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Objective: This study aimed to standardize a quantitative polymerase chain reaction (qPCR)-based test to identify and quantify the uncultivable bacteria associated with periodontitis., Methods: The standardization of qPCR, the curves for the quantification of Eubacterium saphenum, Eubacterium brachy, Desulfobulbus oralis, and Filifactor alocis were developed by cloning the 16 S rRNA target gene fragment, using the GEMTEasy vector. The qPCRs were validated in 55 subgingival biofilm clinical samples, from different stages of periodontitis and from periodontally healthy/gingivitis individuals, which were previously evaluated by next-generation sequencing (NGS). The results obtained by the two methods were compared by the concordance of Cohen's Kappa index, and sensitivity, specificity, receiver operating characteristic (ROC) curve, and predictive values were established., Results: obtained by the two methods were compared using the concordance of Cohen's Kappa index, and sensitivity, specificity, predictive values, and ROC curves were generated. The qPCR test was standardized with efficiencies between 90% and 100% and R 2 : 0.997-0.999. Concordance between the qPCR and NSG was moderate to F. alocis (agreement 78.2%; kappa 0.56, p < 0.05) and fair to the other microorganisms (agreement 67.27%-72.73; kappa 0.37-0.38, p < 0.05). qPCR exhibited a high sensitivity (82.2-100%) and specificity (100%) for E. brachy, E. saphenum, and F. alocis. Sensitivity was lower to D. oralis. Conversely, qPCR demonstrated higher sensitivity to E. saphenum than NSG (100 vs. 68.1)., Conclusions: The uncultivable microorganisms associated with periodontitis, D. oralis, E. brachy, E. saphenum, and F. alocis can be detected and quantified with the newly developed and validates qPCR test., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Aim: To characterize the subgingival microbiome in subjects with different periodontal health statuses., Materials and Methods: In this cross-sectional observational study, subgingival samples were harvested from Spanish subjects with different periodontal health statuses, based on the 2018 Classification of Periodontal and Peri-Implant Diseases and Conditions. Samples were processed using high-throughput sequencing technologies (Illumina MiSeq). Taxa differentially abundant were identified using Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC). α- and β-diversity metrics were calculated using q2-diversity in QIIME2. The analyses were adjusted for age, gender and smoking status., Results: The identified subgingival microbiome showed statistically significant differences among subjects, categorized into periodontal health, gingivitis and stages I-II and III-IV periodontitis (p < .05). In patients with severe (stages III-IV) periodontitis, the genera Filifactor and Fretibacterium were detected 24 times more frequently than in periodontally healthy subjects. Similarly, the genera Porphyromonas, Prevotella and Tannerella were detected four times more frequently (p < .05). The genera Granulicatella, Streptococcus, Paracoccus, Pseudomonas, Haemophilus, Actinobacteria, Bergeyella and Capnocytophaga were significantly associated with healthier periodontal status (p < .05)., Conclusions: Significant differences were detected in the subgingival microbiome among periodontal health, gingivitis and stages I-II or III-IV periodontitis, suggesting overlapping, yet distinguishable microbial profiles., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: Culturable and unculturable microorganisms have been associated with periodontitis. Their differential proportions and composition have not been evaluated by their severity and complexity defined by stages in the 2018 AAP-EEP classification., Methods: One hundred eighty subgingival biofilm samples were collected in Spain and Colombia from subjects categorized as health/gingivitis: periodontitis stages I/II periodontitis stages III/IV. Target culturable microorganisms (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Treponema denticola, and Eubacterium nodatum) and target unculturable microorganisms (Filifactor alocis, Eubacterium saphenum, Eubacterium brachy, Desulfobulbus oralis) were evaluated by quantitative PCR analysis. In addition, their differences and association with periodontal status were analyzed by ANCOVA and logistic regression models once adjusted to age, current smoking, and country., Results: P. gingivalis was significantly associated with periodontitis stages I/II, OR 2.44 (CI 95% 1.08-5.47) and stages III/V, OR 6.43 (CI 95% 2.43-16.9). T forsythia, OR 7.53 (CI 95% 2.07-27.4); D. oralis, OR 5.99 (CI 95% 2.71-13.23); F. alocis, OR 10.9 (CI 95% 4.56-23.2); E. brachy, 3.57 (CI 95% 1.40-9.11); and E. saphenum, 4.85 (CI 95% 1.99-11.7) were significantly associated only with stages III/IV periodontitis. P. gingivalis evidenced significant differences with the increase in the severity of the periodontal lesion: 2.97 colony forming unit (CFU)/μL (CI 95% 2.32-3.54) health/gingivitis, and 4.66 CFU/μL (CI 95% 4.03-5.30) and 5.90 CFU/μL (CI 95% 5.20-6.48) in stages I/II and III/IV respectively (p < 0.0001). Unculturable microorganisms only evidenced differences in concentration in stages III/IV compared with health-gingivitis (p ≤ 0.001)., Conclusion: Culturable and unculturable are strongly associated with stages III/IV periodontitis. Classic culturable microorganisms are more sensitive to differentiate between stages of periodontitis in the quantitative analysis., Clinical Relevance: Future interventional studies of periodontal disease should include Filifactor alocis, Eubacterium saphenum, Eubacterium brachy, and Desulfobulbus oralis as possible markers of therapy response and as indicators of progressive disease., (© 2023. The Author(s).)

No microbiological criteria were included in the 2018 EFP-AAP classification of periodontal diseases that could be used to differentiate between stages and grades. Furthermore, differences in the subgingival microbiome depending on stage and grade have not been established. Sixty subgingival biofilm samples were collected in Spain (n = 30) and Colombia (n = 30) from three distinct patient categories: those with periodontal health/gingivitis (n = 20), those with stage I-II periodontitis (n = 20), and those with stage III-IV periodontitis (n = 20). Patients were evaluated by 16S rRNA gene amplification sequencing. Amplicon sequence variants were used to assign taxonomic categories compared to the Human Oral Microbiome Database (threshold ≥97% identity). Alpha diversity was established by Shannon and Simpson indices, and principal coordinate analysis, ANOSIM, and PERMANOVA of the UNIFRAC distances were performed using QIIME2. Although differences in the alpha diversity were observed between samples according to country, Filifactor alocis, Peptostreptococcaceae [XI][G-4] bacterium HMT 369, Fretibacterium fastidiosum, Lachnospiraceae [G-8] bacterium HMT 500, Peptostreptococcaceae [XI][G-5] [Eubacterium] saphenum, Peptostreptococcus stomatis, and Tannerella forsythia were associated with periodontitis sites in all stages. However, only F. alocis, Peptostreptococcaceae [XI][G-4] bacterium HMT 369, Peptostreptococcaceae [XI][G-9] [Eubacterium] brachy, Peptostreptococcaceae [XI][G-5] [Eubacterium] saphenum, and Desulfobulbus sp. HMT 041 were consistent in stage III-IV periodontitis in both countries. Porphyromonas gingivalis and Tannerella forsythia were differentially expressed in severe lesions in the countries studied. Although some non-cultivable microorganisms showed differential patterns between the different stages of periodontitis, they were not the same in the two countries evaluated. Further studies using larger samples with advanced next-generation techniques for high-throughput sequencing of phyla and non-cultivable bacteria within the subgingival microbiome could provide more insight into the differences between stages of periodontitis., Competing Interests: The authors have declared that no competing interests exist.

Background: The objective was to qualitatively and quantitatively describe the subgingival cultivable bacteria in Albanian subjects and to compare it with a similar Spanish population., Materials and Methods: Consecutive patients, diagnosed as periodontitis in stages I-II or III-IV, and as periodontally healthy or with gingivitis, were studied clinically and microbiologically by means of microbiological culture, including total anaerobic counts, proportions, and frequency of detection of target species. Outcome variables were analysed by Mann-Whitney, Kruskal-Wallis, ANOVA, ANCOVA and Chi-square tests., Results: In this cross-sectional study, 83 (Albania) and 90 (Spain) subjects were included. No statistically significant differences were observed between test and control populations regarding demographic variables or smoking habit. Significantly higher total anaerobic counts in the Albanian population (p = 0.022) were observed, especially in the periodontal health/gingivitis group (p = 0.001). In the test population, the proportions of the cultivable bacteria of Fusobacterium nucleatum were significantly lower in both the healthy/gingivitis (p = 0.022) and stages I-II periodontitis (p = 0.034) groups., Conclusions: The subgingival cultivable bacteria in both periodontitis and non-periodontitis subjects from Albania showed significantly higher total anaerobic counts and lower proportions of the cultivable bacteria of F. nucleatum than a similar population of subjects from Spain., (© 2022. The Author(s).)

Objectives: To evaluate the efficacy of a 0.03% chlorhexidine (CHX) and 0.05% cetylpyridinium chloride (CPC) mouth rinse, as an adjunct to professional plaque removal (PPR) and mechanical hygiene, in the treatment of peri-implant mucositis (PiM) and gingivitis., Material and Methods: Patients displaying PiM in, at least, one implant were included in this randomized, double-blinded, clinical trial. Subjects received PPR (at baseline and 6-month visits) and were instructed to rinse, twice daily, during 1 year with the tested mouth rinse or a placebo. Clinical and patient-reported outcomes were recorded at baseline and 6 and 12 months., Results: Fifty-four patients were included in the study and 46 attended the final visit. In the teeth and implants with inflammation, a higher reduction in BOP was observed in the test group. Statistically significant differences between groups were only observed in the lingual sites of the teeth with gingivitis (mean difference = 11.96%; 95% confidence interval [1.09; 22.83]; p = 0.03). Overall, compliance and satisfaction were good, even though staining were higher for the test group (p < 0.05)., Conclusions: The combined use of mechanical debridement with a 0.03% CHX and 0.05% CPC mouth rinse may have adjunctive benefits in the management of gingivitis, and it is associated with a higher degree of staining., Clinical Relevance: The control of gingivitis can be improved, after professional mechanical debridement, with toothbrushing and the supplementary use of a 0.03% CHX and 0.05% CPC mouth rinse at home., Clinical Trial Registration Number: NCT03533166.

Background: The recently introduced 2017 World Workshop on the classification of periodontitis, incorporating stages and grades of disease, aims to link disease classification with approaches to prevention and treatment, as it describes not only disease severity and extent but also the degree of complexity and an individual's risk. There is, therefore, a need for evidence-based clinical guidelines providing recommendations to treat periodontitis., Aim: The objective of the current project was to develop a S3 Level Clinical Practice Guideline (CPG) for the treatment of Stage I-III periodontitis., Material and Methods: This S3 CPG was developed under the auspices of the European Federation of Periodontology (EFP), following the methodological guidance of the Association of Scientific Medical Societies in Germany and the Grading of Recommendations Assessment, Development and Evaluation (GRADE). The rigorous and transparent process included synthesis of relevant research in 15 specifically commissioned systematic reviews, evaluation of the quality and strength of evidence, the formulation of specific recommendations and consensus, on those recommendations, by leading experts and a broad base of stakeholders., Results: The S3 CPG approaches the treatment of periodontitis (stages I, II and III) using a pre-established stepwise approach to therapy that, depending on the disease stage, should be incremental, each including different interventions. Consensus was achieved on recommendations covering different interventions, aimed at (a) behavioural changes, supragingival biofilm, gingival inflammation and risk factor control; (b) supra- and sub-gingival instrumentation, with and without adjunctive therapies; (c) different types of periodontal surgical interventions; and (d) the necessary supportive periodontal care to extend benefits over time., Conclusion: This S3 guideline informs clinical practice, health systems, policymakers and, indirectly, the public on the available and most effective modalities to treat periodontitis and to maintain a healthy dentition for a lifetime, according to the available evidence at the time of publication., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The oral flagellated protozoan Trichomonas tenax has been associated with patients with periodontal disease. However, no recent studies have been conducted on the prevalence of T. tenax in Chile. The aim of this study was to determine the presence of T. tenax in patients with periodontal disease, admitted to the Dental Clinic of the University of Antofagasta, Chile, through Polymerase Chain Reaction (PCR) amplification of the beta-tubulin gene., Methods: An observational, cross-sectional study was conducted on 50 patients diagnosed with periodontal disease, 20 of them with gingivitis and 30 with periodontitis. T. tenax was identified by PCR amplification of the beta-tubulin gene. Associations between the protozoan and periodontal disease or the presence of risk factors to establish T. tenax infection were determined using the chi-square test and binary logistic regression analysis., Results: T. tenax was present in 28 out of 50 (56%) of patients with periodontal disease, and was more prevalent when associated with periodontitis (21 out of 30; 70%) than dental plaque-induced gingivitis (7 out of 20; 35%). Non-statistically-significant associations were observed between the presence of T. tenax and age, gender, smoking habit or diabetes. Statistically significant associations were observed between the presence of T. tenax and periodontal disease, and between T. tenax and the Periodontal Screening and Recording (PSR) index., Conclusion: T. tenax showed a high presence in patients with progressive states of periodontal diseases. Consequently, T. tenax detection is strongly recommended in patients with periodontal disease diagnosis and with a PSR index greater than 3.

Aim: The aim of this network meta-analysis (NMA) was to compare the efficacy of different oral hygiene products for chemical biofilm control, in 6-month home-use, randomized clinical trials (RCTs), in terms of changes in gingival index (GI)., Material and Methods: Six-month RCTs assessing GI were identified and screened for inclusion. Relevant information was extracted, and quality and potential risk of bias were estimated. Mean differences between baseline and end were calculated to obtain standardized mean differences (SMDs). NMA protocols were applied to assess direct and indirect comparisons among products using Löe & Silness GI, modified GI and gingival severity index., Results: Fifty-three papers were included, 19 studies for mouth rinses, 32 for dentifrices, comprising data from 5,775 and 2,682 subjects, respectively. When ranking treatments, similar results were observed for all tested dentifrices, with the lowest effect observed for sanguinarine and baking soda. For mouth rinses, essential oils, triclosan-copolymer, chlorhexidine (at concentrations ≥ 0.10%) and cetylpyridinium chloride (>0.05%) demonstrated the greatest effect., Conclusion: Although NMA revealed significant differences when comparing placebo versus some active agents, when comparing among active agents, no differences were found for dentifrices, while mouth rinses containing essential oils showed the greatest effect on GI scores., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: The primary aim of this systematic review was to evaluate the effect of external tooth-bleaching products on plaque indices and to compare it to a placebo or a negative control., Methods: A protocol was developed aimed to answer the next focused question: What is the effect of tooth bleaching compared to no treatment or a placebo, in subjects without periodontitis aged ≥18 years in the levels of plaque and gingivitis? Two electronic databases were used as sources in the search for studies satisfying the inclusion criteria: (a) The National Library of Medicine (MEDLINE via PubMed); (b) Cochrane Central Register of Controlled Trials., Results: Only randomized controlled trials were included. The initial search found 382 potential publications. Seven of them were finally included, and six were used in the meta-analyses. The use of bleaching products showed higher reductions in plaque (n = 6; standardized mean difference [SMD] = 0.47; 95% confidence interval [CI] = 0.06, 0.88; P < 0.001) and gingivitis indices (n = 4; SMD = 0.47; 95% CI = 0.22, 0.73; P < 0.001), when compared to a control group (no treatment or placebo)., Conclusions: External tooth bleaching is associated with statistically significant short-term reductions in plaque and gingivitis indices. However, no data were available to evaluate long-term effects., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To assess the clinical, microbiological, and patient-based effects of using a cetylpyridinium chloride (CPC) toothpaste and mouth rinse in orthodontic patients., Design: Parallel randomized controlled, triple-blinded trial (participants, examiners, outcomes' assessors). A computer-generated list was used to allocate treatments. Central allocation was used for concealment., Participants: Thirty-one placebo (10 males, 21 females; mean age 15.2 ± 2.1) and 32 test patients (15 males, 17 females; mean age 15.0 ± 1.8) with fixed orthodontic appliance were included in the study., Interventions: Patients were randomly assigned to both brush and rinse with placebo or with CPC-based products. Products were purposely prepared in white opaque bottles. After screening and professional prophylaxis, patients received a baseline examination, and started to use the assigned products. Patients were monthly assessed during a 3-month period., Main Outcome Measures: Plaque (PlI) and gingival (GI) indexes. Secondary outcome variables: Calculus index, tooth staining, subgingival microbiological samples, patient-based variables (questionnaire, compliance, and remaining mouth rinse), side-effects (debonded braces or mucosal injuries)., Results: Sixty-three patients were randomized, 13 patients were excluded from analysis because of early dropout, leaving 50 patients for intention to treat analysis. PlI in the upper jaw diminished (0.18; SD = 0.82) after 1 month in the test group, while it increased (0.26; SD = 0.62) in the placebo group (P = 0.024). Statistically significant higher GI values were observed in the placebo group at 1 month (mean increase = 0.05; SD = 0.33 versus mean decrease = 0.03; SD = 0.38) and 3 months (mean = 0.05; SD = 0.43 versus mean = 0.02; SD = 0.34). The taste of the test product was rated higher than the placebo at 2- and 3-month visits (P < 0.05). Non-significant changes were observed in microbiological parameters, overgrowth of opportunistic species or remaining secondary outcome variables, including side-effects., Conclusion: The use of CPC-based toothpaste and mouth rinse in orthodontic patients had limited effect in reducing plaque accumulation and gingival inflammation. Effects were little and highly variable. The use of the test products was not associated with relevant adverse effects., Trial Registration: Trial registration: Local internal reference: P07/133.

Aim: To evaluate the efficacy of a probiotic combination in the treatment of gingivitis and to assess its impact on the subgingival microbiota., Materials and Methods: A placebo-controlled clinical trial was conducted in gingivitis subjects during 6 weeks. Test treatment consisted of the administration of two oral tablets per day containing the probiotic strains Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici; the control group received the same tablets but without live bacteria. The main outcome variable was the changes in gingival index (GI). Subgingival samples were collected and analysed by quantitative polymerase chain reaction (qPCR) for five putative periodontal pathogens. Outcome variables were compared between and within groups, and multiple regression analysis was performed., Results: A total of 59 patients (29 tests, 30 placebos) were included in the analysis. Both treatment groups experienced a statistically significant improvement in mean GI (p < .0001), but no differences between treatment groups were found for any clinical index. A significantly higher reduction in the number of sites with higher GI scores (GI = 3 at baseline) was observed in the test group. In subgingival samples, a significant reduction in T. forsythia was significant only in the test group (p < .008)., Conclusions: The use of probiotic tablets containing L. plantarum, L. brevis and P. acidilactici did not lead to significant changes in mean GI; although a significant reduction occurred in the number of sites with severe inflammation. Furthermore, the adjunctive use of this probiotic promoted a significant microbiological impact., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: To report the evidence on the effect of mechanical and/or chemical plaque control in the simultaneous management of gingivitis and caries., Material and Methods: A protocol was designed to identify randomized (RCTs) and controlled (CCTs) clinical trials, cohort studies and prospective case series (PCS), with at least 6 months of follow-up, reporting on plaque, gingivitis and caries. Relevant information was extracted from full papers, including quality and risk of bias. Meta-analyses were performed whenever possible., Results: After the screening of 1,373 titles, 15 RCTs, 10 CCTs and 2 PCS were included. Low to moderate evidence support that combined professional and self-performed mechanical plaque control significantly reduces standardized plaque index [n = 4; weighted mean difference (WMD) = 1.294; 95% CI (0.445; 2.144); p = 0.003] and gingivitis scores [n = 4; WMD = 1.728; 95% CI (0.631; 2.825); p = 0.002]. The addition of fluoride to mechanical plaque control is relevant for caries management [n = 5; WMD = 1.159; 95% CI (0.145; 2.172); p = 0.025] while chlorhexidine rinses are relevant for gingivitis., Conclusion: Mechanical plaque control procedures are effective in reducing plaque and gingivitis. The addition of fluoride to mechanical plaque control is significant for caries management. Chlorhexidine rinse has a positive effect on gingivitis and inconclusive role in caries., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: The aim of this systematic review was to evaluate the efficacy of anti-plaque chemical formulations for managing gingivitis, in 6-month, home-use, randomised clinical trials (RCTs)., Material and Methods: A protocol was designed, including inclusion and exclusion criteria to identify RCTs assessing gingival and/or bleeding indices. Electronic and hand-searches identified relevant papers, which were screened and evaluated for inclusion. Full-papers were retrieved and relevant information was extracted (also plaque indices), including quality and risk of bias. Mean treatment effects were calculated to obtain weighted mean differences (WMD) and 95% confidence intervals., Results: After the process of screening and selection, 87 articles with 133 comparisons, were included in the review. The additional effects of the tested products were statistically significant in terms of Löe & Silness gingival index (46 comparisons, WMD -0.217), modified gingival index (n = 23, -0.415), gingivitis severity index (n = 26, -14.939%) or bleeding index (n = 23, -7.626%), with significant heterogeneity. For plaque, additional effects were found for Turesky (66 studies, WMD -0.475), Silness & Löe (n = 26, -0.109), and plaque severity (n = 12, -23.4%) indices, with significant heterogeneity., Conclusion: Within the limitations of the present study, formulations with specific agents for chemical plaque control provide statistically significant improvements in terms of gingival, bleeding and plaque indices., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Unlabelled: Periodontitis is a ubiquitous and irreversible inflammatory condition and represents a significant public health burden. Severe periodontitis affects over 11% of adults, is a major cause of tooth loss impacting negatively upon speech, nutrition, quality of life and self-esteem, and has systemic inflammatory consequences. Periodontitis is preventable and treatment leads to reduced rates of tooth loss and improved quality of life. However, successful treatment necessitates behaviour change in patients to address lifestyle risk factors (e.g. smoking) and, most importantly, to attain and sustain high standards of daily plaque removal, lifelong. While mechanical plaque removal remains the bedrock of successful periodontal disease management, in high-risk patients it appears that the critical threshold for plaque accumulation to trigger periodontitis is low, and such patients may benefit from adjunctive agents for primary prevention of periodontitis., Aim: The aims of this working group were to systematically review the evidence for primary prevention of periodontitis by preventing gingivitis via four approaches: 1) the efficacy of mechanical self-administered plaque control regimes; 2) the efficacy of self-administered inter-dental mechanical plaque control; 3) the efficacy of adjunctive chemical plaque control; and 4) anti-inflammatory (sole or adjunctive) approaches., Methods: Two meta-reviews (mechanical plaque removal) and two traditional systematic reviews (chemical plaque control/anti-inflammatory agents) formed the basis of this consensus., Results: Data support the belief that professionally administered plaque control significantly improves gingival inflammation and lowers plaque scores, with some evidence that reinforcement of oral hygiene provides further benefit. Re-chargeable power toothbrushes provide small but statistically significant additional reductions in gingival inflammation and plaque levels. Flossing cannot be recommended other than for sites of gingival and periodontal health, where inter-dental brushes (IDBs) will not pass through the interproximal area without trauma. Otherwise, IDBs are the device of choice for interproximal plaque removal. Use of local or systemic anti-inflammatory agents in the management of gingivitis has no robust evidence base. We support the almost universal recommendations that all people should brush their teeth twice a day for at least 2 min. with fluoridated dentifrice. Expert opinion is that for periodontitis patients 2 min. is likely to be insufficient, especially when considering the need for additional use of inter-dental cleaning devices. In patients with gingivitis once daily inter-dental cleaning is recommended and the adjunctive use of chemical plaque control agents offers advantages in this group., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: To assess the efficacy of a 0.07% cetylpyridinium chloride (CPC) mouth rinse in the control of plaque and gingival inflammation during a 6-month period., Material and Methods: Adult subjects with moderate gingivitis were selected [≥40% bleeding on marginal probing (BOMP)]. After retrieving microbiological samples and evaluating the clinical parameters (plaque, BOMP and stain indexes), a professional prophylaxis was performed and subjects were randomly assigned to the test (CPC mouth rinse) or to the placebo group. Subjects were re-assessed after 3 and 6 months., Results: A total of 67 patients (35 test, 32 placebo) were included in the analysis. At 6 months, intra-group significant plaque reductions were observed in the test group (0.691, p < 0.001), but not in the placebo (0.181, p = 0.653). At 6 months, the mean BOMP values were lower in the test group (p = 0.052). Changes between baseline and 6 months were significantly higher in the test group both for plaque (p = 0.002) and BOMP (p = 0.037) when compared with the placebo. A microbiological impact was observed in the test group, especially for Prevotella intermedia., Conclusion: The evaluated 0.07% CPC-based mouth rinse, used three times per day adjunctively to mechanical tooth cleaning, prevents plaque accumulation and gingival inflammation, as compared to the placebo, for at least 6 months., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: To obtain an overall quantitative estimate of the association between pregnancy and gingival inflammation., Material and Methods: Medline and EMBASE databases were searched through August 2011. Prospective cohort or cross-sectional studies assessing the effect of pregnancy on gingival inflammation evaluated by the gingival index (GI) and/or bleeding on probing were included. Meta-analyses were performed if possible., Results: Forty-four articles representing 33 studies (14 cohort and 19 cross-sectional) were included. Meta-analyses, performed whenever possible, revealed (1) a significantly lower GI in pregnant women in the first term compared with those in their second or third term of pregnancy; (2) a lower mean GI score in post-partum women compared with women in their second [WMD = 0.143; 95% CI (0.031; 0.255); p = 0.012] or third term [WMD = 0.256; 95% CI (0.151; 0.360); p < 0.001] of pregnancy, when considering cohort studies; (3) Non-pregnant women had lower mean GI values than women in their second or third term of pregnancy. Small changes in plaque levels were reported., Conclusion: Despite the limited number of studies included in the meta-analyses, the present systematic review confirms the existence of a significant increase in GI throughout pregnancy and between pregnant versus post-partum or non-pregnant women, without a concomitant increase in plaque levels., (© 2012 John Wiley & Sons A/S.)

Dentifrices are a general term used to describe preparations that are used together with a toothbrush with the purpose to clean and/or polish the teeth. Active toothpastes were first formulated in the 1950s and included ingredients such as urea, enzymes, ammonium phosphate, sodium lauryl sarcosinate and stannous fluoride. Later, therapeutic agents were included. Today's toothpastes have two objectives: to help the toothbrush in cleaning the tooth surface and to provide a therapeutic effect. The therapeutic effect may have an antiplaque or anti-inflammatory basis when the nature of the agents is antimicrobial. Plaque inhibitory and antiplaque activity of toothpastes used for chemical plaque control is evaluated in distinct consecutive stages, the last being home use randomized clinical trials of at least 6 months' duration. In this chapter, the scientific evidence supporting the use of the most common antiplaque agents, included in toothpaste formulations, is reviewed, with a special emphasis on 6-month clinical trials, and systematic reviews with meta-analyses of the mentioned studies. Among the active agents, the following have been included in toothpastes: enzymes, amine alcohols, herbal or natural products, triclosan, bisbiguanides (chlorhexidine), quaternary ammonium compounds (cetylpyridinium chloride) and different metal salts (zinc salts, stannous fluoride, stannous fluoride with amine fluoride). Dentifrices are the ideal vehicles for any active ingredient used as an oral health preventive measure since they are used in combination with toothbrushing, which is the most frequently employed oral hygiene method. The most important indications of dentifrices with active ingredients are associated with long-term use to prevent bacterial biofilm formation, mostly in gingivitis patients or in patients on supportive periodontal therapy., (Copyright © 2013 S. Karger AG, Basel.)

Objectives: This study was performed to evaluate the microbiological profile and the calprotectin expression in gingival crevicular fluid (GCF) in spontaneous and experimentally induced gingival inflammation., Materials and Methods: Thirty-seven periodontally healthy subjects were evaluated in real life conditions (N-O gingivitis) as well as after 21 days of experimental gingivitis trial (E-I gingivitis). During the experimental gingivitis trial, in one maxillary quadrant (test quadrant), gingival inflammation was induced by oral hygiene abstention, while in the contralateral (control) quadrant, oral hygiene was routinely continued., Results: The results of the study showed that (1) the microbiological profile of quadrants where gingival inflammation was experimentally induced (i.e., E-I test quadrants) differed significantly from that of either quadrants where gingival inflammation was controlled by proper plaque control (i.e., E-I control quadrants) or quadrants with N-O gingivitis, and (2) GCF calprotectin was significantly higher at E-I test quadrants compared to either E-I control quadrants or quadrants with N-O gingivitis. A positive intrasubject correlation was found between GCF concentration of calprotectin at sites presenting N-O and E-I gingivitis., Conclusions: N-O and E-I gingivitis showed a different microbiological profile of the subgingival environment. GCF calprotectin is a reliable marker of gingival inflammation, and its concentration in N-O gingivitis is correlated with its expression in E-I gingivitis., Clinical Relevance: The modality of plaque accumulation seems to affect the subgingival microbiological profile associated with a gingivitis condition. Calprotectin levels in GCF may be regarded as a promising marker of the individual susceptibility to develop gingival inflammation in response to experimentally induced plaque accumulation.

Objective: To investigate the effects of an orally administered probiotic on the oral microbiota., Methods: A placebo-controlled, parallel study was conducted in 40 gingivitis subjects during 8 weeks. Treatment consisted on the administration of a daily tablet, either containing Lactobacillus reuteri or placebo. Unstimulated saliva and subgingival samples were collected and analysed by culture and PCR. Clinical and microbiological outcome variables were compared between and within groups., Results: There were no significant changes between and within the groups in the clinical variables. In saliva, total anaerobic counts after 4 weeks (p = 0.021) and counts of Prevotella intermedia after 8 weeks (p = 0.030), showed reductions in the test group. In subgingival samples, significant reductions in the changes baseline to 4 weeks were observed for P. gingivalis counts (p = 0.008). With PCR, L. reuteri ATCC-PTA-5289 was more frequently detected than L. reuteri DSM-17938., Conclusions: The effect of L. reuteri administered in tablets resulted in a reduction in the number of selected periodontal pathogens in the subgingival microbiota, without an associated clinical impact., (© 2012 John Wiley & Sons A/S.)

Objectives: the aim of this study was to evaluate the effects of the use of a mouth rinse and dentifrice with cetylpyridinium chloride (CPC) in patients with gingivitis., Methods: the study was designed as a 1-month, double-blind, parallel, randomized clinical trial comparing a negative control regimen (minus active ingredients dentifrice and mouth rinse) with the test products (dentifrice and mouth rinse with 0.05% CPC) in terms of plaque and gingival indexes (PI, GI), patient-based and microbiological outcome variables. The comparisons in relation to the main outcome variables (PI and GI) were made by means of the t-test, either unpaired or paired for the intergroup and intragroup comparisons, respectively., Results: no differences were detected at baseline. Both groups showed statistically significant decreases in GI (0.17-0.19), without intergroup differences. The PI demonstrated a significant decrease of -0.12 in the test group and minor changes in the negative control group (increase of +0.01). Differences between groups showed a tendency towards statistical significance. A limited impact was observed for microbiological variables in both groups., Conclusion: the results of this study show limited benefits of the evaluated formulations as adjuncts to unsupervised oral hygiene in reducing plaque accumulation, and no effect on gingivitis., (© 2011 John Wiley & Sons A/S.)

Aims: To identify predictor variables involved in exacerbated gingival inflammation associated with pregnancy., Material and Methods: In this cohort study, 48 pregnant and 28 non-pregnant women without periodontitis were included. The pregnant women were evaluated in the first, second and third trimester and at 3 months postpartum, whilst the non-pregnant women were evaluated twice, with a 6-month interval. At each visit, clinical [plaque index (PlI) and gingival index (GI)], hormonal (salivary progesterone and estradiol), immunological [gingival crevicular fluid interleukin-1β, interleukin-6, tumour necrosis factor-α (TNF-α) and prostaglandin-E(2) ] and microbiological (periodontal pathogens culture) evaluations were performed. Statistical analysis was undertaken using exhaustive chi-square automatic interaction detection (exhaustive CHAID) to analyse the predictive value of the independent outcomes to develop pregnancy GI., Results: PlI was the strongest predictor implicated in the GI throughout pregnancy and after delivery. During the second and third trimesters the presence of Porphyromonas gingivalis significantly contributed to the worsening of gingival inflammation. When compared with the non-pregnant group, significant differences were found in TNF-α amounts and concentrations and in the third trimester site-specific GI., Conclusions: Bacterial challenge to the gingival tissues, both quantitatively (PlI) and qualitatively (harbouring P. gingivalis) appears to affect the level of gingival inflammation observed during pregnancy., (© 2011 John Wiley & Sons A/S.)

Aim: To determine whether the exacerbated gingival inflammation that develops in pregnant women is related to a change in the subgingival biofilm induced by the increase in hormone levels during pregnancy., Material and Methods: This open cohort study included 48 pregnant and 28 non-pregnant women without periodontitis. Pregnant women were evaluated in the first, second and third trimester and at 3 months after delivery. Non-pregnant women were evaluated twice, with a 6-month interval, assessing microbiological, clinical and hormonal variables at each visit. Total anaerobic counts and frequency of detection and proportions were calculated. The Friedman test with the Bonferroni correction was used for intra-group comparisons and Mann-Whitney U-tests for inter-group assessment. Correlations were analysed by means of Spearman's rank correlation coefficient., Results: Proportions of the subgingival periodontal pathogens did not differ throughout pregnancy, although significant differences were found for all the pathogens after delivery. Porphyromonas gingivalis-positive patients presented an increase in gingival inflammation (p<0.001) that was not related to plaque. Correlations were found between maternal hormone levels and P. gingivalis and Prevotella intermedia., Conclusion: Qualitative differences in periodontal pathogens were found from pregnancy to post-partum. Patients harbouring P. gingivalis presented and increased gingival inflammatory status.

Aim: To test whether exacerbated gingival inflammation in pregnancy is associated with increased salivary hormone levels and changes in gingival crevicular fluid (GCF) interleukin-1beta (IL-1beta) and prostaglandin-E2 (PGE2) levels., Material and Methods: In this cohort study, 48 pregnant women without periodontitis were evaluated in the first, second, and third trimesters and at 3 months postpartum. Twenty-eight non-periodontitis non-pregnant women were evaluated twice, with a 6-month interval. Plaque and gingival indices (PlI, GI), salivary progesterone and estradiol and GCF IL-1beta and PGE2 levels were determined. anova for repeated measures or Friedman's test were used for intragroup analyses. Inter-group comparisons were analysed with t-test or Mann-Whitney U-test. Correlations were evaluated with Pearson's and Spearman's test., Results: Pregnant women showed an increase in GI (p<0.05) despite maintaining low PlI values. No changes in IL-1beta and PGE2 levels were observed during pregnancy. No significant correlation was found between the GI increase and salivary hormone levels. GI (p<0.05) and IL-1beta levels (p<0.001) were lower in non-pregnant than in pregnant women., Conclusions: This study confirms the presence of an exacerbated gingival inflammation during pregnancy, but this phenomenon could not be associated with an increase in progesterone or estradiol or with changes in PGE2 or IL-1beta.

Background and Objective Bacteria in the oral cavity grow in the form of biofilms; these structures are subject to constant saliva or gingival crevicular fluid flow conditions. The aims of this study were: (i) to develop and to characterize an in-vitro biofilm model with oral bacteria growing under flow and shear conditions; and (ii) to demonstrate the usefulness of the model for evaluating the activity of three antiplaque agents. Material and Methods We used a bioreactor to grow the oral bacteria Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis under planktonic conditions. Biofilms were established using a modified Robbins device on hydroxyapatite ( HAP) discs. Three- to 7-d-old biofilms were analysed using culture methods, scanning electron microscopy, Live/ Dead staining and fluorescence in-situ hybridization (confocal laser scanning microscopy). Finally, we assessed the antimicrobial activity of three mouthrinses [0.12% chlorhexidine ( CHX), 0.12% chlorhexidine and sodium fluoride ( CHX+ Na F) and 0.12% chlorhexidine and 0.05% cetylpyridinium chloride ( CHX+ CPC)] using a planktonic test (short interval-killing test) and in our 4-d biofilm model. Results The viable cell counts showed that each species was consistently found in the biofilms throughout the study. The architecture and cell distribution were similar to those described for biofilms in situ, with the exception of a thin layer of living cells that was found close to the HAP. The effectiveness test of the mouthwashes demonstrated that cells in biofilms showed more tolerance compared with planktonic cells. Moreover, it was observed that in 4-d biofilm formed in vitro, CHX+ CPC caused significantly higher mortality compared with CHX ( p = 0.003) and CHX+ Na F ( p < 0.001). Conclusion Our results suggest that we have a highly reproducible system for multispecies oral biofilm formation and that it is a useful tool for assessing antibacterial molecules before their clinical evaluation. It also has great potential to be used in basic research on supragingival and subgingival biofilms. [ABSTRACT FROM AUTHOR]