SRP + metronidazole (Mtz), and for SRP + Mtz + amoxicillin (Amx) (MD: 0.45 mm, MD: 0.53 mm, respectively; p < 0.00001) than SRP alone/placebo. Bayesian network meta-analysis showed additional benefits in CA gain and PD reduction when SRP was associated with systemic antibiotics., Conclusions: SRP plus systemic antibiotics led to an additional clinical effect compared with SRP alone in the treatment of AgP. Of the antibiotic protocols available for inclusion into the Bayesian network meta-analysis, Mtz and Mtz/Amx provided to the most beneficial outcomes., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The aim of this study was to compare subgingival bacterial recolonization patterns after scaling and root planing in current smokers and non-smokers., Methods: 15 smokers and 15 non-smokers with chronic periodontitis received scaling and root planing in six visits lasting one hour each, over a period of 21 days. Clinical monitoring was performed at baseline and 180 days, and microbiological monitoring was performed at baseline, immediately after scaling and root planing (Day 0) and at 42, 63 and 180 days post-therapy. Subgingival plaque samples were analysed by checkerboard DNA-DNA hybridization., Results: An improvement in clinical condition was observed for smokers and non-smokers; however, non-smokers showed a greater reduction in mean clinical attachment level in intermediate sites in comparison with smokers (p < 0.05). At Day 0, there was a significant reduction in the mean counts of the three pathogens from the red complex, Eubacterium nodatum and Parvimonas micra only in non-smokers (p < 0.05). There was a significant increase in the proportion of host-compatible species in non-smokers and smokers from baseline to 180 days post-therapy (p < 0.05). However, a significant decrease in the pathogenic species was observed only in non-smokers., Conclusions: Smokers were more susceptible to the re-establishment of a pathogenic subgingival biofilm than non-smokers., (© 2015 Australian Dental Association.)

Chronic periodontitis is one of the most prevalent human diseases and is caused by dysbiosis of the subgingival microbiota. Treatment involves primarily mechanical disruption of subgingival biofilms and, in certain cases, adjunctive use of systemic antibiotic therapy. In vitro biofilm models have been developed to study antimicrobial agents targeting subgingival species. However, these models accommodate a limited number of taxa, lack reproducibility, and have low throughput. We aimed to develop an in vitro multispecies biofilm model that mimics subgingival plaque, to test antimicrobial agents. Biofilms were cultivated using the Calgary Biofilm Device and were exposed to amoxicillin (AMX), metronidazole (MTZ), azithromycin (AZM), and AMX-MTZ at four different concentrations for 12, 24, or 36 h. Chlorhexidine (CHX) (0.12%) was used as the positive control. The compositions of the biofilms were analyzed by checkerboard DNA-DNA hybridization, and the percent reduction in biofilm metabolic activity was determined using 2,3,5-triphenyltetrazolium chloride and spectrophotometry. Thirty-five of the 40 species used in the inoculum were consistently recovered from the resulting in vitro biofilms. After 36 h of exposure at the 1:27 dilution, AMX-MTZ reduced metabolic activity 11% less than CHX (q = 0.0207) but 54% more than AMX (q = 0.0031), 72% more than MTZ (q = 0.0031), and 67% more than AZM (q = 0.0008). Preliminary evidence of a synergistic interaction between AMX and MTZ was also observed. In summary, we developed reproducible biofilms with 35 subgingival bacterial species, and our results suggested that the combination of AMX and MTZ had greater antimicrobial effects on these in vitro multispecies biofilms than expected on the basis of the independent effects of the drugs., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Objective: The aim of this systematic review was to compare the clinical effectiveness of systemic antibiotics administered in the active stage of periodontal treatment or after the healing phase., Material and Methods: An electronic search was performed in the databases EMBASE, MEDLINE and Cochrane Central Register of Controlled Trials (CENTRAL), in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) statement. A manual search of the reference list of selected studies and of review articles was also performed up to November 2013. Randomized Clinical Trials (RCT) that evaluated the systemic administration of antibiotics as adjuvants to scaling and root planning (SRP) at different phases of periodontal treatment were included. Systematic reviews and studies that evaluated subjects with systemic diseases and those that used subantimicrobial doses of antibiotics were excluded., Results: The initial search identified 1,039 articles, of which seven were selected, and only one met the inclusion criteria. This study showed that subjects taking metronidazole and amoxicillin at the initial phase of treatment exhibited statistically significantly greater reduction in pocket depth and gain in clinical attachment level in initially deep sites (PD≥7 mm) than subjects taking antibiotics after healing (p<0.05). This comparison was conducted 2 months after antibiotic intake, at the healing phase., Conclusion: To date, only one short-term RCT has directly compared different moments of systemic antibiotics administration, as adjuncts to SRP, in the treatment of periodontitis. Although the results of this study suggested some benefits for antibiotics intake during the active phase of therapy, these findings need to be confirmed by larger placebo-controlled randomized clinical trials with longer follow-up periods.

Host DNA may adversely affect metagenomic studies focusing on the prokaryotic microbiota. This study compared the levels of host DNA in subgingival plaque collected by paper points and curette, using quantitative PCR. Lower proportions of host DNA and higher proportions of bacterial DNA were recovered from samples collected with curettes., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Class II malocclusions are generally characterized by mandible retrusion. For this reason, forward bite jumping appliances, also known as functional appliances were originally designed to enhance mandibular forward projection. However, there is still insufficient evidence to support the effectiveness, predictability and stability of functional appliances in modifying mandibular growth. This article was aimed at presenting evidences and hypotheses that mandibular growth may be enhanced through the use of adjunctive methods in conjunction with functional appliances. In formulating our hypothesis, we considered relevant data, mostly derived from animal studies, concerning alternative methods, such as low-intensity ultrasound and light-emitting diode, as well as their related cellular and molecular mechanisms. According to the evidences covered in this article, we suggest that both methods are potentially effective, and theoretically able to act in synergistic way to enhance functional appliances treatment on mandibular and condylar additional growth. The rationale for the use of these methods as adjunctive therapies for mandibular underdevelopment is attributed to their abilities on stimulating angiogenesis, cell differentiation, proliferation, and hypertrophy, as well as enhancing matrix production and endochondoral bone formation, especially on the condyle of growing animals. This article also proposed a study design which would be able to either prove or refute our hypothesis. If ratified, it would represent a significant scientific accomplishment which provides support for further investigations to be carried out on well-designed clinical trials., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Aim: To assess the failure and bone-to-implant contact rate of dental implants placed on osteoporotic subjects., Methods: Extensive examination strategies were created to classify studies for this systematic review. MEDLINE (via PubMed) and EMBASE database were examined for studies in English up to and including May 2014. The examination presented a combination of the MeSH words described as follow: "osteoporosis" or "osteopenia" or "estrogen deficiency" AND "implant" or "dental implant" or "osseointegration". Assessment of clinical and/or histological peri-implant conditions in osteoporosis subjects treated with titanium dental implants. The examination included a combination of the MeSH terms described as follow: "osteoporosis" or "osteopenia" or "estrogen deficiency" AND "implant" or "dental implant" or "osseointegration"., Results: Of 943 potentially eligible articles, 12 were included in the study. A total of 133 subjects with osteoporosis, 73 subjects diagnosed with osteopenia and 708 healthy subjects were assessed in this systematic review. In these subjects were installed 367, 205, 2981 dental implants in osteoporotic, osteopenic and healthy subjects, respectively. The failure rate of dental implant was 10.9% in osteoporotic subjects, 8.29% in osteopenic and 11.43% in healthy ones. Bone-to-implant contact obtained from retrieved implants ranged between 49.96% to 47.84%, for osteoporosis and non-osteoporotic subjects., Conclusion: Osteoporotic subjects presented higher rates of implant loss, however, there is a lower evidence to strengthen or refute the hypothesis that osteoporosis may have detrimental effects on bone healing. Consequently, final conclusions regarding the effect of osteoporosis in dental implant therapy cannot be made at this time. There are no randomized clinical trial accessible for evaluation and the retrospective nature of the evaluated studies shall be taken in account when interpreting this study.

Background and Objective: Microbiological and immunological hypotheses have been raised to explain the differences in the clinical manifestations of aggressive periodontitis and chronic periodontitis. However, studies comparing the cytokine/chemokine profiles in gingival crevicular fluid between these two clinical conditions have so far not been compiled. This systematic review aimed to answer the following question: "Do subjects with aggressive periodontitis and chronic periodontitis have a different profile of cytokines/chemokines in the gingival crevicular fluid?", Material and Methods: An electronic database search of MEDLINE/PubMed and Embase was performed from 1990 up to and including August 2013, using MeSH terms and other keywords. Titles and abstracts were screened and the papers that satisfied eligibility criteria were assessed., Results: Of 1954 titles, 17 studies reporting the levels of 21 different cytokines/chemokines were included. Most studies did not find any significant differences in the gingival crevicular fluid levels of cytokines/chemokines between aggressive periodontitis and chronic periodontitis. Some studies demonstrated that the levels of specific proinflammatory and anti-inflammatory cytokines/chemokines were higher (n = 5) and lower (n = 3), respectively, in aggressive periodontitis than in chronic periodontitis. The studies differed in the manner in which they reported the results (e.g. concentrations or total amounts). It was not clear in some studies whether the sample sites from both groups were matched for disease severity. Some studies did not take into account confounders, such as smoking., Conclusion: The current weight of evidence is not sufficient to prove that there are distinct gingival crevicular fluid cytokine/chemokine profiles for patients with aggressive periodontitis and chronic periodontitis., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Despite the fact that several clinical studies have shown additional benefits when certain systemic antibiotics are used as adjuncts to periodontal treatment, clear guidelines for the use of these agents in the clinical practice are not yet available. Basic questions concerning the use of systemic antibiotics to treat periodontitis remain unanswered, such as: which drug(s) should be used; which patients would most benefit from treatment; which are the most effective protocols (i.e. doses and durations); and in which phase of the mechanical therapy should the drug(s) be administered? Although not all of those questions have been directly addressed by controlled randomized clinical trials, recent concepts related to the ecology of periodontal diseases, as well as the major advances in laboratory and clinical research methods that have occurred in the past decade, have significantly broadened our knowledge in this field. This article endeavored to provide a 'state of the art' overview on the use of systemic antibiotics in the treatment of periodontitis, based on the most recent literature on the topic as well as on a compilation of data from studies conducted at the Center of Clinical Trials at Guarulhos University (São Paulo, Brazil) from 2002 to 2012., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)