interleukin-8 and Dental-Plaque

Several microbiologic and epidemiologic studies have suggested an association between dental plaque, poor oral health, and respiratory diseases such as nosocomial pneumonia and chronic obstructive pulmonary disease (COPD). A number of hypotheses are suggested to help explain how oral bacteria may participate in the pathogenesis of respiratory infection. Resident bacteria in oral secretions are likely aspirated along with respiratory pathogens and may affect the adhesion of the later organisms to the respiratory epithelium. Preliminary studies performed in our laboratory suggest that oral bacteria may modulate the adhesion of respiratory pathogens to epithelial cell lines. In addition, oral bacterial products or cytokines in oral/pharyngeal aspirates may stimulate cytokine production from respiratory epithelial cells, resulting in recruitment of inflammatory cells. The resulting inflamed epithelium may be more susceptible to respiratory infection. Further preliminary data are presented that some species of oral bacteria may induce the release of proinflammatory cytokines from epithelial cell lines to an extent similar to that seen for respiratory pathogens.

Topics: Aggregatibacter actinomycetemcomitans; Analysis of Variance; Bacterial Adhesion; Cell Line; Chemotaxis, Leukocyte; Cytokines; Dental Plaque; Epithelial Cells; Haemophilus influenzae; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Linear Models; Mouth; Pneumonia, Bacterial; Porphyromonas gingivalis; Pulmonary Disease, Chronic Obstructive; Respiratory Mucosa; Respiratory Tract Infections; Streptococcus; Streptococcus pneumoniae; Tumor Necrosis Factor-alpha

To determine the preventive effect of a periodontal dressing containing colophony, zinc oxide and magnesium oxide applied after scaling and root planing on clinical variables, subgingival bacteria and local immune response in patients with chronic periodontitis.. In this randomised prospective clinical study, 28 volunteers with generalised moderate chronic periodontitis were treated with full-mouth scaling in a split-mouth design. In the test quadrants, the periodontal dressing was applied during the first three days. At baseline and after 6 and 12 weeks, probing pocket depth (PD), attachment level (AL) and bleeding on probing (BOP) were recorded, and subgingival plaque samples were taken for laboratory analysis.. In both groups, PD, AL and BOP were significantly reduced (p=0.001). BOP was significantly lower in the control than the test group after 6 weeks (p=0.046). Significantly reduced bacterial counts of Porphyromonas gingivalis were found in the control group after 12 weeks (p=0.013). No differences were found for the microbiological results between the groups. After 12 weeks, interleukin (IL)-8 and matrix metalloproteinase (MMP)-8 were significantly higher in the test group (p=0.023 and p=0.003, respectively).. The adjunctive application of a periodontal dressing had no additional preventive effect on clinical data 12 weeks after scaling and root planing.

Topics: Adult; Aged; Bacterial Load; Chronic Periodontitis; Combined Modality Therapy; Dental Plaque; Dental Scaling; Female; Follow-Up Studies; Humans; Interleukin-8; Magnesium Oxide; Male; Matrix Metalloproteinase 8; Middle Aged; Periodontal Attachment Loss; Periodontal Dressings; Periodontal Index; Periodontal Pocket; Pinus; Porphyromonas gingivalis; Prospective Studies; Resins, Plant; Root Planing; Tars; Treatment Outcome; Zinc Oxide

A controlled clinical trial was conducted to evaluate the effects of oral prophylaxis on halitosis-associated, immunological and microbiological parameters.. Thirty subjects were included in this controlled clinical trial (patients with generalized chronic periodontitis and controls without clinical attachment loss; each n = 15). Before oral prophylaxis and 14 days after (including tongue cleaning) volatile sulphur compounds (VSC), organoleptic scores and a tongue coating index were evaluated. The levels of IL-1β, IL-8, IL-10 and MMP-8 were measured in GCF, and also major periodontal pathogens were detected. Data were statistically analysed using anova and paired t-test.. Supragingival plaque and calculus removal with combined tongue cleaning was able to reduce significantly (P < 0.05) the VSC values in both groups (no significant differences between both groups). Two weeks after periodontal debridement, the VSC values were observed in the periodontitis group, but not in the control group, similar to the baseline values. The difference between the groups was statistically significant (P < 0.05). Only a repeated prophylaxis session in the periodontitis group was able to reduce VSC values significantly in comparison with baseline (P < 0.05). Organoleptic scores (10 and 30 cm) were significantly different (P < 0.05) between both groups before and after the treatment. Periodontal pathogens and host-derived markers were not significantly affected by a single prophylaxis session.. Oral prophylaxis may result in a significant decrease in VSC values. However, in periodontal diseases, a more complex treatment seems to be necessary.

Topics: Adult; Aged; Aggregatibacter actinomycetemcomitans; Bacteroides; Chronic Periodontitis; Dental Calculus; Dental Plaque; Dental Prophylaxis; Female; Follow-Up Studies; Gingival Crevicular Fluid; Halitosis; Humans; Interleukin-10; Interleukin-1beta; Interleukin-8; Male; Matrix Metalloproteinase 8; Middle Aged; Oral Hygiene; Periodontal Debridement; Porphyromonas gingivalis; Sulfur Compounds; Tongue; Treponema denticola; Volatile Organic Compounds; Young Adult

To examine microbiological and immunological alterations following two periodontal surgical techniques, over a 6-month period.. A total of 30 chronic periodontitis patients participated in the present randomized controlled clinical trial and were randomized in two groups. Modified Widman flap (MWF) was applied in the control group and apically positioned flap (APF), without intervention to the bone, in the experimental group. Gingival crevicular fluid samples and subgingival plaque samples from the operated sites were collected at baseline, 6th, 12th and 24th post-operative week.. No major differences were noticed in immunological and microbiological profile of patients receiving either modified MWF or APF, for a period of 6 months.. The choice of the periodontal surgical procedure does not seem to affect the immunological and the microbiological profile of patients with chronic periodontitis.

Topics: Actinomyces; Adult; Aged; Bacteroides; Chronic Periodontitis; Dental Plaque; Dental Plaque Index; Dental Scaling; Female; Follow-Up Studies; Gingival Crevicular Fluid; Humans; Interleukin-10; Interleukin-1beta; Interleukin-4; Interleukin-6; Interleukin-8; Male; Middle Aged; Periodontal Index; Porphyromonas gingivalis; Prospective Studies; Root Planing; Streptococcus mitis; Surgical Flaps; Treponema denticola; Tumor Necrosis Factor-alpha

Investigate short-term effects of power brushing following experimental induction of biofilm overgrowth in periodontal disease states.. Overall, 175 subjects representing each of five biofilm-gingival interface (BGI) periodontal groups were enrolled in a single-blind, randomized study. After stent-induced biofilm overgrowth for 21 days subjects received either a manual or a power toothbrush to use during a 4 weeks resolution phase. At baseline and during induction and resolution, standard clinical parameters were measured. Subclinical parameters included multikine analysis of 13 salivary biomarkers and 16s Human Oral Microbe Identification Microarray (HOMIM) probe analysis of subgingival plaque samples.. All groups exhibited significantly greater reductions in bleeding on probing (BOP) (p = 0.002), gingival index (GI) (p = 0.0007), pocket depth (PD) (p = 0.04) and plaque index (p = 0.001) with power brushing compared to manual. When BGI groups were combined to form a shallow PD (PD ≤ 3 mm) and a deep PD group (PD > 4 mm) power brushing reduced BOP and GI in subjects with both pocket depths. Power brushing significantly reduced IL-1β levels at resolution while changes in bacterial levels showed non-significant trends between both brushing modalities.. Short-term changes in select clinical parameters and subclinical salivary biomarkers may be useful in assessing efficacy of power brushing interventions in a spectrum of periodontal disease states.

Topics: Acute-Phase Proteins; Adult; Bacteria; Biofilms; Biomarkers; Dental Plaque; Electrical Equipment and Supplies; Female; Gingival Hemorrhage; Gingivitis; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1beta; Interleukin-8; Lipocalin-2; Lipocalins; Male; Matrix Metalloproteinases; Microarray Analysis; Periodontal Diseases; Periodontal Pocket; Proto-Oncogene Proteins; Saliva; Single-Blind Method; Tissue Inhibitor of Metalloproteinases; Toothbrushing

A low-grade systemic inflammatory status originating from periodontal infection has been proposed to explain the association between periodontal disease and systemic conditions, including adverse obstetric outcomes. The aim of this study was to evaluate the effect of periodontal therapy during pregnancy on the gingival crevicular fluid and serum levels of six cytokines associated with periodontal disease and preterm birth.. A subsample of 60 women (18-35 years of age) up to 20 gestational weeks, previously enrolled in a larger randomized clinical trial, was recruited for the present study. Participants were randomly allocated to receive either comprehensive nonsurgical periodontal therapy before 24 gestational weeks (n = 30, test group) or only one appointment for supragingival calculus removal (n = 30, control group). Clinical data, and samples of blood and gingival crevicular fluid, were collected at baseline, at 26-28 gestational weeks and 30 d after delivery. The levels of interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70 and tumor necrosis factor-α were analyzed by flow cytometry.. After treatment, a major reduction in periodontal inflammation was observed in the test group, with bleeding on probing decreasing from 49.62% of sites to 11.66% of sites (p < 0.001). Periodontal therapy significantly reduced the levels of IL-1β and IL-8 in gingival crevicular fluid (p < 0.001). However, no significant effect of therapy was observed on serum cytokine levels. After delivery, the levels of IL-1β in the gingival crevicular fluid of the test group were significantly lower than were those in the control group (p < 0.001), but there were no significant differences between test and control groups regarding serum cytokine levels.. Although periodontal therapy during pregnancy successfully reduced periodontal inflammation and gingival crevicular fluid cytokine levels, it did not have a significant impact on serum biomarkers.

Topics: Adolescent; Adult; Biomarkers; Cytokines; Dental Calculus; Dental Plaque; Dental Scaling; Female; Gingival Crevicular Fluid; Humans; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Oral Hygiene; Periodontal Attachment Loss; Periodontal Diseases; Periodontal Index; Periodontal Pocket; Postpartum Period; Pregnancy; Pregnancy Complications, Infectious; Pregnancy Outcome; Pregnancy Trimester, Second; Premature Birth; Root Planing; Tumor Necrosis Factor-alpha; Young Adult

The aim of this human investigation is to explore the relationship of gingivitis with salivary biomarkers, periodontal pathogens, and interleukin (IL)-1 polymorphism after a transient inflammatory burden.. Thirty healthy human participants were randomized by IL-1 genotype status to control for potential influences of this particular single nucleotide polymorphism on the inflammatory profile. Oral hygiene practices ceased for 21 days to induce gingivitis (induction), after which home care was reinstated until 35 days (resolution). Clinical parameters included plaque (PI) and gingival (GI) indices and papillary bleeding score (PBS). Levels and proportions of 40 subgingival bacteria were determined using checkerboard DNA-DNA hybridization. Saliva was analyzed using a multiplex protein array for 30 biomarkers associated with host defense, inflammation, tissue destruction, and angiogenesis.. Mean PI, GI, and PBS values were significantly increased during induction and decreased during resolution as measured at 35 days (P <0.01), although no differences were observed between IL-1 groups. Participants were stratified as either "high" or "low" responders based on inflammatory response (high: GI >1.5; low: GI ≤1.5). Baseline levels of salivary IL-6 and IL-8 demonstrated the highest ability to discriminate between high and low responders (area under the curve [AUC] of 0.81 and 0.72, respectively). Salivary biomarkers, matrix metalloproteinases (MMPs), and bacterial biofilm were combined to generate receiver operating characteristic curves. High levels of IL-6 and MMP-1 at baseline demonstrated the strongest ability to predict high responders (AUC of 0.89; odds ratio of 17.0; 95% confidence interval, 1.7 to 171.7).. In this proof-of-concept investigation, we identified specific biomarker and microbial signatures that are associated with gingival inflammation (ClinicalTrials.gov number NCT00980525).

Topics: Adolescent; Adult; Biomarkers; Chi-Square Distribution; Dental Plaque; DNA, Bacterial; Female; Genetic Predisposition to Disease; Gingivitis; Humans; Inflammation Mediators; Interleukin-1; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 8; Multiplex Polymerase Chain Reaction; Nucleic Acid Hybridization; Periodontal Index; Polymorphism, Single Nucleotide; Protein Array Analysis; ROC Curve; Saliva; Young Adult

Bacterial plaque is an etiologic factor in the development of gingival inflammation and periodontitis. The presence of orthodontic bands and brackets influences plaque growth and maturation. The purposes of this research were to monitor microbiologic and periodontal changes after placement of orthodontic attachments over a 1-year period and to link these changes to alterations in cytokine concentrations in the gingival crevicular fluid (GCF).. This longitudinal split-mouth trial included 24 patients. Supragingival and subgingival plaque composition, probing depth, bleeding on probing, and GCF flow and composition were assessed at baseline (Tb) and after 1 year (T52). A statistical comparison was made over time and between the banded and bonded sites. Prognostic factors for the clinical reaction at T52 in the GCF at Tb were determined.. Between Tb and T52, the pathogenicity of the plaque and all periodontal parameters increased significantly, but intersite differences were not seen, except for bleeding on probing. The cytokine concentrations in the GCF did not differ significantly between the sites or between Tb and T52. The interleukin-6 concentration in the GCF at Tb was a significant predictive value for the GCF flow at T52 (P <0.05). The same relationship was found between the interleukin-8 concentration at Tb and the increase in probing depth at T52 (P <0.05).. Interleukin-6 and interleukin-8 concentrations before orthodontic treatment were shown to be significant predictive factors for some potential inflammatory parameters during treatment.

Topics: Adolescent; Bacteria; Bacterial Load; Chemokine CCL2; Chemokine CXCL10; Cytokines; Dental Plaque; Extraoral Traction Appliances; Female; Follow-Up Studies; Gingival Crevicular Fluid; Gingival Hemorrhage; Humans; Interferon-gamma; Interleukin-6; Interleukin-8; Longitudinal Studies; Male; Oral Hygiene; Orthodontic Appliances; Orthodontic Brackets; Orthodontic Wires; Patient Education as Topic; Periodontal Index; Periodontal Pocket; Predictive Value of Tests; Prospective Studies

Experimental gingivitis has been studied extensively as a well-controlled laboratory model of gingivitis. It is unclear, however, how experimental gingivitis compares with persistent plaque and gingivitis in more naturalistic settings. The present study compares both conditions in a randomized controlled design.. Twenty-six students suffering from plaque and gingivitis were randomly assigned to either a persistent gingivitis or an experimental gingivitis condition. Subjects with persistent gingivitis continued their habitual (i.e. insufficient) oral hygiene behaviour, resulting in persistence of plaque and gingivitis. Experimental gingivitis consisted of initial prophylaxis and subsequent total neglect of oral hygiene. Crevicular interleukin-1beta and interleukin-8 and clinical data were assessed weekly.. After 4 wk, subjects with experimental gingivitis showed significantly more plaque accumulation (p = 0.005), higher interleukin-1beta (p = 0.037), and lower interleukin-8 (p = 0.043) concentrations than subjects with persistent gingivitis. Whereas in experimental gingivitis we observed considerable fluctuations in clinical and immunological parameters over the 4-wk period, persistent gingivitis was characterized by little fluctuation, indicating that we were monitoring an inflammatory steady state.. The data indicate that conditions observed after 4 wk of experimental gingivitis are not comparable with persistent gingival inflammation in a naturalistic setting. Results are discussed with respect to current studies, indicating that chronic inflammation may reflect a stage of down-regulated pro-inflammatory response.

Topics: Adult; Dental Plaque; Epidemiologic Methods; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-1beta; Interleukin-8; Male; Oral Hygiene; Periodontal Index; Sex Factors

To compare in persons aged 70 years or older the clinical and inflammatory changes occurring around implants and natural teeth during and after a phase of undisturbed plaque accumulation.. Twenty partially edentulous participants with titanium implants refrained from oral hygiene practices while being clinically monitored in weekly intervals for 21 days. Teeth and implants were then cleaned, oral hygiene resumed, and the participants were further monitored for 3 weeks. Twelve biomarkers were assessed in gingival and peri-implant crevicular fluid (GCF, PCF).. During 3 weeks of oral hygiene abstention, the gingival index (GI) continuously increased. On day 21, there were significantly more sites with GI >1 at implants than at teeth. After restarting oral hygiene, the GI decreased markedly in both groups. Throughout the experiment, the plaque index was significantly higher on teeth than on implants. The different biomarkers reacted variably. IL-1β increased significantly with plaque accumulation. IL-1β, GM-CSF, TNF-α, and IFN-γ were significantly higher in GCF compared to PCF at day 21. IL-8 decreased significantly in GCF up to day 14. MIP-1β decreased significantly in GCF, but not in PCF. At the 3-week follow-up, the levels of all biomarkers assessed in GCF and PCF had returned to baseline values.. In an elderly cohort, plaque accumulation induced an inflammatory reaction around both teeth and implants. Although there was less plaque accumulation on implants, the peri-implant mucosa showed a stronger clinical response than gingiva.

Topics: Aged; Aged, 80 and over; Biomarkers; Chemokine CCL4; Dental Implants; Dental Plaque; Female; Gingival Crevicular Fluid; Gingivitis; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-1beta; Interleukin-8; Male; Periodontal Index; Stomatitis; Tumor Necrosis Factor-alpha

It has been shown that toll-like receptor (TLR) 2- and TLR4-stimulating abilities of supragingival plaque (SPP) are associated with periodontal conditions. It is hypothesized that SPP might affect the periodontium through its influence on subgingival plaque (SBP). This study investigates relationships between TLR2- and TLR4-stimulating abilities of SBP and periodontal conditions.. One hundred thirteen SBP samples were collected from the deepest pockets in patients with chronic periodontitis. TLR2- and TLR4-stimulating abilities were measured using genetically engineered nuclear factor-kappa B reporter cells. Numbers of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in each plaque sample were determined by real-time polymerase chain reaction. Peripheral blood mononuclear cells (PBMCs) were stimulated with SBP samples in presence or absence of TLR4 or TLR2 inhibitor. Production of tumor necrosis factor (TNF)-α and interleukin (IL)-8 was analyzed by enzyme-linked immunosorbent assay.. TLR4-stimulating ability of SBP was associated with plaque index (PI), but not with other clinical parameters at sampling sites. TLR2-stimulating ability of SBP was associated with none of the parameters. Number of P. gingivalis and A. actinomycetemcomitans in each plaque sample was not associated with TLR2- or TLR4-stimulating ability of SBP. PBMCs stimulated with SBP samples produced TNF-α and IL-8, which was inhibited by TLR4 but not by TLR2 inhibitor.. TLR4- but not TLR2-stimulating ability of SBP is associated with PI. Enhanced TLR4-stimulating ability at sites with accumulated plaque may mediate gingival inflammation.

Topics: Dental Plaque; Humans; Interleukin-8; Leukocytes, Mononuclear; Toll-Like Receptor 2; Toll-Like Receptor 4

To test the following two hypotheses: 1) different types of retainers result in distinct levels of biomarkers in gingival crevicular fluid (GCF) and 2) the retainer bonded to all mandibular anterior teeth induces more detrimental outcomes to the periodontium.. The Department of Orthodontics at the University of Florida. The population consisted of individuals in the retention phase of orthodontic treatment.. This was a cross-sectional study that enrolled 36 individuals. Subjects in group 1 had retainers bonded to the mandibular canines only. Group 2 consisted of individuals having retainers bonded to all mandibular anterior teeth. Group 3 included patients using mandibular removable retainers. After clinical examination, GCF was collected from the mandibular incisor and biomarker levels were compared between the groups.. Plaque accumulation and gingivitis differed significantly among groups, with the highest median values in group 2 subjects. Pairwise comparison of the groups with respect to gingivitis showed significant differences between groups 1 and 2. Significant differences among groups were detected for RANKL, OPG, OPN, M-CSF, MMP-3, and MMP-9. The ratio RANKL/OPG was significantly higher in group 2 subjects, with pairwise comparisons indicating that groups 1 and 2 differed from group 3.. An association was found between orthodontic retention groups and GCF biomarker levels, which should be further explored in longitudinal studies. The presence of retainers bonded to all anterior teeth seems to increase plaque accumulation and gingivitis.

Topics: Adolescent; Adult; Biomarkers; Cross-Sectional Studies; Cuspid; Dental Bonding; Dental Plaque; Dental Plaque Index; Female; Gingival Crevicular Fluid; Gingival Recession; Gingivitis; Humans; Incisor; Interleukin 1 Receptor Antagonist Protein; Interleukin-1beta; Interleukin-6; Interleukin-8; Macrophage Colony-Stimulating Factor; Male; Mandible; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Middle Aged; Orthodontic Appliance Design; Orthodontic Retainers; Osteopontin; Osteoprotegerin; Periodontal Index; RANK Ligand

To histologically and immunologically assess experimental peri-implant mucositis at surface enhanced modified (mod) hydrophilic titanium implants.. In a split-mouth design (n = 6 foxhounds), four different implants were inserted on each side of the maxilla: three titanium-zirconium alloy implants (TiZr) with either modSLA (sand-blasted, acid etched and chemically mod), modMA (machined, acid etched and chemically mod), or M (machined) surfaces in the transmucosal portion, and one titanium implant with a machined transmucosal portion (TiM). Experimental mucositis was induced at one randomly assigned side (NPC), whereas the contra-lateral maxillary side received mechanical plaque removal three times per week (PC). At 16 weeks, tissue biopsies were processed for histological (primary outcome: apical extension of the inflammatory cell infiltrate measured from the mucosal margin - PM-aICT) and immunohistochemical (CD68 antigen reactivity) analyses. Peri-implant sulcus fluid was analysed for interleukin (IL)-1β, IL-8, matrix metalloproteinase (MMP)-8 and myeloperoxidase (MPO).. Mean PM-aICT values varied between 1.86 (TiZrmodSLA) and 3.40 mm (TiM) in the UPC group, and between 0.88 (TiZrmodSLA) and 2.08 mm (TiZrM) in the PC group. Mean CD68, IL-1β, IL-8, MMP-8 and MPO values were equally distributed between mod- and control implants in both NPC and PC groups.. The progression of experimental mucositis was comparable at all implant surfaces investigated.

Topics: Acid Etching, Dental; Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Dental Alloys; Dental Etching; Dental Implants; Dental Plaque; Dental Prosthesis Design; Disease Models, Animal; Dogs; Female; Hydrophobic and Hydrophilic Interactions; Interleukin-1beta; Interleukin-8; Macrophages; Male; Matrix Metalloproteinase 8; Peroxidase; Random Allocation; Stomatitis; Surface Properties; Titanium; Zirconium

The aim of this study was to investigate the effect of non-surgical treatment of periodontitis on the levels of periodontopathogens and clinical parameters in patients with different genetic backgrounds produced by polymorphisms in the Interleukin ( IL8) gene. Thirty patients grouped according to IL8 ATC/TTC or AGT/TTC haplotypes were submitted to non-surgical periodontal treatment. Levels of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were determined in 240 subgingival plaque samples by qPCR. The association between IL8 haplotypes and the levels of periodontopathogens and clinical parameters was investigated by multilevel analysis accounting for the clustering of diseased sites analyzed within patients. It was observed that neither levels of periodontopathogens nor non-surgical treatment was associated with the IL8 haplotype. The clinical parameters after periodontal treatment were similar in diseased and healthy sites, independently of the IL8 haplotype. Nonetheless, in the same period, diseased sites of AGT/TTC patients harbored higher levels of P. gingivalis, T. denticola, T. forsythia, and red complex than those of ATC/TTC patients. However, the non-surgical periodontal therapy decreased the levels of these periodontopathogens and of the tested clinical parameters of diseased sites in both groups. Non-surgical therapy is equally effective in improving clinical parameters and decreasing the levels of periodontopathogens, independent of the genotype groups produced by the IL8 haplotype.

Topics: Bacterial Load; Dental Plaque; Genetic Predisposition to Disease; Haplotypes; Humans; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Real-Time Polymerase Chain Reaction; Tannerella forsythia; Treatment Outcome; Treponema denticola

A number of oral diseases, including periodontitis, derive from microbial biofilms and are associated with increased antimicrobial resistance. Despite the widespread use of mouthwashes being used as adjunctive measures to control these biofilms, their prolonged use is not recommended due to various side effects. Therefore, alternative broad-spectrum antimicrobials that minimise these effects are highly sought after. Carbohydrate derived fulvic acid (CHD-FA) is an organic acid which has previously demonstrated to be microbiocidal against Candida albicans biofilms, therefore, the aims of this study were to evaluate the antibacterial activity of CHD-FA against orally derived biofilms and to investigate adjunctive biological effects.. Minimum inhibitory concentrations were evaluated for CHD-FA and chlorhexidine (CHX) against a range of oral bacteria using standardised microdilution testing for planktonic and sessile. Scanning electron microscopy was also employed to visualise changes in oral biofilms after antimicrobial treatment. Cytotoxicity of these compounds was assessed against oral epithelial cells, and the effect of CHD-FA on host inflammatory markers was assessed by measuring mRNA and protein expression.. CHD-FA was highly active against all of the oral bacteria tested, including Porphyromonas gingivalis, with a sessile minimum inhibitory concentration of 0.5%. This concentration was shown to kill multi-species biofilms by approximately 90%, levels comparable to that of chlorhexidine (CHX). In a mammalian cell culture model, pretreatment of epithelial cells with buffered CHD-FA was shown to significantly down-regulate key inflammatory mediators, including interleukin-8 (IL-8), after stimulation with a multi-species biofilm.. Overall, CHD-FA was shown to possess broad-spectrum antibacterial activity, with a supplementary function of being able to down-regulate inflammation. These properties offer an attractive spectrum of function from a naturally derived compound, which could be used as an alternative topical treatment strategy for oral biofilm diseases. Further studies in vitro and in vivo are required to determine the precise mechanism by which CHD-FA modulates the host immune response.

Topics: Analysis of Variance; Bacteria; Benzopyrans; Biofilms; Cell Line, Transformed; Chlorhexidine; Colony Count, Microbial; Dental Plaque; Down-Regulation; Epithelial Cells; Gene Expression; Humans; Immunomodulation; Inflammation Mediators; Interleukin-8; Periodontitis; Statistics, Nonparametric

Regardless of gingival health and subgingival microbiology, production of cytokines within peri-implant tissues may be different from that of teeth. The objective of this study was to describe the peri-implant levels of pro-inflammatory cytokines and subgingival microbiology in clinically healthy sites.. Subgingival plaque and gingival crevicular fluid (GCF) were obtained from 28 clinically healthy implants and 26 teeth selected from 24 individuals. Microbial composition was determined by selective anaerobic culture techniques. Pro-inflammatory cytokines were quantified by flow cytometry analysis of GCF. The concentration of cytokines between implants and teeth were compared with the independent t-test.. The concentration of cytokines was higher in GCF from healthy implants than in teeth. The profile of cytokines was characteristic of an innate immune response. A more frequent detection of periodontopathic bacteria was observed in teeth than implants. Cultivable levels of periodontopathic bacteria were similar between implants and teeth.. Despite gingival tissue health and scarce plaque accumulation, the profile of inflammatory cytokines in implant crevicular fluid was distinctive of an innate immune response and in higher concentration than in teeth. Other than bacterial stimulus, intrinsic factors related to implants may account for more cytokine production than teeth.

Topics: Bacteriological Techniques; Bacteroides; Campylobacter; Cytokines; Dental Implants; Dental Plaque; Female; Fusobacterium; Gingiva; Gingival Crevicular Fluid; Humans; Immunity, Innate; Inflammation Mediators; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Jaw, Edentulous, Partially; Male; Middle Aged; Osseointegration; Porphyromonas gingivalis; Tooth; Tumor Necrosis Factor-alpha

To assess the pattern of early bacterial colonization at implants and teeth in patients with a history of chronic periodontitis compared with a group of healthy subjects. Furthermore, the presence of host-derived markers at teeth and implants in the two subject groups was determined.. Subgingival and submucosal plaque and gingival crevicular fluid samples from 37 nonsubmerged healing dental implants and the deepest tooth sites per quadrant were analyzed 2 to 5 months after implant insertion. The presence of periodontal pathogens was assessed by means of real-time polymerase chain reaction. Further, the levels of interleukin (IL)-1Β, IL-8, and IL-10; secretory leukocyte protease inhibitor; and the neutrophil elastase activity were determined.. Eleven patients with chronic periodontitis and 13 subjects without periodontitis were recruited for this study. Bacterial species associated with periodontitis were detectable at both the teeth and implants. The presence was always higher in the chronic periodontitis group; the difference was significant for Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans at both the implants and teeth. The levels of IL-1Β were higher at teeth than at implants; in contrast, more IL-10 was measured at the implants.. The present results indicate that (1) dental implants inserted in periodontally compromised patients are colonized with periodontal pathogens within the first weeks of healing; (2) inflammatory markers (IL-1Β) are present in higher levels at teeth as compared with implants, whereas at implants, anti-inflammatory cytokines (IL-10) might play the important role; and (3) the importance of periodontal treatment prior to implant insertion to reduce bacterial load and inflammation should be emphasized.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Bacteria; Bacterial Load; Bacteroides; Biomarkers; Chronic Periodontitis; Dental Implants; Dental Plaque; Female; Follow-Up Studies; Fusobacterium nucleatum; Gingival Crevicular Fluid; Gingival Hemorrhage; Humans; Inflammation Mediators; Interleukin-10; Interleukin-1beta; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Pocket; Porphyromonas gingivalis; Secretory Leukocyte Peptidase Inhibitor; Tooth

In a cross-sectional study design we test the hypothesis of whether obesity in adolescence is associated with periodontal risk indicators or disease.. Obese adolescents (n=52) and normal weight subjects (n=52) with a mean age of 14.5 years were clinically examined with respect to dental plaque, gingival inflammation, periodontal pockets and incipient alveolar bone loss. The subjects answered a questionnaire concerning medical conditions, oral hygiene habits, smoking habits and sociodemographic background. Body mass index (BMI) was calculated and adjusted for age and gender (BMI-SDS). Samples of gingival crevicular fluid (GCF) were analyzed for the levels of adiponectin, plasminogen activator inhibitor-1 (PAI-1), interleukin-1β (IL-β), interleukin-8 (IL-8) and tumor necrosis factor α (TNF-α).. Obese subjects exhibited more gingival inflammation (P<0.001) and more pathological periodontal pockets (>4 mm) (P<0.001) but not incipient alveolar bone loss compared with the normal weight subjects. Higher levels of IL-1β (P<0.001) and IL-8 (P=0.002) were measured in GCF from obese subjects compared with the controls. In a multivariate logistic regression analysis, adjusted BMI-SDS (P=0.03; Odds Ratio [OR]=1.87) was significantly associated with the occurrence of pathological periodontal pockets.. The study demonstrates an association between obesity and periodontal risk indicators in adolescents that in the long term may lead to oral morbidity. This result further strengthens obesity's negative effect on teenagers' periodontal health and highlights the importance of a close collaboration between dentists and pediatricians in the prevention and treatment of obesity.

Topics: Adiponectin; Adolescent; Alveolar Bone Loss; Analysis of Variance; Body Mass Index; Case-Control Studies; Chi-Square Distribution; Child; Cross-Sectional Studies; Dental Plaque; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Logistic Models; Male; Obesity; Odds Ratio; Periodontal Diseases; Periodontal Pocket; Plasminogen Activator Inhibitor 1; Risk Assessment; Risk Factors; Surveys and Questionnaires; Sweden; Tumor Necrosis Factor-alpha

Our goal was to examine differences in clinical, microbiologic, and immunologic responses to non-surgical mechanical therapy in patients with generalized chronic periodontitis (GCP) and generalized aggressive periodontitis (GAgP).. Twenty patients with GCP and 14 patients with GAgP were evaluated. Clinical data, gingival crevicular fluid (GCF), and subgingival plaque samples were collected at baseline and 3 months after non-surgical periodontal treatment. Levels of 40 subgingival species were measured using checkerboard DNA-DNA hybridization. GCF interleukin (IL)-1β, -4, and -8 and interferon-γ (IFN-γ) were analyzed using a multiplexed bead immunoassay, and elastase activity was measured using an enzymatic assay. The significance of changes with time was examined using the Wilcoxon rank sum test. Changes in clinical, microbiologic, and immunologic parameters after therapy were compared between groups using the Mann-Whitney U test.. After periodontal therapy, we found significant improvements for all clinical parameters in both groups. We also observed significant reductions in elastase activity in shallow and deep sites from the GAgP group and in deep sites from the GCP group. Microbiologic data showed significant reductions in proportions of orange and red complexes and an increase in proportions of Actinomyces species in both clinical groups. When the clinical, microbiologic, and immunologic responses after therapy were compared between groups, only minor differences were found.. This study fails to show any significant differences between severe forms of GCP and GAgP in response to non-surgical periodontal treatment.

Topics: Actinomyces; Adult; Aggressive Periodontitis; Bacteroides; Chronic Periodontitis; Dental Plaque; Dental Scaling; Eubacterium; Female; Follow-Up Studies; Fusobacterium; Fusobacterium nucleatum; Gingival Crevicular Fluid; Humans; Interferon-gamma; Interleukin-1beta; Interleukin-4; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Peptostreptococcus; Periodontal Attachment Loss; Periodontal Pocket; Porphyromonas gingivalis; Root Planing; Smoking; Treatment Outcome; Treponema denticola

Inflammatory responses of host cells to oral pathogenic bacteria, such as Porphyromonas gingivalis, are crucial in the development of periodontitis. Host cells, such as periodontal ligament and gingival fibroblasts, from periodontitis patients may respond to P. gingivalis in a different manner compared with cells from healthy persons. The aim of this study was to investigate inflammatory responses to viable P. gingivalis by periodontal ligament and gingival fibroblasts from periodontitis patients and healthy control subjects.. Primary periodontal ligament and gingival fibroblasts from periodontitis patients (n=14) and healthy control subjects (n=8) were challenged in vitro with viable P. gingivalis. Gene expression of Toll-like receptors (TLRs) 1, 2, 4, 6, 7 and 9, CD14, nuclear factor-κB1 and its putative inhibitor NF-κB inhibitor-like protein1, and of interleukin-1β, interleukin-6, interleukin-8, tumour necrosis factor-α, monocyte chemotactic protein-1 and regulated upon activation, normal T-cel expressed, and secreted, were assessed by real-time PCR..   Periodontal ligament fibroblasts from periodontitis patients had a higher mRNA expression of TLR1, TLR4, TLR7 and CD14, and a lower expression of NFKBIL1, both before and after P. gingivalis challenge. In contrast, gingival fibroblasts from periodontitis patients had stronger induction of TLR1, TLR2 and TLR7 by P. gingivalis. Cytokine responses were not different between patients and control subjects. Interestingly, periodontal ligament, but not gingival, fibroblasts from P. gingivalis culture-positive persons responded more strongly to P. gingivalis than periodontal ligament fibroblasts from P. gingivalis-negative persons.. Periodontal ligament and gingival fibroblasts respond to P. gingivalis in a different manner and may play different roles in periodontitis. Both subsets of fibroblasts from patients appear more active in interaction with P. gingivalis. Moreover, periodontal ligament fibroblasts from P. gingivalis-positive donors are more responsive to an in vitro P. gingivalis challenge.

Topics: Adaptor Proteins, Signal Transducing; Cells, Cultured; Chemokine CCL2; Dental Plaque; Female; Fibroblasts; Gingiva; Histocompatibility Antigens Class II; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharide Receptors; Major Histocompatibility Complex; Male; Middle Aged; NF-kappa B; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; T-Lymphocytes; Toll-Like Receptor 1; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptor 6; Toll-Like Receptor 7; Toll-Like Receptor 9; Tumor Necrosis Factor-alpha

Previous studies have shown that Porphyromonas gingivalis is found in the amniotic fluid and placentae of pregnant women with some obstetric diseases. However, the biological effects of P. gingivalis on intrauterine tissues remain unclear. The aim of this study was to investigate the presence of P. gingivalis in chorionic tissues from hospitalized high-risk pregnant women, and the effects of P. gingivalis lipopolysaccharide on the production of proinflammatory molecules in human chorion-derived cells.. Twenty-three subjects were selected from Japanese hospitalized high-risk pregnant women. The presence of P. gingivalis in chorionic tissues was analyzed by PCR. Cultured chorion-derived cells or Toll-like receptor-2 (TLR-2) gene-silenced chorion-derived cells were stimulated with P. gingivalis lipopolysaccharide. Real-time PCR was performed to evaluate TLR-2 and Toll-like receptor-4 (TLR-4) mRNA expression in the cells. Levels of interleukin-6 and interleukin-8 in culture supernatants of the chorion-derived cells were measured by ELISA.. P. gingivalis DNA was detected in chorionic tissues from two women with threatened preterm labor, two with multiple pregnancy and two with placenta previa. Stimulation of chorion-derived cells with P. gingivalis lipopolysaccharide significantly increased TLR-2 mRNA expression, whereas TLR-4 mRNA expression was not changed. P. gingivalis lipopolysaccharide induced interleukin-6 and interleukin-8 production in chorion-derived cells, but the P. gingivalis lipopolysaccharide-induced interleukin-6 and interleukin-8 production was reduced in TLR-2 gene-silenced chorion-derived cells.. Our results suggest that P. gingivalis can be detected in chorionic tissues of hospitalized high-risk pregnant women, and that P. gingivalis lipopolysaccharide induces interleukin-6 and interleukin-8 production via TLR-2 in chorion-derived cells.

Topics: Adult; Cells, Cultured; Chorion; Dental Plaque; Escherichia coli; Female; Gene Silencing; Gingivitis; Hospitalization; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Lipopolysaccharides; Periodontal Attachment Loss; Periodontal Diseases; Periodontal Pocket; Periodontitis; Placenta Previa; Porphyromonas gingivalis; Pregnancy; Pregnancy, High-Risk; Pregnancy, Multiple; Premature Birth; Saliva; Toll-Like Receptor 2; Toll-Like Receptor 4; Young Adult

To evaluate the gingival crevicular fluid (GCF) contents of interleukin-6 (IL-6) and interleukin-8 (IL-8) and the clinical parameters of the teeth supporting fixed partial denture (FPD) and the contralateral teeth and to assess the effect of scaling and root planning (SRP) on clinical parameters and the GCF levels of cytokines.. The study population included 23 patients. Probing depth (PD), clinical attachment level (CAL), plaque index (PI), and gingival index (GI) were recorded, and GCF samples were collected for analysis of cytokine levels from the teeth with FPD (Test Group), the contralateral teeth (Control Group) of each participant at baseline. After initial measurements, all participants received primary phase of non-surgical treatment including oral hygiene instruction and scaling and root planning (SRP). At the 1st month and the 3rd month after SRP, these procedures were repeated.. In both groups, all clinical parameters and the total amount of IL-8 showed decreases from initial to the 3rd month (P < 0.05), but from the 1st month to the 3rd month; PD, PI, and GI values significantly increased in the test group (P < 0.05).. The non-surgical periodontal treatment reduced the total amount of IL-8, not IL-6, and the clinical parameters of the teeth with FPD and contralateral teeth. But, there was a trend to the higher levels of PD, PI, and GI in the teeth with FPD. Therefore, a regular program for dental prophylaxis is also important for the maintenance of periodontal health in patients with FPD.

Topics: Adult; Case-Control Studies; Chronic Periodontitis; Dental Abutments; Dental Plaque; Dental Scaling; Denture, Partial, Fixed; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Statistics, Nonparametric

To compare gingival crevicular fluid (GCF) biomarker levels and microbial distribution in plaque biofilm (SP) samples for subjects with type 1 diabetes (T1DM) versus healthy subjects without diabetes during experimental gingivitis (EG).. A total of nine T1DM patients and nine healthy controls of age and gender similar to the T1DM patients were monitored for 35 days during EG. Hygiene practices were stopped for 3 weeks, and GCF, SP, plaque index (PI) and gingival index were determined. IL-1beta, IL-8, MMP-8 and MMP-9 were quantified by enzyme-linked immunosorbent assay, and SP samples were assessed by DNA-DNA hybridization for a panel of 40 subgingival microbial species.. IL-1beta levels in T1DM patients were elevated compared with healthy individuals, and showed differences between groups at 7-21 days while healthy patients showed IL-1beta increases from baseline to 14-21 days (p<0.05). Differences were observed in MMP-9 levels between patients with and without T1DM at 7-14 days (p<0.05). Orange complex species and PI measurements displayed a superior correlation with biomarker levels when compared with other complexes or clinical measurements during EG.. The mean GCF biomarker levels for IL-1beta and MMP-8 were most significantly elevated in T1DM subjects compared with healthy individuals during EG, not resulting from differences in the mean PI or microbial composition.

Topics: Adolescent; Adult; Bacteria; Biofilms; Biomarkers; Case-Control Studies; Cohort Studies; Dental Plaque; Dental Plaque Index; Diabetes Mellitus, Type 1; Follow-Up Studies; Gingival Crevicular Fluid; Gingivitis; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Nucleic Acid Hybridization; Periodontal Index; Prospective Studies; Young Adult

The dental plaque is comprised of numerous bacterial species, which may or may not be pathogenic. Human gingival epithelial cells (HGECs) respond to perturbation by various bacteria of the dental plaque by production of different levels of inflammatory cytokines, which is a putative reflection of their virulence. The aim of the current study was to determine responses in terms of interleukin (IL)-1beta, IL-6, IL-8 and IL-10 secretion induced by Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Streptococcus gordonii in order to gauge their virulence potential.. HGECs were challenged with the four bacterial species, live or heat killed, at various multiplicity of infections and the elicited IL-1beta, IL-6, IL-8 and IL-10 responses were assayed by enzyme-linked immunosorbent assay.. Primary HGECs challenged with live P. gingivalis produced high levels of IL-1beta, while challenge with live A. actinomycetemcomitans gave high levels of IL-8. The opportunistic pathogen F. nucleatum induces the highest levels of pro-inflammatory cytokines, while the commensal S. gordonii is the least stimulatory.. We conclude that various dental plaque biofilm bacteria induce different cytokine response profiles in primary HGECs that may reflect their individual virulence or commensal status.

Topics: Aggregatibacter actinomycetemcomitans; Bacteria; Bacteriological Techniques; Cells, Cultured; Colony Count, Microbial; Dental Plaque; Epithelial Cells; Fusobacterium nucleatum; Gingiva; Humans; Inflammation Mediators; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Interleukins; Porphyromonas gingivalis; Streptococcus gordonii; Virulence

Mucosal inflammatory responses are orchestrated largely by pro-inflammatory chemokines. The chemokine granulocyte chemotactic protein 2 (CXCL6) is involved in neutrophil recruitment and migration. Previous studies have shown that granulocyte chemotactic protein 2 is up-regulated during mucosal inflammation (e.g. in inflammatory bowel disease), similarly to the functionally and structurally related chemokine interleukin-8. Nevertheless, unlike interleukin-8, a role of granulocyte chemotactic protein 2 in gingival inflammation has not been yet demonstrated. In this study we aimed to evaluate the expression of the chemokine granulocyte chemotactic protein 2 in clinically healthy vs. diseased gingival tissues and to explore possible correlations with clinical and microbiological markers of periodontitis.. Gene expression in 184 'diseased' and 63 'healthy' gingival tissue specimens from 90 patients with periodontitis was analyzed using Affymetrix U133Plus2.0 arrays. The expression of granulocyte chemotactic protein 2 was further confirmed by real-time reverse transcription-polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay, while the localization of granulocyte chemotactic protein 2 in gingival tissues was analyzed by immunohistochemistry. Plaque samples from the adjacent periodontal pockets were collected and evaluated for 11 species of periodontal bacteria using checkerboard DNA-DNA hybridizations.. Among all known chemokines, GCP-2 expression was the most up-regulated (3.8-fold, p < 1.1 x 10(-16)), in 'diseased' vs. 'healthy' tissue as compared to a 2.6-fold increased expression of interleukin-8 mRNA (p < 1.2 x 10(-15)). Increased expression of granulocyte chemotactic protein 2 correlated with higher levels of 'red' and 'orange' complex pathogens and with increased probing depth, but not with attachment loss. Immunohistochemistry showed that granulocyte chemotactic protein 2 was expressed in gingival vascular endothelium.. The level of expression of granulocyte chemotactic protein 2 correlates with the severity of periodontitis and appears to act as a hitherto unrecognized functional adjunct to interleukin-8 in diseased gingival tissues.

Topics: Actinomyces; Adolescent; Adult; Aged; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Bacteroides; Campylobacter rectus; Chemokine CXCL16; Chemokines, CXC; Chronic Periodontitis; Dental Plaque; Eikenella corrodens; Endothelium, Vascular; Female; Fusobacterium nucleatum; Gingiva; Humans; Inflammation Mediators; Interleukin-8; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Pocket; Porphyromonas gingivalis; Prevotella intermedia; Receptors, Scavenger; Treponema denticola; Up-Regulation; Veillonella; Young Adult

The purpose of this study was to compare the levels of the cytokines interleukin-1beta (IL-1beta), IL-4, and IL-8 in the gingival crevicular fluid (GCF) of adolescents and young adults.. Twenty-five adolescents aged between 14 and 16 years (Group A) and 20 periodontally healthy young adults aged between 25 and 35 years (Group B) were selected from two private dental clinics limited to pedodontics and periodontics respectively in Piraeus Greece. All subjects were systemically healthy. Clinical examination included probing pocket depth (PPD), presence or absence of plaque, and bleeding on probing (BOP). GCF was collected from four sites per subject. IL-1beta, IL-4, and IL-8, measured as total amounts (pg/30 s), were evaluated in 180 samples using a commercially available sandwich enzyme-linked immunosorbent assay.. IL-1beta mean levels of Groups A and B were adjusted for BOP and PPD. Differences of IL-1beta mean levels between the two age groups were statistically significant (F = 50.245, P < 0.001) in favour of Group A. Adolescents showed statistically significantly lower mean levels of IL-4 than young adults in the presence of BOP (F = 10.690, P = 0.001). There was no statistically significant difference between adolescents and adults for the means of IL-8 adjusted for BOP and plaque presence (F = 2.032, P = 0.161).. Within the limits of this study the differences reported in mean levels of IL-1beta and IL-4 may be attributed to the different age status.

Topics: Adolescent; Adult; Age Factors; Case-Control Studies; Cross-Sectional Studies; Dental Plaque; Female; Gingival Crevicular Fluid; Humans; Interleukin-1beta; Interleukin-4; Interleukin-8; Male; Periodontal Index

The aim of this study was to describe the development and validation of the checkerboard immunoblotting (CBIB) technique for the high-throughput quantification of multiple inflammatory mediators in gingival crevicular fluid (GCF) samples.. Monoclonal antibodies were used to bind GCF interleukin (IL)-1beta and -8 and matrix metalloproteinase (MMP)-8 to the surface of membranes. Biotinylated antibodies were used to detect bound antigens in a checkerboard format. Signals were developed using chemiluminescence, captured on film, and quantified using software for array analysis. The assay was tested for potential cross-reactions among the three pairs of antibodies. Eleven CBIBs were processed to determine the analytical sensitivity of the assay. Forty GCF samples were analyzed using CBIB and enzyme-linked immunosorbent assay (ELISA) in parallel, and the significance of the correlations among the results was tested using the Pearson correlation coefficient. Nine hundred thirty-one GCF samples were collected from 20 periodontally healthy subjects and 20 periodontitis subjects and analyzed using CBIB to test the assay's sensitivity and dynamic ranges using clinical samples.. The CBIB was capable of distinguishing among the three analytes. The sensitivity and dynamic ranges of the assay were suitable for the detection of the three targets in the majority of GCF samples. There were highly statistically significant (P <0.0001) positive correlations between CBIB and ELISA data for all three biomarkers. The periodontitis subjects had statistically significantly higher mean levels of IL-1beta and -8 compared to healthy subjects.. The CBIB technique is a sensitive and specific assay for the high-throughput quantification of MMP-8 and IL-8 and -1beta in GCF.

Topics: Adult; Antibodies, Immobilized; Biomarkers; Chronic Periodontitis; Cross Reactions; Dental Plaque; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Gingival Hemorrhage; Gingival Recession; Gingivitis; Humans; Immunoblotting; Interleukin-1beta; Interleukin-8; Luminescence; Male; Matrix Metalloproteinase 8; Membranes, Artificial; Middle Aged; Periodontal Pocket; Periodontium; Polyvinyls; Sensitivity and Specificity; Software

In our previous study, we found that the ability of supragingival plaque to induce Toll-like receptor (TLR)4-mediated stimulation was positively associated with plaque score and bleeding on probing (BOP) at the sampled sites and that the ability to induce TLR2-mediated stimulation was negatively associated with probing depth (PD) and clinical attachment level (CAL). Because signaling from TLR leads to the induction of pro- and anti-inflammatory cytokines, we further analyzed the influence of the ability of supragingival plaque to induce TLR2-/TLR4-mediated stimulation of cytokine production by peripheral blood mononuclear cells (PBMCs).. The abilities of 125 plaque samples to induce TLR2- or TLR4-mediated stimulation were determined using genetically engineered Chinese hamster ovary reporter cells that express a reporter molecule upon activation of nuclear factor-kappa B through TLR2 or TLR4. PBMCs were stimulated with each plaque sample, and the production of proinflammatory cytokines (tumor necrosis factor-alpha and interleukin [IL]-6 and -8) and an anti-inflammatory cytokine (IL-10) was analyzed by enzyme-linked immunosorbent assay.. The levels of the cytokines produced by PBMCs all correlated with the ability of supragingival plaque to induce TLR4-mediated stimulation but not with its ability to induce TLR2-mediated stimulation. Cytokine production was inhibited by an anti-TLR4 monoclonal antibody and a TLR4 antagonist, compound 406. The levels of cytokines were associated with plaque index, BOP, PD, and CAL at the sampled sites.. The production of pro-/anti-inflammatory cytokines by PBMCs was associated with the ability of supragingival plaque to induce TLR4-mediated stimulation. The cytokines induced by supragingival plaque via TLR4 might modulate periodontal status.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antibodies, Monoclonal; CHO Cells; Cricetinae; Cricetulus; Cytokines; Dental Plaque; Dental Plaque Index; Female; Gingival Hemorrhage; Glycolipids; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Lipid A; Male; Middle Aged; NF-kappa B; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Toll-Like Receptor 2; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha; Young Adult

To our knowledge, changes in the patterns of whole-transcriptome gene expression that occur during the induction and resolution of experimental gingivitis in humans were not previously explored using bioinformatic tools.. Gingival biopsy samples collected from 14 subjects during a 28-day stent-induced experimental gingivitis model, followed by treatment, and resolution at days 28 through 35 were analyzed using gene-expression arrays. Biopsy samples were collected at different sites within each subject at baseline (day 0), at the peak of gingivitis (day 28), and at resolution (day 35) and processed using whole-transcriptome gene-expression arrays. Gene-expression data were analyzed to identify biologic themes and pathways associated with changes in gene-expression profiles that occur during the induction and resolution of experimental gingivitis using bioinformatic tools.. During disease induction and resolution, the dominant expression pathway was the immune response, with 131 immune response genes significantly up- or downregulated during induction, during resolution, or during both at P <0.05. During induction, there was significant transient increase in the expression of inflammatory and oxidative stress mediators, including interleukin (IL)-1 alpha (IL1A), IL-1 beta (IL1B), IL8, RANTES, colony stimulating factor 3 (CSF3), and superoxide dismutase 2 (SOD2), and a decreased expression of IP10, interferon inducible T-cell alpha chemoattractant (ITAC), matrix metalloproteinase 10 (MMP10), and beta 4 defensin (DEFB4). These genes reversed expression patterns upon resolution in parallel with the reversal of gingival inflammation.. A relatively small subset (11.9%) of the immune response genes analyzed by array was transiently activated in response to biofilm overgrowth, suggesting a degree of specificity in the transcriptome-expression response. The fact that this same subset demonstrates a reversal in expression patterns during clinical resolution implicates these genes as being critical for maintaining tissue homeostasis at the biofilm-gingival interface. In addition to the immune response pathway as the dominant response theme, new candidate genes and pathways were identified as being selectively modulated in experimental gingivitis, including neural processes, epithelial defenses, angiogenesis, and wound healing.

Topics: Adolescent; Adult; Aged; beta-Defensins; Biofilms; Chemokine CCL5; Chemokine CXCL10; Chemokine CXCL11; Colony-Stimulating Factors; Computational Biology; Dental Plaque; Female; Follow-Up Studies; Gene Expression Profiling; Genes, MHC Class II; Gingiva; Gingivitis; Humans; Inflammation Mediators; Interleukin-1alpha; Interleukin-1beta; Interleukin-8; Male; Matrix Metalloproteinase 10; Middle Aged; Oxidative Stress; Superoxide Dismutase; Young Adult

Antimicrobial peptides such as human beta-defensin-2 (hBD-2), psoriasin (PSO), and ribonuclease 7 (RNase 7) play an important role in innate immunity. The aim of the present study was to test the hypothesis that epithelial cells show a differential gene expression pattern of antimicrobial peptides (hBD-2, PSO, RNase 7) and inflammatory mediators such as interleukin-8 (IL-8) and 5-lipoxygenase (5-LO) in response to different stages of naturally formed biofilms.. Epithelial cells were cultured from biopsies obtained from five healthy individuals. Native bacterial biofilms were taken from the same subjects that donated the gingival biopsies. To obtain different stages of biofilm formation, polymer disks were attached to prostheses and carried intraorally for 1, 3, 5, and 9 days. The expression of genes for hBD-2, PSO, RNase 7, 5-LO, and IL-8 was examined using semi-quantitative reverse transcription-polymerase chain reaction. The bacterial composition of the individual biofilms was defined using a microarray system (Parocheck), which showed the presence of 20 different bacterial species that are associated with plaque formation.. The expression of the messenger RNAs of hBD-2, RNase 7, and 5-LO was upregulated as a result of the exposure to early biofilm stages, whereas the gene expression of IL-8 was increased in response to matured biofilms. Inter-individual differences in the innate immune response were observed.. The results of the present study showed a time-dependent messenger RNA expression of antimicrobial peptides (hBD-2, RNase 7), 5-LO, and IL-8 in oral epithelial cells responding to different stages of biofilm formation.

Topics: Arachidonate 5-Lipoxygenase; Bacteria; beta-Defensins; Biofilms; Calcium-Binding Proteins; Colony Count, Microbial; Dental Plaque; Epithelial Cells; Gene Expression Regulation, Bacterial; Gingiva; Humans; Immunity, Innate; Inflammation Mediators; Interleukin-8; Keratinocytes; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleases; S100 Calcium Binding Protein A7; S100 Proteins; Up-Regulation

This study evaluated whether the biochemical changes associated with type 2 diabetes modulate the expression of interleukin-1beta, interleukin-6, interleukin-8, and interferon-gamma in sites with chronic periodontitis.. Biopsies were harvested and divided into three groups: group 1, systemically and periodontally healthy subjects (n = 10); group 2, systemically healthy subjects with moderate-to-severe chronic periodontitis (probing depth > 6 mm) (n = 20); and group 3, type 2 diabetic subjects with periodontitis (n = 20). Cytokine levels were assessed in the gingival tissues by enzyme-linked immunosorbent assay analysis.. Data analysis demonstrated that the interleukin-1beta, interleukin-6, interleukin-8, and interferon-gamma levels were higher in the presence of periodontal inflammation than in the absence of inflammation, regardless of systemic status. The interleukin-1beta and interleukin-6 levels were higher in diabetic subjects (group 3) than in systemically healthy patients with comparable types of periodontitis (group 2). No difference was observed for the interleukin-8 and interferon-gamma levels between groups 2 and 3.. Within the limits of this study, it was concluded that type 2 diabetes was associated with increased expression of interleukin-1beta and interleukin-6 in periodontally inflamed tissues of diabetic patients, relative to nondiabetic subjects, and that such overexpression may be involved in the mechanisms by which type 2 diabetes enhances periodontal destruction.

Topics: Adult; Chronic Disease; Dental Plaque; Diabetes Mellitus, Type 2; Enzyme-Linked Immunosorbent Assay; Female; Gingiva; Humans; Interferon-gamma; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Periodontal Diseases; Periodontal Index

Threatened premature labor (TPL) often results in preterm birth (PB). The aim of the present study was to evaluate the associations of periodontal and general health conditions with TPL and PB in relation to serum cytokine levels and the composition of subgingival plaque.. Eighty-eight women were enrolled in the study. Systemic conditions were assessed, and subgingival plaque samples obtained for bacterial analysis. Periodontal examinations included assessments of plaque, gingivitis, clinical attachment level, probing depth, and bleeding on probing. Serum cytokine levels also were analyzed. Gestational age at delivery was recorded, and the mothers were divided into a TPL or non-TPL group, and into a non-TPL-TB (term birth), non-TPL-PB, TPL-TB, or TPL-PB group, accordingly.. Forty subjects were classified as TPL and 18 as TPL-PB. There were significant differences between the TPL and non-TPL subjects in several of the systemic and periodontal parameters and serum cytokine levels. Significant differences were observed between the TPL-TB and TPL-PB groups in the percentage of Tannerella forsythensis (Tf, formerly Bacteroides forsythus), and the serum interleukin (IL)-8 and IL-1beta levels. Significant negative correlations between the gestational age at delivery and several periodontal parameters and serum IL-8 and IL-1beta levels, and significant positive correlations between periodontal status and serum IL-8 and IL-1beta levels, were observed.. The TPL women revealed worsened periodontal conditions and elevated serum IL-8 and IL-1beta levels compared to the non-TPL women. The elevated levels of serum IL-8 and IL-1beta could have affected the maintenance of the proper uterine-fetus relationship, resulting in premature uterine contractions.

Topics: Adult; Analysis of Variance; Bacteroides; Cytokines; Dental Plaque; Female; Gestational Age; Health Status; Humans; Infant, Low Birth Weight; Infant, Newborn; Interleukin-1; Interleukin-6; Interleukin-8; Male; Obstetric Labor, Premature; Periodontal Diseases; Periodontal Index; Pregnancy; Statistics, Nonparametric; Tumor Necrosis Factor-alpha

To determine the effect of scaling and root planing (SRP) on the interrelations of subgingival periodontopathogens and both interleukin-8 (IL-8) and granulocyte elastase activity in gingival crevicular fluid (GCF), and to assess their relations to the short-term treatment response in management of chronic periodontitis.. GCF and subgingival plaque were collected from 16 subjects with untreated chronic periodontitis at baseline and 4 weeks after SRP. IL-8 levels were determined by ELISA. Granulocyte elastase activity was analyzed with a specific substrate, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA) was calculated. 5 DNA-probes were used to detect the presence of A. actinomycetemcomitans (A. a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.), with a sensitivity = 103 cells/paper point.. IL-8 and MR-EA levels in GCF decreased significantly after SRP (p < 0.001) with a corresponding reduction of total count of the species. Of the sites with probing depth (PD) >/= 5.0 mm and co-infection by B.f., P.g., P.i. & T.d. at baseline, the sites without persistent co-infection of these species after SRP exhibited a significant reduction of IL-8 levels (p < 0.02), MR-EA levels (p < 0.02) and PD (p < 0.01). No such change was found in the sites where such a co-infection persisted. Moreover, reduction of IL-8 levels in those pocket sites was accompanied by a concomitant reduction of MR-EA (p < 0.02) and PD (p < 0.01), while no significant change in MR-EA levels and PD was noted in those pocket sites that exhibited an increase of IL-8 levels after SRP. At baseline, the former group of sites showed significantly higher IL-8 levels than the latter group of sites (p < 0.02).. IL-8-related granulocyte elastase activity was related to the change in infection patterns of the target periodontopathogens following scaling and root planing. Varying initial IL-8 levels in GCF and a corresponding shifting change of granulocyte elastase activity in GCF may characterize the different short-term treatment responses.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Bacteroides; Chronic Disease; Colony Count, Microbial; Dental Plaque; Dental Scaling; Follow-Up Studies; Gingival Crevicular Fluid; Gram-Negative Bacteria; Humans; Interleukin-8; Leukocyte Elastase; Linear Models; Matched-Pair Analysis; Middle Aged; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Prevotella intermedia; Root Planing; Statistics as Topic; Statistics, Nonparametric; Treponema

A study was undertaken to examine cytokine markers in gingival crevicular fluid (GCF) during the early stages of plaque accumulation.. A panel of five subjects with good oral hygiene went without brushing for 1 or 3 days, after which GCF samples were taken by placing paper strips into the gingival margins of the maxillary premolars and first molar for 30 s. GCF flow rates were determined with a Periotron instrument (Oraflow, Inc., Plainview, New York), and neutrophils (polymorphonuclear leukocytes) were determined as myeloperoxidase activity. Interleukin-1b (IL-1b) and IL-8 were eluted from the paper strips and assayed with enzyme-linked immunosorbent assay (ELISA) systems.. The plaque index rose to 2.7 +/- 0.2 (mean +/- SE) after 3 days without brushing, and the GCF flow rate increased to 146.8% of baseline. PMN and IL-8 concentrations fell but, when corrected for dilution as a result of increased GCF flow, were not statistically different from baseline. IL-1b was slightly elevated after 1 day, and increased to 223.8 +/- 54.3% (from 6.8 +/- 1.7 to 13.8 +/- 3.6 pg/30 s; p = 0.04) after 3 days of plaque accumulation. Resumption of tooth brushing led to a return of IL-1b to baseline (109.1% after 2 days of brushing). When subjects rinsed with 0.12% chlorhexidine during the 3-day no-brushing period, the increases in plaque index, GCF flow rates and IL-1b release rates did not occur.. The results indicate that IL-1b release rates increase in the GCF after 3 days of plaque accumulation, before any clinical signs of inflammation appear.

Topics: Adult; Analysis of Variance; Anti-Infective Agents; Biomarkers; Chlorhexidine; Dental Plaque; Dental Plaque Index; Enzyme-Linked Immunosorbent Assay; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-1; Interleukin-8; Mouthwashes; Neutrophils; Peroxidase

This study aimed to determine the relationships among interleukin (IL)-8 and granulocyte elastase levels in gingival crevicular fluid (GCF) and the concomitant presence of periodontopathogens in untreated adult periodontitis.. GCF and subgingival plaque samples were collected from 16 patients with untreated adult periodontitis and 10 healthy control subjects. IL-8 levels were determined by enzyme-linked immunosorbent assay (ELISA). Granulocyte elastase was analyzed with a neutrophilic granulocyte-specific, low molecular weight and chromogenic substrate, L-pyroglutamyl-L-prolyl-L-valine-p-nitroanilide, and the maximal rate of elastase activity (MR-EA) was calculated. Five DNA probes were used to detect the presence of A. actinomycetemcomitans (A.a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.).. Lower IL-8 concentrations and higher granulocyte elastase activities were found in patients than in healthy controls as well as in diseased conditions co-infected with B.f., P.g., P.i., and T.d. as compared to healthy conditions without the target species (P <0.05). IL-8 concentrations were positively correlated with MR-EA levels in the periodontitis conditions co-infected with B.f., P.g., P.i., and T.d. (P <0.05). A wide range of IL-8 concentrations was found among 15 patients when the periodontitis condition was characterized by co-infection with B.f., P.g., P.i., and T.d. MR-EA levels in the high IL-8 group of subjects were significantly higher than those in the low IL-8 group of subjects (P <0.01).. The present study shows that the local host-bacteria interactions in untreated periodontitis are diverse in terms of the intensity of inflammatory responses measured by IL-8-related granulocyte elastase activity in GCF. This might reflect different phases of the inflammatory response due to shifts in host-bacteria interactions and therefore be indicative of a range of periodontal disease activity levels.

Topics: Actinobacillus Infections; Adult; Aggregatibacter actinomycetemcomitans; Analysis of Variance; Bacteria; Bacteroidaceae Infections; Bacteroides; Bacteroides Infections; Dental Plaque; Gingival Crevicular Fluid; Humans; Interleukin-8; Leukocyte Elastase; Linear Models; Middle Aged; Periodontitis; Porphyromonas gingivalis; Prevotella intermedia; Statistics, Nonparametric; Treponema; Treponemal Infections

We used flow cytometry to analyze the expression of adhesion molecules and the oxidative burst of whole-blood polymorphonuclear neutrophils (PMN) from 26 patients with periodontitis. Three different clinical entities were studied: adult periodontitis (AP), localized juvenile periodontitis (LJP), and rapidly progressive periodontitis (RPP). Unstimulated PMN from the patients showed reduced Lewis x, sialyl-Lewis x, and L-selectin expression relative to those from healthy control subjects. These alterations were present whatever the severity of periodontal disease. However, PMN from RPP patients showed increased basal H2O2 production and decreased L-selectin shedding. These latter impairments, which correlated with increased IL-8 plasma levels, could contribute to initial vascular damage. In addition, decreased IL-8 priming of H2O2 production by PMN from RPP patients could account for a lower bactericidal capacity of PMN, leading to the large number of bacteria in the subgingival region of RPP patients. Soluble L-selectin plasma levels were also decreased in the RPP group, indicating more severe or diffuse endothelial damage. These abnormalities were not found in the patients with less destructive forms of periodontitis (AP and LJP). Porphyromonas gingivalis, a bacterial pathogen known to increase IL-8 production by PMN, was found in the periodontal pockets of RPP patients only. These results show links among PMN abnormalities, the clinical form of periodontitis, and the gingival bacterial flora.

Topics: Adolescent; Adult; Aggressive Periodontitis; Cell Adhesion Molecules; Cytokines; Dental Plaque; Disease Progression; Female; Gingiva; Humans; Hydrogen Peroxide; Interleukin-8; L-Selectin; Male; Membrane Proteins; Middle Aged; Mouth Mucosa; Neutrophils; Periodontitis; Severity of Illness Index; Solubility

Periodontitis, which is widespread in the adult population, is a persistent bacterial infection associated with Porphyromonas gingivalis. Gingival epithelial cells are among the first cells encountered by both P. gingivalis and commensal oral bacteria. The chemokine interleukin 8 (IL-8), a potent chemoattractant and activator of polymorphonuclear leukocytes, was secreted by gingival epithelial cells in response to components of the normal oral flora. In contrast, P. gingivalis was found to strongly inhibit IL-8 accumulation from gingival epithelial cells. Inhibition was associated with a decrease in mRNA for IL-8. Antagonism of IL-8 accumulation did not occur in KB cells, an epithelial cell line that does not support high levels of intracellular invasion by P. gingivalis. Furthermore, a noninvasive mutant of P. gingivalis was unable to antagonize IL-8 accumulation. Invasion-dependent destruction of the gingival IL-8 chemokine gradient at sites of P. gingivalis colonization (local chemokine paralysis) will severely impair mucosal defense and represents a novel mechanism for bacterial colonization of host tissue.

Topics: Dental Plaque; Humans; Interleukin-8; KB Cells; Periodontal Diseases; Porphyromonas gingivalis; RNA, Messenger

The expression of adhesion molecules and the local production of chemotactic cytokines within the epithelium are considered to be key events in neutrophil (PMN) migration at sites of mucosal infections. In their journey toward the gingival sulcus, PMNs have been shown to selectively migrate through the junctional epithelium. Little, however, is known about the molecular mechanisms involved in this key process aimed at the control of subgingival bacterial plaque. This investigation describes the expression of IL-8 mRNA-positive cells and the establishment of a gradient of intercellular adhesion molecule-1 (ICAM-1) receptors within the junctional epithelium of clinically healthy gingiva. Expression of ICAM-1 and IL-8 was topographically associated with the area of PMN migration; i.e., the junctional epithelium. Levels of ICAM-1 expression increased from the basal cells toward the surface of the junctional epithelium and thus toward areas exposed to bacterial challenges. IL-8 mRNA-positive cells were also present at highest density in the most superficial junctional epithelial layers. The combination of the haptotactic stimuli, resulting from the interaction of the PMN's beta2 integrin receptors with the gradient of ICAM-1 expression, and the location of IL-8 mRNA-positive cells, consistent with the establishment of a discrete PMN chemotactic source, may play an important physiologic role in efficiently routing PMNs to the gingival sulcus. This process contributes to the maintenance of a local host-parasite equilibrium and to the limitation of PMN-associated tissue damage.

Topics: CD18 Antigens; Cell Movement; Cells, Cultured; Chemotaxis, Leukocyte; Coloring Agents; Connective Tissue; Dental Plaque; E-Selectin; Epithelial Attachment; Epithelium; Gene Expression Regulation; Gingiva; Haptens; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Leukocyte Count; Neutrophils; Receptors, Leukocyte-Adhesion; RNA, Messenger

Polymorphonuclear leukocytes (PMN) play a critical role in the host's response to the subgingival microflora. Interleukin-8 (IL-8) is a potent chemotactic and activating factor for PMN. In this study, the presence of IL-8 in gingival crevicular fluid (GCF) was examined in relation to the PMN indicator beta-glucuronidase (beta G), as well as clinical parameters of chronic inflammatory periodontal disease. Data was obtained from 30 patients with periodontitis and 14 healthy controls. For the control group, GCF and clinical data were obtained only once. For the periodontitis patients, clinical data and GCF samples were collected prior to treatment, and GCF samples were again collected 2 weeks after scaling and root planing. Comparing control and periodontitis patients prior to treatment, IL-8 concentration was lower in the patients with periodontitis. Scaling and root planing resulted in either an increase or a decrease in total IL-8 and IL-8 concentration GCF. A reduction in total IL-8 or IL-8 concentration was accompanied by a corresponding reduction in beta G activity. An increase in total IL-8 or IL-8 concentration after scaling and root planing was associated with an increase in beta G activity in some patients and a reduction in beta G activity in other patients. The periodontitis patients who did not demonstrate a linkage between IL-8 and beta G activity in GCF were those individuals with the highest beta G activity prior to treatment. As elevated beta G activity in GCF has been associated with an increased risk for probing attachment loss, the absence of a direct relationship between IL-8 in GCF and PMN recruitment into the gingival crevice may characterize individuals at risk for progression of periodontitis.

Topics: Adult; Biomarkers; Chemotaxis, Leukocyte; Chronic Disease; Cross-Sectional Studies; Dental Plaque; Dental Scaling; Disease Progression; Disease Susceptibility; Follow-Up Studies; Gingiva; Gingival Crevicular Fluid; Gingival Hemorrhage; Glucuronidase; Humans; Interleukin-8; Longitudinal Studies; Neutrophils; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Risk Factors; Root Planing