interleukin-8 and Periodontal-Diseases

This paper's goal is to review the relationship between infections and chronic respiratory disease, with particular reference to periodontal disease. The link between oral diseases in general, periodontal disease, and respiratory disease remains somewhat controversial. However, with cooperation between dentistry and medicine, the nature of the connection between dental and medical pathology can be better defined. An overview of respiratory disease and some of the factors that can contribute to respiratory infection is presented below, with special reference to infections related to aspiration.

Topics: Bacteria; Cough; Deglutition; Humans; Interleukin-8; Lung; Periodontal Diseases; Pneumonia, Aspiration; Pneumonia, Bacterial; Pulmonary Disease, Chronic Obstructive; Respiratory Physiological Phenomena; Respiratory Tract Diseases; Respiratory Tract Infections; Risk Factors; Saliva; Smoking

Soluble proteins that serve as mediators of cell function and are produced by various cell types, such as structural and inflammatory cells, are collectively called cytokines. Several lines of evidence have revealed that cytokines play important roles not only in tissue homeostasis but also in the pathogenesis of many infectious diseases. Recent research on biological activities in normal periodontium and the pathogenesis of periodontal diseases has clarified the involvement of various cytokines in the biological activities observed in the sites. Cytokines play crucial roles in the maintenance of tissue homeostasis, a process which requires a delicate balance between anabolic and catabolic activities. In particular, growth factors--such as fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), transforming growth factor-beta (TGF-beta)--are thought to play important roles in modulating the proliferation and/or migration of structural cells in the periodontium and the production of various extracellular matrices by these cells. On the other hand, there is little doubt that excessive and/or continuous production of cytokines in inflamed periodontal tissues is responsible for the progress of periodontitis and periodontal tissue destruction. Particularly, inflammatory cytokines--such as IL-1 alpha, IL-1 beta, IL-6, and IL-8--are present in the diseased periodontal tissues, and their unrestricted production seems to play a role in chronic leukocyte recruitment and tissue destruction. It is possible that monitoring cytokine production or its profile may allow us to diagnose an individual's periodontal disease status and/or susceptibility to the disease. In addition, although the hypothesis is still controversial, it has been suggested that discrete T-cell subsets (Th1 and Th2) with different cytokine profiles play specific roles in the immunopathogenesis of periodontal diseases.

Topics: Cell Division; Cell Movement; Cytokines; Disease Susceptibility; Extracellular Matrix; Fibroblast Growth Factors; Gene Expression Regulation; Homeostasis; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Leukocytes; Periodontal Diseases; Periodontitis; Periodontium; Platelet-Derived Growth Factor; Somatomedins; T-Lymphocyte Subsets; Transforming Growth Factor beta

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

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

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

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

Billions of individuals worldwide suffer from periodontal disease, an inflammatory disease that results in hard-tissue and soft-tissue destruction. A viable therapeutic option to treat periodontal disease may be via cannabinoids that exert immunomodulatory effects, and the endocannabinoid system (ECS) is readily present in periodontal tissues that exhibit cannabinoid type 1 and 2 receptors (CB1R and CB2R). Phytocannabinoids (pCBs), which are a part of a heterogeneous group of molecules acting on cannabinoid receptors (CBR) derived from the cannabis plants, have been attributed to a wide variety of effects including anti-inflammatory activity and some pro-inflammatory effects depending on the cell type. Thus, this study aims to examine the effects of pCBs on primary human gingival fibroblasts (HGFs) in IL-1β stimulated (simulated periodontal disease) HGFs.. The effective inhibition of IL-1β-stimulated production of PGE2 and cytokines by the pCB in HGFs suggests that targeting the endocannabinoid system may lead to the development of therapeutic strategies for periodontal therapy. However, each pCB has its unique anti-inflammatory profile, in which certain pro-inflammatory activities are also exhibited. The pCBs alone or in combination may benefit and aid in improving public oral health.

Topics: Anti-Inflammatory Agents; Cannabinoids; Cells, Cultured; Cytokines; Dinoprostone; Endocannabinoids; Fibroblasts; Gingiva; Humans; Inflammation; Interleukin-10; Interleukin-13; Interleukin-1beta; Interleukin-2; Interleukin-4; Interleukin-6; Interleukin-8; Periodontal Diseases; Receptors, Cannabinoid; Tumor Necrosis Factor-alpha

Fusobacterium nucleatum is associated with many conditions and diseases, including periodontal diseases that affect tooth-supporting tissues. The aim of the present study was to investigate the effects of a cocoa extract (Theobroma cacao L.) on F. nucleatum with respect to growth, biofilm formation, adherence, and hydrogen sulfide (H2S) production. The anti-inflammatory properties and the effect on epithelial barrier function of the cocoa extract were also assessed. The cocoa extract, whose major phenolic compound is epicatechin, dose-dependently inhibited the growth, biofilm formation, adherence properties (basement membrane matrix, oral epithelial cells), and H2S production of F. nucleatum. It also decreased IL-6 and IL-8 production by F. nucleatum-stimulated oral epithelial cells and inhibited F. nucleatum-induced NF-κB activation in monocytes. Lastly, the cocoa extract enhanced the barrier function of an oral epithelial model by increasing the transepithelial electrical resistance. We provide evidence that the beneficial properties of an epicatechin-rich cocoa extract may be useful for preventing and/or treating periodontal diseases.

Topics: Biofilms; Cacao; Catechin; Cell Adhesion; Epithelial Cells; Fusobacterium nucleatum; Gene Expression Regulation; Humans; Hydrogen Sulfide; Interleukin-6; Interleukin-8; Monocytes; Periodontal Diseases; Phenols; Plant Extracts

Topics: Anti-Inflammatory Agents; Cell Line; Chalcones; Fibroblasts; Gingiva; Humans; Interleukin-8; Lead; Lipopolysaccharides; Periodontal Diseases; Porphyromonas gingivalis

Topics: Anti-Inflammatory Agents; Cell Line; Cell Movement; Cell Proliferation; Cell Survival; Fibroblasts; Gingiva; Humans; Hyaluronic Acid; Inflammation; Interleukin-10; Interleukin-1beta; Interleukin-4; Interleukin-6; Interleukin-8; MAP Kinase Signaling System; Molecular Weight; NF-kappa B; Periodontal Diseases; Porphyromonas gingivalis; Wound Healing

The oral pathogen Tannerella forsythia possesses a unique surface (S-) layer with a complex O-glycan containing a bacterial sialic acid mimic in the form of either pseudaminic acid or legionaminic acid at its terminal position. We hypothesize that different T. forsythia strains employ these stereoisomeric sugar acids for interacting with the immune system and resident host tissues in the periodontium. Here, we show how T. forsythia strains ATCC 43037 and UB4 displaying pseudaminic acid and legionaminic acid, respectively, and selected cell surface mutants of these strains modulate the immune response in monocytes and human oral keratinocytes (HOK) using a multiplex immunoassay. When challenged with T. forsythia, monocytes secrete proinflammatory cytokines, chemokines and vascular endothelial growth factor (VEGF) with the release of interleukin-1β (IL-1β) and IL-7 being differentially regulated by the two T. forsythia wild-type strains. Truncation of the bacteria's O-glycan leads to significant reduction of IL-1β and regulates macrophage inflammatory protein-1. HOK infected with T. forsythia produce IL-1Ra, chemokines and VEGF. Although the two wild-type strains elicit preferential immune responses for IL-8, both truncation of the O-glycan and deletion of the S-layer result in significantly increased release of IL-8, granulocyte-macrophage colony-stimulating factor and monocyte chemoattractant protein-1. Through immunofluorescence and confocal laser scanning microscopy of infected HOK we additionally show that T. forsythia is highly invasive and tends to localize to the perinuclear region. This indicates, that the T. forsythia S-layer and attached sugars, particularly pseudaminic acid in ATCC 43037, contribute to dampening the response of epithelial tissues to initial infection and hence play a pivotal role in orchestrating the bacterium's virulence.

Topics: Cell Membrane; Chemokines; Cytokines; Humans; Intercellular Signaling Peptides and Proteins; Interleukin 1 Receptor Antagonist Protein; Interleukin-1beta; Interleukin-7; Interleukin-8; Keratinocytes; Macrophage Inflammatory Proteins; Membrane Glycoproteins; Monocytes; Mutation; N-Acetylneuraminic Acid; Periodontal Diseases; Polysaccharides; Sialic Acids; Sugar Acids; Tannerella forsythia; Vascular Endothelial Growth Factor A; Virulence

Psoralen and angelicin are two effective compounds isolated from psoraleae, a traditional Chinese medicine. They have a wide range of applications for bone disease treatment and immune modulation. In this study, we explored their new applications for the treatment of periodontal diseases. This study aimed to investigate the effects of psoralen and angelicin on

Topics: Alkaline Phosphatase; Alveolar Bone Loss; Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Biofilms; Bone Morphogenetic Proteins; Cell Survival; Core Binding Factor Alpha 1 Subunit; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Combinations; Ficusin; Furocoumarins; Gene Expression; Homeodomain Proteins; Humans; Inflammation; Interleukin-1beta; Interleukin-8; Lipopolysaccharides; Male; Medicine, Chinese Traditional; Mice; Mice, Inbred C57BL; Microbial Sensitivity Tests; Osteogenesis; Osteopontin; Periodontal Diseases; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; RNA, Messenger; THP-1 Cells; Transcription Factors; Up-Regulation

It is well established that aging is associated with increased susceptibility to infectious diseases. Fusobacterium nucleatum is a well-known bacterial species that plays a central bridging role between early and late colonizers in the human oral cavity. Further, the ability of F. nucleatum to invade gingival fibroblasts (GFs) is critical to the development of periodontal diseases. However, the mechanisms underlying the age-related infection of GFs by F. nucleatum remain unknown. We used young (fourth passage) and senescent (22nd passage) GFs to investigate the mechanisms of F. nucleatum infection in aged GFs and first observed increased invasion of F. nucleatum in senescent GFs. We also found that the co-localization of caveolin-1 (Cav-1), a protein marker of aging, with F. nucleatum and the knockdown of Cav-1 in GFs reduced F. nucleatum invasion. Additionally, F. nucleatum infection triggered the production of reactive oxygen species (ROS) through activation of NADPH oxidase in GFs, but senescent GFs exhibited significantly lower levels of NADPH oxidase activity and ROS production compared with young GFs in both the uninfected and infected conditions. Also, senescent GFs exhibited a decline in proinflammatory cytokine production and extracellular signal regulated kinase (ERK) phosphorylation following F. nucleatum infection. Interestingly, the knockdown of Cav-1 in senescent GFs increased NADPH oxidase activity and caused the upregulation of interleukin-6 and interleukin-8 and the phosphorylation of ERK. Collectively, the increased expression of Cav-1 might play a critical role in F. nucleatum invasion and could hinder the host response in senescent GFs.

Topics: Caveolin 1; Cells, Cultured; Cellular Senescence; Cytokines; Fibroblasts; Gingiva; Humans; Interleukin-6; Interleukin-8; Mitogen-Activated Protein Kinase 3; NADPH Oxidases; Periodontal Diseases; Phosphorylation; Reactive Oxygen Species; Signal Transduction; Up-Regulation

Porphyromonas gingivalis produces different LPS isoforms with significant structural variations of their lipid A and O-antigen moieties that can affect its pro-inflammatory and bone-resorbing potential. We show here, for the first time, that P. gingivalis LPS isolated from W83 strain is highly sialylated and possesses significantly reduced inflammatory potential compared with less sialylated ATCC 33277 strain LPS. Nevertheless, the reduction in the endotoxin activity is not mediated by the presence of sialic acid LPS moieties as the sialic acid-free LPS produced by the mutant W83 strain exhibits a similar inflammatory potential to the wild type strain. Furthermore, our findings suggest that the interaction between the sialic acid LPS moieties and the inhibitory CD33 receptor is prevented by endogenously expressed sialic acid on the surface of THP-1 cells that cannot be out-competed by sialic acid containing P. gingivalis LPS. The present study also highlights the importance of endogenous sialic acid as a 'self-associated molecular pattern' and CD33 receptors in modulation of innate immune response as human gingival fibroblasts, which do not express CD33 receptors, and desialylated THP-1 cells have both been found to have much higher spontaneous IL-8 production than naïve THP-1 cells.

Topics: Bacteroidaceae Infections; Cell Line; Fibroblasts; Gingiva; Host-Pathogen Interactions; Humans; Immunity, Innate; Interleukin-8; Lipid A; Lipopolysaccharides; Monocytes; Mutation; N-Acetylneuraminic Acid; O Antigens; Periodontal Diseases; Porphyromonas gingivalis; Protein Processing, Post-Translational; Sialic Acid Binding Ig-like Lectin 3

Cholesterol deposits and pro-inflammatory cytokines play an essential role in the pathogenesis of atherosclerosis, a predominant cause of cardiovascular disease (CVD). Epidemiological evidence has linked periodontal disease (PD) with atherosclerotic CVD. Accordingly, viable periodontal pathogens, including Porphyromonas gingivalis, have been found in atherosclerotic plaques in humans and mice. We aimed to determine whether cholesterol crystals (CHCs) and oral bacteria synergize in the stimulation of human monocytes. Incubation of human monocytes with CHCs induced secretion of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, and IL-8. Moreover, CHCs markedly enhanced secretion of IL-1β by monocytes stimulated with the toll-like receptor (TLR) 4 agonist Escherichia coli lipopolysaccharide (LPS), and the TLR2 agonist Staphylococcus aureus lipoteichoic acid. Notably, CHCs also enhanced IL-1β secretion induced by P. gingivalis LPS and IL-1β secretion induced by whole P. gingivalis bacteria. This enhancement was abrogated by the NLRP3 inflammasome inhibitors Z-YVAD-FMK and glibenclamide. CHCs had no effect on cytokine production induced by P. gingivalis gingipains. Taken together, our findings support that CHCs, via stimulation of NLRP3 inflammasomes, act in synergy with the periodontal pathogen P. gingivalis to promote monocyte secretion of pro-atherogenic cytokines.

Topics: Animals; Atherosclerosis; Cholesterol; Humans; Inflammasomes; Interleukin-1beta; Interleukin-6; Interleukin-8; Mice; Monocytes; NLR Family, Pyrin Domain-Containing 3 Protein; Periodontal Diseases; Plaque, Atherosclerotic; Porphyromonas gingivalis; Staphylococcus aureus; Toll-Like Receptor 2; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Smoking is an important risk factor for periodontal disease and effects the pathogenesis of the disease. This study evaluated the impact of smoking on gingival crevicular fluid interleukin-8 (IL-8) and lipoxin A4 (LxA4 ) levels in patients with and without periodontal disease.. A total of 122 participants were grouped as follows: smokers with generalized aggressive periodontitis (S-GAgP, n = 15); smokers with chronic periodontitis (S-CP, n = 17); smokers with gingivitis (SG, n = 15); smokers classified as periodontally healthy (SH, n = 15); nonsmokers with generalized aggressive periodontitis (N-GAgP, n = 15); nonsmokers with chronic periodontitis (N-CP, n = 15); nonsmokers with gingivitis (NG, n = 15); and nonsmokers classified as periodontally healthy (NH, n = 15). Gingival index, plaque index, probing pocket depth and clinical attachment level were recorded. Gingival crevicular fluid IL-8 and LxA4 levels were analyzed by ELISA.. Gingival crevicular fluid IL-8 levels varied among groups, as follows: S-GAgP>S-CP>SG>SH and N-GAgP>N-CP>NG>NH. The gingival crevicular fluid IL-8 levels were significantly higher in the S-GAgP group compared with the N-GAgP group and in the S-CP group compared with the N-CP group (p < 0.05); differences between the SG and NG and the SH and NH groups were not statistically significant (p > 0.05). Gingival crevicular fluid LxA4 levels also varied among groups, but in an inverse direction when compared with the IL-8 levels, as follows: S-GAgP 0.05).. The study findings suggest that the observed increases in gingival crevicular fluid IL-8 levels and decreases in gingival crevicular fluid LxA4 levels reflect changes in immune and inflammatory responses that occur as a result of smoking.

Topics: Adult; Cross-Sectional Studies; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Lipoxins; Male; Periodontal Diseases; Periodontal Index; Smokers; Smoking

Farrerol, a new type of 2,3-dihydro-flavonoid isolated from rhododendron, has been shown to have anti-bacterial and anti-inflammatory activities. In the present study, we investigated the anti-inflammatory effects of farrerol on the production of IL-6 and IL-8 in human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS).. The cytotoxicity of farrerol was determined using the MTT assay. The production of IL-6 and IL-8 was measured using ELISA and qRT-PCR. The effects of farrerol on PI3K, Akt phosphorylation, and NF-κB activation were detected using western blotting analyses.. These results showed that farrerol inhibited LPS-induced IL-6 and IL-8 production in a dose dependent manner. LPS-induced NF-κB activation was suppressed by farrerol. Furthermore, farrerol suppressed LPS-induced PI3K and Akt phosphorylation, which are upstream molecules of NF-κB.. These results indicated that farrerol attenuated IL-6 and IL-8 production by inhibition of PI3K and AKT phosphorylation, resulting in an inhibition of NF-κB activation. Farrerol may be a therapeutic agent for the treatment of periodontal disease.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents; Cell Survival; Cells, Cultured; Chromones; Fibroblasts; Gingiva; Humans; Interleukin-6; Interleukin-8; Lipopolysaccharides; NF-kappa B; Periodontal Diseases; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Signal Transduction

Microvesicles (MVs) are extracellular vesicles secreted by various cell types that are involved in intercellular communication. We hypothesized that in human periodontal disease, the pocket epithelium releases MVs, which then modulate gene expression in the underlying fibroblasts to control periodontal inflammation. MVs were isolated from culture medium of gingival epithelial cells (GECs) treated with oral bacterial biofilm extract or left untreated. Biofilm treatment significantly increased MV release from the GECs. Mass spectrometry of GEC-MVs identified a total of 2,173 proteins, of which about 80% were detected in MVs from both control and biofilm-treated GECs. Among 80 signature genes of human gingival fibroblasts, 20 were significantly regulated (P < 0.05) by MVs from control and biofilm-treated GECs in a similar manner. Matrix metalloproteinase 1 and 3 and interleukin 6 and 8 showed the strongest regulation at the mRNA and protein levels. Several cellular signaling pathways were activated by GEC-MVs in human gingival fibroblasts, including Smad and mitogen-activated protein kinase-associated pathways ERK1/2, JNK, and p38. However, ERK1/2 signaling dominated in the MV-induced gene expression changes. The results demonstrate that GEC-MVs have a strong regulatory effect on the expression of fibroblast genes associated with inflammation and matrix degradation and that bacterial biofilm stimulates the generation of GEC-MVs. This suggests that bacterial biofilms can contribute to the initiation and progression of periodontal disease by promoting a tissue-destructive phenotype in gingival fibroblasts via the enhanced secretion of epithelial MVs.

Topics: Biofilms; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Extracellular Vesicles; Fibroblasts; Gene Expression Regulation; Gingiva; Humans; Inflammation; Interleukin-6; Interleukin-8; Mass Spectrometry; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Microscopy, Electron; Microscopy, Fluorescence; Periodontal Diseases; Phenotype; Signal Transduction

Shikonin, which is derived from Lithospermum erythrorhizon, a herb used in traditional medicine, has long been considered to be a useful treatment for various diseases in traditional oriental medicine. Shikonin has recently been reported to have several pharmacological properties, e.g., it has anti-microbial, anti-tumor, and anti-inflammatory effects. The aim of this study was to examine whether shikonin is able to influence the production of interleukin (IL)-6, IL-8, and/or chemokine C-C motif ligand (CCL)20, which contribute to the pathogenesis of periodontal disease, in human periodontal ligament cells (HPDLC). The production levels of IL-6, IL-8, and CCL20 in HPDLC were determined using an ELISA. Western blot analysis was used to detect nuclear factor kappa B (NF-κB) pathway activation in HPDLC. Shikonin prevented IL-1β- or tumor necrosis factor (TNF)-α-mediated IL-6, IL-8, and CCL20 production in HPDLC. Moreover, we found that shikonin suppressed the phosphorylation and degradation of inhibitor of kappa B-alpha (IκB-α) in IL-1β- or TNF-α-stimulated HPDLC. These findings suggest that shikonin could have direct beneficial effects against periodontal disease by reducing IL-6, IL-8, and CCL20 production in periodontal lesions.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cells, Cultured; Chemokine CCL20; Cytokines; Humans; Inflammation; Interleukin-6; Interleukin-8; Naphthoquinones; Periodontal Diseases; Periodontal Ligament

"Perioceutics" including antimicrobial therapy and host modulatory therapy has emerged as a vital adjunctive treatment of periodontal disease. Melatonin level was significantly reduced in patients with periodontal diseases suggesting melatonin could be applied as a potential "perioceutics" treatment of periodontal diseases. This study aims to investigate the effects of melatonin receptor agonists (melatonin and ramelteon) on Porphyromonas gingivalis virulence and Porphyromonas gingivalis-derived lipopolysaccharide (Pg-LPS)-induced inflammation.. Effects of melatonin receptor agonists on Porphyromonas gingivalis planktonic cultures were determined by microplate dilution assays. Formation, reduction, and viability of Porphyromonas gingivalis biofilms were detected by crystal violet staining and MTT assays, respectively. Meanwhile, biofilms formation was also observed by confocal laser scanning microscopy (CLSM). The effects on gingipains and hemolytic activities of Porphyromonas gingivalis were evaluated using chromogenic peptides and sheep erythrocytes. The mRNA expression of virulence and iron/heme utilization was assessed using RT-PCR. In addition, cell viability of melatonin receptor agonists on human gingival fibroblasts (HGFs) was evaluated by MTT assays. After pretreatment of melatonin receptor agonists, HGFs were stimulated with Pg-LPS and then release of cytokines (IL-6 and lL-8) was measured by enzyme-linked immunosorbent assay (ELISA).. Melatonin and ramelteon did exhibit antimicrobial effects against planktonic culture. Importantly, they inhibited biofilm formation, reduced the established biofilms, and decreased biofilm viability of Porphyromonas gingivalis. Furthermore, they at sub-minimum inhibitory concentration (sub-MIC) concentrations markedly inhibited the proteinase activities of gingipains and hemolysis in a dose-dependent manner. They at sub-MIC concentrations significantly inhibited the mRNA expression of virulence factors (kgp, rgpA, rgpB, hagA, and ragA), while increasing the mRNA expression of ferritin (ftn) or hemolysin (hem). They did not show obvious cytotoxicity toward HGFs. They inhibited Pg-LPS-induced IL-6 and IL-8 secretion, which was reversed by luzindole, the melatonin receptor antagonist.. Melatonin receptor agonists can inhibit planktonic and biofilm growth of Porphyromonas gingivalis by affecting the virulent properties, as well as Pg-LPS-induced inflammatory response. Our study provides new evidence that melatonin receptor agonists might be useful as novel "perioceutics" agents to prevent and treat Porphyromonas gingivalis-associated periodontal diseases.

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Biofilms; Cells, Cultured; Fibroblasts; Gene Expression Regulation, Bacterial; Hemolysis; Humans; Indenes; Interleukin-6; Interleukin-8; Melatonin; Periodontal Diseases; Porphyromonas gingivalis; Receptors, Melatonin; Sheep

Recent studies point to the clinical utility of using peri-implant sulcular fluid (PISF) as a valuable diagnostic aid for monitoring peri-implant tissue health. The objectives of this study are to determine the levels of key biomarkers in PISF in periodontal maintenance participants and compare them with their corresponding levels in gingival crevicular fluid (GCF) obtained from the same participants.. PISF and GCF were collected from an implant and a contralateral natural tooth after the clinical examination of 73 participants. The levels of interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-10, IL-12, IL-17A, tumor necrosis factor (TNF)-α, C-reactive protein, osteoprotegerin, leptin, and adiponectin were determined using multiplex proteomic immunoassays. The correlation of biomarker concentrations between GCF versus PISF, within GCF or PISF, and with several covariates (age, brushing frequency, days since professional cleaning, probing depth [PD], and plaque index) were also determined.. Significantly higher levels of IL-17A (P = 0.02) and TNF-α (P = 0.03) were noted in PISF when compared with their levels in GCF. Significant positive correlations were noted between the concentrations of cytokines in PISF versus their levels in GCF. Among the covariates, a significant positive correlation was noted between mean PDs around implants and levels of IL-1β (P <0.05) and IL-8 (P <0.05) in PISF.. The results of this study point to the differential expression of specific biomarkers in GCF versus their levels in PISF in periodontal maintenance patients, which is critical information before establishing PISF as a diagnostic fluid to monitor peri-implant health.

Topics: Adiponectin; Adult; Aged; Aged, 80 and over; Biomarkers; C-Reactive Protein; Cross-Sectional Studies; Dental Implants; Dental Plaque Index; Dental Prophylaxis; Female; Gingival Crevicular Fluid; Humans; Interleukin-10; Interleukin-12; Interleukin-17; Interleukin-1alpha; Interleukin-1beta; Interleukin-6; Interleukin-8; Leptin; Male; Middle Aged; Osteoprotegerin; Periodontal Diseases; Periodontal Pocket; Toothbrushing; Tumor Necrosis Factor-alpha; Young Adult

Protocatechuic acid (PCA), a major metabolite of anthocyanins, has been shown to have antioxidant, antitumoral, and anti-inflammatory activities. In this study, we investigated the anti-inflammatory effects of PCA on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). The effects of PCA on LPS-induced inflammatory cytokines IL-6 and chemokines IL-8 in HGFs were detected by ELISA. The expression of NF-κB and PPAR-γ was detected by Western blot analysis. Our results demonstrated that PCA suppressed IL-6 and IL-8 production in LPS-stimulated HGFs. We also found that PCA inhibited LPS-induced NF-κB activation. Furthermore, the inhibition of PCA on LPS-induced IL-6 and IL-8 production can be reversed by PPAR-γ antagonist GW9662. In conclusion, the anti-inflammatory mechanism of PCA is associated with activating PPAR-γ, thereby inhibiting LPS-induced inflammatory response.

Topics: Anilides; Anti-Inflammatory Agents; Antioxidants; Cell Survival; Enzyme Activation; Gingiva; Humans; Hydroxybenzoates; Inflammation; Interleukin-6; Interleukin-8; Lipopolysaccharides; NF-kappa B; Periodontal Diseases; PPAR gamma

Periodontitis that affects the underlying structures of the periodontium, including the alveolar bone, is a multifactorial disease, whose etiology involves interactions between specific bacterial species of the subgingival biofilm and the host immune components. In the present study, we investigated the effects of myricetin, a flavonol largely distributed in fruits and vegetables, on growth and virulence properties of Porphyromonas gingivalis as well as on the P. gingivalis-induced inflammatory response in host cells. Minimal inhibitory concentration values of myricetin against P. gingivalis were in the range of 62.5 to 125 μg/ml. The iron-chelating activity of myricetin may contribute to the antibacterial activity of this flavonol. Myricetin was found to attenuate the virulence of P. gingivalis by reducing the expression of genes coding for important virulence factors, including proteinases (rgpA, rgpB, and kgp) and adhesins (fimA, hagA, and hagB). Myricetin dose-dependently prevented NF-κB activation in a monocyte model. Moreover, it inhibited the secretion of IL-6, IL-8 and MMP-3 by P. gingivalis-stimulated gingival fibroblasts. In conclusion, our study brought clear evidence that the flavonol myricetin exhibits a dual action on the periodontopathogenic bacterium P. gingivalis and the inflammatory response of host cells. Therefore, myricetin holds promise as a therapeutic agent for the treatment/prevention of periodontitis.

Topics: Collagenases; Colorimetry; Fibroblasts; Flavonoids; Gene Expression Regulation, Bacterial; Gingiva; Host-Pathogen Interactions; Humans; Inflammation; Interleukin-6; Interleukin-8; Iron Chelating Agents; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Microbial Sensitivity Tests; NF-kappa B; Periodontal Diseases; Porphyromonas gingivalis; Real-Time Polymerase Chain Reaction; RNA, Messenger; Siderophores; Virulence Factors

The alpha 7 nicotinic receptor (α7nAChR) is expressed by oral keratinocytes. α7nAChR activation mediates anti-inflammatory responses. The objective of this study was to determine if α7nAChR activation inhibited pathogen-induced interleukin-8 (IL-8) expression by oral keratinocytes.. Periodontal tissue expression of α7nAChR was determined by real-time PCR. OKF6/TERT-2 oral keratinocytes were exposed to Porphyromonas gingivalis in the presence and absence of a α7nAChR agonist (PHA-543613 hydrochloride) alone or after pre-exposure to a specific α7nAChR antagonist (α-bungarotoxin). Interleukin-8 (IL-8) expression was measured by ELISA and real-time PCR. Phosphorylation of the NF-κB p65 subunit was determined using an NF-κB p65 profiler assay and STAT-3 activation by STAT-3 in-cell ELISA. The release of ACh from oral keratinocytes in response to P. gingivalis lipopolysaccharide was determined using a GeneBLAzer M3 CHO-K1-bla cell reporter assay.. Expression of α7nAChR mRNA was elevated in diseased periodontal tissue. PHA-543613 hydrochloride inhibited P. gingivalis-induced expression of IL-8 at the transcriptional level. This effect was abolished when cells were pre-exposed to a specific α7nAChR antagonist, α-bungarotoxin. PHA-543613 hydrochloride downregulated NF-κB signalling through reduced phosphorylation of the NF-κB p65-subunit. In addition, PHA-543613 hydrochloride promoted STAT-3 signalling by maintenance of phosphorylation. Furthermore, oral keratinocytes upregulated ACh release in response to P. gingivalis lipopolysaccharide.. These data suggest that α7nAChR plays a role in regulating the innate immune responses of oral keratinocytes.

Topics: Acetylcholine; alpha7 Nicotinic Acetylcholine Receptor; Animals; Bridged Bicyclo Compounds, Heterocyclic; Bungarotoxins; CHO Cells; Cricetulus; Humans; Interleukin-8; Keratinocytes; Lipopolysaccharides; Mouth Mucosa; Periodontal Diseases; Porphyromonas gingivalis; Quinuclidines; RNA, Messenger; STAT3 Transcription Factor; Transcription Factor RelA

Periodontitis is an inflammatory infectious disease that destroys the tooth-supporting tissues. It is caused by multi-species subgingival biofilms that colonize the tooth surface. Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia (i.e. 'red complex' bacteria) are characteristic subgingival biofilm species. The triggering receptor expressed on myeloid cells 1 (TREM-1) is a cell surface receptor of the immunoglobulin superfamily, with a role in the amplification of proinflammatory cytokine production during infection. This study aimed to investigate TREM-1 mRNA expression in gingival tissues from patients with chronic periodontitis, generalized aggressive periodontitis and healthy subjects and its correlation with the levels of periodontal pathogens in the tissue. A further aim was to investigate the regulation of TREM-1 in human monocytic cells (MM6) challenged with an in-vitro subgingival biofilm model. Gingival tissue TREM-1 expression was increased in both chronic and aggressive periodontitis, compared to health, and correlated with the levels of the 'red complex' species in the tissue. No significant differences were detected between the two forms of periodontitis. Biofilm-challenged MM6 cells exhibited higher TREM-1 expression and secretion compared to controls, with partial involvement of the 'red complex'. Engagement or inhibition of TREM-1 affected the capacity of the biofilms to stimulate interleukin (IL)-1β, but not IL-8, secretion by the cells. In conclusion, this study reveals that TREM-1 tissue expression is enhanced in periodontal disease, and correlates with the level of periodontal pathogens. It also provides a mechanistic insight into the regulation of TREM-1 expression and the associated IL-1β production in biofilm-challenged monocytes.

Topics: Adult; Aggressive Periodontitis; Biofilms; Cells, Cultured; Chronic Periodontitis; Gingiva; Humans; Interleukin-1beta; Interleukin-8; Membrane Glycoproteins; Middle Aged; Monocytes; Periodontal Diseases; Receptors, Immunologic; RNA, Messenger; Triggering Receptor Expressed on Myeloid Cells-1

Inflammation within the oral cavity occurs due to dysregulation between microbial biofilms and the host response. Understanding how different oral hygiene products influence inflammatory properties is important for the development of new products. Therefore, creation of a robust host-pathogen biofilm platform capable of evaluating novel oral healthcare compounds is an attractive option. We therefore devised a multi-species biofilm co-culture model to evaluate the naturally derived polyphenol resveratrol (RSV) and gold standard chlorhexidine (CHX) with respect to anti-biofilm and anti-inflammatory properties.. An in vitro multi-species biofilm containing S. mitis, F. nucleatum, P. gingivalis and A. actinomycetemcomitans was created to represent a disease-associated biofilm and the oral epithelial cell in OKF6-TERT2. Cytotoxicity studies were performed using RSV and CHX. Multi-species biofilms were either treated with either molecule, or alternatively epithelial cells were treated with these prior to biofilm co-culture. Biofilm composition was evaluated and inflammatory responses quantified at a transcriptional and protein level.. CHX was toxic to epithelial cells and multi-species biofilms at concentrations ranging from 0.01-0.2%. RSV did not effect multi-species biofilm composition, but was toxic to epithelial cells at concentrations greater than 0.01%. In co-culture, CHX-treated biofilms resulted in down regulation of the inflammatory chemokine IL-8 at both mRNA and protein level. RSV-treated epithelial cells in co-culture were down-regulated in the release of IL-8 protein, but not mRNA.. CHX possesses potent bactericidal properties, which may impact downstream inflammatory mediators. RSV does not appear to have bactericidal properties against multi-species biofilms, however it did appear to supress epithelial cells from releasing inflammatory mediators. This study demonstrates the potential to understand the mechanisms by which different oral hygiene products may influence gingival inflammation, thereby validating the use of a biofilm co-culture model.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Anti-Infective Agents, Local; Anti-Inflammatory Agents; Biofilms; Cell Line; Chlorhexidine; Coculture Techniques; Down-Regulation; Epithelial Cells; Fusobacterium nucleatum; Host-Pathogen Interactions; Humans; Inflammation Mediators; Interleukin-8; Keratinocytes; Microbial Consortia; Periodontal Diseases; Porphyromonas gingivalis; Resveratrol; RNA, Messenger; Saliva, Artificial; Stilbenes; Streptococcus mitis

To investigate effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats.. Eighty SD rats were randomly divided into 4 groups, blank control group (group A) with 5 rats, treatment normal group (group B) with 25 rats, inflammation control group (group (group C) with 25 rats, inflammation treatment group (group D) with 25 rats. Immunohistochemistry and histomorphometric analysis was performed to measure the expression of IL-6, IL-8 and the first molar to the recent movement in the distance.. The expression of IL-8 reached a maximum on day 5 and declined thereafter in group B; the expression of IL-6 reached a maximum on day 5 in group B. The expression of IL-6 and IL-8 was gradually weakened with time in group C. The expression of IL-6 and IL-8 were high, and reached a maximum on day 5 and declined thereafter in group D. AD of positive cells in group D were higher than group B at each time point (P<0.05). The time which 0.49 N orthodontic force was loaded was longer, orthodontic tooth movement distance was greater. Movement distance in group D were longer than group B (P<0.05).. Orthodontic force as well as inflammatory stimulus can evoke the expression of IL-6 and IL-8. Under the combined effects of inflammation and orthodontic force, the expression of IL-6, IL-8 will increase.

Topics: Animals; Biomechanical Phenomena; Humans; Interleukin-6; Interleukin-8; Male; Molar; Periodontal Diseases; Rats; Rats, Sprague-Dawley; Stress, Mechanical; Tooth Migration; Tooth Movement Techniques

The antimicrobial peptide LL-37 is known to have a potent lipopolysaccharide (LPS)-neutralizing activity in various cell types. Because of observed heterogeneity within periodontopathogenic LPS, the authors hypothesized that LL-37 had specificity to neutralize such LPS activity. The present study, therefore, aims to investigate the LPS-neutralizing activity of LL-37 to various periodontopathogenic LPS in interleukin-8 (IL-8) production after challenging them in human oral fibroblasts.. Human periodontal ligament fibroblasts (PDLF) and gingival fibroblasts (GF) were cultured from biopsies of periodontal ligament and gingival tissues. After cell confluence in 24-well plates, LPS (10 μg/mL) from Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans were added with or without LL-37 (10 μg/mL). After 18 hours, the supernatant was collected and analyzed in IL-8 production by enzyme-linked immunosorbent assay.. All periodontopathogenic LPS statistically significantly induced IL-8 production in both PDLF and GF (P <0.01). After neutralization with LL-37, both PDLF and GF showed a statistically significant reduction in IL-8 production compared with LPS-treated groups without LL-37 (P <0.01), and the percentage of reduction in IL-8 production in PDLF appeared to be higher than in GF. In addition, the percentage of reduction in IL-8 production varied considerably according to each periodontopathogenic LPS.. The antimicrobial peptide LL-37 had an ability to suppress periodontopathogenic LPS-induced IL-8 production in both PDLF and GF. Its LPS-neutralizing activity revealed specificity to periodontopathogenic LPS and seemed to be dependent on the heterogeneity within LPS between different genera.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Antimicrobial Cationic Peptides; Cathelicidins; Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Fibroblasts; Fusobacterium nucleatum; Gingiva; Humans; Interleukin-8; Limulus Test; Lipopolysaccharides; Periodontal Diseases; Periodontal Ligament; Porphyromonas gingivalis; Prevotella intermedia; Species Specificity; Young Adult

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

Primary gingival epithelial cells were cultured in multilayers as a model for the study of interactions with oral bacteria associated with health and periodontal disease. Multilayers maintained at an air-liquid interface in low-calcium medium displayed differentiation and cytokeratin properties characteristic of junctional epithelium. Multilayers were infected with fluorescently labeled Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum or Streptococcus gordonii, and bacterial association was determined by confocal microscopy and quantitative image analysis. Porphyromonas gingivalis invaded intracellularly and spread from cell to cell; A. actinomycetemcomitans and F. nucleatum remained extracellular and showed intercellular movement through the multilayer; whereas S. gordonii remained extracellular and predominantly associated with the superficial cell layer. None of the bacterial species disrupted barrier function as measured by transepithelial electrical resistance. P. gingivalis did not elicit secretion of proinflammatory cytokines. However, A. actinomycetemcomitans and S. gordonii induced interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6 and IL-8 secretion; and F. nucleatum stimulated production of IL-1β and TNF-α. Aggregatibacter actinomycetemcomitans, F. nucleatum and S. gordonii, but not P. gingivalis, increased levels of apoptosis after 24 h infection. The results indicate that the organisms with pathogenic potential were able to traverse the epithelium, whereas the commensal bacteria did not. In addition, distinct host responses characterized the interaction between the junctional epithelium and oral bacteria.

Topics: Aggregatibacter actinomycetemcomitans; Apoptosis; Bacteria; Cell Culture Techniques; Epithelial Attachment; Epithelial Cells; Fusobacterium nucleatum; Gingiva; Host-Pathogen Interactions; Humans; Image Processing, Computer-Assisted; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Keratin-13; Keratin-9; Microscopy, Confocal; Mouth Mucosa; Periodontal Diseases; Porphyromonas gingivalis; Streptococcus gordonii; Time Factors; Tumor Necrosis Factor-alpha

Chronic parodontit inflammation in elderly patients was found to be accompanied by inbalance of complement system; IgA (sIgA), IgA, IgG, IgM hyperproduction; the increase of IL-8, IL-1α concentration and decrease of IL-4 level; increase Hsp-70 antibodies.

Topics: Aged; Chronic Disease; Complement System Proteins; Female; HSP70 Heat-Shock Proteins; Humans; Immunoglobulin A, Secretory; Immunoglobulin G; Immunoglobulin M; Interleukin-1alpha; Interleukin-8; Male; Middle Aged; Myocardial Ischemia; Periodontal Diseases

A-type cranberry proanthocyanidins (AC-PACs) have recently been reported to be beneficial for human health, especially urinary tract health. The effect of these proanthocyanidins on periodontitis, a destructive disease of tooth-supporting tissues, needs to be investigated. The purpose of this study was to investigate the effects of AC-PACs on various virulence determinants of Porphyromonas gingivalis as well as on the inflammatory response of oral epithelial cells stimulated by this periodontopathogen. We examined the effects of AC-PACs on P. gingivalis growth and biofilm formation, adherence to human oral epithelial cells and protein-coated surfaces, collagenase activity, and invasiveness. We also tested the ability of AC-PACs to modulate the P. gingivalis-induced inflammatory response by human oral epithelial cells. Our results showed that while AC-PACs neutralized all the virulence properties of P. gingivalis in a dose-dependent fashion, they did not interfere with growth. They also inhibited the secretion of interleukin-8 (IL-8) and chemokine (C-C motif) ligand 5 (CCL5) but did not affect the secretion of IL-6 by epithelial cells stimulated with P. gingivalis. This anti-inflammatory effect was associated with reduced activation of the nuclear factor-kappaB (NF-kappaB) p65 pathway. AC-PACs may be potentially valuable bioactive molecules for the development of new strategies to treat and prevent P. gingivalis-associated periodontal diseases.

Topics: Anti-Inflammatory Agents; Bacterial Adhesion; Bacteroidaceae Infections; Biofilms; Cells, Cultured; Chemokine CCL5; Epithelial Cells; Extracellular Matrix Proteins; Humans; Interleukin-6; Interleukin-8; Mouth Mucosa; Nuclear Magnetic Resonance, Biomolecular; Periodontal Diseases; Porphyromonas gingivalis; Proanthocyanidins; Vaccinium macrocarpon

The prevalence of chronic renal disease in industrialized countries is increasing, and chronic renal disease and periodontitis can have significant, reciprocal effects. The aim of this study was to evaluate the associations between specific clinical parameters and the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) in the gingival crevicular fluid of hemodialysis (HD) patients with periodontal disease.. Forty-three HD patients and 43 systemically healthy subjects were enrolled in this study. Plaque index (PI), gingival index (GI) and pocket depth were used to determine periodontal status. Venous blood samples were obtained from each patient in the morning before the dialysis session and analyzed to determine the levels of inflammatory, biochemical and hematological parameters. Gingival crevicular fluid was collected from all subjects, and the levels of TNF-alpha and IL-8 were determined in the gingival crevicular fluid samples.. The following results were obtained from HD patients and controls: TNF-alpha (pg/mL), 31.40 +/- 1.46 and 3.06 +/- 0.15 (p < 0.001); IL-8 (pg/mL), 90.98 +/- 94.03 and 35.05 +/- 16.86 (p < 0.001); PI, 1.69 +/- 1.02 and 0.04 +/- 0.02 (p < 0.001); GI, 0.82 +/- 0.06 and 0.04 +/- 0.02 (p < 0.001); and pocket depth, 2.23 +/- 0.63 and 1.51 +/- 0.05 (p < 0.001), respectively. In addition, there were positive correlations between TNF-alpha and PI (r = 0.642, p < 0.001), between TNF-alpha and GI (r = 0.565, p < 0.001), between TNF-alpha and pocket depth (r = 0.522, p < 0.001), between IL-8 and PI (r = 0.402, p = 0.002), between IL-8 and GI (r = 0.396, p = 0.002), and between IL-8 and pocket depth (r = 0.326, p = 0.012). There were negative correlations between albumin and PI (r = -0.491, p < 0.001), albumin and GI (r = -0.406, p < 0.001), albumin and pocket depth (r = -0.464, p < 0.001) and albumin and CRP (r = -0.467, p = 0.002) and between the gingival crevicular fluid levels of TNF-alpha and IL-8, TNF-alpha and hemoglobin (r = -0.745, p < 0.001; r = -0.285, p < 0.05) (respectively).. The levels of TNF-alpha and IL-8 in gingival crevicular fluid were significantly higher in HD patients than in controls. There were strong, positive correlations between clinical periodontal parameters and the levels of inflammatory cytokines in gingival crevicular fluid from the HD patients.

Topics: Adolescent; Adult; Aged; C-Reactive Protein; Case-Control Studies; Creatinine; Cross-Sectional Studies; Dental Plaque Index; Female; Gingival Crevicular Fluid; Hemoglobins; Humans; Interleukin-8; Kidney Failure, Chronic; Leukocyte Count; Male; Middle Aged; Periodontal Diseases; Periodontal Index; Periodontal Pocket; Renal Dialysis; Serum Albumin; Tumor Necrosis Factor-alpha; Urea; Young Adult

Desulfovibrio are sulfate-reducing anaerobic gram-negative rods that have been proposed as potential periodontopathogens. We investigated the capacity of Desulfovibrio to invade epithelial cells and induce cytokine secretion from these cells. Desulfovibrio strains were co-cultured with KB cells and counts of intracellular bacteria evaluated up to 3 days after infection. Desulfovibrio desulfuricans and Desulfovibrio fairfieldensis were able to survive within epithelial cells. Intracytoplasmic location of both bacterial species was confirmed by confocal laser scanning microscopy and transmission electron microscopy. Invasion was sensitive to nocodazole, an inhibitor of microtubule polymerization, but not to cytochalasin D, a microfilament inhibitor, suggesting that microtubule rearrangements were involved in the internalization of Desulfovibrio strains by KB cells. Infection by Desulfovibrio resulted in increased production of IL-6 and IL-8 by KB cells. The ability of D. desulfuricans and D. fairfieldensis to survive within oral epithelial cells and to modulate the epithelial immune response may contribute to the initiation and progression of periodontal diseases.

Topics: Antibodies, Bacterial; Coculture Techniques; Cytochalasin D; Cytoplasm; Desulfovibrio; Endocytosis; Epithelial Cells; Host-Pathogen Interactions; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; KB Cells; Microscopy, Confocal; Microscopy, Electron; Microtubules; Nocodazole; Nucleic Acid Synthesis Inhibitors; Periodontal Diseases; Tubulin Modulators

The antimicrobial peptide LL-37 is known to have a potent LPS-neutralizing activity in monocytes and macrophages. Recently, LL-37 in gingival crevicular fluids is suggested to be the major protective factor preventing infection of periodontogenic pathogens. In this study, we tried to address the effect of LL-37 on proinflammatory responses of human gingival fibroblasts (HGFs) stimulated with Toll-like receptor (TLR)-stimulant microbial compounds. LL-37 potently suppressed LPS-induced gene expression of IL6, IL8 and CXCL10 and intracellular signaling events, degradation of IRAK-1 and IkappaBalpha and phosphorylation of p38 MAPK and IRF3, indicating that the LPS-neutralizing activity is also exerted in HGFs. LL-37 also suppressed the expression of IL6, IL8 and CXCL10 induced by the TLR3 ligand poly(I:C). LL-37 modestly attenuated the expression of IL6 and IL8 induced by the TLR2/TLR1 ligand Pam(3)CSK(4), but did not affect the expression induced by the TLR2/TLR6 ligand MALP-2. Interestingly, LL-37 rather upregulated the expression of IL6, IL8 and CXCL10 induced by another TLR2/TLR6 ligand FSL-1. Thus, the regulatory effect of LL-37 is differently exerted towards proinflammatory responses of HGFs induced by different microbial stimuli, which may lead to unbalanced proinflammatory responses of the gingival tissue to infection of oral microbes.

Topics: Antimicrobial Cationic Peptides; Bacterial Infections; Cathelicidins; Cells, Cultured; Chemokine CXCL10; Fibroblasts; Gene Expression Regulation; Gingiva; Humans; Immunity, Innate; Interleukin-1 Receptor-Associated Kinases; Interleukin-6; Interleukin-8; Ligands; p38 Mitogen-Activated Protein Kinases; Periodontal Diseases; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 3; Toll-Like Receptor 4

Whereas periodontal disease is ultimately of bacterial etiology, from multispecies biofilms of gram-negative anaerobic microorganisms, much of the deleterious effects are caused by the resultant epithelial inflammatory response. Hence, development of a treatment that combines anti-biofilm antibiotic activity with anti-inflammatory activity would be of great utility. Antimicrobial peptides (AMPs) such as defensins are naturally occurring peptides that exhibit broad-spectrum activity as well as a variety of immunomodulatory activities. Furthermore, bacteria do not readily develop resistance to these agents. However, clinical studies have suggested that they do not represent optimal candidates for exogenous therapeutic agents. Small-molecule mimetics of these AMPs exhibit similar activities to the parent peptides, in addition to having low toxicity, high stability and low cost. To determine whether AMP mimetics have the potential for treatment of periodontal disease, we examined the activity of one mimetic, mPE, against biofilm cultures of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Metabolic assays as well as culture and biomass measurement assays demonstrated that mPE exhibits potent activity against biofilm cultures of both species. Furthermore, as little as 2 μg ml(-1) mPE was sufficient to inhibit interleukin-1β-induced secretion of interleukin-8 in both gingival epithelial cells and THP-1 cells. This anti-inflammatory activity is associated with a reduction in activation of nuclear factor-κB, suggesting that mPE can act both as an anti-biofilm agent in an anaerobic environment and as an anti-inflammatory agent in infected tissues.

Topics: Aggregatibacter actinomycetemcomitans; Alkynes; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antimicrobial Cationic Peptides; Bacterial Load; Biofilms; Biomimetic Materials; Cell Culture Techniques; Cell Line; Epithelial Cells; Gingiva; Humans; I-kappa B Proteins; Interleukin-1beta; Interleukin-8; Keratinocytes; Microbial Viability; Monocytes; NF-kappa B; Periodontal Diseases; Phenethylamines; Porphyromonas gingivalis

Periodontal diseases are a group of inflammatory disorders initiated by specific Gram-negative periodontopathogenic bacteria that lead to the destruction of tooth-supporting tissues. In this study, we tested whether a carbon dioxide-supercritical extract of Glycyrrhiza uralensis (licorice) can reduce the periodontopathogen-induced inflammatory response.. Monocyte-derived macrophages were treated with various concentrations of the licorice extract prior to being stimulated with Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) and Porphyromonas gingivalis lipopolysaccharide (LPS). The capacity of the licorice extract to mediate the inflammatory response was also tested in an ex vivo whole blood model stimulated with P. gingivalis LPS. The secretion of interleukin (IL)-1beta, -6, and -8 and tumor necrosis factor-alpha (TNF-alpha) in both models was assessed by enzyme-linked immunosorbent assays. Changes in the phosphorylation state of macrophage intracellular kinases induced by A. actinomycetemcomitans LPS and the licorice extract in the macrophage model were characterized by immunoblotting.. The licorice extract exhibited potent anti-inflammatory properties, inhibiting the periodontopathogen LPS-induced IL-1beta, -6, and -8 and TNF-alpha responses of macrophages. The licorice extract inhibited the phosphorylation of important macrophage intracellular signaling proteins, including nuclear factor-kappa B p65 nuclear transcription factor and Jun proto-oncogene-encoded activator protein (AP) 1 transcription factor, which are involved in inflammatory signaling pathways. The licorice extract was also a potent inhibitor of the proinflammatory cytokine response in the ex vivo human whole blood model.. This CO(2)-supercritical licorice extract is a potential candidate for the development of a new therapy to prevent and/or treat periodontitis-associated tissue destruction.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Inflammatory Agents; Cytokines; Glycyrrhiza; Humans; Immunoblotting; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Intracellular Signaling Peptides and Proteins; Lipopolysaccharides; Macrophages; Mitogen-Activated Protein Kinase 7; Periodontal Diseases; Phosphorylation; Plant Extracts; Porphyromonas gingivalis; Proto-Oncogene Mas; Transcription Factor AP-1; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Baicalin is a flavonoid compound purified from the medicinal plant Scutellaria baicalensis Georgi and has been reported to possess anti-inflammatory activity through its ability to complex with chemokines and thus reduces the capacity of chemokines to bind and activate their receptors. In the present study, we investigated whether baicalin could block polymorphonuclear leukocytes (PMNs) degranulation induced by interleukin (IL)-8, a CXC chemokine. Human PMNs were isolated from the peripheral blood of periodontal healthy donors and incubated with various concentrations of IL-8 (preincubated with or without baicalin). The morphology of PMNs was examined by transmission electron microscopy and extracellular concentration of granule component matrix metalloproteinase-8 (MMP-8) was detected by enzyme-linked immunosorbent assay (ELISA). Results showed that IL-8 could significantly induce MMP-8 release from PMNs when compared to control, and its inductive activity was concentration-dependent. But when preincubated with various concentrations of baicalin, the amount of MMP-8 release from PMNs decreased significantly. The present study concludes that baicalin could block MMP-8 release from PMNs induced by IL-8, which suggests that it may play an important role in the prevention and treatment of periodontal disease.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Degranulation; Cells, Cultured; Dose-Response Relationship, Drug; Flavonoids; Humans; Interleukin-8; Microscopy, Electron, Transmission; Neutrophils; Periodontal Diseases

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

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

Oral treponemes are well-known as causative agents of periodontal diseases; however, the details have not been fully clarified. Here, we examined the effects of Treponema medium glycoconjugate on the activation of human gingival fibroblasts using phenol-water extracts from Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, and Actinobacillus actinomycetemcomitans. The phenol-water extracts activated human gingival fibroblasts to mediate IL-8 production, as well as IL-8 mRNA expression, phosphorylation of p38 mitogen-activated protein kinase, and expression of intercellular adhesion molecule-1. T. medium glycoconjugate exhibited no activation of human gingival fibroblasts, while phenol-water extract-induced activation of human gingival fibroblasts was clearly inhibited by T. medium glycoconjugate. Furthermore, binding of biotinylated phenol-water extracts to CD14 in the presence of LPS-binding protein was blocked with T. medium glycoconjugate. These results suggest that T. medium glycoconjugate has an inhibitory effect on host cell activation by periodontopathic bacteria caused by binding to CD14- and LPS-binding protein.

Topics: Acute-Phase Proteins; Aggregatibacter actinomycetemcomitans; Carrier Proteins; Cell Extracts; Cells, Cultured; Fibroblasts; Fusobacterium nucleatum; Gingiva; Glycoconjugates; Gram-Negative Bacteria; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Lipopolysaccharide Receptors; Lipopolysaccharides; Membrane Glycoproteins; p38 Mitogen-Activated Protein Kinases; Periodontal Diseases; Phenols; Phosphorylation; Porphyromonas gingivalis; Prevotella intermedia; Treponema; Water

Host-mediated immunoinflammatory pathways activated by bacteria lead to destruction of the periodontal connective tissues and alveolar bone. The objective of this study was to elucidate the activation of the inflammatory processes in periodontal disease by quantitative assessment of cytokines and periodontopathogens. Gingival crevicular fluids (GCF) and subgingival plaque samples were collected from patients with chronic periodontitis and gingivitis and from periodontally healthy sites. Vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), and interleukin 8 (IL-8) in GCF were analyzed by enzyme-linked immunosorbent assay. Periodontopathogens, including Bacteroides forsythus, Campylobacter rectus, Porphyromonas gingivalis and Prevotella intermedia, were analyzed by immunofluorescence and dark-field microscopy. There was significantly more VEGF and IL-8 in chronic periodontitis and gingivitis sites than in periodontally healthy sites. There were significant positive correlations between the concentrations and total amounts of VEGF and IL-8 in chronic periodontitis and gingivitis sites, and between the levels of periodontopathogens and the total amounts of VEGF, MCP-1 and IL-8. These data indicate that inflammatory processes induced by periodontopathogens and the activation of certain cytokines (VEGF, MCP-1, IL-8) in periodontal diseases may be relevant to host-mediated destruction in chronic periodontitis.

Topics: Adult; Chemokine CCL2; Endothelial Growth Factors; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Lymphokines; Middle Aged; Periodontal Diseases; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

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

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

We previously demonstrated that butyric acid, an extracellular metabolite from periodontopathic bacteria, induces cytotoxicity and apoptosis in murine thymocytes, splenic T cells, and human Jurkat T cells. In this study, we used a cell-to-cell interaction system to examine the contribution of gingival fibroblasts to the regulation of T-cell death induced by butyric acid. Butyric acid slightly suppressed fibroblast viability in a concentration-dependent fashion. However, DNA fragmentation assays indicated that butyric acid did not induce apoptosis for up to 21 h in human gingival fibroblasts (Gin 1, F41-G, and H. pulp cells). The culture supernatants were assayed for interleukin 1alpha (IL-1alpha), IL-1beta, IL-6, IL-8, IL-11, tumor necrosis factor alpha, and transforming growth factor beta, but only the IL-6, IL-8, and IL-11 levels were significantly increased by addition of butyric acid. Butyric acid- or Fas-induced Jurkat-cell apoptosis was attenuated when Jurkat cells were cocultured with either F41-G or Gin 1 cells that had been preincubated for 6 h with butyric acid. IL-8 slightly stimulated butyric acid- or Fas-induced Jurkat-cell apoptosis in a dose-dependent manner, although a low dose of IL-8 had a mildly inhibitory effect on apoptosis. In contrast, IL-6 and IL-11 significantly suppressed butyric acid- or Fas-induced apoptosis in a dose-dependent fashion. Furthermore, the addition of monoclonal antibodies against human IL-6 and IL-11 to cocultures of gingival fibroblasts and Jurkat cells partially eliminated T-cell recovery. These results suggest that the proinflammatory cytokines such as IL-6 and IL-11, produced in fibroblasts stimulated with butyric acid, are involved in the attenuation of T-cell apoptosis by gingival fibroblasts.

Topics: Apoptosis; Butyric Acid; Cell Communication; Cell Division; Cell Line; Cytokines; Fibroblasts; Gingiva; Humans; Interleukin-11; Interleukin-6; Interleukin-8; Periodontal Diseases; T-Lymphocytes

In the inflammatory response elicited by bacterial colonization in periodontal pockets, pocket epithelial cells not only serve as a barrier to isolate the pocket microenvironment from external stimuli but also regulate the functions of neighboring cells including fibroblasts and inflammatory cells. To elucidate this mechanism, we characterized the effects of periodontopathic bacterium Eikenella corrodens 1073 components on the production of some inflammatory mediators in a human oral epithelial cell line (KB). In enzyme-linked immunosorbent assay (ELISA), the E. corrodens supernatant induced interleukin-6 (IL-6), IL-8 and prostaglandin E2 but not interferon-gamma (IFN-gamma) production by KB cells. After incubation with E. corrodens supernatant, KB cells showed a marked increase in the levels of IL-6, IL-8 and PG G/H synthase (cyclooxygenase)-2, but not IFN-gamma, gene expression by reverse-transcriptase polymerase chain reaction. All these E. corrodens products responsible for production of these inflammatory mediators resisted freezing and boiling and were present in a 10-kDa filtrate. These results imply that these soluble small-molecular-mass products from E. corrodens stimulate various inflammatory mediator productions by human oral epithelial cells and may play a role in the initiation of periodontal inflammation and subsequently perpetuate the inflammatory response during chronic infection.

Topics: Bacterial Adhesion; Culture Media, Conditioned; Cyclooxygenase 2; Cytokines; Dinoprostone; Eikenella corrodens; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gene Expression; Humans; Immunoblotting; Inflammation Mediators; Interleukin-6; Interleukin-8; Isoenzymes; KB Cells; Membrane Proteins; Periodontal Diseases; Prostaglandin-Endoperoxide Synthases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Up-Regulation

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

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

Periodontal diseases result from the interaction of bacterial pathogens with the host gingival tissues. The role of gingival epithelial cells in the initiation of host defense mechanisms after encountering oral bacteria has not been investigated. Actinobacillus actinomycetemcomitans is a key periodontal pathogen that adheres to and invades oral epithelial cells. Thus, we examined whether gingival epithelial cells increase secretion of the potent neutrophil chemoattractant interleukin-8 (IL-8) following A. actinomycetemcomitans challenge. Normal human oral keratinocytes (NHOK), isolated from gingival tissue, and 3 oral epithelial cell lines (HOK-18A, HOK-16B-BaP-T1, and HEp-2) were co-cultured with A. actinomycetemcomitans for 2 hours to allow bacteria-epithelial cell interactions. The epithelial cells were then washed, and fresh medium with gentamicin was added to kill extracellular bacteria. Cell cultures were further incubated for 24 hours before the supernatant was collected for IL-8 detection with ELISA. The results showed that IL-8 secretion increased 2- to 7-fold 24 hours after bacterial challenge. The highest IL-8 secretion was at the multiplicity of infection (MOI) of 1,000:1 in bacterial dose response studies using HOK-16B-BaP-T1 cells. Time-course studies revealed that IL-8 secretion rapidly reached a maximum level 6 hours after bacterial challenge and subsequently decreased to basal levels. These data indicate that gingival epithelial cells are capable of upregulating IL-8 expression rapidly in response to A. actinomycetemcomitans challenge and thus may facilitate the recruitment of neutrophils as a host defense mechanism.

Topics: Aggregatibacter actinomycetemcomitans; Bacterial Adhesion; Cells, Cultured; Chemotaxis, Leukocyte; Colony Count, Microbial; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gene Expression Regulation; Gingiva; Humans; Interleukin-8; Keratinocytes; Neutrophil Activation; Neutrophils; Periodontal Diseases; Time Factors; Up-Regulation

Gingipains are the major cysteine proteinases synthesized by Porphyromonas gingivalis which, in soluble form, are able to initially convert IL-8 (77 amino acid residues) to a more potent species truncated at the amino terminus, followed by slow degradation and destruction of chemokine biological activity. In contrast, the same enzymes when associated with bacterial outer-membrane blebs (vesicles), instantly degrade this chemokine. This division of enhancing and inactivating activity between soluble and membrane-bound gingipains can cause the compartmentalization of pro- and anti-inflammatory reactions to distal and proximal positions from bacterial plaque, respectively, which may explain why, despite the massive neutrophil accumulation at periodontitis sites, there is no elimination of infection.

Topics: Adhesins, Bacterial; Cysteine Endopeptidases; Gingipain Cysteine Endopeptidases; Hemagglutinins; Humans; In Vitro Techniques; Interleukin-8; Neutrophil Activation; Neutrophils; Peptide Hydrolases; Periodontal Diseases; Porphyromonas gingivalis; Recombinant Proteins

Periodontal diseases are inflammatory disorders caused by microorganisms of dental plaque that colonize the gingival sulcus and, subsequently, the periodontal pocket. As in other mucosal infections, the host response to plaque bacteria is characterized by an influx of polymorphonuclear leukocytes (PMNs) to the gingival crevice. Neutrophil migration through the epithelial lining of the gingival pocket is thought to be the first line of defense against plaque bacteria. In order to model this phenomenon in vitro, we used the oral epithelial cell line KB and human PMNs in the Transwell system and examined the impact of Porphyromonas gingivalis-epithelial cell interactions on subsequent PMN transepithelial migration. We demonstrate here that P. gingivalis infection of oral epithelial cells failed to trigger transmigration of PMNs. Furthermore, it significantly inhibited neutrophil transmigration actively induced by stimuli such as N-formylmethionyl leucyl phenylalanine, interleukin-8 (IL-8), and the intestinal pathogen enterotoxigenic Escherichia coli. The ability of P. gingivalis to block PMN transmigration was strongly positively correlated with the ability to adhere to and invade epithelial cells. In addition, P. gingivalis attenuated the production of IL-8 and the expression of intercellular adhesion molecule 1 by epithelial cells. The ability of P. gingivalis to block neutrophil migration across an intact epithelial barrier may critically impair the potential of the host to confront the bacterial challenge and thus may play an important role in the pathogenesis of periodontal disease.

Topics: Bacterial Adhesion; Bacteroidaceae Infections; CD18 Antigens; Cell Communication; Chemotaxis, Leukocyte; Diffusion Chambers, Culture; Down-Regulation; Epidermal Cells; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Mouth Mucosa; Neutrophils; Periodontal Diseases; Porphyromonas gingivalis; Tumor Cells, Cultured

Topics: Animals; Humans; Interleukin-1; Interleukin-8; Periodontal Diseases; Porphyromonas gingivalis