In-vitro effects of novel periodontal scalers with a planar ultrasonic piezoelectric transducer on periodontal biofilm removal, dentine surface roughness, and periodontal ligament fibroblasts adhesion.

OBJECTIVES: To compare ultrasonic scaler prototypes based on a planar piezoelectric transducer with different working frequencies featuring a titanium (Ti-20, Ti-28, and Ti-40) or stainless steel (SS-28) instrument, with a commercially available scaler (com-29) in terms of biofilm removal and reformation, dentine surface roughness and adhesion of periodontal fibroblasts. MATERIALS AND METHODS: A periodontal multi-species biofilm was formed on specimens with dentine slices. Thereafter specimens were instrumented with scalers in a periodontal pocket model or left untreated (control). The remaining biofilms were quantified and allowed to reform on instrumented dentine slices. In addition, fibroblasts were seeded for attachment evaluation after 72 h of incubation. Dentine surface roughness was analyzed before and after instrumentation. RESULTS: All tested instruments reduced the colony-forming unit (cfu) counts by about 3 to 4 log10 and the biofilm quantity (each p < 0.01 vs. control), but with no statistically significant difference between the instrumented groups. After 24-hour biofilm reformation, no differences in cfu counts were observed between any groups, but the biofilm quantity was about 50% in all instrumented groups compared to the control. The attachment of fibroblasts on instrumented dentine was significantly higher than on untreated dentine (p < 0.05), with the exception of Ti-20. The dentine surface roughness was not affected by any instrumentation. CONCLUSIONS: The planar piezoelectric scaler prototypes are able to efficiently remove biofilm without dentine surface alterations, regardless of the operating frequency or instrument material. CLINICAL RELEVANCE: Ultrasonic scalers based on a planar piezoelectric transducer might be an alternative to currently available ultrasonic scalers.

Quercetin in the Prevention of Induced Periodontal Disease in Animal Models: A Systematic Review and Meta-Analysis.

BACKGROUND: Periodontitis is an inflammatory condition initiated by oral bacteria and is associated with several systemic diseases. Quercetin is an anti-inflammatory and anti-bacterial poly-phenol present in various foods. The aim of this meta-analysis was the evaluation of the effects of quercetin administration in animal models of experimental periodontitis. METHODS: A systematic search was performed in electronic databases using the following search terms: "periodontitis" or "periodontal disease" or "gingivitis" and "quercetin" or "cyanidanol" or "sophoretin" or "pentahydroxyflavone". In vivo preclinical animal models of experimental periodontal disease with a measurement of alveolar bone loss were included in the analysis. The risk of bias of the included studies was assessed using the SYRCLE tool. RESULTS: The systematic search yielded 335 results. Five studies were included, four of them qualified for a meta-analysis. The meta-analysis showed that quercetin administration decreased alveolar bone loss (τ2 = 0.31, 1.88 mm 95%CI: 1.09, 2.67) in experimental periodontal disease animal models. However, the risk of bias assessment indicated that four SYRCLE domains had a high risk of bias. CONCLUSIONS: Quercetin diminishes periodontal bone loss and prevents disease progression in animal models of experimental periodontal disease. Quercetin might facilitate periodontal tissue hemostasis by reducing senescent cells, decreasing oxidative stress via SIRT1-induced autophagy, limiting inflammation, and fostering an oral bacterial microenvironment of symbiotic microbiota associated with oral health. Future research will show whether and how the promising preclinical results can be translated into the clinical treatment of periodontal disease.

Effect of sodium hypochlorite gel on bacteria associated with periodontal disease.

OBJECTIVES: An adjunct in non-surgical periodontal therapy might be sodium hypochlorite (NaOCl)-based agents. The purpose of the present in vitro study was to get deeper knowledge on the influence of different parameters as time after mixing, pH, and chemical composition of an amino acid 0.475% NaOCl (AA-NaOCl) gel consisting of two components on its anti-biofilm activity. MATERIALS AND METHODS: Six-species biofilms were cultured for 5 days, before AA-NaOCl gel was applied. In the different series, the influence of the time after mixing of the two components before application, of the concentration of NaOCl in the gel mixture, of the pH of the gel mixture, and of an exchange of the amino acid component by hyaluronic acid (HA), was analyzed. RESULTS: Mixing time point experiments showed that the AA-NaOCl gel is capable of statistically significantly reducing colony-forming unit (cfu) counts up to 30 min after mixing, but only up to 20 min after mixing the reduction was more than 2 log10 cfu. The pH experiments indicate that a reduced pH results in a reduced activity of the NaOCl formulation. NaOCl concentrations in the formulation in the range from 0.475 to 0.2% provide adequate activity on biofilms. A HA/NaOCl gel was equally active against the biofilm as the AA-NaOCl gel. CONCLUSION: Mixing of the components should be made in a timeframe of 20 min before applications. An optimization of the composition of the NaOCl formulation might be possible and should be a topic in further in vitro studies. CLINICAL RELEVANCE: The AA-NaOCl gel formulation can be mixed up to 20 min before application. Further, the study indicates that the composition of the NaOCl gel formulation can be optimized.

Glucose-6-Phosphatase-Dehydrogenase activity as modulative association between Parkinson's disease and periodontitis.

The association between periodontitis (PD) and Parkinson's disease (PK) is discussed due to the inflammatory component of neurodegenerative processes. PK severity and affected areas were determined using the following neuropsychological tests: Unified Parkinson's Disease Rating Score (UPDRS) and Hoehn and Yahr; non-motoric symptoms by Non-Motor Symptoms Scale (NMSS), and cognitive involvement by Mini-Mental State Examination (MMSE). Neuroinflammation and the resulting Glucose-6-Phosphatase-Dehydrogenase (G6PD) dysfunction are part of the pathophysiology of PK. This study aimed to evaluate these associations in periodontal inflammation. Clinical data and saliva-, serum-, and RNA-biobank samples of 50 well-characterized diametric patients with PK and five age- and sex-matched neurologically healthy participants were analyzed for G6PD function, periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Campylobacter rectus, Fusobacterium nucleatum, and Filifactor alocis), monocyte chemoattractant protein (MCP) 1, and interleukin (IL) 1-beta. Regression analysis was used to identify associations between clinical and behavioral data, and t-tests were used to compare health and disease. Compared with PK, no pathogens and lower inflammatory markers (p < 0.001) were detectible in healthy saliva and serum, PK-severity/UPDRS interrelated with the occurrence of Prevotella intermedia in serum as well as IL1-beta levels in serum and saliva (p = 0.006, 0.019, 0.034), Hoehn and Yahr correlated with Porphyromonas gingivalis, Prevotella intermedia, RNA IL1-beta regulation, serum, and saliva IL1-beta levels, with p-values of 0.038, 0.011, 0.008, <0.001, and 0.010, while MMSE was associated with Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, serum MCP 1 levels, RNA IL1-beta regulation and G6PD serum activity (p = 0.036, 0.003, 0.045, <0.001, and 0.021). Cognitive and motor skills seem to be important as representative tests are associated with periodontal pathogens and oral/general inflammation, wherein G6PD-saliva dysfunction might be involved. Clinical trial registration: https://www.bfarm.de/DE/Das-BfArM/Aufgaben/Deutsches-Register-Klinischer-Studien/_node.html, identifier DRKS00005388.

Clinical evaluation of a novel protocol for supportive periodontal care: A randomized controlled clinical trial.

BACKGROUND: The aim of this study was to compare the clinical efficacy and the patient perception of subgingival debridement with either guided biofilm management (GBM) or conventional scaling and root planing (SRP) during supportive periodontal care (SPC). METHODS: Forty-one patients in SPC were randomly assigned to either treatment with GBM or SRP every 6 months. The primary outcome was the percentage of bleeding on probing (BoP) at 1 year. Moreover, pocket probing depths (PPD), recession, and furcation involvements were also measured. Full-mouth and specific site analyzes were performed at baseline, 6 and 12 months of SPC. Patient comfort was evaluated using a visual analogue scale (VAS) at 12 months. RESULTS: At 1 year, mean BoP percentage decreased from 12.2% to 9.0% (p = 0.191) and from 14.7% to 7.9% (p = 0.004) for the GBM and SRP groups, respectively. Furcation involved multirooted teeth but no through-and-through lesions were significantly fewer in the GBM than in the SRP group after 12 months (p = 0.015). The remaining parameters showed slight improvement in both groups without any statistically significant differences between the two groups after 1 year. Pain evaluation as patient reported outcome measures (pain evaluation) was in favor (p = 0.347) of the SRP group, while overall satisfaction was similar for both groups. Treatment time was not statistically significantly different between the two groups (p = 0.188). CONCLUSION: In well-maintained SPC patients, SRP protocols resulted in significant clinical improvements in terms of BoP; however, for the other clinical improvements, similar efficacy for both GBM and SRP was observed.

Non-Invasive Physical Plasma Reduces the Inflammatory Response in Microbially Prestimulated Human Gingival Fibroblasts.

Non-invasive physical plasma (NIPP), an electrically conductive gas, is playing an increasingly important role in medicine due to its antimicrobial and regenerative properties. However, NIPP is not yet well established in dentistry, although it has promising potential, especially for periodontological applications. The aim of the present study was to investigate the effect of NIPP on a commercially available human gingival fibroblast (HGF) cell line and primary HGFs in the presence of periodontitis-associated bacteria. First, primary HGFs from eight patients were characterised by immunofluorescence, and cell numbers were examined by an automatic cell counter over 5 days. Then, HGFs that were preincubated with Fusobacterium nucleatum (F.n.) were treated with NIPP. Afterwards, the IL-6 and IL-8 levels in the cell supernatants were determined by ELISA. In HGFs, F.n. caused a significant increase in IL-6 and IL-8, and this F.n.-induced upregulation of both cytokines was counteracted by NIPP, suggesting a beneficial effect of physical plasma on periodontal cells in a microbial environment. The application of NIPP in periodontal therapy could therefore represent a novel and promising strategy and deserves further investigation.

Antimicrobial Efficiency of Pistacia lentiscus L. Derivates against Oral Biofilm-Associated Diseases-A Narrative Review.

Pistacia lentiscus L. (PlL) has been used for centuries in traditional medicine. The richness in antimicrobial biomolecules of Pll derivates can represent an alternative to chemically formulated agents used against oral infections. This review summarizes the knowledge on the antimicrobial activity of PlL essential oil (EO), extracts, and mastic resin against microorganisms being of relevance in oral biofilm-associated diseases. Results demonstrated that the potential of PlL polyphenol extracts has led to increasing scientific interest. In fact, the extracts are a significantly more effective agent than the other PlL derivates. The positive findings regarding the inhibition of periodontal pathogens and C. albicans, together with the antioxidant activity and the reduction of the inflammatory responses, suggest the use of the extracts in the prevention and/or reversal of intraoral dysbiosis. Toothpaste, mouthwashes, and local delivery devices could be effective in the clinical management of these oral diseases.

In vitro activity of hyaluronic acid and human serum on periodontal biofilm and periodontal ligament fibroblasts.

OBJECTIVES: A beneficial effect of cross-linked hyaluronic acid (cHA) on periodontal wound healing and regeneration has recently been demonstrated. The present in vitro study was designed to obtain deeper knowledge on the effect of cHA when applied in the gingival sulcus (serum-rich environment) during non-surgical periodontal therapy. MATERIALS AND METHODS: The influence of cHA, human serum (HS), and cHA/HS on (i) a 12-species biofilm formation, (ii) the adhesion of periodontal ligament fibroblasts (PDLF) to dentine surface, (iii) the expression and secretion of interleukin-8, and (iv) the expression of receptors of HA in PDLF and gingival fibroblasts (GF) were evaluated. RESULTS: At 4 h of biofilm formation, cHA and HS in combination (cHA/HS) slightly decreased the colony-forming unit counts in biofilm whereas the metabolic activity of biofilm was reduced in all test groups (cHA, HS, cHA/HS) vs. control. At 24 h, the quantity of biofilm was reduced in all test groups vs. untreated control. The test substances did not affect adhesion of PDLF to dentin. HS increased the expression of IL-8 by PDLF and GF which was partially downregulated by cHA. HS and/or cHA promoted the expression of the HA receptor RHAMM in GF but not in PDLF. CONCLUSIONS: In summary, the present data indicate that serum neither negatively affect the activity of cHA against periodontal biofilm nor had any unwanted influence on the activity of PDLF. CLINICAL RELEVANCE: These findings lend additional support for the positive effects of cHA on cells involved in periodontal wound healing, thus pointing to its potential use in non-surgical periodontal therapy.

Antibiotic Resistance among Fusobacterium, Capnocytophaga, and Leptotrichia Species of the Oral Cavity.

PURPOSE: Antibiotics play an important role in treating periodontal diseases. Due to the effectiveness of antibiotic therapies, their usage in dentistry has significantly increased. The aim of this study focused on the in-vitro susceptibility of different gram-negative oral bacteria species - which are associated with periodontal diseases (Fusobacterium spp., Capnocytophaga spp. and Leptotrichia buccalis) and have different geographical origins (Asia and Europe) - against antimicrobials that are clinically relevant in dental therapy. MATERIALS AND METHODS: A total of 45 strains were tested (29 Fusobacterium spp., 13 Capnocytophaga spp. and 3 L. buccalis) that were either isolated from Chinese patients or were obtained from different strain collections. Their antimicrobial susceptibility to the antimicrobial agents benzylpenicillin, amoxicillin, amoxicillin-clavulanic acid, ciprofloxacin, moxifloxacin, clindamycin, doxycycline, tetracycline and metronidazole was tested using the E-Test. Strains with particular resistance to penicillin, clindamycin and metronidazole were further analysed for resistance genes. RESULTS: All tested bacterial isolates were sensitive to amoxicillin, amoxicillin-clavulanic acid, doxycycline and tetracycline, but showed variable sensitivity towards other antibiotics such as benzylpenicillin, ciprofloxacin, moxifloxacin, clindamycin and metronidazole. CONCLUSION: The results of the present study suggest that certain periodontal disease-related bacterial strains can be resistant towards antimicrobial agents commonly used in adjuvant periodontal therapy.

Apelin Enhances the Effects of Fusobacterium nucleatum on Periodontal Ligament Cells In Vitro.

This study aimed to explore effects of Fusobacterium nucleatum with or without apelin on periodontal ligament (PDL) cells to better understand pathomechanistic links between periodontitis and obesity. First, the actions of F. nucleatum on COX2, CCL2, and MMP1 expressions were assessed. Subsequently, PDL cells were incubated with F. nucleatum in the presence and absence of apelin to study the modulatory effects of this adipokine on molecules related to inflammation and hard and soft tissue turnover. Regulation of apelin and its receptor (APJ) by F. nucleatum was also studied. F. nucleatum resulted in elevated COX2, CCL2, and MMP1 expressions in a dose- and time-dependent manner. Combination of F. nucleatum and apelin led to the highest (p < 0.05) expression levels of COX2, CCL2, CXCL8, TNF-α, and MMP1 at 48 h. The effects of F. nucleatum and/or apelin on CCL2 and MMP1 were MEK1/2- and partially NF-κB-dependent. The combined effects of F. nucleatum and apelin on CCL2 and MMP1 were also observed at protein level. Moreover, F. nucleatum downregulated (p < 0.05) the apelin and APJ expressions. In conclusion, obesity could contribute to periodontitis through apelin. The local production of apelin/APJ in PDL cells also suggests a role of these molecules in the pathogenesis of periodontitis.

Bovine Milk and Yogurt Affect Oral Microorganisms and Biofilms In-Vitro.

PURPOSE: To evaluate the effect of bovine milk and yogurt on selected oral microorganisms and different oral biofilms. MATERIALS AND METHODS: Milk was prepared from 0.5% fat (low-fat) and 16% fat (high fat) milk powder. For yogurt preparation, the strains Lactobacillus delbrueckii ssp. bulgarcius and Streptococcus thermophilus were added to the milk. Minimal inhibitory concentrations (MIC) and minimal microbiocidal concentrations (MMC) of the test compounds were measured against various microorganisms by the microbroth dilution technique. Cariogenic periodontal biofilms and one containing Candida were created on plastic surfaces coated with test substances. Further, preformed biofilms were exposed to the test substances at a concentration of 100% for 10 min and thereafter 10% for 50 min. Both colony forming units (cfu) and metabolic activity were quantified in the biofilms. RESULTS: Neither high-fat milk, low-fat milk nor casein inhibited the growth of any species. Yogurt and L. delbrueckii ssp. bulgaricus at low MIC and MMC suppressed the growth of Porphyromonas gingivalis and other bacteria associated with periodontal disease. High-fat yogurt decreased cfu in the forming periodontal biofilm by 90%. Both low- and high-fat yogurts reduced metabolic activity in newly forming and preformed periodontal and Candida biofilms, but not in the cariogenic biofilm. CONCLUSIONS: Yogurt and L. delbru eckii ssp. bulgaricus, but not milk, were bactericidal against periodontopathogenic bacteria. Yoghurt reduced the metabolic activity of a Candida biofilm and a periodontal biofilm. Yogurt and L. delbrueckii ssp. bulgaricus may have potential in prevention and therapy of periodontal diseases and Candida infections.

Periodontal microbiology and microbial etiology of periodontal diseases: Historical concepts and contemporary perspectives.

This narrative review summarizes the collective knowledge on periodontal microbiology, through a historical timeline that highlights the European contribution in the global field. The etiological concepts on periodontal disease culminate to the ecological plaque hypothesis and its dysbiosis-centered interpretation. Reference is made to anerobic microbiology and to the discovery of select periodontal pathogens and their virulence factors, as well as to biofilms. The evolution of contemporary molecular methods and high-throughput platforms is highlighted in appreciating the breadth and depth of the periodontal microbiome. Finally clinical microbiology is brought into perspective with the contribution of different microbial species in periodontal diagnosis, the combination of microbial and host biomarkers for this purpose, and the use of antimicrobials in the treatment of the disease.

In Vitro Study on Green Propolis as a Potential Ingredient of Oral Health Care Products.

Propolis is increasingly being discussed as an alternative to commonly used antiseptics. This in vitro study focused on the ethanolic extract of green Brazilian propolis (EEPg) as an additive in an oral health care product. We investigated (i) a potential inflammation-modulation activity of EEPg when a periodontal or Candida biofilm was exposed to monocytic (MONO-MAC-6) cells, (ii) the adhesion of oral pathogens to gingival keratinocytes and (iii) the antimicrobial and antibiofilm effect of different toothpaste formulations. EEPg decreased the levels of interleukin (IL)-1ß and increased IL-10 in MONO-MAC cells challenged with a periodontal biofilm. In contact with TIGK cells, EEPg reduced the numbers of adherent Porphyromonas gingivalis to 0.5% but did not affect the adhesion of Candida albicans. The frequent brushing of a cariogenic biofilm with a toothpaste supplemented with EEPg reduced the surface microhardness loss of enamel specimens. Mixing an experimental erythritol toothpaste with 25 and 50 mg/mL of EEPg confirmed the antibacterial activity of EEPg against oral bacteria and particularly inhibited periodontal biofilm formation. The suggested toothpaste formulations seem to have potential in the prevention of caries, gingivitis and periodontitis and should be evaluated in further in vitro research and in clinical trials.

In vitro activity of anti-rheumatic drugs on release of pro-inflammatory cytokines from oral cells in interaction with microorganisms.

Periodontitis patients suffering concomitantly from rheumatoid arthritis (RA) often present with less inflamed periodontal tissues due to the ongoing anti-rheumatic therapy. This in vitro study was aimed to analyze whether anti-inflammatory drugs used in the therapy of RA can modulate the release of IL-8 and IL-1ß by professional and non-professional immune cells stimulated with microorganisms. Periodontal ligament (PDL) fibroblasts, monocytic MONO-MAC-6-cells, and gingival keratinocytes were exposed to ibuprofen, prednisolone, and methotrexate with and without lysates of Fusobacterium nucleatum or Candida albicans. Supernatants were obtained and the levels of interleukin(IL)-8 and IL-1ß (only MONO-MAC-6) were quantified. The addition of F. nucleatum lysate resulted in the strongest release of proinflammatory cytokines by PDL fibroblast and MONO-MAC-6 cells, while the modification by the tested anti-rheumatic drugs was only minor. After stimulation of the MONO-MAC-cells with F. nucleatum, prednisolone increased the release of IL-8, whereas methotrexate decreased the level. Anti-inflammatory drugs increased the adherence of C. albicans to epithelial cells. In patients with RA, the reduction of the microbial load in subgingival biofilm (biofilm removal) is of major importance; however, the intake of inflammatory drugs may interfere with the inflammatory response.

Clinical, microbiological, and immunological effects of 3- or 7-day systemic antibiotics adjunctive to subgingival instrumentation in patients with aggressive (Stage III/IV Grade C) periodontitis: A randomized placebo-controlled clinical trial.

AIM: To evaluate the clinical non-inferiority of a 3-day protocol of systemic antibiotics adjunctive to subgingival instrumentation (SI) compared with a 7-day-protocol in patients with Stage III/IV Grade C periodontitis. MATERIALS AND METHODS: Fifty systemically healthy patients (32.7 ± 4.3 years) with aggressive periodontitis (AgP; Stage III/IV Grade C periodontitis) were treated by SI and adjunctive amoxicillin and metronidazole and were randomly assigned to Group A: (n = 25) 500 mg antibiotics (AB) 3 times a day for 3 days, followed by placebo 3 times a day for 4 days, or Group B: (n = 25) 500 mg AB 3 times a day for 7 days. Clinical, microbial, and immunological parameters were assessed at baseline, 3 months, and 6 months, and patient-related outcomes were assessed after 2 weeks. The primary outcome variable was the number of residual sites with pocket depth (PD) ≥6 mm at 6 months. RESULTS: For the primary outcome variable (the number of residual sites with PD ≥6 mm at 6 months), the null hypothesis was rejected and non-inferiority of the 3-day AB protocol compared with the 7-day AB protocol was demonstrated (the upper limits of the 95% confidence interval for intention to treat analysis: [-2.572; 1.050] and per protocol analysis: [-2.523; 1.318] were lower than the assumed margin of Δ = 3.1). Comparable clinical improvements were obtained for all parameters with both antibiotic protocols (p > .05). All investigated periodontopathogens and pro-inflammatory host-derived markers were statistically significantly reduced without differences between the treatments (p > .05). CONCLUSIONS: These findings indicate that in patients with AgP (Stage III/IV Grade C periodontitis), a 3-day systemic administration of amoxicillin and metronidazole adjunctive to SI may lead to non-inferior clinical outcomes after 6-months with fewer adverse events compared with a 7-day-protocol.

Non-surgical mechanical therapy of peri-implantitis with or without repeated adjunctive diode laser application. A 6-month double-blinded randomized clinical trial.

OBJECTIVES: The objective of this study is to investigate the outcomes following non-surgical therapy of peri-implantitis (PI) with or without adjunctive diode laser application. MATERIALS AND METHODS: A double-blinded randomized controlled clinical trial was carried out in 25 subjects with 25 implants diagnosed with PI. Following curettage of granulation tissue, test implants (T) were treated with adjunctive application of a diode laser for 90 s (settings: 810 nm, 2.5 W, 50 Hz, 10 ms), while at control implants (C) non-activated adjunctive diode laser was applied. The entire treatment procedure was performed at days 0 (i.e., baseline), 7 and 14. The primary outcome measure was change in mean pocket probing depth (PPD). Clinical and microbiological outcomes, as well as host-derived inflammatory markers were evaluated at baseline, 3 and 6 months, while radiographic outcomes were assessed at baseline and at the 6-month follow-up. RESULTS: No statistically significant differences with respect to baseline patient characteristic were observed. After 6 months, both test and control implants yielded statistically significant PPD changes compared with baseline (T: 1.28 and C: 1.47 mm) but without statistically significant difference between groups (p = .381). No statistically significant changes in peri-implant marginal bone levels were detected (p = .936). No statistically significant differences between test and control implants were observed with respect to microbiological and host-derived parameters (p > .05). At the 6-month follow-up, treatment success was observed in 41.7% (n = 5) of test and 46.2% (n = 6) of control patients, respectively (p = .821). CONCLUSION: Repeated adjunctive application of diode laser in the non-surgical management of PI failed to provide significant benefits compared with mechanical instrumentation alone.

One-Year Clinical, Microbiological and Immunological Results of Local Doxycycline or Antimicrobial Photodynamic Therapy for Recurrent/Persisting Periodontal Pockets: A Randomized Clinical Trial.

We evaluated, in this study, the clinical, microbiological and immunological effects of local drug delivery (LDD) or photodynamic therapy (PDT), adjunctive to subgingival instrumentation (SI) in persistent or recurrent periodontal pockets in patients enrolled in supportive periodontal therapy (SPT) after one year. A total of 105 patients enrolled in SPT with persistent/recurrent pockets were randomly treated with SI +PDT or SI + LDD or SI (control). The number of treated sites with bleeding on probing (n BOP+), probing pocket depths (PPD), clinical attachment level (CAL), full-mouth plaque and bleeding scores (gingival bleeding index, %bleeding on probing-BOP) was evaluated at baseline and after 12 months. Additionally, eight periodontopathogens and the immunomarkers IL-1ß (interleukin)and MMP-8 (matrix metalloprotease) were quantitatively determined using real-time PCR and ELISA, respectively. All three treatments resulted in statistically significant clinical improvements (p < 0.05) without statistically significant intergroup differences (p > 0.05), which were maintained up to 12 months. The presence of BOP negatively affected the PPD and CAL. Moreover, statistically significantly fewer bleeding sites at 12 months were observed in the test groups (p = 0.049). Several periodontopathogens were reduced after 12 months. In conclusion, the present data indicate that in periodontal patients enrolled in SPT, treatment of persistent/recurrent pockets with SI alone or combined with either PDT or LDD may lead to comparable clinical, microbiological and immunological improvements, which are maintained up to 12 months. Secondly, the presence of BOP directly impacts the PPD and CAL.

The In-Vitro Activity of a Cold Atmospheric Plasma Device Utilizing Ambient Air against Bacteria and Biofilms Associated with Periodontal or Peri-Implant Diseases.

Due to its antimicrobial and healing-promoting effects, the application of cold atmospheric plasma (CAP) appears to be a promising modality in various fields of general medicine and dentistry. The aim of the present study was to evaluate the antibacterial and anti-biofilm activity of a handheld device utilizing ambient air for plasma generation. Suspensions of 11 oral bacteria (among them Fusobacterium nucleatum, Porphyromonas gingivalis, Parvimonas micra, Streptococcus gordonii, and Tannerella forsythia) were exposed to CAP for 10, 30, 60, and 120 s. Before and after treatment, colony forming unit (CFU) counts were determined. Then, 12-species biofilms were cultured on dentin and titanium specimens, and CAP was applied for 30, 60, and 120 s before quantifying CFU counts, biofilm mass, and metabolic activity. A reduction of ≥3 log10 CFU, was found for ten out of the eleven tested species at 30 s (except for T. forsythia) and for all species at 60 s. For biofilm grown on dentin and titanium specimens, the log10 reductions were 2.43 log10 CFU/specimen and by about 4 log10 CFU/specimen after 120 s of CAP. The CAP application did not reduce the biomass significantly, the metabolic activity of the biofilms on dentin and titanium decreased by 98% and 95% after 120 s of CAP. An application of 120 s of CAP had no cytotoxic effect on gingival fibroblasts and significantly increased the adhesion of gingival fibroblasts to the titanium surface. These results are promising and underline the potential of CAP for implementation in periodontal and peri-implantitis therapy.

TLR2 Activation by Porphyromonas gingivalis Requires Both PPAD Activity and Fimbriae.

Porphyromonas gingivalis, a keystone oral pathogen implicated in development and progression of periodontitis, may also contribute to the pathogenicity of diseases such as arthritis, atherosclerosis, and Alzheimer's. P. gingivalis is a master manipulator of host immune responses due to production of a large variety of virulence factors. Among these, P. gingivalis peptidilarginine deiminase (PPAD), an enzyme unique to P. gingivalis, converts C-terminal Arg residues in bacterium- and host-derived proteins and peptides into citrulline. PPAD contributes to stimulation of proinflammatory responses in host cells and is essential for activation of the prostaglandin E2 (PGE2) synthesis pathway in gingival fibroblasts. Since P. gingivalis is recognized mainly by Toll-like receptor-2 (TLR2), we investigated the effects of PPAD activity on TLR2-dependent host cell responses to P. gingivalis, as well as to outer membrane vesicles (OMVs) and fimbriae produced by this organism. Using reporter cell lines, we found that PPAD activity was required for TLR2 activation by P. gingivalis cells and OMVs. We also found that fimbriae, an established TLR2 ligand, from wild-type ATCC 33277 (but not from its isogenic PPAD mutant) enhanced the proinflammatory responses of host cells. Furthermore, only fimbriae from wild-type ATCC 33277, but not from the PPAD-deficient strains, induced cytokine production and stimulated expression of genes within the PGE2 synthesis pathway in human gingival fibroblasts via activation of the NF-ĸB and MAP kinase-dependent signaling pathways. Analysis of ten clinical isolates revealed that type I FimA is preferable for TLR2 signaling enhancement. In conclusion, the data strongly suggest that both PPAD activity and fimbriae are important for TLR2-dependent cell responses to P. gingivalis infection.

Effect of Bacterial Infection on Ghrelin Receptor Regulation in Periodontal Cells and Tissues.

The effect of bacterial infection on the expression of growth hormone secretagogue receptor (GHS-R) was investigated in periodontal cells and tissues, and the actions of ghrelin were evaluated. GHS-R was assessed in periodontal tissues of rats with and without periodontitis. Human gingival fibroblasts (HGFs) were exposed to Fusobacterium nucleatum in the presence and absence of ghrelin. GHS-R expression was determined by real-time PCR and immunocytochemistry. Furthermore, wound healing, cell viability, proliferation, and migration were evaluated. GHS-R expression was significantly higher at periodontitis sites as compared to healthy sites in rat tissues. F. nucleatum significantly increased the GHS-R expression and protein level in HGFs. Moreover, ghrelin significantly abrogated the stimulatory effects of F. nucleatum on CCL2 and IL-6 expressions in HGFs and did not affect cell viability and proliferation significantly. Ghrelin stimulated while F. nucleatum decreased wound closure, probably due to reduced cell migration. Our results show original evidence that bacterial infection upregulates GHS-R in rat periodontal tissues and HGFs. Moreover, our study shows that ghrelin inhibited the proinflammatory actions of F. nucleatum on HGFs without interfering with cell viability and proliferation, suggesting that ghrelin and its receptor may act as a protective molecule during bacterial infection on periodontal cells.