Combination of Neovestitol and Vestitol Modifies the Profile of Periodontitis-Related Subgingival Multispecies Biofilm.

The aim of this study was to evaluate the effect of the combination of neovestitol-vestitol (CNV) compounds obtained from Brazilian red propolis on the microbiological profile of a mature multispecies subgingival biofilm. The biofilm with 32 bacterial species associated with periodontitis was formed for seven days using a Calgary device. Treatment with CNV (1600, 800, 400, and 200 µg/mL), amoxicillin (54 µg/mL), and vehicle control was performed for 24 h on the last day of biofilm formation. Biofilm metabolic activity and DNA-DNA hybridization (checkerboard) assays were performed. The groups treated with CNV 1600 and amoxicillin reduced 25 and 13 species, respectively, compared to the control vehicle treatment (p ≤ 0.05); both reduced P. gingivalis, while only CNV reduced T. forsythia. When the data from the two treatments (CNV and AMOXI) were compared, a statistically significant difference was observed in 13 species, particularly members of Socransky's orange complex. Our results showed that CNV at 1600 µg/mL showed the best results regarding the metabolic activity of mature biofilms and obtained a reduction in species associated with the disease, such as T. forsythia, showing a better reduction than amoxicillin. Therefore, CNV seems to be a promising alternative to eradicate biofilms and reduce their pathogenicity.

Influence of gender on periodontal outcomes: A retrospective analysis of eight randomized clinical trials.

AIMS: To explore the influence of gender on periodontal treatment outcomes in a dataset of eight RCTs conducted in Brazil, United States, and Germany. METHODS: Clinical parameters were compared between men and women with stages III/IV grades B/C generalized periodontitis at baseline and 1-year post-therapy, including scaling and root planing with or without antibiotics. RESULTS: Data from 1042 patients were analyzed. Men presented a tendency towards higher probing depth (p = .07, effect size = 0.11) and clinical attachment level (CAL) than women at baseline (p = .01, effect size = 0.16). Males also presented statistically significantly lower CAL gain at sites with CAL of 4-6 mm at 1-year post-therapy (p = .001, effect size = 0.20). Among patients with Grade B periodontitis who took antibiotics, a higher frequency of women achieved the endpoint for treatment (i.e., ≤4 sites PD ≥5 mm) at 1 year than men (p < .05, effect size = 0.12). CONCLUSION: Men enrolled in RCTs showed a slightly inferior clinical response to periodontal therapy in a limited number of sub-analyses when compared to women. These small differences did not appear to be clinically relevant. Although gender did not dictate the clinical response to periodontal treatment in this population, our findings suggest that future research should continue to explore this topic.

Antimicrobial Activity of Methylene Blue Associated with Photodynamic Therapy: In Vitro Study in Multi-Species Oral Biofilm.

The control of infectious diseases caused by biofilms is a continuing challenge for researchers due to the complexity of their microbial structures and therapeutic implications. Photodynamic therapy as an adjunctive anti-infective treatment has been described as a possible valid approach but has not been tested in polymicrobial biofilm models. This study evaluated the effect of photodynamic therapy in vitro with methylene blue (MB) 0.01% and red LEDs (λ = 660 nm, power density ≈ 330 mW/cm2, 2 mm distance from culture) on the metabolic activity and composition of a multispecies subgingival biofilm. Test Groups LED and MB + LED showed a more significant reduction in metabolic activity than the non-LED application group (~50 and 55%, respectively). Groups LED and MB equally affected (more than 80%) the total bacterial count in biofilms. No differences were noted in the bacterial biofilm composition between the groups. In vitro LED alone or the MB + LED combination reduced the metabolic activity of bacteria in polymicrobial biofilms and the total subgingival biofilm count.

Evaluation of the Microbial Profile on the Polydioxanone Membrane and the Collagen Membrane Exposed to Multi-Species Subgingival Biofilm: An In Vitro Study.

Dehiscence in surgeries involving membranes often leads to bacterial contamination, hindering the healing process. This study assessed bacterial colonization on various membrane materials. Polydioxanone (PDO) membranes, with thicknesses of 0.5 mm and 1 mm, and a collagen membrane were examined. Packages containing polystyrene pins were crafted using these membranes, attached to 24-well plates, and exposed to oral bacteria from supra and subgingival biofilm. After a week's anaerobic incubation, biofilm formation was evaluated using the DNA-DNA hybridization test. Statistical analysis employed the Kruskal-Wallis test with Dunn's post hoc test. The biofilm on the polystyrene pins covered by the 0.5 mm PDO membrane showed a higher count of certain pathogens. The collagen membrane had a greater total biofilm count on its inner surface compared to both PDO membranes. The external collagen membrane face had a higher total biofilm count than the 0.5 mm PDO membrane. Furthermore, the 1 mm PDO membrane exhibited a greater count of specific pathogens than its 0.5 mm counterpart. In conclusion, the collagen membrane presented more biofilm and pathogens both internally and on its inner surface.

Antimicrobial activity of Desplac® oral gel in the subgingival multispecies biofilm formation.

Natural products are well-known due to their antimicrobial properties. This study aimed to evaluate the antimicrobial effect of Desplac® product (composed of Aloe Vera, Propolis Extract, Green Tea, Cranberry, and Calendula) on the subgingival biofilm. Two different protocols were used to treat the 33-species biofilms: (A) 2×/day (12/12 h) for 1 min with Desplac® or Noplak Toothpaste (Chlorhexidine + Cetylpyridinium Chloride) or Oral B ProGengiva (stannous Fluoride) or a placebo gel; (B) a 12-h use of the Desplac® product or 0.12% chlorhexidine gel or a placebo gel. After 7 days of biofilm formation, the metabolic activity (MA) and biofilm profile were determined by 2,3,5-triphenyltetrazolium chloride and Checker-board DNA-DNA hybridization, respectively. Statistical analysis used the Kruskal-Wallis test followed by Dunn's post-hoc. In protocol A, all treatments presented reduced MA compared to the placebo (p ≤ 0.05). The Desplac®-treated biofilm showed a similar microbial profile to other antimicrobials, although with higher bacterial total counts. In protocol B, MA of Desplac®-treated biofilms was lower than the placebo's MA but higher than chlorhexidine-treated biofilms (p ≤ 0.05). Pathogen levels in Desplac®-treated biofilms were lower than in placebo-treated biofilms and elevated compared to the chlorhexidine-treated biofilms (p ≤ 0.05). Desplac® inhibited the biofilm development and disrupted the mature subgingival biofilm, highlighting its effect on Tannerella forsythia counts.

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

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

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

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

Systemic Antibiotics and Chlorhexidine Associated with Periodontal Therapy: Microbiological Effect on Intraoral Surfaces and Saliva.

The effect of systemic antibiotics on the microbial profile of extracrevicular sites after periodontal treatment is currently the subject of research. This study evaluated the microbiological effects on different oral cavity sites of scaling and root planing (SRP) combined with antimicrobial chemical control in the treatment of periodontitis. Sixty subjects were randomly assigned to receive SRP alone or combined with metronidazole (MTZ) + amoxicillin (AMX) for 14 days, with or without chlorhexidine mouth rinse (CHX) for 60 days. Microbiological samples were evaluated by checkerboard DNA-DNA hybridization until 180 days post therapy. The adjunctive use of antibiotics plus CHX significantly reduced the mean proportions of red complex species from subgingival biofilm and saliva (p < 0.05). Furthermore, the analysis of all intraoral niches showed a significantly lower mean proportion of the red complex species in the same group. In conclusion, the concomitant use of antimicrobial chemical control (systemic and local) demonstrated a beneficial effect on the composition of the oral microbiota.

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

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

Probiotic consumption can modify the resilience of periodontal tissues in rats under experimental periodontitis.

BACKGROUND: This study evaluated the effects of systemic administration of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) on experimental periodontitis (EP) in rats. METHODS: Thirty-two rats were allocated to groups C (control), C-HN019 (probiotic), EP (EP only), and EP-HN019 (EP+probiotic). From day 0, the animals of C-HN019 and EP-HN019 groups received B. lactis HN019 (1 × 109 CFU/ml) daily. On the 14th day, the animals of EP and EP-HN019 groups received silk ligature around mandibular molars. Animals were euthanized on the 28th day. Samples of oral biofilm, gingival tissues, blood serum, and mandible were obtained for microtomographic, histomorphometric, microbiological, and immunological analyses. Data were statistically analyzed (p < 0.05). RESULTS: Group EP-HN019 presented significantly less alveolar bone loss when compared with Group EP in histomorphometric and microtomographic analyses. In gingival tissue and serum, Group EP-HN019 presented lower proinflammatory/anti-inflammatory cytokines ratios than Group EP. Group EP-HN019 showed higher expression of beta-defensins and less TRAP-positive cells than Group EP. Group EP presented higher gene expression of Ifng and lower gene expression of Foxp3 when compared with Group EP-HN019 in gingival tissue. In oral biofilm, EP-HN019 group presented a lower percentage of species similar to Fusobacterium periodonticum and a higher percentage of species similar to Actinomyces gereneseriae, Actinomyces israelli, and Streptococcus gordonii when compared with Group EP. There was a significant increase of B. lactis HN019 after administration of probiotic therapy in oral biofilm of Group EP-HN019. CONCLUSION: The consumption of B. lactis HN019 promotes a protective effect against alveolar bone loss by modifying local and systemic microbiological and immunoinflammatory parameters.

Microbiological effects of amoxicillin plus metronidazole in the treatment of young patients with Stages III and IV periodontitis: A secondary analysis from a 1-year double-blinded placebo-controlled randomized clinical trial.

BACKGROUND: Despite the body of evidence supporting the clinical benefits of metronidazole (MTZ) and amoxicillin (AMX) in the treatment of young patients with periodontitis, the microbiological outcomes of this antibiotic protocol have been less explored. This study evaluated the microbiological effects of adjunctive MTZ+AMX in the treatment of young patients with periodontitis. METHODS: Subjects with periodontitis Stages III or IV and ≤30 years old were randomly allocated to receive scaling and root planing (SRP) with placebo (n = 15) or with MTZ (400 mg) and AMX (500 mg) three times a day for 14 days (n = 15). Nine subgingival biofilm samples per subject (three samples from each probing depth (PD) category: ≤3, 4-6, and ≥7 mm) were collected at baseline and 3-, 6-, and 12-months post-treatment and individually analyzed for 40 bacterial species by checkerboard DNA-DNA hybridization. RESULTS: Thirty subjects (15/group) with mean ages 27.6 ± 3.5 (control) and 26.8 ± 3.9 (test) were included. At 12 months post-therapy, the antibiotic group harbored lower proportions of red complex (1.3%) than the placebo group (12.5%) (p < 0.05). SRP + MTZ+AMX was more effective than mechanical treatment in reducing levels/proportions of several pathogens and increasing proportions of Actinomyces species (p < 0.05). Levels/proportions of Aggregatibacter actinomycetemcomitans were only reduced in the antibiotic group (p < 0.05). This group also exhibited greater reduction in the number of sites with PD ≥5 mm and higher percentage of subjects reaching the clinical end point for treatment (≤4 sites with PD ≥5 mm) than the control group (p < 0.05). CONCLUSION: SRP+MTZ+AMX allowed for establishing a long-term healthier subgingival biofilm community and periodontal clinical condition, than SRP only.

Propolis, Aloe Vera, Green Tea, Cranberry, Calendula, Myrrha and Salvia Properties against Periodontal Microorganisms.

The oral cavity harbors hundreds of microorganisms that may be uncontrolled and provoke several diseases. In this sense, periodontitis is a complex multifactorial disease with an essential microbial component in its etiology. Periodontal treatment involves mechanical control of the supra- and subgingival biofilm, but not all patients respond predictably to treatment. In this way, the biofilm chemical control helps in the reduction of periodontal pathogens during treatment or in the delay of bacterial re-colonization after scaling and root planning. Several products have been studied as adjunctive therapy and have shown promising results. Therefore, the present article reviews the biological effects of propolis, aloe vera, green tea, cranberry, calendula, myrrha and salvia that may support their use in the control of subgingival biofilm in patients with periodontitis. All the natural products cited above showed exciting results against microorganisms related to oral diseases, mainly periodontitis. These substances also have anti-inflammatory and antioxidant activities. The natural agents propolis, aloe vera, green tea, cranberry, calendula, myrrha and salvia demonstrated potential to be used as oral hygiene products, based on their antimicrobial and anti-inflammatory actions.

Do the progression of experimentally induced gingivitis and peri-implant mucositis present common features? A systematic review of clinical human studies.

This systematic review evaluated the features of the progression of experimentally induced gingivitis and peri-implant mucositis in humans. Included were studies that evaluated clinical, immunological, or microbiological responses between experimentally induced gingivitis and peri-implant mucositis in periodontally healthy patients. A total of 887 articles were initially identified, but only 12 were included in the final analysis. Implants accumulate less biofilm and suffer the most heterogeneous alterations in the microbiota, in the abstinence of oral hygiene, compared with the tooth. Interestingly, although dental implants presented less biofilm accumulation, the peri-implant mucosa showed a more exacerbated clinical response than the gingival tissue. The risk of bias of the selected studies was moderate to low, with one study presenting serious risk. The progression events of peri-implant mucositis were similar to those of experimental gingivitis but led to a different host response. This review was registered in the PROSPERO database CRD420201 123360.

In vitro antimicrobial effect of titanium anodization on complex multispecies subgingival biofilm.

Anodization is a routine industrial galvanic method that produces a titanium oxide layer on the surface of titanium. Considering the possibility that this technique could influence microbial adsorption and colonization, this in vitro study was conducted to evaluate the impact of a process of anodization applied to a titanium surface on the microbial profile of multispecies subgingival biofilm. Titanium discs produced by using two different processes-conventional and Anodization-were divided into two groups: conventional titanium discs with machined surface (cpTi) Control Group and titanium discs with anodic oxidation treatment (anTi) Test Group. Subgingival biofilm composed of 33 species was formed on the titanium discs that were positioned vertically in 96-well plates, for 7 days. The proportions and the counts of microbial species were determined using a DNA-DNA hybridization technique, and data were evaluated using Mann-Whitney test (p < 0.05). Mean total bacterial counts were lower in Test Group in comparison with Control Group (p < 0.05). Nine bacterial species differed significantly, and were found in higher levels in Control Group in comparison with Test Group, including T. forsythia, E. nodatum, and F. periodonticum. In conclusion, titanium discs with anodization could alter the microbial profile of the biofilm formed around them. Further clinical studies should be conducted to confirm the clinical impact of these findings.

Adverse events of metronidazole and amoxicillin: Retrospective analysis of a large data set of five randomized clinical trials.

AIM: To evaluate the frequency of side effects associated with intake of metronidazole (MTZ) + amoxicillin (AMX) in periodontal treatment, and to explore associations between these events and patients' features. MATERIALS AND METHODS: Data of five randomized clinical trials testing MTZ + AMX adjunctive to mechanical therapy were evaluated. Volunteers answered an adverse event questionnaire. RESULTS: Information from 656 subjects was assessed. The frequency of side effects in the antibiotic- and placebo-treated groups ranged from 1.0% to 17.7% and 0.9% to 13.7%, respectively. The events more frequently observed in the antibiotic than in the placebo group were diarrhoea and metallic taste (p < .05). Diabetes significantly raised the odds of a patient reporting discomfort (odds ratio [OR] = 2.6), diarrhoea (OR = 4.0), weakness (OR = 6.0) and excessive sleepiness (OR = 2.9). In systemically healthy volunteers, using antibiotics 3 months post-mechanical treatment (healing phase) (OR = 3.0), being a woman (OR = 3.9) and aged ≤49 (OR = 4.5) significantly increased the chances of reporting adverse events. CONCLUSIONS: The occurrence of side effects during MTZ + AMX treatment ranged from uncommon (1%) to very common (17.7%). The main factors raising the chances of a patient reporting adverse events were diabetes and taking antibiotics in the healing phase, instead of in the active phase of treatment. Patients ≤ 49 years old and females also tend to report more side effects.

Systematic and scoping reviews to assess biological parameters.

INTRODUCTION: An evidence synthesis approach compiling biological/laboratory data is effective in advancing health-related knowledge. However, this approach is still underused in the oral health field. METHODS: This commentary discusses the opportunities and challenges of systematic and scoping reviews of laboratory data in dentistry. Special focus is on the potential of these reviews to elucidate etiological and treatment concepts of oral diseases, such as periodontitis and peri-implantitis. RESULTS: The following difficulties associated with such studies are discussed: (i) selection of ideal study design, (ii) assessment of "risk of bias" and definition of "certainty of evidence", (iii) evidence assembly and summary, and (iv) the paper review process. DISCUSSION: Despite those challenges, high-quality reviews integrating laboratory data may generate relevant scientific information and help identify new avenues for future investigations. Experts in different oral health topics should build a process capable of helping researchers assemble and interpret these types of data.

Recent Updates on Microbial Biofilms in Periodontitis: An Analysis of In Vitro Biofilm Models.

The development of oral biofilm models has been extremely important to study the specific role of most microbial species at the early stages of periodontitis. The current knowledge on monospecies or multispecies biofilms originates mainly from the observation of in vitro dynamic or static biofilm model systems, which were engineered to mimic clinical oral conditions. In the last few decades, mounting evidence has confirmed that biofilms are the major form of bacterial lifestyle, and more importantly, that microorganisms dwelling in sessile mixed-species aggregates display completely different phenotypes and physiological characteristics than when living in planktonic pure cultures. Interspecies interactions within these communities, mediated by chemical communication systems, have been shown to affect biofilm physiology and increase antimicrobial resistance by up to 1000 fold. These aspects reinforce the importance of developing multispecies biofilm models to better understand and control biofilms. Literature reports demonstrate that while monospecies models are still most commonly used in caries research, authors have used different multispecies models to study periodontal diseases. Periodontitis is a polymicrobial biofilm-dependent disease mainly associated with Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Interestingly, these species hardly adhere to substrates commonly used for biofilm formation, which makes multispecies models essential for an accurate analysis of periodontitis-related biofilms. The multispecies models currently available are generally composed of 6-10 species, but a more recent 34-species model was developed to better examine the dynamics within oral biofilms. The complexity of such polymicrobial biofilm models mimics more consistently the oral microbiome and different aspects of the oral environment. Collectively, the evidence on multispecies biofilm models described herein may support future studies on the use of antimicrobials for biofilm control as well as provide research opportunities to expand the current knowledge on interspecies interactions. The present manuscript reviews the most recent updates on in vitro biofilm model systems for periodontitis.

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

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

Decontamination and Biomodification of Periodontally Affected Root Surface for Successful Regeneration: Is There Room for Improvement?

Periodontitis is a multifactorial inflammatory condition associated with an oral microbiome dysbiosis that results in gingival inflammation and clinical attachment loss. Periodontal therapies are based on scaling and root planing to disturb the bacterial biofilm mechanically and remove calculus and contaminated cementum. Research does not support the use of root modifiers for decontamination and biomodification of periodontally affected root surfaces. Standardized clinical trials in large populations, assessing biological and patient-reported outcome measures, are necessary to evaluate candidate biomaterials for decontamination and biomodification of periodontally affected root surfaces.

Clinical, microbiological, and immunological effects of systemic probiotics in periodontal treatment: study protocol for a randomized controlled trial.

BACKGROUND: The association of scaling and root planing (SRP) with systemic metronidazole (MTZ) plus amoxicillin (AMX) has shown to be an effective treatment protocol, particularly for periodontitis stages III and IV, generalized. More recently, probiotics have also been suggested as a promising adjunctive treatment for periodontal diseases due to their antimicrobial and anti-inflammatory properties. Therefore, the aim of this randomized clinical trial (RCT) is to evaluate the clinical, microbiological, and immunological effects of probiotics as adjuncts to SRP alone or with MTZ+AMX in the treatment of periodontitis. METHODS: Subjects with periodontitis are being randomly assigned to receive (i) SRP alone, or with (ii) two probiotic lozenges/day for 90 days (Prob), (iii) MTZ (400 mg) and AMX (500 mg) thrice a day (TID) for 14 days (MTZ+AMX), or (iv) Prob and MTZ+AMX. Subjects are being monitored for up to 12 months post-treatment. Nine subgingival plaque samples per patient are being collected at baseline and at 3, 6, and 12 months post-therapy and analyzed by checkerboard DNA-DNA hybridization for 40 bacterial species. Peripheral blood and gingival crevicular fluid (GCF) of four randomly selected periodontal sites will be analyzed by means of a multiplex fluorescent bead-based immunoassay for 17 cyto/chemokines. STATISTICAL ANALYSES: The significance of differences in each group (over the course of the study) will be sought using repeated measures ANOVA or Friedman tests and among groups (at each time point) using either ANOVA/ANCOVA or Kruskal-Wallis tests, depending on normality of the data. The chi-square test will be used to compare differences in the frequency of subjects achieving the clinical endpoint for treatment (≤ 4 sites with PD ≥ 5 mm) at 1 year and of self-perceived adverse effects. A stepwise forward logistic regression analysis will be performed in order to investigate the impact of different predictor variables on the percentage of patients achieving the clinical endpoint for treatment. The Number Needed to Treat (NNT) with different treatment protocols will be also calculated. Statistical significance will be set at 5%. TRIAL REGISTRATION: ClinicalTrials.gov NCT03733379. Registered on November 7, 2018.