interleukin-8 and Gingivitis

In the oral cavity, the epithelial surface is constantly exposed to a number of different microorganisms that are organized in a well-structured biofilm. The aim of this study was to monitor gingival expression of antimicrobial peptides (AMPs) and interleukin-8 (IL-8) in an early gingivitis model.. Experimental gingivitis was allowed to develop in healthy volunteers (n = 17). Bleeding on probing (BOP%) and gingival crevicular fluid volume (GCF) were assessed at baseline and day 1, 3, 5, 7 and 14. Expression of AMPs (human beta-defensin-2, hBD-2; CC-chemokine ligand 20, CCL20; psoriasin, pso/S100A7) and IL-8 was analyzed by immunohistochemistry in gingival biopsies. In addition, hBD-2 and IL-8 protein expression was monitored in GCF using the ELISA technology.. Experimental gingivitis gradually developed with an increase in BOP scores and GCF volume over time. In GCF, elevated concentrations of hBD-2 and IL-8 were monitored at day 1, 5 and 7 (p ≤ 0.0002). Immunohistochemical analysis of gingival sections demonstrated increased staining for hBD-2 at day 3, whereas the CCL20, pso/S100A7, and IL-8 expression was increased at later time points (p < 0.05).. For the first time, this study showed the time-dependent regulation of AMPs, following clinical signs of experimentally induced gingival inflammation. Differential temporal expression for AMPs may ensure a constant antimicrobial activity against changes in the bacterial composition of the growing dental biofilm.

Topics: Adult; Antimicrobial Cationic Peptides; Biopsy; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression Profiling; Gingiva; Gingivitis; Healthy Volunteers; Humans; Immunohistochemistry; Interleukin-8; Male; Prospective Studies; Young Adult

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

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

To analyze in a randomized controlled study whether acute psychological stress alters local proinflammatory signals in a human model of chronic inflammation, i.e., gingivitis. Chronic inflammation represents a crucial factor in a variety of diseases and factors that contribute to the onset and progression of disease. Psychological stress is assumed to represent such a factor. However, experimental human research in this area is rare.. A total of 25 students (n = 11 females, 14 males) suffering from gingivitis were subjected to a stress (public-speaking task) and to a control condition in randomized order. Local concentrations of interleukin (IL)-8 were quantified as an indicator of proinflammatory activity at sites of inflammation. IL-8 is a strong proinflammatory mediator and involved in a variety of disease processes. Samples were taken at sites of inflammation before stress versus control condition and 0, 45, and 90 minutes afterward.. A significant main effect (p = .03) of acute stress on local IL-8 was found. Stress induced an increase of IL-8-concentrations; univariate effect sizes varied between d = 0.23 and d = 0.36.. This is the first human experimental in vivo study demonstrating that psychological stress alters the local concentrations of IL-8 under conditions of chronic inflammation. It provides direct evidence acute stress is involved in the regulation of local proinflammatory responses in chronic inflammation. Future studies should now explore the effects of more enduring stress conditions and the factors mediating stress effects on inflammatory signals.

Topics: Adult; Arousal; Chronic Disease; Cross-Over Studies; Female; Gingivitis; Humans; Inflammation Mediators; Interleukin-8; Male; Signal Transduction; Stress, Psychological

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

Cigarette smoking is a significant risk factor in the pathogenesis of periodontal disease, able to influence both the subgingival microbiota and host responses.. The aim of the present study was to determine the influence of smoking on the amount of IL-1beta, IL-4 and IL-8 in gingival crevicular fluid (GCF) during experimental gingivitis in humans.. Twenty-two healthy subjects, 10 smokers and 12 non-smokers, participated in the study. After professional cleaning, they performed optimal hygiene to reach perfect clinical gingival health. Oral hygiene measures were ceased for a period of 10 days. Clinical indices, including plaque index (PI), gingival index (GI), probing pocket depth (PPD) and bleeding on probing (BOP), were assessed 2 days before (day -2), at the beginning (day 0) and at the end of the experimental gingivitis period (day 10). At the same time, GCF was collected from 12 sites in each patient, by means of durapore filter membranes. Total amounts of IL-1beta, IL-4 and IL-8 were determined by enzyme-linked immunoadsorbent assay.. Clinical data revealed that both smokers and non-smokers showed an increase in PI, GI and BOP scores during the experiment. Although no differences were noted with regard to PI at day 10, the GI and BOP were significantly less pronounced in smokers than non-smokers (p < 0.005). Non-smokers showed higher total amounts of IL-4 but lower amounts of IL-8 than smokers, throughout the experiment. Total amounts of IL-1beta and IL-8 increased significantly during plaque accumulation in both groups. IL-4 remained stable for the smoker group and decreased for the non-smoker group.. The present results indicate that smoking interferes with cytokine production. When performing studies regarding the pathogenesis of periodontitis, the smoking status of the participants needs to be taken into consideration.

Topics: Adult; Dental Plaque Index; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-1; Interleukin-4; Interleukin-8; Interleukins; Linear Models; Male; Periodontal Index; Reference Values; Reproducibility of Results; Smoking

Human saliva is a complex fluid containing proteins such as salivary cytokines, which can be used for diagnostic purposes, particularly among the pediatric population. This study aimed to assess the concentrations of salivary cytokines in healthy children and adolescents and determine their associations with age, sex, and oral and dental findings. Healthy children and adolescents aged 4-18 years were enrolled in this cross-sectional study. The concentrations of the following salivary cytokines were measured by Luminex technology: IFN-γ, IL-1α, IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IP-10, TNF-α, and VEGF-A. Additionally, oral and dental parameters were recorded using a standardized protocol. A total of 128 participants (mean age, 10.7 years; males, 50.8%) were enrolled. The levels of 1β, IL-6, IL-8, and IL-10 were significantly higher in those with gingivitis. Increased salivary flow rates were negatively correlated with IL-1α, IL-1β, IL-6, IL-8, IL-10, TNF-α, and VEGF-A concentrations. The findings of this study showed that the concentrations of most of the salivary cytokines were positively correlated with age and the presence of oral pathologies (such as gingivitis and caries) and negatively correlated with salivary flow rate.

Topics: Adolescent; Chemokine CXCL10; Child; Cross-Sectional Studies; Cytokines; Gingivitis; Humans; Interleukin-10; Interleukin-13; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Male; Oral Health; Saliva; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

The aim of the study is to compare the levels of Gingival Crevicular Fluid (GCF) interleukin 8 (IL-8), matrix metalloproteinase 8 (MMP-8) and advanced glycated-end products (AGEs) in a cohort of type 1 diabetic (T1D) subjects and healthy controls.. GCF samples and periodontal examination were assessed in 50 subjects with T1D (30 males and 20 females; mean age: 35.2 years) recruited from the Diabetology Unit of the Geneva University Hospitals and in 50 control subjects matched for gender, age and smoking status. Samples were assessed for IL-8 and MMP-8 using a bead array multianalyte detection system and for AGEs the ELISA. The two groups were compared using the Wilcoxon signed rank test.. The mean HbA1c differed significantly between the groups (8.3% for the T1D group vs. 5.2% for the control group, p < 0.001). T1D subjects had significantly more plaque and gingival inflammation and presented more sites with bleeding on probing compared to the controls. The GCF levels of IL-8, MMP-8 and AGEs did not differ significantly between the groups. Further analysis of the GCF markers in younger (<40 years) and older (≥40 years) cohorts, revealed no significant differences between younger diabetics and controls or between older diabetics and controls. When the groups were divided according to their glycemic status (HbA1c 6.1-8, and > 8%), again no significant differences could be identified for any of the biochemical markers.. T1D subjects, particularly the younger ones, exhibited more inflammation compared to the matched healthy controls. Results on the GCF expression of IL-8, MMP-8 and AGEs did not differ between the groups. The diabetic population of our cohort was for the most part fairly-controlled, with little if any complications and with presence of only mild type of periodontal disease, as 68% had gingivitis.

Topics: Adult; Biomarkers; Case-Control Studies; Diabetes Mellitus, Type 1; Female; Gingival Crevicular Fluid; Gingivitis; Glycated Hemoglobin; Humans; Inflammation; Interleukin-8; Male; Matrix Metalloproteinase 8

We sought to compare the characteristics and clinical significance of neutrophil extracellular traps in gingival samples from patients with periodontitis and those with gingivitis. The clinical indexes of gingival samples from patients with periodontitis and gingivitis were measured; the expression of TNF-alpha and IL-8 was measured by real-time fluorescent quantitative PCR; and the expression of TLR-8 and MMP-9 was measured by western blotting assays. Chemotaxis, phagocytosis and phagocytic activity of neutrophils were measured. Compared with the healthy group, the expression of TNF-α and IL-8 in the periodontitis group and the gingivitis group increased significantly (p < 0.05), and TNF-α in the gingivitis group was significantly lower than that in the healthy group (p < 0.05). The expression of IL-8 in the periodontitis group was significantly higher than that in the periodontitis group (p < 0.05). Furthermore, the expression of TLR-8 and MMP-9 in the periodontitis group was different from that in the gingivitis group and the healthy group, and the expression of TLR-8 and MMP-9 in the gingivitis group was significantly different from that in the healthy group (p < 0.05). In addition, the neutrophil mobility index in healthy people was 3.02 ± 0.53, that in the periodontitis group was 2.21 ± 0.13, and that in the gingivitis group was 2.31 ± 0.12. In conclusion, the chemotaxis of neutrophils in gingival samples of patients with periodontitis and gingivitis was decreased, the phagocytotic ability and activity of neutrophils were reduced, and the release of the extracellular trap-releasing inducible factors TNF-alpha and IL-8 also declined.

Topics: Actins; Adult; Blotting, Western; Case-Control Studies; Electrophoresis, Agar Gel; Extracellular Traps; Female; Gingivitis; Humans; Interleukin-8; Male; Matrix Metalloproteinase 9; Middle Aged; Neutrophils; Periodontal Index; Periodontitis; Real-Time Polymerase Chain Reaction; Reference Values; RNA; Toll-Like Receptor 8; Tumor Necrosis Factor-alpha; Young Adult

Porphyromonas gingivalis and Tannerella forsythia secrete proteases, gingipains and KLIKK-proteases. In addition, T. forsythia produces a serpin (miropin) with broad inhibitory spectrum. The aim of this pilot study was to determine the level of expression of miropin and individual proteases in vivo in periodontal and peri-implant health and disease conditions. Biofilm and gingival crevicular fluid (GCF)/ peri-implant sulcular fluid (PISF) samples were taken from healthy tooth and implant sites (n = 10), gingivitis and mucositis sites (n = 12), and periodontitis and peri-implantitis sites (n = 10). Concentration of interleukin-8 (IL-8), IL-1β and IL-10 in GCF was determined by enzyme-linked immunosorbent assay. Loads of P. gingivalis and T. forsythia and the presence of proteases and miropin genes were assessed in biofilm by quantitative PCR, whereas gene expression was estimated by quantitative RT-PCR. The presence of P. gingivalis and T. forsythia, as well as the level of IL-8 and IL-1β, were associated with disease severity in the periodontal and peri-implant tissues. In biofilm samples harboring T. forsythia, genes encoding proteases were found to be present at 72.4% for karilysin and 100% for other KLIKK-protease genes and miropin. At the same time, detectable mRNA expression of individual genes ranged from 20.7% to 58.6% of samples (for forsylisin and miropsin-1, respectively). In comparison with the T. forsythia proteases, miropin and the gingipains were highly expressed. The level of expression of gingipains was associated with those of miropin and certain T. forsythia proteases around teeth but not implants. Cumulatively, KLIKK-proteases and especially miropin, might play a role in pathogenesis of both periodontal and peri-implant diseases.

Topics: Bacterial Proteins; Biofilms; Biomarkers; Dental Implants; Gene Expression Regulation, Bacterial; Genes, Bacterial; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-10; Interleukin-1beta; Interleukin-8; Mucositis; Peptide Hydrolases; Peri-Implantitis; Periodontitis; Pilot Projects; Porphyromonas gingivalis; Protease Inhibitors; RNA, Messenger; Serpins; Sweden; Tannerella forsythia

Calprotectin (S100A8/A9) is a heterodimer of S100A8 and S100A9 and is associated with multiple inflammatory diseases, including Crohn's disease, rheumatoid arthritis and periodontitis. Levels of calprotectin are elevated in the gingival crevicular fluid of patients with periodontitis; however, the effects of calprotectin on human gingival fibroblasts (HGFs) remain unknown. This study investigated the proinflammatory activity of calprotectin on HGFs and the functional receptors and signaling pathways engaged by calprotectin.. HGFs were stimulated by equimolar concentrations of S100A8 and/or S100A9, and the expression levels of interleukin (IL)-6 and IL-8 were detected using real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assays. The calprotectin receptors were identified by pre-incubating HGFs with the toll-like receptor (TLR) 4 inhibitor or the antibody targeting the advanced glycation end product receptor (RAGE). The involvement of reactive oxygen species (ROS) and signaling pathways were also investigated by treating HGFs with ROS inhibitor or specific pathway inhibitors, respectively.. S100A9 and S100A8/A9 significantly upregulated IL-6 and IL-8 expression, which was inhibited upon treatment with the TLR4 inhibitor TAK242. Pretreatment with RAGE-blocking antibodies did not affect cytokine expression. Additionally, S100A9 promoted the production of IL-6 and IL-8 from HGFs via different signaling pathways. IL-6 expression was upregulated via the NF-κB, c-Jun amino-terminal kinase (JNK) 1/2 and p38 mitogen-activated protein kinase (MAPK) pathways, and IL-8 expression was upregulated via NF-κB, p38, JNK1/2 and extracellular-regulated kinase 1/2 MAPK pathways. The release of both cytokines was dependent upon the production of ROS.. Our findings suggest that calprotectin exerts proinflammatory effects on HGFs via the S100A9 subunit and TLR4-mediated NF-κB and MAPK signaling pathways.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Female; Fibroblasts; Gingiva; Gingivitis; Humans; Interleukin-6; Interleukin-8; Leukocyte L1 Antigen Complex; Male; MAP Kinase Signaling System; Signal Transduction; Toll-Like Receptor 4; Young Adult

Evidence of increased apoptosis is observed in periodontitis and may be associated with destruction of the periodontal tissue caused by the increased cell death, with the release of danger signals and subsequent stimulation of the proinflammatory processes. However, the exact mechanisms associated with these processes remain unclear. This study aimed to investigate the presence of the periodontal pathogen Treponema denticola, apoptosis, high mobility group box 1 as a damage-associated molecular pattern, and several inflammatory markers in periodontitis and gingivitis subjects.. Soft tissue specimens from gingival tissues of periodontitis and gingivitis patients were used for immunohistochemical and immunofluorescence staining of T. denticola chymotrypsin-like proteinase (CTLP), apoptosis markers, high mobility group box 1, Toll-like receptor 4, inflammatory cell markers, and proinflammatory cytokines.. Treponema denticola was detected in all periodontitis-affected tissues. This was associated with a significant increase in the number of apoptotic cells, including macrophages, alterations in the expression of high mobility group box 1 and its receptor, and increased levels of proinflammatory cytokines compared with gingivitis.. In summary, the presence of T. denticola (especially its CTLP), apoptosis, high mobility group box 1, and inflammatory markers suggests their potential involvement in the pathogenesis of periodontitis.

Topics: Adult; Aged; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Apoptosis; Caspase 3; Female; Gingivitis; HMGB1 Protein; Humans; Immunohistochemistry; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Peptide Hydrolases; Periodontitis; Toll-Like Receptor 4; Treponema denticola

Polyhexamethylene guanidine phosphate (PHMG-P) was compared to chlorhexidine (CHX) in order to determine potential cytotoxic and immune-modulatory effects on human gingival fibroblasts.. Cytotoxic effects of PHMG-P and CHX on human gingival fibroblasts were assessed using cell viability assay at various time points and concentrations. The effects of PHMG-P and CHX on the secretion of prostaglandin (PG) E. PHMG-P concentration 0.00009% led to the total loss of fibroblast viability within 24 h, whereas inhibition of fibroblast viability by CHX occurred at significantly higher concentrations of 0.0009% (p < .001). Short-term exposure to 0.005% PHMG-P led to loss of fibroblast viability after 5 min, whilst cells exposed to 0.005% CHX survived 30 min of treatment (p < .001). IL-1β stimulation induced an inflammatory response with a significant increase in the secretion of PGE. Cytotoxic effects on gingival fibroblasts were triggered by both PHMG-P and CHX at concentrations below those used in clinical practice. The tested antiseptics did not cause inflammation and reduced IL-1β-induced secretion of inflammatory mediators and collagenase by gingival fibroblasts, which suggests anti-inflammatory properties.

Topics: Anti-Infective Agents, Local; Cells, Cultured; Chlorhexidine; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gingiva; Gingivitis; Guanidines; Humans; Interleukin-6; Interleukin-8

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

To evaluate the relation among gingival inflammation, salivary osmolality, levels of IL-1β, IL-6, IL-8, TNF-α, and s-IgA concentrations in children with spastic CP with or without cervical motor control in a cross-sectional study.. Unstimulated whole saliva and the gingival index were collected in 37 and 34 CP children with and without cervical motor control, respectively. The data were dichotomized as follows: (=0) absence of gingival inflammation and (≥0.1) presence of gingival inflammation.. The group without cervical control presented statistically higher mean values of salivary osmolality, s-IgA, and cytokines. In addition, statistically positive correlation between the gingival index and salivary cytokines was observed in the group with cervical control. Salivary osmolality, salivary cytokines, and s-IgA from both groups presented a significant positive correlation. Significant differences (P = 0.00336) in the values of salivary osmolality were observed between the CP individuals with (93.9 ± 32.7) and without gingival inflammation (74.4 ± 16.6). ROC analysis was performed, and values of salivary osmolality >80 indicated a sensitivity of 0.54 and a specificity of 0.79.. Children without cervical motor control presented a more pronounced oral inflammatory status that was characterized by higher levels of cytokines.

Topics: Adolescent; Biomarkers; Brazil; Cerebral Palsy; Child; Cross-Sectional Studies; Cytokines; Female; Gingivitis; Humans; Immunoglobulin A; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Osmolar Concentration; Periodontal Index; Rehabilitation; Saliva; Tumor Necrosis Factor-alpha

Veratric acid, one of the major benzoic acid isolated from vegetables and fruits, has been reported to have anti-inflammatory activity. The purpose of this study was to investigate the anti-inflammatory effects of veratric acid on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). HGFs were pretreated with veratric acid 1 h before LPS stimulation. The productions of IL-6 and IL-8 were detected by ELISA. The expression of iNOS, COX-2, PI3K, AKT, and NF-κB were detected by western blotting. The results showed that veratric acid inhibited LPS-induced IL-6 and IL-8 production, as well as iNOS and COX-2 expression. Veratric acid also inhibited LPS-induced NF-κB activation. In addition, veratric acid was found to suppress LPS-induced PI3K and AKT phosphorylation. In conclusion, the anti-inflammatory mechanism of veratric acid is due to its ability to inhibit PI3K/Akt/NF-κB signaling pathway.

Topics: Anti-Inflammatory Agents; Cell Survival; Cells, Cultured; Cyclooxygenase 2; Fibroblasts; Gingiva; Gingivitis; Humans; Inflammation; Interleukin-6; Interleukin-8; Lipopolysaccharides; MAP Kinase Signaling System; NF-kappa B; Nitric Oxide Synthase Type II; Periodontitis; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Vanillic Acid

The gingival epithelium plays an important role in protecting against the invasion of periodontal pathogens, and the permeability of gingival epithelial cells has been implicated in the initiation of periodontitis. Azithromycin (AZM) has been used in the treatment of chronic inflammatory airway diseases because it regulates cell-cell contact in airway epithelial cells. Therefore, AZM may also regulate barrier function in gingival epithelial cells. In the present study, we examined the effects of AZM on the permeability of human gingival epithelial cells (HGEC) under inflammatory conditions in vitro.. HGEC were stimulated by tumor necrosis factor-α (TNF-α) in the presence of AZM or p38 MAP kinase and ERK inhibitors. Permeability was assessed based on transepithelial electrical resistance (TER). The expression of E-cadherin, phosphorylated p38 MAP kinase, and ERK was analyzed by Western blotting.. TNF-α decreased TER in HGEC, and AZM and the p38 MAP kinase and ERK inhibitors recovered this decrease. AZM inhibited the phosphorylation of ERK and p38 MAP kinase in TNF-α-stimulated HGEC. Furthermore, AZM recovered the decrease in E-cadherin expression in HGEC stimulated with TNF-α.. These results suggested that AZM regulated gingival epithelial permeability through p38 MAP kinase and ERK signaling, and may contribute to suppress the inflammation in gingival tissue.

Topics: Azithromycin; Cadherins; Cell Line; Cell Membrane Permeability; Cells, Cultured; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gingiva; Gingivitis; Humans; Interleukin-8; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha

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

The aim of this cross-sectional study is to compare the local and systemic levels of soluble receptor activator of nuclear factor-κB ligand (sRANKL), osteoprotegerin (OPG), a proliferation-inducing ligand (APRIL), B-cell activating factor (BAFF), interleukin (IL)-6, and IL-8 in biofluids of patients with thalassemia major (TM) with or without gingivitis.. Seventy-seven patients are included in this study (TM, n = 29; systemically healthy, n = 48). Gingival crevicular fluid (GCF), saliva, and serum levels of IL-6, IL-8, sRANKL, OPG, BAFF, and APRIL were determined by enzyme-linked immunosorbent assay. Data were analyzed by appropriate non-parametric or parametric statistical tests.. Median GCF, serum, and saliva levels for BAFF (P <0.001) and IL-6 and IL-8 (P <0.005) were higher in TM gingivitis than in systemically healthy gingivitis (P <0.001). GCF, serum, and saliva levels for APRIL, sRANKL, IL-6, and IL-8 were higher in TM than in systemically and periodontally healthy comparison groups (P <0.05). Positive correlations were found between bleeding on probing (BOP), plaque index (PI) scores, and GCF APRIL, serum sRANKL, serum OPG, and sRANKL concentrations in TM groups (P <0.05). Several significant positive correlations were found between BOP, PI scores, and biofluid parameters also in systemically healthy groups.. TM may have a role in the underlying systemic hematologic condition and potentially affect gingival inflammation via dysregulation of lymphocytes and increased activation of osteoclasts.

Topics: Adolescent; Adult; B-Cell Activating Factor; beta-Thalassemia; Cross-Sectional Studies; Dental Plaque Index; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Osteoprotegerin; Periodontal Index; RANK Ligand; Saliva; Tumor Necrosis Factor Ligand Superfamily Member 13; Young Adult

Pregnancy-associated gingivitis is a bacterial-induced inflammatory disease with a remarkably high prevalence ranging from 35% to 100% across studies. Yet little is known about the attendant mechanisms or diagnostic biomarkers that can help predict individual susceptibility for rational personalized medicine. We aimed to define inflammatory proteins in saliva, induced or inhibited by estradiol, as early diagnostic biomarkers or target proteins in relation to pregnancy-associated gingivitis. An in silico gene/protein interaction network model was developed by using the STITCH 3.1 with "experiments" and "databases" as input options and a confidence score of 0.700 (high confidence). Salivary estradiol, interleukin (IL)-1β and -8, myeloperoxidase (MPO), matrix metalloproteinase (MMP)-2, -8, and -9, and tissue inhibitor of matrix metalloproteinase (TIMP)-1 levels from 30 women were measured prospectively three times during pregnancy and twice during postpartum. In silico analysis revealed that estradiol interacts with IL-1β and -8 by an activation link when the "actions view" was consulted. In saliva, estradiol concentrations associated positively with TIMP-1 and negatively with MPO and MMP-8 concentrations. When the gingival bleeding on probing percentage (BOP%) was included in the model as an effect modifier, the only association, a negative one, was found between estradiol and MMP-8. Throughout gestation, estradiol modulates the inflammatory response by inhibiting neutrophilic enzymes, such as MMP-8. The interactions between salivary degradative enzymes and proinflammatory cytokines during pregnancy suggest promising ways to identify candidate biomarkers for pregnancy-associated gingivitis, and for personalized medicine in the field of dentistry. Finally, we call for greater investments in, and action for biomarker research in periodontology and dentistry that have surprisingly lagged behind in personalized medicine compared to other fields, such as cancer research.

Topics: Biomarkers; Computer Simulation; Dentistry; Estradiol; Female; Gingivitis; Humans; Interleukin-1beta; Interleukin-8; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Peroxidase; Precision Medicine; Pregnancy; Pregnancy Complications; Prospective Studies; Protein Interaction Maps; Tissue Inhibitor of Metalloproteinase-1

We previously demostrated that EMMPRIN participates in the periodontitis and its interaction with Cyclophilin A possibly exists in animal periodontitis models. This study is aimed to address the expression and potential role of cyclophilin A (CypA) in human periodontitis.. Gingival tissues and peripheral blood were collected from patients with moderate to severe periodontitis or from healthy donors. Western blotting and immunohistochemistry were performed to detect the expression and distribution of CypA in the gingival tissues. Peripheral blood mononuclear cells (PBMCs) and neutrophils were isolated from the peripheral blood by Ficoll-Paque density-gradient centrifugation. Chemotaxis assays were applied to evaluate the effects of different concentrations of CypA (100, 300 and 500 ng/mL) on the migration of PBMCs and neutrophils. Supernatants of human THP-1 cells were collected after treatment with 200 ng/mL of CypA for different periods of time (1, 3, 6, 12 and 24 h) to detect the levels of interleukin (IL)-1β, IL-8 and tumor necrosis factor alpha (TNF-α) by ELISA.. Western blot analyses revealed an increase of CypA expression in inflamed gingival tissues compared with healthy tissues. Immunohistochemistry identified that the over-expressed CypA was localized in the infiltrating cells and/or in the extracellular matrix in the inflamed gingival connective tissues. The positive infiltrating cells contained mononuclear cells and lobulated-nuclei neutrophils. Chemotactic assays showed that 300 ng/mL of CypA apparently facilitated the chemotaxis of PBMCs/neutrophils from healthy donors, compared with the no-treatment control (p < 0.01 for PBMCs, p < 0.05 for neutrophils), whereas 100 and 500 ng/mL of CypA only weakly enhanced the chemotaxis of PBMCs/neutrophils (p > 0.05 for PBMCs/neutrophils, not significant). The PBMCs/neutrophils from patients with periodontitis exhibited a stronger ability to migrate when stimulated with 300 ng/mL of CypA than did PBMCs/neutrophils from healthy donors (p < 0.05 for PBMCs, p < 0.01 for neutrophils). ELISA revealed that the level of TNF-α secreted by THP-1 cells was elevated after treatment with 200 ng/mL of CypA for 12 h compared with the no-treatment 0-h control (p < 0.05). The IL-8 level was sharply raised after 3 h of stimulation with 200 ng/mL of CypA (p < 0.01 compared with 0 h), but no significant change was observed at the other time points (p > 0.05). There was no statistical difference at any of the treatment time points for the secretion of IL-1β (p > 0.05 for 1, 3, 6, 12 and 24 h compared with 0 h).. CypA participates in the pathogenesis of human periodontitis. It may be involved in the inflammatory response of periodontal tissues through inducing the chemotaxis of PBMCs/neutrophils and the secretion of TNF-α/IL-8.

Topics: Cell Line; Cell Movement; Chemotaxis, Leukocyte; Cyclophilin A; Extracellular Matrix; Gingiva; Gingivitis; Humans; Interleukin-1beta; Interleukin-8; Leukocytes, Mononuclear; Monocytes; Neutrophils; Periodontitis; Time Factors; Tumor Necrosis Factor-alpha

Gingival epithelial cells (GECs) represent a physical barrier against bacteria and are involved in the processes of innate immunity. Recently, an anti-inflammatory and immune-modulatory effect of the amino acid glycine has been demonstrated. However, there is only little information about the immune-modulatory effects of glycine in oral tissues. This study aimed to investigate the existence and role of the glycine receptor in gingival tissue analyzing tissues/cells from extracted human molars via immunohistochemical analysis. In vitro, GECs were challenged by inflammatory conditions with IL-1 β alone or in combination with glycine and analyzed for cytokine expression of IL6/IL8 via real-time PCR. On protein level, the effect of nuclear translocalization of NF κ B protein p65 was analyzed using immunofluorescence analysis. A distinct proof of the GlyR in oral gingival tissue and keratinocytes could be demonstrated. Isolated challenge of the keratinocytes with IL-1 β as well as with glycine resulted in an upregulation of IL6 and IL8 mRNA expression and activation of NF κ B pathway. The presence of glycine in combination with the inflammatory stimulus led to a significant decrease in inflammatory parameters. These results indicate a possible anti-inflammatory role of glycine in gingival inflammation and encourage further research on the utility of glycine in the prevention or therapy of inflammatory periodontitis.

Topics: Epithelial Cells; Gene Expression Regulation; Gingivitis; Glycine; Humans; Immunity, Innate; Immunologic Factors; Interleukin-1beta; Interleukin-6; Interleukin-8; Keratinocytes; Receptors, Glycine; RNA, Messenger; Signal Transduction; Transcription Factor RelA

The aim of this study was to assess gingival fluid (GCF) cytokine messenger RNA (mRNA) levels, subgingival bacteria, and clinical periodontal conditions during a normal pregnancy to postpartum.. Subgingival bacterial samples were analyzed with the checkerboard DNA-DNA hybridization method. GCF samples were assessed with real-time PCR including five proinflammatory cytokines and secretory leukocyte protease inhibitor.. Nineteen pregnant women with a mean age of 32 years (S.D. ± 4 years, range 26-42) participated in the study. Full-mouth bleeding scores (BOP) decreased from an average of 41.2% (S.D. ± 18.6%) at the 12th week of pregnancy to 26.6% (S.D. ± 14.4%) at the 4-6 weeks postpartum (p < 0.001). Between week 12 and 4-6 weeks postpartum, the mean probing pocket depth changed from 2.4 mm (S.D. ± 0.4) to 2.3 mm (S.D. ± 0.3) (p = 0.34). Higher counts of Eubacterium saburreum, Parvimonas micra, Selenomonas noxia, and Staphylococcus aureus were found at week 12 of pregnancy than at the 4-6 weeks postpartum examinations (p < 0.001). During and after pregnancy, statistically significant correlations between BOP scores and bacterial counts were observed. BOP scores and GCF levels of selected cytokines were not related to each other and no differences in GCF levels of the cytokines were observed between samples from the 12th week of pregnancy to 4-6 weeks postpartum. Decreasing postpartum counts of Porphyromonas endodontalis and Pseudomonas aeruginosa were associated with decreasing levels of Il-8 and Il-1β.. BOP decreased after pregnancy without any active periodontal therapy. Associations between bacterial counts and cytokine levels varied greatly in pregnant women with gingivitis and a normal pregnancy outcome. Postpartum associations between GCF cytokines and bacterial counts were more consistent.. Combined assessments of gingival fluid cytokines and subgingival bacteria may provide important information on host response.

Topics: Adult; Bacterial Load; Cytokines; Eubacterium; Female; Gingiva; Gingival Crevicular Fluid; Gingival Hemorrhage; Gingivitis; Humans; Inflammation Mediators; Interleukin-1alpha; Interleukin-1beta; Interleukin-8; Peptostreptococcus; Periodontal Index; Periodontal Pocket; Periodontitis; Porphyromonas endodontalis; Postpartum Period; Pregnancy; Pseudomonas aeruginosa; RNA, Messenger; Secretory Leukocyte Peptidase Inhibitor; Selenomonas; Staphylococcus aureus; Tumor Necrosis Factor-alpha

Cytokines produced by various cells are strong local mediators of inflammation. Mucosa-associated epithelial chemokine (CCL28), interleukin-8 (IL-8), interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) are major cytokines that play important roles in the periodontal inflammatory process. In this study we aimed to compare the levels of CCL28, IL-8, IL-1β and TNF-α in the gingival crevicular fluid of both periodontally healthy subjects and in subjects diagnosed with gingivitis, chronic periodontitis and generalized aggressive periodontitis.. A total of 84 subjects participated in the study: 21 subjects had gingivitis, 21 subjects had chronic periodontitis, 21 subjects had generalized aggressive periodontitis and 21 were periodontally healthy. The levels of CCL28, IL-8, IL-1β and TNF-α were analyzed using enzyme-linked immune sorbent assay (ELISA).. The total levels of CCL28 and IL-8 in the gingival crevicular fluid of the generalized aggressive periodontitis group (324.74 ± 42.62 pg/30 s, 487.62 ± 49.21 pg/30 s) were significantly higher than those of the chronic periodontitis group (268.81 ± 28.64 pg/30 s, 423.65 ± 35.24 pg/30 s), the gingivitis group (146.35 ± 17.46 pg/30 s, 310.24 ± 48.20 pg/30 s) and the periodontally healthy group (92.46 ± 22.04 pg/30 s, 148.41 ± 24.64 pg/30 s). Similarly, the total levels of IL-1β and TNF-α in the generalized aggressive periodontitis group (110.23 ± 9.20 pg/30 s, 1284.46 ± 86.32 pg/30 s) were significantly higher than those in the chronic periodontitis group (423.65 ± 35.24 pg/30 s, 82.64 ± 9.12 pg/30 s), the gingivitis group (52.10 ± 7.15 pg/30 s, 824.24 ± 44.68 pg/30 s) and the periodontally healthy group (36.44 ± 8.86 pg/30 s, 628.26 ± 34.61 pg/30 s).. CCL28, IL-8, IL-1β and TNF-α may play key roles in the host response to inflammation in periodontal diseases. As the severity of periodontal diseases increases, destruction of periodontal tissues also increases. Inflammation is one among many factors that trigger periodontal tissue destruction. Identification of the mediators that influence the development and progression of inflammation in periodontal diseases may be very important in understanding the prognoses of periodontal diseases.

Topics: Adolescent; Adult; Aggressive Periodontitis; Alveolar Bone Loss; Chemokines, CC; Chronic Periodontitis; Female; Gingival Crevicular Fluid; Gingival Hemorrhage; Gingivitis; Humans; Immunity, Mucosal; Interleukin-1beta; Interleukin-8; Male; Periodontal Attachment Loss; Periodontal Pocket; Periodontium; Tumor Necrosis Factor-alpha; Young Adult

In periodontitis, tissue damage results mainly from aberrant host responses to oral microorganisms. Fibroblasts can play an important role in this. Gingival fibroblasts do not develop tolerance against the lipopolysaccharide of Porphyromonas gingivalis, a keystone pathogen in periodontitis, which may partly explain the persistence of inflammation. However, besides lipopolysaccharide, live P. gingivalis possess numerous virulence traits to impair host-responses. We hypothesized that fibroblast-responsiveness to a bacterial challenge could be affected by live P. gingivalis. We investigated if inflammatory responses of gingival fibroblasts to P. gingivalis were altered, when the fibroblasts had encountered P. gingivalis previously. On consecutive days, primary human gingival fibroblasts were challenged twice for 6 h with live P. gingivalis, or fibroblasts were preincubated for 24 h with a lower concentration of live P. gingivalis and re-challenged for 6 h with a higher concentration. As the P. gingivalis capsule and proteases are involved in modulating host responses, we used encapsulated P. gingivalis W83 and a non-encapsulated mutant, and P. gingivalis ATCC33277 and a lys-gingipain and arg-gingipain mutant, to challenge fibroblasts. With all P. gingivalis-strains, interleukin-8 and monocyte chemoattractant protein-1 responses to the second challenge were less strong in fibroblasts that had been challenged with P. gingivalis before. These lower responses might correspond with higher interleukin-1 receptor agonist expression. Fibroblast responses to a second challenge were not influenced by 24 h preincubation. Reduced chemokine responses after consecutive potent P. gingivalis challenges indicate that gingival fibroblast responsiveness is affected by a previous bacterial encounter. In periodontitis, such reduced chemokine responses may impair chemotaxis and clearance of oral microorganisms, thereby leading to prolonged inflammatory responses and tissue damage.

Topics: Adhesins, Bacterial; Adult; Chemokine CCL2; Chemokines; Chemotaxis; Cysteine Endopeptidases; Female; Fibroblasts; Gingipain Cysteine Endopeptidases; Gingiva; Gingivitis; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Periodontitis; Porphyromonas gingivalis; Statistics, Nonparametric; Virulence Factors

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

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

High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of high mobility group (HMG) family, play important role in inflammation. The purpose of this study was to investigate the expression of HMGB1 and HMGN2 in periodontistis.. The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalised aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1β, IL-6, IL-8, TNF-α and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA.. HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1β, IL-6, IL-8 proinflammaory cytokines.. To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.

Topics: Adolescent; Adult; Aged; Aggressive Periodontitis; Alveolar Bone Loss; Chronic Periodontitis; Dental Calculus; Dental Implants; Dental Plaque Index; Female; Gingiva; Gingival Crevicular Fluid; Gingival Hemorrhage; Gingivitis; HMGB1 Protein; HMGN2 Protein; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Peri-Implantitis; Periodontal Pocket; Periodontitis; Periodontium; Tumor Necrosis Factor-alpha; 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

Hydrogen sulfide (H(2)S), a volatile sulfur compound, is implicated as a cause of inflammation, especially when it is produced by bacteria colonizing gastrointestinal organs. However, it is unclear if H(2)S produced by periodontal pathogens affects the inflammatory responses mediated by oral/gingival epithelial cells. Therefore, the aims of this study were (1) to compare the in vitro production of H(2)S among 14 strains of oral bacteria and (2) to evaluate the effects of H(2)S on inflammatory response induced in host oral/gingival epithelial cells. Porphyromonas gingivalis (Pg) produced the most H(2)S in culture, which, in turn, resulted in the promotion of proinflammatory cytokine IL-8 from both gingival and oral epithelial cells. The up-regulation of IL-8 expression was reproduced by the exogenously applied H(2)S. Furthermore, the mutant strains of Pg that do not produce major soluble virulent factors, i.e. gingipains, still showed the production of H(2)S, as well as the promotion of epithelial IL-8 production, which was abrogated by H(2)S scavenging reagents. These results demonstrated that Pg produces a concentration of H(2)S capable of up-regulating IL-8 expression induced in gingival and oral epithelial cells, revealing a possible mechanism that may promote the inflammation in periodontal disease.

Topics: Cell Line; Epithelial Cells; Gingiva; Gingivitis; Humans; Hydrogen Sulfide; Interleukin-8; Porphyromonas gingivalis

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

Our aim was to assess the modalities of use and the anti-inflammatory activity of enoxolone included in toothpaste and in a mouthwash solution.. We used gingival fragments kept alive during 3 days at 37 degrees C. To induce inflammation, inflammatory mediators (SP and LPS) were applied to culture medium on contact with corium. The toothpaste versus placebo was applied on epithelium, in double blind. Histological analysis was then performed on hematoxylin and eosin stained slides. Edema was evaluated with semi-quantitative scores. Vasodilatation was studied by counting the percentage of dilated vessels according to scores and the surface of these dilated vessels by morphometrical image analysis. An inflammatory cytokine, IL8, was measured in culture supernatants. Dosing IL1alpha tested the mouth solution.. The toothpaste induced a significant decrease of edema, vasodilatation, and IL8 excretion. The enoxolone solution induced a decrease of IL1alpha.. Enoxolone demonstrated an anti-inflammatory property whatever the carrier was.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Dentifrices; Diffusion Chambers, Culture; Gingivitis; Glycyrrhetinic Acid; Humans; Interleukin-1alpha; Interleukin-8; Mouth Mucosa; Mouthwashes; Polysaccharides, Bacterial; Substance P; Tissue Culture Techniques

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

To compare salivary IL-1beta, IL-6, IL-8, and TNF-alpha levels between patients with burning mouth syndrome (BMS) and controls.. Forty female patients with BMS (mean age: 61.6+/-10.1 years) and 20 female control subjects (mean age: 65.1+/-9.0 years) were included in the study. Unstimulated (UWS) and stimulated whole saliva samples (SWS) were collected and their flow rates were determined. Salivary IL-1beta, IL-6, IL-8, and TNF-alpha levels and total protein concentration were also determined. Salivary transferrin level was determined to investigate the level of blood contamination in saliva samples. Gingival index of the subjects was also examined. Student's t-test, Pearson's correlation analysis, and analysis of covariance were used.. No significant differences were found in the salivary levels of IL-1beta, IL-6, IL-8, and TNF-alpha in BMS patients compared with controls. Salivary flow rates and their total protein concentrations did not differ significantly between the groups. The levels of salivary cytokines and total protein concentration correlated significantly with the level of blood contamination in both UWS and SWS.. There were no differences in the salivary levels of IL-1beta, IL-6, IL-8, and TNF-alpha in BMS patients compared with controls. Cytokine levels in whole saliva were affected mainly by the amount of blood contamination.

Topics: Aged; Blood; Burning Mouth Syndrome; Case-Control Studies; Female; Gingival Hemorrhage; Gingivitis; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Middle Aged; Periodontal Index; Saliva; Salivary Proteins and Peptides; Secretory Rate; Transferrin; Tumor Necrosis Factor-alpha

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

In the present study, we investigated the anti-inflammatory effects of a Kampo medicine Shosaikoto (TJ-9) using in vitro periodontal disease model, in which human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS) from Porphyromonas gingivalis (PgLPS) produce IL-6, IL-8 and prostaglandin E2 (PGE2). Treatment with PgLPS (10 ng/ml), TJ-9 (up to 1 mg/ml) and their combinations for 24 h did not affect the viability of HGFs. Moreover, TJ-9 did not alter LPS-induced IL-6 and IL-8 productions. However, TJ-9 significantly suppressed LPS-induced PGE2 production in a dose-dependent manner but TJ-9 alone did not affect basal PGE2 level. Western blotting demonstrated that TJ-9 decreased cyclooxygenase-2 (COX-2) expression in a dose-dependent manner but not phospholipase A2. Moreover, TJ-9 selectively and dose-dependently inhibited COX-2 activity. These results suggest that TJ-9 decreased PGE2 production by inhibition of both COX-2 expression and activity and that TJ-9 may be useful to improve gingival inflammation in periodontal disease.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Blotting, Western; Cell Survival; Cyclooxygenase 1; Cyclooxygenase 2; Dinoprostone; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gingiva; Gingivitis; Humans; Indicators and Reagents; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages; Medicine, Kampo; Monocytes; Porphyromonas gingivalis; Tetrazolium Salts; Thiazoles

The aim of this study was to investigate if coherence length is of importance in laser phototherapy. Twenty patients with moderate periodontitis were selected. After oral hygiene instructions, scaling and root planing (SRP), one side of the upper jaw was randomly selected for HeNe (632.8 nm, 3 mW) or InGaAlP (650 nm, 3 mW) laser irradiation. One week after SRP, the following parameters were measured: pocket depth, gingival index, plaque index, gingival crevicular fluid volume, matrix metalloproteinase (MMP-8), interleukin (IL-8) and subgingival microflora. The irradiation (180 s per point, energy 0.54 J) was then performed once a week for 6 weeks. At the follow up examination, all clinical parameters had improved significantly in both groups. A more pronounced decrease of clinical inflammation was observed after HeNe treatment. MMP-8 levels were considerably reduced on the HeNe side, while there was no difference for IL-8 or microflora. Coherence length appears to be an important factor in laser phototherapy.

Topics: Adult; Female; Gingiva; Gingivitis; Health Status Indicators; Humans; Inflammation; Interleukin-8; Lasers, Gas; Low-Level Light Therapy; Male; Matrix Metalloproteinase 8; Middle Aged; Periodontitis; Phototherapy; Pilot Projects; Prospective Studies

The aim of the present study was to compare concentrations of cytokines, matrix metalloproteinases (MMPs) and a metalloproteinase inhibitor (TIMP-1) in gingival crevicular fluids (GCF) from sites with gingival inflammation in 28 young patients with Papillon-Lefèvre syndrome (PLS), and in age- and gender-matched controls. Each group consisted of 17 females and 11 males with a mean age of 11.0 years (range 4-22 years). In both groups, anterior upper sites with a clinical diagnosis of gingival inflammation and with pockets < or = 3 mm were selected for sampling of GCF, which was carried out with filter disks inserted into the gingival crevice until saturated. The concentrations of cytokines (IL-1alpha, IL-1beta, TNF-alpha, and IL-8), matrix metalloproteinases (MMP-1, MMP-3, MMP-8, and MMP-9), and their tissue inhibitor (TIMP-1) were analysed using commercial ELISA kits. Significantly higher levels of IL-1beta (P < 0.001) and MMP-8 (P < 0.05) were disclosed among the PLS patients compared with their controls, while the opposite was found for IL-8 (P < 0.05) and MMP-1 (P < 0.001). The individual variations were considerable in both groups. When comparing the expression of cytokines, MMPs, and TIMP-1 in PLS patients with clinically active and non-active periodontitis, the non-active PLS patients showed significantly higher values of IL-1beta than the patients with active periodontal disease (ANOVA, P < 0.01). In conclusion, this study was unable to demonstrate a clear-cut pathognomonic expression of cytokines or MMPs in patients with PLS, but further studies on cytokine and MMP output are warranted.

Topics: Adolescent; Adult; Child; Child, Preschool; Cytokines; Female; Gingival Crevicular Fluid; Gingival Pocket; Gingivitis; Humans; Interleukin-1; Interleukin-8; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Papillon-Lefevre Disease; Periodontitis; Tissue Inhibitor of Metalloproteinase-1; Tumor Necrosis Factor-alpha

In the inflammatory gingival tissues of patients with periodontitis, cytokines such as interleukin (IL)-1 alpha, IL-1 beta, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha have been detected. Gingival fibroblasts are the major constituents of gingival tissue. We recently demonstrated that lipopolysaccharide (LPS) from periodontopathic bacteria induces inflammatory reactions in various tissues via CD14 and/or Toll-like receptors (TLRs) in gingival tissues [Biochem. Biophys. Res. Commun. 273 (2000) 1161]. To confirm this, we examined the expression of IL-1 alpha, IL-1 beta, IL-6, IL-8, TNF-alpha, CD14, TLR2, and TLR4 in human gingival fibroblasts (HGFs) obtained from patients with healthy or inflammatory gingiva using DNA microarray analysis. We also studied the expression levels of these proteins by flow cytometric analysis (FACS). The expression levels of all eight genes in the HGFs of the Inflammatory group were significantly higher than those in the Healthy group on DNA microarray analysis. FACS revealed that the expression levels of all eight proteins on the HGFs of the Inflammatory group were higher than those on the Healthy group. Our data indicated that these eight proteins in HGFs are involved in inflammatory conditions in the gingiva, including periodontal disease. Our results suggested that these eight proteins, in turn, act directly or indirectly on the immune response by activating host cells involved in inflammatory processes.

Topics: Cells, Cultured; Cytokines; Fibroblasts; Flow Cytometry; Gene Expression Profiling; Gingiva; Gingivitis; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Lipopolysaccharide Receptors; Membrane Glycoproteins; Oligonucleotide Array Sequence Analysis; Receptors, Cell Surface; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptors; Tumor Necrosis Factor-alpha

The diseased periodontium appears to express features of a systemic and a mucosal immune response. Our aims were to determine differences in immunoglobulin expression between gingivitis and periodontitis lesions and to ascertain whether immune and inflammatory cells were recruited into the diseased periodontium by the mucosal addressin adhesion molecule (MAdCAM-1).. In situ hybridization and immunohistochemistry were used to detect the expression of chemokines, adhesion molecules and immunoglobulins in tissue sections of gingival and granulation tissues excised from periodontitis-affected sites and of healthy tissue and gingivitis-affected tissue excised during crown-lengthening procedures.. Greater numbers of plasma cells were observed in periodontitis gingival/granulation tissue lesions compared with gingivitis lesions. While IgA1 were predominant in all lesions, IgA2 and J-chain expressing plasma cells were present in increased proportions in gingival tissues compared with granulation tissue. Intracellular adhesion molecule-1 (ICAM-1) was higher in periodontitis than in gingivitis and interleukin-8 mRNA was higher in lesions with a pronounced neutrophil infiltrate. Vascular cell adhesion molecule-1 (VCAM-1) localized to the deep connective tissue and indicated the presence of a systemic type of immune response in this region. Periodontal tissues (n=71 biopsies) did not appear to express MAdCAM-1, in positive control sections of small intestine where it was detected.. Overall, the systemic-type immune response is predominant, and although the mucosal immune response is minor and limited to the superficial tissues it may have an important role in the host defense to periodontal pathogens.

Topics: Adult; Cell Adhesion Molecules; Connective Tissue; Gingiva; Gingivitis; Granulation Tissue; Humans; Immunity, Mucosal; Immunoglobulin A; Immunoglobulin J-Chains; Immunoglobulins; Intercellular Adhesion Molecule-1; Interleukin-8; Middle Aged; Mucoproteins; Neutrophils; Periodontitis; Periodontium; Plasma Cells; Receptors, Lymphocyte Homing; Vascular Cell Adhesion Molecule-1

Our aim was to evaluate the effect of a gingival gel containing chlorhexidine and Rheum Palmatum extract on gingival fragments stimulated by SP (substance P) and lipopolysaccharides (LPS).. Gingival fragments were maintained in survival for 3 days at 37 degrees C. To induce inflammation, SP and LPS were applied to the culture medium in contact with the corium. The gingival gel was applied on epithelium. Histological analysis was then performed on hematoxylin and eosin stained slides. Edema was evaluated with semi-quantitative scores. Vasodilation was studied by counting the percent of dilated vessels according to scores and the surface of these dilated vessels by morphometrical image analysis. An inflammatory cytokine, IL8, was measured in culture supernatants. Immunohistochemical expression of metalloproteinase type 9 (MMP9) implicated in inflammatory processes, was also studied (% of positive cells).. Edema, vasodilation and IL8 were significantly increased after application of SP and LPS. Application of gingival gel showed a significant decrease of these parameters. A significant decrease of MMP9 on fibroblasts and mononuclear cells was observed after use of gingival gel.

Topics: Anti-Infective Agents, Local; Capillaries; Cells, Cultured; Chlorhexidine; Coloring Agents; Dilatation, Pathologic; Gels; Gingiva; Gingivitis; Humans; Image Processing, Computer-Assisted; Interleukin-8; Lipopolysaccharides; Matrix Metalloproteinase 9; Phytotherapy; Plant Extracts; Rheum; Substance P

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

Periodontopathic bacteria induce inflammation of periodontal tissues. The cytokines and nitric oxide released in periodontal lesions have been reported to play a protective role in bacterial infection and to relate to the process of inflammation. To clarify the relationship between colonization of periodontopathic bacteria and cytokines, we evaluated profiles of inflammatory cytokines, chemokine, anti-inflammatory cytokines, and inducible nitric oxide synthase (iNOS) and colonization by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, which are major pathogens of periodontitis.. mRNA expression of cytokines and iNOS in inflamed and healthy gingival tissue was determined using reverse transcription-polymerase chain reaction (RT-PCR), and the relationship between their profiles and the detection of specific bacteria was analyzed.. The relative expression of interleukin (IL)-6 and iNOS mRNAs in periodontal lesions was significantly higher than those in healthy individuals. IL-6 mRNA expression was also significantly higher at bleeding on probing (BOP)-positive sites than at BOP-negative sites. The expressions of IL-1alpha and IL-8 increased, but IL-10 expression decreased at sites where A. actinomycetemcomitans was detected. We found no correlation between the expression of cytokine and iNOS mRNA and infection by P. gingivalis.. The expression of IL-6 may reflect inflammation in gingival tissue, and iNOS may be involved in the inflammatory process in periodontitis. The presence of A. actinomycetemcomitans or P. gingivalis might relate to the different cytokine profiles of IL-1alpha, IL-8, and IL-10.

Topics: Actinobacillus Infections; Adolescent; Adult; Aged; Aggregatibacter actinomycetemcomitans; Bacteroidaceae Infections; Chemokines; Child; Cytokines; Female; Gene Expression Regulation; Gene Expression Regulation, Enzymologic; Gingival Hemorrhage; Gingivitis; Humans; Inflammation Mediators; Interleukin-1; Interleukin-10; Interleukin-6; Interleukin-8; Male; Middle Aged; Nitric Oxide Synthase; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Statistics as Topic; Statistics, Nonparametric

Habitual use of smokeless tobacco leads to accumulation of inflammatory leukocytes at the site of placement, which may contribute to tissue damage. Recruitment of leukocytes is facilitated by the endothelial lining of blood vessels, which can be activated to express adhesion molecules and to produce chemoattractants. The ability of aqueous extracts of chewing tobacco, dry snuff, and moist snuff to stimulate such changes was investigated using cultured human umbilical vein endothelial cells (HUVEC). All three extracts caused HUVEC to express the adhesion molecule E-selectin and to produce the chemokines interleukin-8 and monocyte chemoattractant protein-1. Neutrophils migrated avidly across HUVEC monolayers that had been previously exposed to the extracts, whereas migration across unstimulated monolayers was negligible. The smokeless tobacco extracts contained relatively high concentrations of bacterial lipopolysaccharide (LPS). Although LPS appeared to be the major stimulatory component in extracts of chewing tobacco, it accounted for only part of the activity found in extracts of moist and dry snuffs. These observations suggest that smokeless tobacco may induce inflammatory changes in vivo by activating endothelium in a manner that promotes recruitment of leukocytes.

Topics: Blood Proteins; Cell Movement; Cells, Cultured; Chemokine CCL2; E-Selectin; Endothelium, Vascular; Gingivitis; Humans; Interleukin-8; Lipopolysaccharides; Neutrophils; Plant Extracts; Plants, Toxic; Tobacco, Smokeless; Umbilical Veins

The substance P (SP) level in human gingival crevicular fluid (GCF) was studied in relation to clinical periodontal variables and to various indicators of host response in the GCF.. GCF was collected from periodontal sites with gingival inflammation and shallow or moderately deep pocket in 48 subjects. The total amount of SP and the substances based on host response factors in a 30-s sample were determined by ELISA and enzymatic methods.. Significant correlation was found between SP and probing depth (r= 0.637, p<0.001), while correlation was weak between SP and either gingival (r= 0.177, p=0.23) or plaque index (r=0.008, p=0.96). SP also showed significant correlation with the indicators of host response: prostaglandin E2, aspartate aminotransferase, alkaline phosphatase, myeloperoxidase, interleukin-1beta, tumor necrosis factor-alpha, interleukin-8 and monocyte chemoattractant protein-1 (r=0.434-0.867, p<0.01-0.001).. These results indicate that neuropeptide SP in GCF may have a potential as an indicator of periodontal inflammation and the host response.

Topics: Alkaline Phosphatase; Aspartate Aminotransferases; Biomarkers; Chemokine CCL2; Dental Plaque Index; Dinoprostone; Enzyme-Linked Immunosorbent Assay; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Periodontal Index; Periodontal Pocket; Periodontitis; Peroxidase; Substance P; Tumor Necrosis Factor-alpha

This report describes our findings regarding the potential contribution of periodontitis to atherosclerotic processes using a nonhuman primate model. The goal of the investigations was to target general mechanisms which could describe the association of these disease processes, including: (i) systemic translocation of bacteria/products during periodontitis; (ii) alterations in systemic inflammatory biomarkers during periodontitis; and (iii) the relationship of periodontitis to serum lipids/lipoproteins. Increases in serum endotoxin (e.g. LPS) during ligature-induced periodontitis were observed in these animals. We determined serum levels of various acute phase reactants and chemokines (e.g. CRP, alpha 1-antitrypsin, haptoglobin, fibrinogen, IL-8). A number of these host factors were significantly increased during gingivitis and/or periodontitis. Finally, we observed specific changes in serum lipid levels (cholesterol, triglycerides, HDL, LDL) and lipoproteins (apoA-I) during periodontitis, which were exacerbated by exposure of the animals to a diet with elevated fat content. Thus, we have described systemic manifestations of periodontitis that include detection of bacterial products, inflammatory biomarkers, and dyslipoproteinemia consistent with an increased atherogenic risk.

Topics: alpha 1-Antitrypsin; Animals; Apolipoprotein A-I; Arteriosclerosis; Bacterial Translocation; Biomarkers; C-Reactive Protein; Chemokines; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Disease Models, Animal; Endotoxins; Fibrinogen; Gingivitis; Haptoglobins; Hyperlipoproteinemias; Inflammation Mediators; Interleukin-8; Lipopolysaccharides; Macaca fascicularis; Periodontitis; Risk Factors; Triglycerides

The objective of this study was to determine and compare concentrations and ratios of 3 proinflammatory cytokines, interleukin (IL) IL-1beta, IL-6, and IL-8 within gingival tissue biopsies adjacent to < or = 3, 4 to 6, or >6 mm sulci. All gingiva adjacent to > or = 4 mm sulci had clinical evidence of active inflammation. Factorial analysis of variance suggested significant effects of sulcus depth on the type and concentration of the three cytokines in the adjacent gingiva (P < 0.001). IL-8 concentrations were highest in gingiva adjacent to < or = 3 and lowest adjacent to >6 mm sulci (P < 0.001). In contrast, IL-6 concentrations were lowest in gingiva adjacent to < or = 6 mm and highest adjacent to >6 mm sites. IL-1beta concentrations were highest in gingiva adjacent to >6 mm and lowest adjacent to 4 to 6 mm sites; they were also higher adjacent to < or = 3 mm than adjacent to 4 to 6 mm sites (P < 0.01). Multiple regression analysis suggested that sulcular depth, type of cytokine, and cytokine concentration were significantly correlated (P < 0.001). Ratios of gingival cytokines changed with increased sulcular depth. In gingiva adjacent to < or = 6 mm sites, IL-8 was the most and IL-6 the least prevalent. In gingiva adjacent to > or = 6 mm sites, IL-8 was the least and IL-1-beta the most prevalent. The data suggest that the characteristics of the gingival cytokine network are affected by adjacent sulcular depth. These data could be used to design adjunct diagnostic tests for progression of periodontal diseases.

Topics: Analysis of Variance; Biopsy; Collagen; Disease Progression; Gingiva; Gingival Pocket; Gingivitis; Humans; Inflammation Mediators; Interleukin-1; Interleukin-6; Interleukin-8; Neutrophils; Plasma Cells; Regression Analysis

Inflamed human gingival fibroblasts (HGF) of patients with chronic periodontal diseases have less active interleukin-8 (IL-8) production compared with normal HGF of volunteers with healthy gingival tissues, after stimulation with Porphyromonas gingivalis surface components such as fimbriae, lipopolysaccharide (LPS) and its lipid A, but not LPS or lipid A from other bacterial species. A decrease in number of specific binding sites for P. gingivalis fimbrial molecules in inflamed HGF is also observed by Scatchard plot analysis. A short exposure (6 h) to P. gingivalis LPS resulted in significant potentiation of the LPS-dependent IL-8 production in normal HGF, whereas a long exposure (48 h) to the LPS significantly reduced IL-8 production. Tyrosine phosphorylation of proteins of 127 kDa and 186 kDa in inflamed HGF stimulated with P. gingivalis fimbriae or its LPS was observed by immunoblotting, and these two phosphoproteins were termed tolerance-induced protein, TIP. Protein bands of 45 kDa which bound to radioiodinated P. gingivalis fimbriae in the presence and absence of fetal bovine serum (FBS), and major 73-kDa and minor 30-kDa and 45-kDa bands which bound to radioiodinated P. gingivalis LPS in the presence of FBS in normal and inflamed HGF were observed by using photocrosslinking. These findings suggest that the hyporesponsiveness of HGF induced by a prolonged exposure to P. gingivalis may emerge because of HGF damage or result from host defense in chronic periodontal lesions.

Topics: Adult; Bacterial Proteins; Benzoquinones; Blotting, Western; Cells, Cultured; Chronic Disease; Enzyme Inhibitors; Fibroblasts; Fimbriae, Bacterial; Gingiva; Gingivitis; Humans; Interleukin-8; Lactams, Macrocyclic; Peptides; Periodontitis; Porphyromonas gingivalis; Protein-Tyrosine Kinases; Quinones; Rifabutin

The objective of this study was to examine the age-dependent relationships between levels of inflammatory cytokines and collagen in human gingival inflammation. The gingival biopsies were obtained from 142 patients, divided into the following age groups: 6 to 14 years (prepubertal children); 18 to 35 years (young adults); 36 to 54 years (mature adults); and 55 years or above. The patients were also divided according to the severity of gingivitis. The tissues were analyzed for the contents of the inflammatory cytokines interleukin (IL)-1 beta, IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) using specific ELISA kits, and interstitial collagen type I and type III using the ELISA method and specific antibodies. We found that in young adults, levels of IL-1 beta and IL-6 were significantly higher in inflamed than in non-inflamed gingiva. Total collagen in the young adults, however, was lower in inflamed than in non-inflamed gingiva. There was no significant difference in the levels of either IL-8 or TNF-alpha between inflamed and non-inflamed gingiva independent of age. No difference in the level of collagen type I between the inflamed and non-inflamed gingiva was found in any age groups. The level of collagen type III was lower in inflamed than in non-inflamed gingiva in both children and > or = 55 year group. The results indicate a disparity in the effect of age on the levels of cytokines and of collagen type I and type III in both clinically normal and inflamed gingiva.

Topics: Adolescent; Adult; Aging; Collagen; Cytokines; Enzyme-Linked Immunosorbent Assay; Gingiva; Gingivitis; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Middle Aged; Periodontal Index; Proteins; Tumor Necrosis Factor-alpha

Periodontitis and gingivitis are chronic inflammatory diseases of the periodontium and adjacent tissues. This site-specific inflammation is characterized by a local infiltration of polymorphonuclear leukocytes and T lymphocytes. Interleukin-8 is a low molecular-weight cytokine that is thought to be responsible for the induction and maintenance of localized inflammation. We hypothesized that locally produced interleukin-8 plays a central role in chronic inflammation of periodontitis by regulating the recruitment and activation of leukocytes in the gingival tissues. To test this hypothesis, we determined whether the interleukin-8 antigen is present locally and is cell-associated. Inflamed and control tissues were analyzed: 1) for the interleukin-8 antigen; 2) by molecular weight; 3) for location; and 4) for the messenger RNA (mRNA) of interleukin-8. The conclusions from these data were that: 1) interleukin-8 antigen and mRNA was elevated in chronically inflamed gingiva; and 2) the major interleukin-8 antigen was detected only in the epithelial cell layer. These results support that interleukin-8 may play a crucial role in the recruitment and activation of neutrophils and T lymphocytes in periodontitis.

Topics: Adolescent; Adult; Animals; Binding Sites, Antibody; Chickens; Connective Tissue; Epithelial Attachment; Epithelial Cells; Epithelium; Fibroblasts; Gingiva; Gingivitis; Humans; Immunoenzyme Techniques; Interleukin-8; Middle Aged; Molecular Weight; RNA, Messenger; Statistics, Nonparametric

This study was undertaken in an effort to understand the role of cytokines on human gingival fibroblasts and T lymphocyte trafficking into inflamed gingival tissue. Using flow cytometry we examined gingival fibroblasts to determine the level of cell surface expression and the percentage of cells positive for intercellular adhesion molecule 1 (ICAM-1), the HLA-DR antigen, lymphocyte function-associated antigen 3 (LFA-3), and the CD44 molecule, which are involved in antigen presentation. The following cytokines were used: interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), IL-6, and IL-8. The levels of ICAM-1 expression were enhanced in a dose- and time-dependent manner by IL-1 beta, TNF-alpha, or IFN-gamma, but not by IL-6 or IL-8. HLA-DR surface expression was induced only by IFN-gamma in a dose- and time-dependent manner, but not by the other cytokines tested. In contrast, the expression of LFA-3 and the CD44 molecule could be detected without the stimulation of any cytokine, but the levels of their expression were not significantly changed by any cytokines. The enhanced ICAM-1 expression by cytokines was reduced in a time-dependent manner following the removal of cytokines from the reaction mixture, while IFN-gamma-induced HLA-DR expression was maintained even 7 days after the removal of IFN-gamma. These data support an interactive role for inflammatory cytokines and the expression of adhesion molecules on gingival fibroblasts in the pathogenesis of gingival inflammation in periodontal disease.

Topics: Antigen-Presenting Cells; Antigens, CD; Carrier Proteins; CD58 Antigens; Cell Adhesion Molecules; Cell Communication; Cells, Cultured; Cytokines; Fibroblasts; Flow Cytometry; Fluorescent Antibody Technique; Gingiva; Gingivitis; HLA-DR Antigens; Humans; Hyaluronan Receptors; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-1; Interleukin-6; Interleukin-8; Lymphocyte Activation; Membrane Glycoproteins; Receptors, Cell Surface; Receptors, Lymphocyte Homing; Recombinant Proteins; T-Lymphocytes; Tumor Necrosis Factor-alpha; Up-Regulation

Human Gingival Fibroblasts (HGF) may have an important role in the orchestration of immuno-participant cells infiltrating the gingiva in response to continuously recurring bacterial infection. To examine the cytokine network regulating HGF-derived interleukin (IL)-8, a potent neutrophil chemotactic cytokine, we analyzed the effects of inflammatory cytokines alone and in combination on IL-8 production by HGF. IL-1 beta, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), IL-6, and IL-8 were used as stimulants. HGF secreted IL-8 in a dose-dependent manner after stimulation with either IL-1 beta or TNF-alpha, but not with IFN-gamma or IL-6. Furthermore, IL-8 itself did not affect IL-8 mRNA accumulation in HGF in an autocrine manner. The combination of IL-1 beta and TNF-alpha synergistically enhanced the secretion of IL-8, whereas IFN-gamma suppressed IL-8 secretion by IL-1 beta- or TNF-alpha-stimulated HGF. These effects were also observed at each level of IL-8 mRNA expression in HGF. IL-8 secretion by cytokine-stimulated HGF was not influenced by the inhibition of PGE2 synthesis with indomethacin, indicating that endogenous PGE2 was not involved in IL-8 production by HGF. These results indicate that IL-8 production by HGF is synergistically stimulated by specific cytokines, IL-1 beta and TNF-alpha, and suggest that these stimulatory effects are down-regulated by IFN-gamma at the transcriptional level through PGE2-independent pathways. Thus, neutrophil-mediated processes in periodontal disease may be regulated in part by HGF in the cytokine network of immuno-participant cells.

Topics: Chemotaxis, Leukocyte; Cytokines; Dinoprostone; Fibroblasts; Gingiva; Gingivitis; Humans; Indomethacin; Interferon-gamma; Interleukin-1; Interleukin-8; Neutrophil Activation; Neutrophils; RNA, Messenger; Tumor Necrosis Factor-alpha

In bacterial infections, mononuclear and polymorphonuclear phagocytes are key components of host defenses. Recent investigations have indicated that chemokines are able to recruit and activate phagocytes. In particular, interleukin-8 (IL-8) attracts polymorphonuclear leukocytes (PMNs), while monocyte chemoattractant protein-1 (MCP-1) is selective for cells of the monocyte/macrophage lineage. In this investigation, we analyzed the in situ expression of IL-8 and MCP-1 mRNAs in human periodontal infections. Specific mRNA was detected by in situ hybridization using 35S-labeled riboprobes in frozen tissue sections. Phagocytes (PMNs and macrophages) were specifically detected as elastase-positive or CD68+ cells by a three-stage immunoperoxidase technique. Results indicated that expression of phagocyte-specific cytokines was confined to selected tissue locations and, in general, paralleled phagocyte infiltration. In particular, IL-8 expression was maximal in the junctional epithelium adjacent to the infecting microorganisms; PMN infiltration was more prominent in the same area. MCP-1 was expressed in the chronic inflammatory infiltrate and along the basal layer of the oral epithelium. Cells of the monocyte/macrophage lineage were demonstrated to be present in the same areas. The observed expression pattern may be the most economic way to establish a cell-type-selective chemotactic gradient within the tissue that is able to effectively direct polymorphonuclear phagocyte migration toward the infecting microorganisms and modulate mononuclear phagocyte infiltration in the surrounding tissues. This process may optimize host defenses and contribute to containing leukocyte infiltration to the infected and inflamed area, thus limiting tissue damage.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Chemokine CCL2; Chemotactic Factors; Gingivitis; Humans; Interleukin-8; Neutrophils; Periodontitis; Phagocytes; RNA, Messenger

In the last few years several bacteriological and immunological studies have investigated the role of bacteria and immune defects in order to establish the etiopathogenesis of periodontal disease. With regard to the immune system, a defect in polymorphonuclear neutrophil (PMN) chemotaxis has been frequently reported in patients with rapidly progressive or juvenile periodontitis. The purpose of this study was to investigate in five patients with rapidly progressive periodontitis and normal chemotaxis of peripheral blood PMNs the presence of chemotaxis inhibitory activity in gingival fluid and to relate such activity to three types of bacteria, often involved in rapidly evolving periodontal lesions, that are able to inhibit in vitro PMN chemotaxis: Bacteroides gingivalis, Capnocytophaga sp., and Actinobacillus actinomycetemcomitans. We found strong inhibitory activity in three of these patients. This activity was consistently associated with the finding of B. gingivalis in gingival pockets. We cannot rule out, however, that other substances not of bacterial origin could be responsible for such inhibitory activity. The strict association with B. gingivalis, known to secrete blocking factors, is highly suggestive, although this data must be considered preliminary.

Topics: Actinobacillus; Adolescent; Adult; Bacteroides; Capnocytophaga; Chemotactic Factors; Chemotaxis, Leukocyte; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-8; Lymphokines; Male; Neutrophils; Periodontitis