Bacteria In Periodontal Pockets Research Articles

α-MANGOSTIN/CLOVE OIL-INCORPORATED LAURIC ACID-BASED SOLVENT REMOVAL-INDUCED IN SITU FORMING MATRICES

Typically, the bacteria in periodontal pocket of periodontitis disease are capable of producing a number of virulence factors, which are directly toxic to host tissues or immune cells, or indirectly damage host tissues via induction of inflammatory cytokines. The eradication of these microbes from periodontal pocket is one of crucial processes of periodontitis treatment. The α-mangostin from Garcinia mangostana Linn has the most potent antibacterial activity with less tendency to acquire resistance; therefore, it is interesting for using as active compound for periodontitis treatment. This study focuses on the development of in situ forming matrices (ISM) involving 50% w/w lauric acid, 5% w/w clove oil and different α-mangostin amount in NMP for periodontal drug delivery. Physicochemical properties of ISM were investigated including pH, density, viscosity, surface tension, in vitro matrix formation, and injectability. Antimicrobial activities of formulated ISM against three standard microbes were evaluated using agar cup diffusion methods. The pH and density of all α-mangostin and clove oil-loaded 50% w/w lauric acid matrices was 4.40 ± 0.04 to 4.78 ± 0.20 and 0.960 ± 0.000 to 0.963 ± 0.001 g/cm3, respectively. The increasing α-mangostin amounts significantly increased the viscosity of prepared formulation and injection force. All formulations had low viscosity that it was easy to injection and spreadable in the periodontal pockets. After contact PBS pH 6.8, they transformed as matrix and the hydrophobic manner of clove oil retarded a solvent exchange. These prepared formulations showed the antimicrobial activities against Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 25922, and Candida albicans ATCC 17011. Hence, they are the potential systems for periodontal drug delivery.

Obesity Is Associated with the Severity of Periodontal Inflammation Due to a Specific Signature of Subgingival Microbiota.

The aim of this study was to analyze the link between periodontal microbiota and obesity in humans. We conducted a cohort study including 45 subjects with periodontitis divided into two groups: normo-weighted subjects with a body mass index (BMI) between 20 and 25 kg/m2 (n = 34) and obese subjects with a BMI > 30 kg/m2 (n = 11). Our results showed that obesity was associated with significantly more severe gingival inflammation according to Periodontal Inflamed Surface Area (PISA index). Periodontal microbiota taxonomic analysis showed that the obese (OB) subjects with periodontitis were characterized by a specific signature of subgingival microbiota with an increase in Gram-positive bacteria in periodontal pockets, associated with a decrease in microbiota diversity compared to that of normo-weighted subjects with periodontitis. Finally, periodontal treatment response was less effective in OB subjects with persisting periodontal inflammation, reflecting a still unstable periodontal condition and a risk of recurrence. To our knowledge, this study is the first exploring both salivary and subgingival microbiota of OB subjects. Considering that OB subjects are at higher periodontal risk, this could lead to more personalized preventive or therapeutic strategies for obese patients regarding periodontitis through the specific management of oral microbiota of obese patients.

Abstract 12122: Impact of Periodontal Bacteria Detected in Periodontal Pockets in Patients with Congestive Heart Failure on Long-Term Clinical Outcomes

Backgrounds: Periodontal disease is known as a risk of cardiovascular events, but the impact of periodontal bacteria on the clinical outcomes has not been elucidated in patients with congestive heart failure (CHF). Aim: We sought to investigate the association between the presence of periodontal bacteria in periodontal pockets. Methods: Between May 2012 and August 2015, consecutive 149 patients who admitted in our department due to non-ischemic CHF were retrospectively included. Periodontal examinations including the presence of major periodontal bacteria (MPB) were performed by the independent trained periodontists. MPB consisted of Porphyromonas gingivalis , Aggregatibacter actinomycetemcomitans and Prevotella intermedia , known as inflammatory bacteria of severe periodontitis. The clinical events of these patients were assessed, and major adverse cardiovascular cerebrovascular event (MACCE) was defined as the composite of cardiovascular death, non-fetal myocardial infarction, non-fetal stroke. Results: A total of 135 patients composed of 98 patients with MPB in the periodontal pocket and 37 patients without that were identified. Median follow up duration was 4.6 years (interquartile: 1.8 - 6.6 years). The Kaplan-Meier curve analysis about MACCE suggested that the free survival rate in MPB group was significantly worse than non-MPB group (p for log-rank test = 0.01). Regarding Kaplan-Meier curve analysis of each MACCE component, only that of cardiovascular death showed MPB group had significantly worse free survival rate (p = 0.02). Multivariate Cox hazard regression model analysis revealed that MPB group was an independent risk of MACCE (HR 9.70; 95% CI 1.27 - 74.39, p = 0.03), and this finding appeared to be obtained from mainly cardiovascular death (HR 8.74; 95% CI 1.13 - 67.71, p = 0.04). Conclusions: In non-ischemic CHF patients with MPB in periodontal pocket were associated with an increased risk of long-term cardiovascular events.

Antimicrobial photodynamic chemotherapy mediated by PapaMBlue on chronic periodontal disease: Study protocol for a randomized, blind, controlled trial.

Background:The elimination of the pathogenic microorganisms of the periodontal pocket is one of the main points for success in periodontal treatment. The objective of this study is to investigate the clinical and antimicrobial effect of papain-mediated photodynamic therapy in the clinical treatment of periodontal disease.Methods:Twenty patients with chronic periodontitis will be selected. Patients will be randomly divided into 2 groups (n = 10). Group 1 will receive conventional periodontal treatment and group 2 will receive conventional treatment and antimicrobial photodynamic therapy (PACT). Conventional treatment will consist of oral hygiene guidance, with brushing technique instructions and recommendation of daily flossing. The calculus deposits on the teeth will be removed with ultrasound equipment and curettes for scraping and root planning. The PACT will be performed at the end of each periodontal treatment session, at sites with bags ≥4 mm. PapaMblue photosensitizer will be deposited in the periodontal pockets with a syringe and a pre-irradiation time of 1 minute will be adopted. Then, the laser emitting wavelength of 660 nm, with power of 100 mW, for 2 minutes, radiant exposure of 30 J/cm2 and power density of 250 mW/cm2 will be applied. Patients will undergo clinical evaluations before treatment (day 1) at 30, 60, and 90 days after the end of treatment; and microbiological evaluations before and immediately after treatment. The distribution of the data within each group and the homogeneity of the variances will be verified. With this information, the most appropriate statistical test in each evaluation will be used. The sample calculation is based on the literature and the significance level of 5% will be adopted.Discussion:The combination of PACT with methylene blue in a papain gel and the conventional treatment may increase the reduction of bacteria in periodontal pockets.

Antimicrobial activity of amixicile against Treponema denticola and other oral spirochetes associated with periodontal disease.

Periodontal disease is a polymicrobial infection characterized by inflammation of the gingiva, alveolar bone resorption and tooth loss. As periodontal disease progresses, oral treponemes (spirochetes) become dominant bacteria in periodontal pockets. Oral treponemes are anaerobes and all encode the enzyme pyruvate-ferredoxin oxidoreductase (PFOR) which catalyzes the oxidative decarboxylation of pyruvate to acetyl-CoA. Here we assess the susceptibility of oral treponemes to amixicile (AMIX), a novel inhibitor of PFOR. The minimum inhibitory concentration (MIC) of AMIX against several oral treponeme species was determined. The impact of AMIX on processes relevant to virulence including motility, H2 S production, and complement evasion were determined. The growth of all oral treponeme species tested was inhibited by AMIX with MIC concentrations (MIC) ranging from 0.5-1.5 μg/mL. AMIX significantly reduced motility, caused a dose-dependent decrease in hydrogen sulfide production and increased sensitivity to killing by human complement (i.e., serum sensitivity). AMIX is effective in vitro in inhibiting growth and other processes central to virulence. AMIX could serve could serve as a new selective therapeutic tool for the treatment of periodontal disease.

Effect of Combination of Water Jet and Sonic Wave Toothbrush on Bacteria in Periodontal Pockets

Effect of Combination of Water Jet and Sonic Wave Toothbrush on Bacteria in Periodontal Pockets

Isolation, identification and growth characteristics of Porphyromonas gingivalis clinical strains

Objective To isolate and identify Porphyromonas gingivalis(P.gingivalis) from clinical subjects,and analyse the growth characteristics of P.gingivalis. Methods Subgingival plaque samples were collected from patients with chronic periodontitis and periodontally healthy subjects,and anaerobic culture was performed.P.gingivalis strains were identified by PCR and 16S rDNA sequencing,and detection rate was analysed.Bacteria colonies and growth characteristics of the clinical strains of P.gingivalis were observed,and the biological properties were analysed. Results Six P.gingivalis clinical strains named as L2,L3,L4,L5,L11 and L12 were isolated from 146 subgingival plaque samples collected from 35 subjects.The detection rate of P.gingivalis was 27.4% in subgingival plaque samples of periodontally healthy subjects,86.7% in periodontally healthy sites of patients with chronic periodontitis,95.6% in periodontal pockets with probing depth of 4-6 mm,and 96.0% in periodontal pockets with probing depth 6 mm.Strains L2,L5,L11 and L12 formed black colonies on BHI plates after culture for 1 week.Strains L3 and L4 formed grey colonies 1 week after culture for 1 week,and formed black colonies after culture for 10 d.Strains L2,L4,L5 and L12 grew to exponential phase 30 h after inoculation,whereas strains L3 and L11 grew to exponential phase 60 h after inoculation. ConclusionP.gingivalis is one of the dominant pathogenic bacteria in periodontal pockets of patients with chronic periodontitis.The detection rate of P.gingivalis in healthy sites of patients with chronic periodontitis is significantly higher than that of periodontally healthy subjects,and the detection rates of P.gingivalis are higher in deeper periodontal pockets.The growth characteristics of P.gingivalis clinical strains vary from each other,indicating that the biological and pathogenic properties of P.gingivalis may be strain-specific.

162* Identical obligate anaerobic bacteria in periodontal pockets and sputum of cystic fibrosis patients

162* Identical obligate anaerobic bacteria in periodontal pockets and sputum of cystic fibrosis patients

Capacity of photodynamic therapy for microbial reduction in periodontal pockets

Practitioners are not successful in implemented treatments due to the great difficulty in completely removing bacterial deposits and their endotoxins. This study aimed to evaluate the capacity of photodynamic therapy to reduce the numbers of viable bacteria in periodontal pockets. Microbiological samples were collected before and after scaling and after photodynamic therapy. Photodynamic therapy was performed through the insertion of the photosensitizer toluidine blue and Endo PTC into the pocket for 3 min, followed by photosensitization with low-intensity diode of 4 J/cm(2). The results (log(10)) were submitted to a descriptive analysis and a t-test. A reduction of 81.24% in the numbers of bacteria after scaling was observed, as well as 95.90% after photodynamic therapy (P < 0.01). Photodynamic therapy is indicated as an adjuvant treatment to reduce the numbers of viable bacteria in periodontal diseases.

Biofilm-forming bacteria in periodontal pockets screened by measurement of viscosity of culture media and observation of cell surface structures

Biofilm-forming bacteria in periodontal pockets screened by measurement of viscosity of culture media and observation of cell surface structures

Amine fluoride gel affects the viability and the generation of superoxide anions in human polymorphonuclear leukocytes: an in vitro study.

Amine hydrofluorides are widely used to prevent caries. As an acidulated gel, they were also studied for their applicability to reduce pathogenic bacteria in periodontal pockets. We assessed the toxicity of this pharmaceutical amine hydrofluoride preparation on human polymorphonuclear leukocytes in vitro by measuring Trypan blue exclusion and the generation of superoxide anions (O2) by the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) after a 3-min contact with gel. Depending on the experimental conditions, gel dilutions up to 1.3 x 10(4) resulted in an increase in Trypan blue-colored cells and liberation of beta-glucuronidase. Dilutions between 3 x 10(4) and 1 x 10(5) augmented the fMLP-mediated O2- generation, which could be prevented by Ca2+ chelation with BAPTA-AM (1,2'-bis (o-aminophenoxyethane-N.N.N'.N'-tetraacetic acid tetra (acetoxymethyl) ester) and ethyleneglycoltetraacetic acid (EGTA) or inhibition of protein kinase C (PKC) with staurosporine and bisindolylmaleimide I. respectively. Compared with data published on the minimal inhibitory concentration for periodontal pathogenic bacteria, the cytotoxicity of amine hydrofluorides on eukaryotic cells is much greater and thus of consequence for their clinical use.

Activation of human gingival epithelial cells by cell-surface components of black-pigmented bacteria: augmentation of production of interleukin-8, granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor and expression of intercellular adhesion molecule 1.

Black-pigmented anaerobic bacteria, such as Porphyromonas gingivalis and Prevotella intermedia, are amongst the predominant bacteria in periodontal pockets and have been implicated in periodontal diseases. To elucidate the roles of gingival keratinocytes, which are the first cells encountered by oral bacteria in periodontal diseases, human gingival keratinocytes in primary culture were stimulated with cell-surface components of P gingivalis and Pr. intermedia. A glycoprotein fraction from Pr. intermedia (PGP) clearly augmented the release of interleukin-8, granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor, as determined by enzyme-linked immunosorbent assay. This PGP also induced expression of intercellular adhesion molecule-1 (ICAM-1), as determined by flow cytometry. The augmentation of mRNA expression for these molecules was also confirmed by reverse transcription PCR. In contrast, lipopolysaccharide (LPS) from Pr. intermedia and Escherichia coli was completely inactive in these assays. LPS fraction and purified fimbriae from P gingivalis exhibited weak activities. Cytokine production and ICAM-1 expression by gingival keratinocytes might cause accumulation and activation of neutrophils in the epithelium and, therefore, may be involved in the initiation and development of inflammation in periodontal tissues.

Serum antibody titers to Bacteroides forsythus in elderly subjects with gingivitis or periodontitis.

The risks for periodontal disease appear to increase with age. To determine associations between clinical findings, the presence of specific bacteria in periodontal pockets, and serum antibody titers. 10 older subjects (mean age=73.0 years SD+/-4.9) with confirmed gingivitis only (gingivitis group) and 10 subjects with periodontitis (mean age: 76.1 years, SD+/-10.4) (periodontitis group) were studied. The mean group differences for probing depth and clinical attachment levels were 4.1 mm and 5.6 mm, respectively, and were significantly different (p<0.001). Both groups had high plaque scores (>60% surfaces with plaque). DNA probes demonstrated that B. forsythus was present in 8/10 samples from the periodontitis group and in 7/10 samples from the gingivitis group. The B. forsythus isolates studied were found in four of the subjects with periodontitis and from 2 of the subjects with gingivitis. Serum antibody titers to 6 ribotypes of B. forsythus were studied. Western blots, gradient gels, and pulsed field gel electrophoresis concurrently demonstrated that the B. forsythus isolates were genotypically, and phenotypically unique for each subject. Antibody titers to two selected B. forsythus isolates were significantly higher in the periodontitis group (p<0.01, Mann-Whitney test). The study confirmed that antibody serum titers to the six different ribotypes of B. forsythus varied greatly between older individuals with gingivitis or periodontitis. Not all strains of B. forsythus elicited higher titers in periodontitis affected subjects. The results of the present study suggest genotype variation of B. forsythus that is unique to the individual and that serotype variation can be expected. It is possible that B. forsythus under specific host conditions can modulate surface antigen factors to evade the host immune response.

Sulfate-reducing bacteria in periodontal pockets and in healthy oral sites.

The aim of this study was to monitor the presence of sulfate-reducing bacteria (SRB) at different sites in the mouths of both healthy individuals and periodontitis patients. In 20 healthy subjects and 21 periodontitis patients, samples were taken from the palate, vestibulum, dorsum of the tongue, supragingival plaque, and periodontal pockets. In order to demonstrate growth of SRB, samples were incubated in an anoxic chamber in a reduced growth-medium for SRB, with an iron-indicator for sulfide production. The SRB were detected throughout the oral cavity. They were found on the mucosa in 10% of both healthy subjects and periodontitis patients. On the tongue and in supragingival plaque, the frequency of detection was slightly higher (22% of the subjects). In contrast, 86% of the periodontitis patients harbored SRB in one or more pockets. In 1/3 of the patients, SRB were present in all 3 pockets that were sampled. The data indicated that SRB belong to the normal oral microbiota, and have a preference for periodontal pockets.

Restriction fragment-length polymorphism analysis of 16S ribosomal RNA genes amplified by polymerase chain reaction for rapid identification of cultivable oral treponemes.

Although oral treponemes are among the most frequently found bacteria in periodontal pockets, identification of these organisms can be difficult. In this study, restriction fragment-length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified 16S ribosomal RNA genes (16S rRNA gene PCR-RFLP) was used to generate restriction profiles of reference strains of oral treponemes including Treponema denticola, Treponema socranskii, Treponema vincentii. Treponema pectinovorum and Treponema medium as well as for Treponema phagedenis and Treponema pailidum and five treponeme strains isolated from human peridontal pockets. Before RFLP analysis, the 16S rRNA gene sequences were obtained from the GenBank database, and the analysis of the theoretical banding patterns for HpaII suggested good species discrimination. 16S rRNA gene sequences were amplified from isolated genomic DNA samples by PCR with spirochete-specific primers. The PCR products were then purified and characterized by single digestion with restriction endonuclease HpaII, and this allowed discrimination between the respective reference strains. Five clinical isolates, four T. denticola and one T. socranskii, were assigned on the basic of their restriction profiles by digestion with HpaII. 16S rRNA gene PCR-RFLP using HpaII is a rapid and reliable method for differentiation of cultivable oral treponemes.

Bactericidal efficacy of metronidazole against bacteria of human periodontal pockets in vitro.

The bactericidal efficacy of metronidazole against bacteria taken from periodontal pockets was estimated in vitro by measuring the difference between bacterial recovery from pocket samples on metronidazole-containing (10μg/ml) BHI-blood agar plates and control plates under strictly anaerobic conditions. More than 105 bacteria were recovered from each of periodontal pocket, whereas more than 99% of the bacteria were not recovered when samples were incubated on metronidazole containing BHI-blood agar plates, indicating that metronidazole effectively disinfected most of the bacteria of periodontal pockets. The results indicate that not only obligate anagerobes but also most facultative bacteria in periodontal pockets were sensitive to metronidazole under anaerobic conditions, beacause the predominant bacteria in periodontal pockets were composed of obligate anaerobes (73%) and facultative bacteria (27%).

Inhibitory Effect of Human Plasma and Saliva on Co-aggregation Between Bacteroides gingivalis and Streptococcus mitis

The effect of human plasma and saliva on co-aggregation between Bacteroides gingivalis and Streptococcus mitis was studied by means of a turbidimetric assay. The co-aggregation activity was obtained from the maximum slope of the absorbance vs. time curve. Its dependence on pH, temperature, and ionic strength was examined, and the number of Bacteroides cells in relation to the number of Streptococcus cells resulting in optimal co-aggregation was established. Co-aggregation inhibition experiments showed that the co-aggregation activity was inhibited by l-arginine and l-lysine, although the activity was unaffected by the sugars tested. Human plasma and saliva were able to inhibit the co-aggregation in a dose-dependent reaction. Plasma exhibited the most potent inhibitory activity in these fluids. Fibrinogen was the most potent inhibitor of the plasma-derived proteins tested. These data suggest the possibility that the oral fluids may modulate the attachment of B. gingivalis to Gram-positive bacteria in periodontal pockets.

Proportions and identity of IgA1-degrading bacteria in periodontal pockets from patients with juvenile and rapidly progressive periodontitis.

Degradation of immunoglobulin A1 (IgAl) has previously been demonstrated in suspected periodontal pathogens. The present study revealed that a considerable proportion of the microbial flora in periodontal pockets of patients with juvenile periodontitis (median 29%) and rapidly progressive periodontitis (median 27%) was capable of degrading IgAl. Four different types of degradation occurred: Complete degradation of IgAl; extensive degradation leaving the Fc part of the molecule intact; traditional IgAl protease activity yielding intact Fc and Fab fragments, and removal of carbohydrate side chains on the IgAl molecule. Apart from species already known to degrade IgAl. IgAl protease was, for the first time, demonstrated in strains of Veillonella spp. The ability to cleave off carbohydrate side chains was a feature of a wide variety of bacteria belonging to the species Streptococcus sanguis, S. mitior, S. milleri, Veillonella spp., Actinomyces naeslundii. A. viscosus. Arachnia propionica, and Bacterionema matruchotii. The ability of subgingivally colonizing bacteria to degrade IgA may be a factor contributing to aggravation and perpetuation of the inflammatory reaction in the periodontal tissues.