interleukin-8 and Periodontitis

IL-8 and its polymorphisms are involved in multiple acute and chronic inflammatory processes including pathological changes to surrounding structures of the teeth called periodontal diseases or periodontitis. The aim of this manuscript was to systematically review studies from 2006 to 2021 on IL-8 polymorphisms and their association with periodontitis. Literature analysis was done following the PRISMA protocol guidance using articles not older than 15 years (2006-2021). The search was carried out using PubMed (MEDLINE), ScienceDirect and Wiley Online Library databases. For the focus question, the PICO (population (P), intervention (I), control (C), and outcome (O)) study design protocol was used, and the following question was formulated: are IL-8 gene polymorphisms associated with periodontitis? A total of 2422 articles were found at the beginning of the search. After applying the inclusion and exclusion criteria, screening, and full-text article exclusion with reasons, 31 studies were included in the analysis. In conclusion, IL-8 and its gene polymorphisms are associated with an increased risk of periodontal diseases.

Topics: Chronic Periodontitis; Humans; Interleukin-8; Periodontitis; Polymorphism, Genetic

Gingival epithelium plays a pivotal role in protecting the underlying periodontium from the microbial colonization found in the gingival sulcus. Having an appropriate phenotype displayed by gingival epithelial cells is a critical host component required for protection against bacterial invasion into gingival tissues. In the present study, gingival epithelial homeostasis associated with the CXCL-8/IL-8 chemokine response was investigated in vitro to determine the mechanisms that gingival epithelial cells utilize for sensing gram-positive and gram-negative microorganisms. The findings of this study have demonstrated, by using Fusobacterium nucleatum, a heterogeneity of gingival epithelial cell response by Toll-like receptor (TLR) 2, a lipoprotein sensor. Notably, however, lipopolysaccharide (LPS), a major virulence factor of gram-negative bacteria, is not recognized by gingival epithelial cells unless the LPS is internalized into the cells. Activation of TLR4 in gingival epithelial cells occurs in the endosome, an intracellular event that requires a vesicular acidification to turn on TLR4 signaling, indicating their stringency for fine-tuning a local LPS response. This study has identified a unique LPS sensing mechanism of the oral epithelium to overcome a periodontal infection associated with LPS derived from gram-negative microbes that arises during dysbiosis.

Topics: Epithelial Cells; Gingiva; Humans; Interleukin-8; Lipopolysaccharides; Periodontitis

Associations between interleukin-8 (IL-8) gene polymorphisms and periodontitis susceptibility have been investigated in many published studies, but the conclusions are still inconsistent. Therefore, we performed this systematic review and meta-analysis to review which polymorphisms have been researched and to obtain a precise result of the same polymorphism from different studies.. Finally 10 publications involving 1938 patients and 1569 controls were yielded, including 12 polymorphisms. Six studies investigated rs4073 polymorphism; two focused on rs2227306 and rs2227307; two referred to rs2227532 and T-738A; one detected rs2230054, rs1126579 and rs1126580; one inspected A2767T, T11722T2 and C1633T, and one for rs2234671 polymorphism. Of them, IL-8 C1633T and rs1126580 polymorphisms showed positive association while the other ten polymorphisms revealed negative results.. A comprehensive literature search from PubMed, Web of Science, and Chinese National Knowledge Infrastructure was conducted for all potentially relevant studies published before January 2, 2017. Two authors selected the studies and extracted data. The pooled analysis was conducted using the RevMan 5.1 software if a polymorphism was reported by two or more studies.. Based on current evidence, the IL-8 rs4073, A2767T, T11722T2, rs2234671, rs2230054, rs1126579, rs2227306, rs2227307, rs2227532, and T-738A polymorphisms were not associated with periodontitis susceptibility; the IL-8 C1633T and rs1126580 polymorphisms were associated with increased risk of periodontitis.

Topics: Alleles; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-8; Odds Ratio; Periodontitis; Polymorphism, Single Nucleotide

This review assesses the associations of interleukin-8 gene (IL-8) -251A/T (rs4073) and -845T/C (rs2227532) polymorphisms with susceptibility to periodontitis.. Several electronic databases were searched for eligible articles. Twelve studies involving 2,233 cases and 2,655 controls were retrieved and analyzed. Odds ratios (ORs) along with 95% confidence intervals (CIs) were calculated to assess the strength of relationship between the IL-8 polymorphisms and periodontitis risk.. No significant association was found for IL-8 -251A/T polymorphism with periodontitis in the overall analysis and stratification by periodontitis type and smoking status. Subgroup analysis by ethnicity revealed that -251A/T T allele and TT genotype were associated with decreased risk of periodontitis in a Brazilian mixed population (T allele versus A allele: OR 0.80, 95% CI 0.68 to 0.94, Pheterogeneity = 0.30; TT versus AA: OR 0.65, 95% CI 0.46 to 0.93, Pheterogeneity = 0.39; TT versus. OR 0.58, 95% CI 0.35 to 0.98, Pheterogeneity = 0.01). In addition, -251A/T T allele was associated with increased periodontitis risk in Asians. Pooled estimates showed that the -845T/C polymorphism was associated with periodontitis susceptibility in overall analysis and the chronic periodontitis subgroup. In addition, marginal associations were observed between -845T/C polymorphism and periodontitis in a Brazilian mixed population. Moreover, this association was also confirmed to be significant in Brazilian non-smokers.. This meta-analysis indicated that both IL-8 -251A/T and -845T/C polymorphisms may be involved in the development of periodontitis in a Brazilian mixed population, whereas the -251A/T allele T appeared to be a risk factor for periodontitis in Asians.

Topics: Adenine; Asian People; Brazil; Cytosine; Ethnicity; Genetic Predisposition to Disease; Humans; Interleukin-8; Periodontitis; Polymorphism, Genetic; Thymine

The pathogenesis of periodontal disease involves the sequential activation of a great variety of components of the host immune response, primarily acting to defend periodontal tissues against bacterial aggression, but also functioning as mediators of tissue destruction. The expression of the disease results from the interaction of host, microbiological agents, and environmental factors. Leukocytes play a critical role in the pathogenesis of the disease, producing different cytokines, chemokines, and other mediators, thus generating a host defense response, as well as inducing tissue inflammation and bone destruction. The aim of this review is to address the role of some inflammatory mediators in response to bacterial aggression in periodontitis.

Topics: Cytokines; Dinoprostone; Humans; Interleukin-1; Interleukin-8; Periodontitis; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

Diabetes mellitus is a systemic disease with several major complications affecting both the quality and length of life. One of these complications is periodontal disease (periodontitis). Periodontitis is much more than a localized oral infection. Recent data indicate that periodontitis may cause changes in systemic physiology. The interrelationships between periodontitis and diabetes provide an example of systemic disease predisposing to oral infection, and once that infection is established, the oral infection exacerbates systemic disease. In this case, it may also be possible for the oral infection to predispose to systemic disease. In order to understand the cellular/molecular mechanisms responsible for such a cyclical association, one must identify common physiological changes associated with diabetes and periodontitis that produce a synergy when the conditions coexist. A potential mechanistic link involves the broad axis of inflammation, specifically immune cell phenotype, serum lipid levels, and tissue homeostasis. Diabetes-induced changes in immune cell function produce an inflammatory immune cell phenotype (upregulation of proinflammatory cytokines from monocytes/polymorphonuclear leukocytes and downregulation of growth factors from macrophages). This predisposes to chronic inflammation, progressive tissue breakdown, and diminished tissue repair capacity. Periodontal tissues frequently manifest these changes because they are constantly wounded by substances emanating from bacterial biofilms. Diabetic patients are prone to elevated low density lipoprotein cholesterol and triglycerides (LDL/TRG) even when blood glucose levels are well controlled. This is significant, as recent studies demonstrate that hyperlipidemia may be one of the factors associated with diabetes-induced immune cell alterations. Recent human studies have established a relationship between high serum lipid levels and periodontitis. Some evidence now suggests that periodontitis itself may lead to elevated LDL/TRG. Periodontitis-induced bacteremia/endotoxemia has been shown to cause elevations of serum proinflammatory cytokines such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha), which have been demonstrated to produce alterations in lipid metabolism leading to hyperlipidemia. Within this context, periodontitis may contribute to elevated proinflammatory cytokines/serum lipids and potentially to systemic disease arising from chronic hyperlipidemia and/o

Topics: Bacteremia; Bacterial Infections; Biofilms; Blood Glucose; Cholesterol, LDL; Diabetes Complications; Diabetes Mellitus; Disease Susceptibility; Down-Regulation; Endotoxemia; Growth Substances; Homeostasis; Humans; Hyperlipidemias; Inflammation; Inflammation Mediators; Insulin Resistance; Interleukin-8; Islets of Langerhans; Periodontitis; Phenotype; Risk Factors; Triglycerides; Tumor Necrosis Factor-alpha; Up-Regulation; Wound Healing

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

Interleukin-8 (IL-8) is a chemoattractant cytokine produced by a variety of tissue and blood cells. Unlike many other cytokines, it has a distinct target specificity for the neutrophil, with only weak effects on other blood cells. Interleukin-8 attracts and activates neutrophils in inflammatory regions. The importance of neutrophil functions has been recognized in periodontal disease for many years. Neutrophils represent the major population of immigrant cells in periodontitis. In diseases with neutrophil dysfunctions periodontal tissue is lost very rapidly. The response of neutrophils to IL-8 is characterized by migration of the cells, the release of granule enzymes, and other intra- and extracellular changes. Connective tissue constituents are efficiently degraded by neutrophil enzymes, released upon activation. Interleukin-8 is a member of the Interleukin-8 supergene family that includes other small chemotactic peptides with structural homology. It also shares with other cytokines DNA sequence features that suggest common regulatory pathways. In vivo intracutaneous application of IL-8 induces local exudation and a massive, long-lasting accumulation of neutrophils. Though IL-8 plays a role in the cytokine network, its major pathophysiological role lies in affecting neutrophils. This article presents a review of literature on the current knowledge of IL-8, its mechanisms of expression, and the effects it exerts on the neutrophil.

Topics: Chemotactic Factors; Humans; Interleukin-8; Neutrophils; Periodontitis

Fibrin sealant (FS) is a biologically derived tissue adhesive for securing flaps. The aim of the present randomized controlled clinical trial was to compare early wound healing by assessing interleukin-1β (IL-1β) and interleukin-8 (IL-8) levels from gingival crevicular fluid (GCF) after using FS and suture for periodontal flap closure.. Thirty selected quadrants in 15 periodontitis patients were randomly assigned to either a test (fibrining) or control group (suturing) for flap closure. IL-1β and IL-8 were assessed in GCF using enzyme-linked immunosorbent assay (ELISA) before and eight days after surgery. Patients were recalled at 7, 14, 21 days and 3 months after surgery for clinical assessment.. There was a statistically significant decrease in IL-1β (84.82 ± 77.18, 29.2 ± 21.97 pg/μl) and IL-8 (57.94 ± 55.47, 21.82 ± 21.93 pg/μl) levels in the test side after fibrining while there was an increase in the control side (IL-1β 31.40 ± 16.82, 128.8 ± 45.14; IL-8 31.40 ± 16.82, 128.83 ± 45.14 pg/μl) (p < 0.05). The change in concentration of IL-1β and IL-8 following intervention correlated significantly in both the sites. Clinical parameters differed significantly only on the seventh day with less plaque and bleeding on the test sites.. Fibrin sealant enhances early wound healing by reducing inflammation after periodontal flap surgery.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Female; Fibrin Tissue Adhesive; Gingival Crevicular Fluid; Humans; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Outcome Assessment, Health Care; Pain, Postoperative; Periodontitis; Suture Techniques; Time Factors; Tissue Adhesives; Wound Healing

The purpose of the present study is to investigate the potential link between maternal periodontitis and pregnancy outcomes, including preterm birth (<37 weeks) and low birth weight (<2,500 g).. Ninety nine pregnant females with mild/moderate periodontitis were randomly allocated to a control (n = 50) or test (n = 49) group. Test group participants received intrapregnancy non-surgical periodontal treatment, whereas this was deferred until after delivery for controls. Demographic and baseline clinical data were obtained for all participants at initial assessment pretreatment. Clinical data were rerecorded for test participants at review 8 weeks after treatment. Birth outcomes were completed at delivery by midwives who also collected cord blood samples when possible; the latter were analyzed to determine the presence/levels of cytokines interleukin (IL)-1β, IL-6, and IL-8. All data were analyzed on an intention-to-treat basis using appropriate statistical tests.. Random allocation of participants resulted in well-balanced control and test groups. All test group participants and all but one control participant gave birth to a live infant. No significant differences were detected between control and test groups with regard to birth outcome measures of birth weight and gestational age or in relation to cytokine prevalence/levels.. Intrapregnancy non-surgical periodontal treatment, completed at 20 to 24 weeks, did not reduce the risk of preterm, low-birth-weight delivery in this population.

Topics: Adult; Birth Weight; Delivery, Obstetric; Dental Plaque Index; Dental Prophylaxis; Dental Scaling; Female; Fetal Blood; Gestational Age; Humans; Infant, Low Birth Weight; Infant, Newborn; Intention to Treat Analysis; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Oral Hygiene; Periodontal Index; Periodontitis; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Premature Birth; Root Planing; Sex Factors

We have previously shown that a single nucleotide polymorphism rs11536889 in the 3'-untranslated region (UTR) of TLR4 was associated with periodontitis. In this study the effects of this single nucleotide polymorphism on Toll-like receptor (TLR) 4 expression were investigated. Monocytes from subjects with the C/C genotype expressed higher levels of TLR4 on their surfaces than those from subjects with the other genotypes. Peripheral blood mononuclear cells (PBMCs) from the C/C and G/C subjects secreted higher levels of IL-8 in response to lipopolysaccharide (LPS), a TLR4 ligand, than the cells from the G/G subjects. However, there was no significant difference in TLR4 mRNA levels in PBMCs from the subjects with each genotype. After stimulation with tripalmitoylated CSK(4) (Pam(3)CSK(4)), TLR4 mRNA levels increased in PBMCs from both the C/C and G/G subjects, whereas TLR4 protein levels increased in PBMCs from the C/C but not G/G subjects. Transient transfection of a series of chimeric luciferase constructs revealed that a fragment of 3'-UTR containing rs11536889 G allele, but not C allele, suppressed luciferase activity induced by LPS or IL-6. Two microRNAs, hsa-miR-1236 and hsa-miR-642a, were predicted to bind to rs11536889 G allele. Inhibition of these microRNAs reversed the suppressed luciferase activity. These microRNA inhibitors also up-regulated endogenous TLR4 protein on THP-1 cells (the G/G genotype) after LPS stimulation. Furthermore, mutant microRNAs that bind to the C allele inhibited the luciferase activity of the construct containing the C allele. These results indicate that genetic variation of rs11536889 contributes to translational regulation of TLR4, possibly by binding to microRNAs.

Topics: 3' Untranslated Regions; Alleles; Asian People; Cell Line; Female; Humans; Interleukin-6; Interleukin-8; Japan; Leukocytes, Mononuclear; Lipopeptides; Lipopolysaccharides; Male; MicroRNAs; Periodontitis; Polymorphism, Single Nucleotide; Protein Biosynthesis; Toll-Like Receptor 4

This split-mouth, single-masked, randomized, controlled clinical trial compares the short-term outcomes of a combined treatment with scaling and root planing (SRP) and neodymium-doped:yttrium, aluminum, and garnet (Nd:YAG)-laser irradiation with treatment with SRP alone.. Thirty patients were recruited. The mandibular left or right side was randomly assigned as the test side (SRP with laser treatment) or control side (SRP alone). The water-cooled Nd:YAG laser was used at 4 W, 80 mJ/pulse, 50 Hz, and with a pulse width of 350 micros. At baseline, gingival crevicular fluid (GCF) samples were taken from the test and control sides, and levels of matrix metalloproteinase (MMP)-8 and interleukin (IL)-1beta, -4, -6, and -8 were measured using standard techniques. The plaque index (PI), gingival index (GI), and probing depth (PD) were measured by calibrated examiners.. At the 1-week follow-up, PD (P <0.001), PI (P <0.05), and GCF volume (P <0.001) showed significant improvement on test sides compared to control sides. At the 3-month follow-up, PD (P <0.01), PI (P <0.01), GI (P <0.01), and GCF volume (P <0.05) also showed significant improvement on test sides compared to control sides. At the 1-week follow up, IL-1beta and MMP-8 levels were significantly reduced on test sides compared to control sides. The 3-month follow-up confirmed that the improvements on test sites had been sustained compared to the treatment outcomes of control sites.. In the short-term, SRP in combination with a single application of a water-cooled Nd:YAG laser significantly improves clinical signs associated with periodontal inflammation compared to treatment with SRP alone.

Topics: Adult; Aged; Combined Modality Therapy; Dental Plaque Index; Dental Scaling; Female; Follow-Up Studies; Gingival Crevicular Fluid; Humans; Interleukin-1beta; Interleukin-4; Interleukin-6; Interleukin-8; Lasers, Solid-State; Male; Matrix Metalloproteinase 8; Middle Aged; Periodontal Index; Periodontal Pocket; Periodontitis; Root Planing; Single-Blind Method; Treatment Outcome; Ultrasonic Therapy

Periodontal disease is the most common multifactorial disease, afflicting a very large proportion of the adult population. Periodontal disease secondarily causes increases in the serum levels of C-reactive protein (CRP) and other markers of inflammation. An increased level of CRP reflects an increased risk for cardiovascular disease. The aim of the current randomized clinical trial was to evaluate the short-term effect of a combination of dipyridamole and prednisolone (CRx-102) on the levels of high-sensitivity (hs)-CRP, proinflammatory markers in blood, and clinical signs of periodontal disease.. Fifty-seven patients with >/=10 pockets with probing depths >/=5 mm were randomized into two groups in this masked single-center placebo-controlled study: CRx-102 (n = 28) and placebo (n = 29). hs-CRP levels, inflammatory markers (interleukin [IL]-6, -1beta, -8, and -12, tumor necrosis factor-alpha, and interferon-gamma [IFN-gamma]), bleeding on probing (BOP), and changes in probing depths were evaluated. The subjects received mechanical non-surgical therapy after 42 days, and the study was completed after 49 days.. At day 42, the differences in the hs-CRP, IFN-gamma, and IL-6 levels between the two groups were statistically significant (P <0.05), whereas no difference was found for the other inflammatory markers. There was no change in probing depth or BOP between the two groups.. The administration of CRx-102 resulted in significant decreases in hs-CRP, IFN-gamma, and IL-6, but it did not significantly change BOP or probing depths.

Topics: Adult; Anti-Inflammatory Agents; C-Reactive Protein; Dental Scaling; Dipyridamole; Drug Combinations; Female; Follow-Up Studies; Gingival Hemorrhage; Humans; Inflammation Mediators; Interferon-gamma; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Periodontal Pocket; Periodontitis; Placebos; Prednisolone; Root Planing; Tumor Necrosis Factor-alpha

The purposes of this study were to determine: 1) if periodontal treatment in pregnant women before 21 weeks of gestation alters levels of inflammatory mediators in serum; and 2) if changes in these mediators are associated with birth outcomes.. A total of 823 pregnant women with periodontitis were randomly assigned to receive scaling and root planing before 21 weeks of gestation or after delivery. Serum obtained between 13 and 16 weeks, 6 days (study baseline) and 29 to 32 weeks of gestation was analyzed for C-reactive protein; prostaglandin E(2); matrix metalloproteinase-9; fibrinogen; endotoxin; interleukin (IL)-1 beta, -6, and -8, and tumor necrosis factor-alpha. Cox regression, multiple linear regression, and the t, chi(2), and Fisher exact tests were used to examine associations among the biomarkers, periodontal treatment, and gestational age at delivery and birth weight.. A total of 796 women had baseline serum data, and 620 women had baseline and follow-up serum and birth data. Periodontal treatment did not significantly alter the level of any biomarker (P >0.05). Neither baseline levels nor the change from baseline in any biomarker were significantly associated with preterm birth or infant birth weight (P >0.05). In treatment subjects, the change in endotoxin was negatively associated with the change in probing depth (P <0.05).. Non-surgical mechanical periodontal treatment in pregnant women, delivered before 21 weeks of gestation, did not reduce systemic (serum) markers of inflammation. In pregnant women with periodontitis, levels of these markers at 13 to 17 weeks and 29 to 32 weeks of gestation were not associated with infant birth weight or a risk for preterm birth.

Topics: Adolescent; Adult; Birth Weight; C-Reactive Protein; Dental Scaling; Dinoprostone; Endotoxins; Female; Fibrinogen; Follow-Up Studies; Gestational Age; Humans; Infant, Newborn; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Matrix Metalloproteinase 9; Periodontitis; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Premature Birth; Risk Factors; Root Planing; Tumor Necrosis Factor-alpha; Young Adult

In order to find a new treatment of periodontitis, this paper studied the effects of Gu Chi Gao on IL-8 in gingival crevicular fluid (GCF) of periodontitis. One tooth was selected from every patient in 24 patients with adult periodontitis. After supragingival scaling, the patients were divided into two groups randomly. One group was treated with Gu Chi Gao, while the other group was untreated as control group. All patients were examined after one month. GCF samples were collected with 2 mm x 20 mm Whatman I filter paper stripes and IL-8 in the GCF samples were detected by the method of ELISA. The result demonstrated that the levels of GCF and IL-8 in the treated group were lower than that of control group (P < 0.01), it suggests that Gu Chi Gao may inhibit the production of IL-8 and GCF.

Topics: Adult; Drugs, Chinese Herbal; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Male; Periodontitis

The purposes of this study were to localize monocyte chemoattractant protein-1-induced protein-1 (MCPIP-1) and its suppressor mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1) in gingival tissues and to profile their protein expression levels in relation to the clinical inflammation, Porphyromonas gingivalis colonization, and interleukin (IL)-8 levels.. Study samples were collected from two independent study populations: (1) Gingival tissues were collected from eight periodontally healthy individuals and eight periodontitis patients to localize MCPIP-1 and MALT-1 immunohistochemically, and (2) forty-one gingival tissue samples with marginal, mild, or moderate to severe inflammation were collected from 20 periodontitis patients to determine MCPIP-1 and MALT-1 levels using immunoblots, P. gingivalis levels with qPCR, P. gingivalis gingipain activities with fluorogenic substrates, and IL-8 levels with multiplex technique.. MCPIP-1 was detectable in the epithelium and in connective tissue, being especially prominent around the blood vessel walls in healthy periodontal tissues. MALT-1 was observed at all layers of gingival epithelium and especially around the accumulated inflammatory cells in connective tissue. No difference in gingival tissue MCPIP-1 and MALT-1 levels was observed in relation to the severity of gingival inflammation. MALT-1 levels were elevated (p = 0.023) with the increase in tissue P. gingivalis levels, and there was an association between MALT-1 and IL-8 levels (β = 0.054, p = 0.001).. Interactions of MALT-1 levels with gingival tissue P. gingivalis counts and IL-8 levels suggest that activation of MALT-1 can take part in P. gingivalis-regulated host immune responses.. Pharmacological targeting the crosstalk between immune response and MCPIP-1/MALT-1 may have benefits in periodontal treatment.

Topics: Gingiva; Humans; Inflammation; Interleukin-8; Periodontitis; Porphyromonas gingivalis

To observe the effect of miR-335-5p derived from human bone marrow mesenchymal stem cell (hBMMSCs) exosomes on osteogenic differentiation of human periodontal ligament stem cell (PDLSCs) model of periodontitis and explore its mechanism.. The exosomes extracted from hBMMSCs were identified by transmission electron microscopy, Western blotting and PKH67 labeling. The human PDLSC model of TNF-α-induced periodontitis were co-cultured with the extracted exosomes, and qRT-PCR was performed to detect the changes in the expressions of miR-335-5p and the mRNA levels of pro-inflammatory cytokines (IL-1β, IL-6, and IL-8) and the osteogenic marker genes (RunX2, OCN and BMP-2). Alizarin red staining and ALP staining were used to detect the formation of calcium nodules in the treated cells, and the expression level of DKK1 protein was detected with Western blotting. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-335-5p and DKK1.. High expressions of CD9 and CD81 were detected in the extracted hBMMSC exosomes (. HBMMSC exosome-derived miR-335-5p promotes osteogenic differentiation of hPDLSCs and inhibits the development of periodontitis by downregulating DKK1.

Topics: Bone Marrow Cells; Calcium; Cell Differentiation; Cells, Cultured; Core Binding Factor Alpha 1 Subunit; Exosomes; Humans; Interleukin-6; Interleukin-8; Mesenchymal Stem Cells; MicroRNAs; Osteogenesis; Periodontal Ligament; Periodontitis; RNA, Messenger; Stem Cells; Tumor Necrosis Factor-alpha

This study aimed to clarify the effects of Foxp3 silencing on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLFs) in an inflammatory environment and on cell proliferation and invasiveness, as well as to explore the role of Foxp3 gene in the development of periodontitis.. An small interfering RNA (siRNA) construct specific for Foxp3 was transfected into hPDLFs. Foxp3 silencing efficiency was verified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the siRNA with the optimum silencing effect of Foxp3 gene was screened. Using lipopolysaccharide to simulate an inflammatory environment. After siRNA transfection, RT-PCR and Western blotting analyses showed that the expression of Foxp3 mRNA in the Foxp3-si3 group decreased significantly (. In an inflammatory environment, Foxp3 gene silencing promoted hPDLFs migration but had no significant effect on hPDLFs proliferation. The expression of inflammatory factors expressed in hPDLFs increased after Foxp3 gene silencing, indicating that Foxp3 gene inhibited inflammation in periodontitis.

Topics: Cell Proliferation; Cells, Cultured; Cytokines; Fibroblasts; Forkhead Transcription Factors; Gene Silencing; Humans; Interleukin-6; Interleukin-8; Periodontal Ligament; Periodontitis; RNA, Small Interfering; Transcription Factors

Non-invasive physical plasma (NIPP), an electrically conductive gas, is playing an increasingly important role in medicine due to its antimicrobial and regenerative properties. However, NIPP is not yet well established in dentistry, although it has promising potential, especially for periodontological applications. The aim of the present study was to investigate the effect of NIPP on a commercially available human gingival fibroblast (HGF) cell line and primary HGFs in the presence of periodontitis-associated bacteria. First, primary HGFs from eight patients were characterised by immunofluorescence, and cell numbers were examined by an automatic cell counter over 5 days. Then, HGFs that were preincubated with

Topics: Cells, Cultured; Cytokines; Fibroblasts; Gingiva; Humans; Interleukin-6; Interleukin-8; Periodontitis

To investigate the pro-inflammatory cytokine profiles in patients with or without cardiovascular disease (CVD) and with or without peri-implantitis.. Serum, peri-implant crevicular fluid (PICF), and gingival crevicular fluid (GCF) were collected from patients with (n = 82) or without CVD (n = 46) at the most severe peri-implantitis site including sites with periodontitis. A panel of proinflammatory molecules including high-sensitivity C-reactive protein (hsCRP), fibrinogen, interleukin-1 beta (IL-1β), IL-6, plasma tumor necrosis factor-alpha (TNF-α), matrix metallo-proteinase-8 (MMP-8), osteoprotegerin (OPG), vascular endothelial growth factor (VEGF), IL-17, IL-8, tissue inhibitor of metalloproteinase-2 (TIMP-2), myeloperoxidase (MPO), and prostaglandin E. Serum IL-1β, TNF-α and fibrinogen were significantly higher in CVD patients than those without. Serum fibrinogen displayed a trend of higher concentration in patients with radiographic bone loss (RBL) ≥2 mm (P = 0.08). PICF TNF-α exhibited a significantly higher detection level in the CVD patients that is coincided with the local peri-implant inflammation. In addition, PICF MMP-8 was significantly higher in the RBL ≥2 mm sites than the healthy implants; whereas IL-1β, IL-8, MMP-8, and TIMP-2 proved to be the significant predictors for peri-implant disease. GCF TNF-α collected from patients with periodontitis was significantly associated with CVD cases.. The augmented expression of local and systemic pro-inflammatory cytokines found in the current study supports the weak association between the chronic peri-implantitis with increasing severity and CVD.

Topics: Cardiovascular Diseases; Dental Implants; Fibrinogen; Gingival Crevicular Fluid; Humans; Interleukin-8; Matrix Metalloproteinase 8; Peri-Implantitis; Periodontitis; Tissue Inhibitor of Metalloproteinase-2; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Implication of human caspase-4 in periodontitis and in sensing periodontal pathogens by gingival epithelial cells (GECs) is unclear. This study aimed to determine caspase-4 and interleukin (IL)-18 expressions in gingival tissues affected with periodontitis and to investigate caspase-4 involvement in mediating innate immune responses in GECs.. Ex vivo, caspase-4 and IL-18 expressions in gingival biopsies, obtained from healthy participants with periodontitis or clinically healthy gingiva (N = 20 each), were determined by immunohistochemistry. In vitro, caspase-4 activation in cultured GECs stimulated with Porphyromonas gingivalis or Fusobacterium nucleatum was analyzed by immunoblotting. mRNA expressions of human β-defensin-2 (hBD-2), IL-8, and IL-18 in stimulated GECs in the presence or absence of a caspase-4 inhibitor were assayed by RT-qPCR.. Ex vivo, compared with healthy gingival epithelium, the epithelium affected with periodontitis displayed a significant decrease in caspase-4 expression (P = 0.015), whereas IL-18 expression was significantly increased (P = 0.012). Moreover, the expression of caspase-4, but not IL-18, was found to be a predictor of periodontitis (P = 0.007). In vitro, caspase-4 was activated in cultured GECs challenged with P. gingivalis, but not F. nucleatum. mRNA upregulations of hBD-2, IL-8, and IL-18 upon P. gingivalis stimulation were significantly reduced when caspase-4 was inhibited (P < 0.05), whereas the inhibitor failed to suppress those inductions by F. nucleatum.. Caspase-4 expression is diminished in the epithelium affected with periodontitis while that of IL-18 is enhanced. Caspase-4 activation in P. gingivalis-infected GECs upregulates the three innate immune effector molecules, suggesting a possible sensing mechanism of caspase-4 in GECs in periodontal disease pathogenesis.

Topics: Bacteroidaceae Infections; Caspases, Initiator; Cells, Cultured; Epithelium; Gingiva; Humans; Interleukin-18; Interleukin-8; Periodontitis; Porphyromonas gingivalis; RNA, Messenger

This study investigated the prevalence and relative abundance of. Participants (age: 18-45 years) diagnosed with stage II-IV periodontitis, gingivitis, or periodontal health underwent periodontal examination and sampling. Subgingival plaque was analyzed by 16S+18S sequencing for. This cross-sectional study included 120 sites from 60 participants. The prevalence and relative abundance of

Topics: Adolescent; Adult; China; Cross-Sectional Studies; Entamoeba; Gingival Crevicular Fluid; Humans; In Situ Hybridization, Fluorescence; Interleukin-1beta; Interleukin-8; Middle Aged; Periodontitis; Porphyromonas gingivalis; Tumor Necrosis Factor-alpha; Young Adult

Peri-implant mucositis and peri-implantitis cases increase in number with the increase of implant applications. Peri-implant mucositis and peri-implantitis are defined as inflammatory diseases with inflammation and loss in soft and hard tissue, similar to the other periodontal diseases. As observed in many diseases, genetic predisposition factors also affect the progress of periodontitis and peri-implantitis.. This study examines if there is any solid genetic predisposition causing periodontitis and peri-implantitis formation in Turkish patients.. In order to evaluate single nucleotide polymorphism (SNP), Interleukin-8 (IL-8) and N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP), playing a role in the chemotaxis of neutrophils, and Fc Gamma Receptor IIA (FcγRIIA) and Fc Gamma Receptor IIIA (FcγRIIIA), playing a role in the antigen-antibody complexes and phagocytosis, were selected. Thirty-two Turkish non-smoking subjects, having periodontitis, thirty-three Turkish non-smoking subjects, having peri-implantitis and thirty-three Turkish non-smoking healthy subjects were selected. In total 98 adults participated in our study. Collected saliva samples from the participants were used for DNA isolation. SNPs were determined in these subgroups of the study by means of genotype-specific polymerase chain reactions.. When IL-8 A-251T, FcγRIIa -H131 and FcγRIIIa -V158 polymorphism were evaluated, no significant difference was found between periodontitis, peri-implantitis and healthy groups. However, this study observed that fMLP Receptor (FPR1) gene polymorphism creates a significant difference in individuals at higher risk of periodontitis or peri-implantitis.. Results show that individuals with the G genotype have a higher risk of periodontitis, while individuals with G / C genotype have higher risk of peri-implantitis.

Topics: Adult; Genetic Predisposition to Disease; Humans; Interleukin-8; Mucositis; Peri-Implantitis; Periodontitis; Polymorphism, Single Nucleotide

Neutrophil granulocytes have been proposed to play a major role in the mediation of periodontitis-associated tissue destruction. Their recruitment and activation are regulated by the chemokine CXCL8. This study aimed to delineate the dependency of CXCL8 expression in gingival crevicular fluid (GCF) and saliva on periodontal status, bacterial infection, and smoking, in patients with periodontitis.. The study cohort comprised 279 subjects with untreated periodontitis. Probing pocket depth (PPD), gingival recession, bleeding on probing (BOP), plaque index, and bone loss were evaluated. CXCL8 was determined in saliva and GCF using flow cytometry.. Considering the entire study sample, CXCL8 levels were correlated with the mean PPD (ρ = 0.131; p = 0.029), severity of periodontitis (ρ = 0.121; p = 0.043), BOP (ρ = 0.204; p = 0.001), and smoking (ρ = -0.219; p < 0.0001) in GCF; and, in whole saliva, with mean PPD (ρ = 0.154; p = 0.010) severity of periodontitis (ρ = 0.140; p = 0.020), gender (ρ = 0.178; p = 0.003), and smoking (ρ = -0.156; p = 0.010). Subgroup analysis among non-smokers revealed significantly higher amounts of CXCL8 in GCF (p = 0.012) and saliva (p = 0.026) comparing subjects with mean PPD ≤3mm and >3mm.. The current study revealed a strong dependency of CXCL8 expression in GCF on the severity and activity of periodontal disease. Smoking causes a significant reduction in CXCL8 expression in saliva and GCF.

Topics: Gingival Crevicular Fluid; Humans; Interleukin-8; Periodontal Attachment Loss; Periodontitis; Smoking

The aim of the present study was to evaluate the expressions of CXCL5, CXCL8, and CXCL10 in periodontal cells and tissues in response to microbial signals and/or biomechanical forces.. Human gingival biopsies from inflamed and healthy sites were used to examine the chemokine expressions and protein levels by real-time PCR and immunohistochemistry. The chemokines were also investigated in gingival biopsies from rats submitted to experimental periodontitis and/or tooth movement. Furthermore, chemokine levels were determined in human periodontal fibroblasts stimulated by the periodontopathogen Fusobacterium nucleatum and/or constant tensile forces (CTS) by real-time PCR and ELISA. Additionally, gene expressions were evaluated in periodontal fibroblasts exposed to F. nucleatum and/or CTS in the presence and absence of a MAPK inhibitor by real-time PCR.. Increased CXCL5, CXCL8, and CXCL10 levels were observed in human and rat gingiva from sites of inflammation as compared with periodontal health. The rat experimental periodontitis caused a significant (p<0.05) increase in alveolar bone resorption, which was further enhanced when combined with tooth movement. In vitro, F. nucleatum caused a significant upregulation of CXCL5, CXCL8, and CXCL10 at 1 day. Once the cells were exposed simultaneously to F. nucleatum and CTS, the chemokines regulation was significantly enhanced. The transcriptional findings were also observed at protein level. Pre-incubation with the MEK1/2 inhibitor significantly (p<0.05) inhibited the stimulatory actions of F. nucleatum either alone or in combination with CTS on the expression levels of CXCL5, CXCL8, and CXCL10 at 1d.. Our data provide original evidence that biomechanical strain further increases the stimulatory actions of periodontal bacteria on the expressions of these chemokines. Therefore, biomechanical loading in combination with periodontal infection may lead to stronger recruitment of immunoinflammatory cells to the periodontium, which might result in an aggravation of periodontal inflammation and destruction.

Topics: Animals; Chemokine CXCL10; Chemokine CXCL5; Fusobacterium nucleatum; Gingiva; Humans; Interleukin-8; Periodontal Ligament; Periodontitis; Periodontium; Rats; Stress, Mechanical

Salivary levels of interleukin-8 (IL-8) are elevated in patients with periodontitis. Caffeic acid phenethyl ester (CAPE) improves the periodontal status in subjects. However, whether CAPE can reduce IL-8 expression is unclear. We collected saliva to determine proinflammatory cytokine levels and used subgingival calculus and surrounding tissues from patients with periodontitis for oral microbiota analysis via 16s ribosomal RNA gene sequencing. THP-1 cells were stimulated with sterile-filtered saliva from patients, and target gene/protein expression was assessed. IL-8 mRNA expression was analyzed in saliva-stimulated THP-1 cells treated with CAPE and the heme oxygenase-1 (HO-1) inhibitor tin-protoporphyrin (SnPP). In 72 symptomatic individuals, IL-8 was correlated with periodontal inflammation (bleeding on probing,

Topics: Anti-Inflammatory Agents; Caffeic Acids; Cytokines; Heme Oxygenase-1; Humans; I-kappa B Proteins; Inflammation; Interleukin-8; Lipopolysaccharides; NF-kappa B; NF-KappaB Inhibitor alpha; Periodontitis; Phenylethyl Alcohol; Phosphorylation; Saliva; THP-1 Cells

Periodontitis is a polymicrobial inflammatory disease often characterized by the excessive colonization of Porphyromonas gingivalis and Fusobacterium nucleatum, which causes alveolar bone resorption and advanced oral inflammation. This study aimed to evaluate the effect of Limosilactobacillus fermentum CCFM1139 on experimental periodontitis induced following ligature and infection with P. gingivalis and F. nucleatum in vivo. The results showed that L. fermentum CCFM1139 significantly reduced weight loss associated with periodontal inflammation (p < 0.05), while decreasing both the P. gingivalis and F. nucleatum populations within the oral cavity of rats (p < 0.05) and regulating the expression of tumor necrosis factor-alpha, interleukin (IL)-1 beta, and IL-8 in the periodontal tissue (p < 0.05). Microcomputed tomography (micro-CT) and histopathological examination revealed that L. fermentum CCFM1139 supplementation reduced the level of alveolar bone loss and bone porosity and increased bone volume (p < 0.05) in the experimental animals. Furthermore, L. fermentum CCFM1139 exhibited promising effects in preventing the deepening of the periodontal pocket and the increase in the gap between adjacent molars. Thus L. fermentum CCFM1139 was shown to have solid potential as an oral probiotic for protection against periodontitis suggesting that this may be a good candidate in the production of a new functional food for improving periodontitis.

Topics: Alveolar Bone Loss; Animals; Disease Models, Animal; Fusobacterium nucleatum; Inflammation; Interleukin-1beta; Interleukin-8; Lactobacillaceae; Male; Mouth; Periodontitis; Porphyromonas gingivalis; Rats; Rats, Wistar; X-Ray Microtomography

Neutrophil plays a critical role in the progression of periodontitis. In general, its chemotaxis and activation are benefit for the host defense of bacterial infection and inflammation. However, previous studies have reported that the hyperactive and reactive neutrophils appear to be one of the reasons for tissue destruction in periodontitis tissues. In this study, we investigated an isoquinoline alkaloid Litcubanine A (LA), which from the Traditional Chinese medicinal plant, Litsea cubeba. We found LA showed significant activity in inhibiting neutrophils chemotaxis in the zebrafish yolk sac microinjection model in vivo and in mouse neutrophils in vitro. Further investigation proved that LA could inhibit the expression levels of neutrophil respiratory burst-related and inflammation-related genes CYBB and NCF2, as well as inhibit the activation of MAPK signaling pathway. Moreover, using LA, we successfully achieved the effect of reducing periodontitis bone loss by regulating neutrophil chemotaxis and related functions in a mouse ligature-induced periodontitis model.

Topics: Alkaloids; Animals; Bone Resorption; Chemotaxis; Gene Expression Regulation; Interleukin-8; Isoquinolines; Male; MAP Kinase Signaling System; Mice, Inbred C57BL; Microinjections; Neutrophil Infiltration; Neutrophils; Periodontitis; Receptors, Chemokine; Respiratory Burst; Yolk Sac; Zebrafish

Natural products constitute a promising class of therapeutics for the treatment of gingivitis and periodontitis as well as the maintenance of oral health. However, the limited understanding behind their potential mechanisms and modes of action have hampered their incorporation into popular western therapeutics. This in vitro study characterizes an Ayurvedic herbal extract mixture, which has been clinically shown to promote gingival health and homeostasis.. Telomerase immortalized gingival keratinocytes (TIGK) were infected with either Fusobacterium nucleatum cell wall, live F. nucleatum, IL-1β or TNF-α for 4 hours with and without the herbal extract. The immunomodulatory effects of the extract on host IL-8 production was measured by ELISA.. It was found that the Ayurvedic herbal extract mixture inhibited gingival epithelial cell IL-8 expression in response to both bacterial and host cytokine agonists. The herbal extract inhibited IL-8 stimulated by F. nucleatum cell wall, live F. nucleatum, IL-1β, and TNF-α in a dose-dependent manner that was not a result of host cell death. Furthermore, the extract showed significantly different ID. In vitro investigation of this herbal extract mixture revealed that it has the ability to modulate gingival epithelial cell IL-8 expression in response to stimulation by bacterial components and host pro-inflammatory signals. This data demonstrates that the reduction in the gingival epithelial cell IL-8 response may in part be responsible for the previously reported ability of the Ayurvedic herbal extract mixture to reduce gingivitis in two separate human clinical studies.

Topics: Cell Line; Epithelial Cells; Fusobacterium nucleatum; Gingiva; Humans; Interleukin-1beta; Interleukin-8; Medicine, Ayurvedic; Periodontitis; Plant Extracts; Tumor Necrosis Factor-alpha

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

Topics: Apoptosis; Bacteroidaceae Infections; Cell Line; Gingiva; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; NF-kappa B; Periodontitis; Porphyromonas gingivalis; RNA, Small Interfering; Signal Transduction; Tumor Necrosis Factor alpha-Induced Protein 3

Periodontitis is an inflammatory disease of the supporting tissue around teeth commonly caused by gram-negative bacterial infections. Interleukin (IL)-1β, a cytokine involved in host immune and inflammatory responses, is known to induce the activation of various intracellular signaling pathways. One of these signaling mechanisms involves the regulation of gene expression by activation of transcription factors (AP-1 and NF-κB). These transcription factors are controlled by mitogen-activated protein kinases (MAPKs), which increase cytokine and matrix metalloproteinase (MMP) expression. We examined the preventive effects of reversine, a 2,6-disubstituted purine derivative, on cytokine and MMP-3 expression in human gingival fibroblasts (HGFs) stimulated with IL-lβ.. Western blot analyses were performed to verify the activities of MAPK, p65, p50, and c-Jun and the expression of MMPs in IL-1β-stimulated HGFs. Cytokine and MMP-3 expression in IL-1β-stimulated HGFs was measured by real-time quantitative polymerase chain reaction.. Reversine decreased the IL-1β-induced expression of proinflammatory cytokines (IL-6 and IL-8) and MMP-3 in HGFs. Furthermore, the mechanism underlying the effects of reversine involved the suppression of IL-1β-stimulated MAPK activation and AP-1 activation.. Reversine inhibits IL-1β-induced MMP and cytokine expression via inhibition of MAPK/AP-1 activation and ROS generation. Therefore, we suggest that reversine may be an effective therapeutic candidate for preventing periodontitis.

Topics: Fibroblasts; Gingiva; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; MAP Kinase Kinase 4; Matrix Metalloproteinase 3; Morpholines; NF-kappa B; Periodontitis; Purines; Reactive Oxygen Species; Transcription Factor AP-1

Although previous studies revealed the potential use of probiotics in the control of periodontitis, little is known about their interactions with gingival epithelial cells (GECs). Since GECs comprise the first defense in the subgingival microenvironment, the aim of this study was to evaluate the effect of probiotic lactobacilli and bifidobacteria strains on OBA-9 cells challenged with Porphyromonas gingivalis.. Immortalized human GECs (OBA-9) were challenged with live P. gingivalis (strains W83 and ATCC33277) and co-infected with one of 12 tested probiotic strains at a multiplicity of infection (MOI) of 1:1000 for 2 hours. Bacterial adhesion and invasion were determined by antibiotic exclusion analysis and CFU counting. OBA-9 viability was assessed by MTT assay, and levels of inflammatory mediators (TNF-α, IL-1β, and CXCL8) in the supernatants were determined by ELISA. The expression of genes encoding Toll-like receptors (TLR2, TLR4) was evaluated by RT-qPCR.. Both strains of P. gingivalis were able to adhere and invade OBA-9 cells, with significant loss in cell viability, increase in the levels of TNF-α and IL-1β, and upregulation of TLR4. However, co-infection with probiotics attenuated these effects in P. gingivalis challenged GECs. Most probiotics maintained OBA-9 viability and reduced pathogens adhesion and invasion. Furthermore, probiotics were able to adhere to GECs, which was enhanced for most strains in the presence of P. gingivalis. The synthesis of IL-1β and TNF-α by P. gingivalis in challenged GECs was reduced in co-culture with most of the tested probiotics, whereas the secretion of CXCL8 increased, and TLR4 was downregulated.. Probiotics can alter the interaction of GECs with P. gingivalis by modulating the pathogen's ability to adhere and invade these cells, as well as by regulating the innate immune response. Such properties are strain-specific and may indicate the most efficient probiotics to control periodontitis.

Topics: Antibiosis; Bifidobacterium; Cells, Cultured; Cellular Microenvironment; Epithelial Cells; Gingiva; Humans; Immunity, Innate; Interleukin-1beta; Interleukin-8; Lactobacillus; Periodontitis; Porphyromonas gingivalis; Probiotics; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Vitamin D [1,25(OH). Determine whether glycated human serum albumin (G-HSA) augments HGF IL-6 and IL-8 production, and whether treatment with 1,25D3 attenuates cytokine production following stimulation with G-HSA + IL-1β and/or IL-17.. HGFs were incubated ±G-HSA or normal human serum albumin (HSA), ±IL-1β and/or IL-17, ±1,25D3. Cytokines were measured by ELISA. Neutralizing anti-RAGE was used to assess AGE-RAGE interaction. Endotoxin was measured using the ToxinSensor™ System. Data were expressed as mean ± standard deviation and analyzed using a one-way analysis of variance (ANOVA) and Scheffe's F procedure for post hoc comparisons.. G-HSA or IL-1β, but not HSA, significantly stimulated IL-6 and IL-8 production. G-HSA or HSA when combined with IL-1β or IL-1β + IL-17 synergistically stimulated IL-6 and IL-8. Neutralizing anti-RAGE inhibited IL-6 and IL-8 produced by cells stimulated with IL-1β + G-HSA but not (+HSA). Synergism caused by HSA did not appear to be mediated by endotoxin since its levels in G-HSA and HSA were not sufficient to stimulate fibroblasts. Vitamin D inhibited IL-6 and IL-8 production stimulated by G-HSA or HSA + IL-1β or IL-1β + IL-17.. Results suggest that the "perioprotective" effects of vitamin D are related to its ability to regulate inflammatory cytokine production by HGFs following AGE-RAGE interaction.

Topics: Calcitriol; Cell Line; Cholecalciferol; Depression, Chemical; Diabetes Mellitus; Endotoxins; Fibroblasts; Gingiva; Glycation End Products, Advanced; Humans; Inflammation Mediators; Interleukin-17; Interleukin-1beta; Interleukin-6; Interleukin-8; Periodontitis; Receptor for Advanced Glycation End Products; Serum Albumin, Human; Stimulation, Chemical

Interleukin (IL)-35 is a novel anti-inflammatory cytokine that is produced by regulatory T cells. IL-35 is reported to suppress IL-17A-producing helper T (Th17) cell activation. IL-17A is related to progression of periodontitis. Furthermore, IL-35 and IL-17A are detected in human gingival crevicular fluid. However, the effect of IL-35 and interaction between IL-35 and IL-17A on pro-inflammatory cytokine production in human periodontal resident cells are still unclear. The aim of this study was to clarify the effect of IL-35 on IL-6 and IL-8 production in human periodontal ligament cells (HPDLCs) stimulated with IL-17A. IL-35 inhibited IL-6 and IL-8 production in IL-17A-stimulated HPDLCs. Moreover, western blot analysis showed that IL-35 suppressed extracellular signal-regulated kinase (ERK) and nuclear factor (NF)-κB p65 phosphorylation in IL-17A-stimulated HPDLCs. Our findings suggested that IL-35 produced from regulatory T cells might inhibit progression of periodontitis by decreasing IL-17A-induced levels of IL-6 and IL-8.

Topics: Cells, Cultured; Extracellular Signal-Regulated MAP Kinases; Humans; Interleukin-17; Interleukin-6; Interleukin-8; Interleukins; NF-kappa B; Periodontal Ligament; Periodontitis; Phosphorylation; Signal Transduction; T-Lymphocytes, Regulatory

Ferritin, an iron-binding protein, is ubiquitous and highly conserved; it plays a crucial role in inflammation, which is the main symptom of periodontitis. Full-length cDNA library analyses have demonstrated abundant expression of ferritin in human periodontal ligament. The aims of the present study were to explore how ferritin is regulated by local inflammation, and to investigate its functions and mechanisms of action in the process of periodontitis.. Human gingival tissues were collected from periodontitis patients and healthy individuals. Experimental periodontitis was induced by ligature of second molars in mice. The expression of ferritin light polypeptide (FTL) and ferritin heavy polypeptide (FTH) were assessed by immunohistochemistry. Meanwhile, after stimulating human periodontal ligament cells (HPDLCs) with. Ferritin is up-regulated by inflammation and exhibits cytokine-like activity in HPDLCs inducing a signaling cascade that promotes expression of pro-inflammatory cytokines associated with periodontitis.

Topics: Animals; Antigens, CD; Apoferritins; Case-Control Studies; Cells, Cultured; Cytokines; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Ferritins; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Mice, Inbred C57BL; Oxidoreductases; p38 Mitogen-Activated Protein Kinases; Periodontal Ligament; Periodontitis; Receptors, Transferrin; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

In periodontitis patients, high levels of several inflammatory markers may be expressed in serum, reflecting the effect of local disease on the general health. The objective of the present analysis was to compare cytokine levels assessed in peripheral blood with those in the gingival crevicular fluid (GCF) and evaluate the impact of nonsurgical periodontal therapy on the incidence of high levels of 12 biomarkers in serum. Twenty-four patients with chronic periodontitis (Group P) contributed with serum and GCF samples at baseline (BL) and 1 and 3 months after periodontal treatment (M1 and M3). Samples were assessed for 12 cytokines using the Bio-Plex bead array multianalyte detection system. For each analyte, peak values were calculated as greater than the mean + 2

Topics: Adult; Case-Control Studies; Chemokine CCL4; Cytokines; Female; Gingival Crevicular Fluid; Humans; Inflammation; Interferon-gamma; Interleukin 1 Receptor Antagonist Protein; Interleukin-6; Interleukin-8; Male; Middle Aged; Periodontitis; Vascular Endothelial Growth Factor A; Young Adult

The aim of this study was to investigate the functionality of ATC/TTC (Hap-1) and ATT/TTC (Hap-2) Interleukin (IL) 8 gene haplotypes in the response of neutrophils to Gram-negative bacteria associated with periodontitis.. Neutrophils were isolated by gradient centrifugation from whole peripheral blood of systemically healthy individuals presenting the two IL8 gene haplotypes. Neutrophils were stimulated with P. gingivalis, A. actinomycetemcomitans and PMA/ionomycin. Cytokine gene expression (RT-qPCR) and migration/chemotaxis (boyden chamber assay) were compared according to the presence of Hap-1 or Hap-2 haplotypes. Protein production was also evaluted in the multiplex assay using the mixed population of leukocytes present in the whole blood from the same individuals. The influence of these two haplotypes on the IL8 promoter activity was assessed in gene-reporter experiments.. Hap-1 haplotype in neutrophils and leukocytes exacerbated the response to stimulation with Gram-negative bacteria, with higher levels of TNF-α (mRNA and protein), IL-1β, IL-2R and IFN-γ (protein) and with increased chemotaxis. Presence of the T allele at the rs4071 polymorphism (alias -251) was associated with increased activity of IL8 proximal promoter.. Neutrophils and leukocytes carrying the Hap-1 haplotype (ATC/TTC) in the IL8 gene present an enhanced response to stimulation with Gram-negative bacteria associated with periodontitis. Presence of the T allele (rs4073) in the IL8 proximal promoter increases transcription activity.

Topics: Cytokines; Genetic Predisposition to Disease; Gram-Negative Bacteria; Haplotypes; Humans; Interleukin-8; Neutrophils; Periodontitis; Pilot Projects; Promoter Regions, Genetic

Periodontal diseases originate from a dysbiosis within the oral microbiota, which is associated with a deregulation of the host immune response. Although little is known about the initiation of dysbiosis, it has been shown that H

Topics: Biofilms; Dysbiosis; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 8; Periodontitis; Polymerase Chain Reaction; Transcriptome; Tumor Necrosis Factor-alpha

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

Periodontitis is a highly prevalent infective and inflammatory disease with an adverse impact on systemic health. Isorhamnetin, a flavonoid mainly isolated from Hippophae fhamnoides L. fruit, has been reported to have anti-inflammatory effect. This study aimed to investigate the anti-inflammatory effects and mechanism of isorhamnetin on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). The production of inflammatory mediators and the expression of proteins were measured by ELISA and western blot analysis. The results demonstrated that isorhamnetin attenuated LPS-induced release of PGE

Topics: Anti-Inflammatory Agents; Cell Survival; Dinoprostone; Fibroblasts; Gingiva; Heme Oxygenase-1; Humans; Inflammation; Interleukin-6; Interleukin-8; Lipopolysaccharides; NF-E2-Related Factor 2; NF-kappa B; Periodontitis; Quercetin; Signal Transduction

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

Topics: Aggressive Periodontitis; Alleles; Case-Control Studies; Chronic Periodontitis; Female; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-8; Iran; Male; Periodontitis; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide

Naturally competent bacteria acquire DNA from their surroundings to survive in nutrient-poor environments and incorporate DNA into their genomes as new genes for improved survival. The secretin HofQ from the oral pathogen Aggregatibacter actinomycetemcomitans has been associated with DNA uptake. Cytokine sequestering is a potential virulence mechanism in various bacteria and may modulate both host defense and bacterial physiology. The objective of this study was to elucidate a possible connection between natural competence and cytokine uptake in A. actinomycetemcomitans. The extramembranous domain of HofQ (emHofQ) was shown to interact with various cytokines, of which IL-8 exhibited the strongest interaction. The dissociation constant between emHofQ and IL-8 was 43 nM in static settings and 2.4 μM in dynamic settings. The moderate binding affinity is consistent with the hypothesis that emHofQ recognizes cytokines before transporting them into the cells. The interaction site was identified via crosslinking and mutational analysis. By structural comparison, relateda type I KH domain with a similar interaction site was detected in the Neisseria meningitidis secretin PilQ, which has been shown to participate in IL-8 uptake. Deletion of hofQ from the A. actinomycetemcomitans genome decreased the overall biofilm formation of this organism, abolished the response to cytokines, i.e., decreased eDNA levels in the presence of cytokines, and increased the susceptibility of the biofilm to tested β-lactams. Moreover, we showed that recombinant IL-8 interacted with DNA. These results can be used in further studies on the specific role of cytokine uptake in bacterial virulence without interfering with natural-competence-related DNA uptake.

Topics: Aggregatibacter actinomycetemcomitans; Bacterial Outer Membrane Proteins; Bacterial Proteins; beta-Lactams; Biofilms; Cytokines; DNA-Binding Proteins; Fimbriae Proteins; Host-Pathogen Interactions; Humans; Interleukin-8; Periodontitis; Protein Interaction Domains and Motifs; Secretin; Virulence

Genetic factors play a role in periodontitis. Here we examined whether the risk haplotype of MHC class III region BAT1-NFKBIL1-LTA and lymphotoxin-α polymorphisms associate with salivary biomarkers of periodontal disease. A total of 455 individuals with detailed clinical and radiographic periodontal health data were included in the study. A 610 K genotyping chip and a Sequenom platform were used in genotyping analyses. Phospholipid transfer protein activity, concentrations of lymphotoxin-α, IL-8 and myeloperoxidase, and a cumulative risk score (combining Porphyromonas gingivalis, IL-1β and matrix metalloproteinase-8) were examined in saliva samples. Elevated IL-8 and myeloperoxidase concentrations and cumulative risk scores associated with advanced tooth loss, deepened periodontal pockets and signs of periodontal inflammation. In multiple logistic regression models adjusted for periodontal parameters and risk factors, myeloperoxidase concentration (odds ratio (OR); 1.37, P = 0.007) associated with increased odds for having the risk haplotype and lymphotoxin-α concentration with its genetic variants rs2857708, rs2009658 and rs2844482. In conclusion, salivary levels of IL-8, myeloperoxidase and cumulative risk scores associate with periodontal inflammation and tissue destruction, while those of myeloperoxidase and lymphotoxin-α associate with genetic factors as well.

Topics: Adaptor Proteins, Signal Transducing; Aged; Bacteroidaceae Infections; DEAD-box RNA Helicases; Female; Genetic Predisposition to Disease; Genotype; Haplotypes; Histocompatibility Antigens Class II; Humans; Interleukin-8; Lymphotoxin-alpha; Male; Matrix Metalloproteinases; Middle Aged; Periodontitis; Polymorphism, Single Nucleotide; Porphyromonas gingivalis; Risk; Saliva; Salivary Glands

Through a study of the molecular mechanism of the effect of resveratrol(RSV) on expression of TLR4 and inflammatory factors in gingival epithelial cells under high glucose environment, the therapeutic effect and molecular mechanism of resveratrol on periodontitis in patients with diabetes mellitus was investigated.. Gingival epithelial cells were cultured in vitro; according to the way of action, the cultured cells were divided into control group, high glucose group(HG) and HG+RSV group. The mRNA expression of TLR4 was detected by PCR; The third generation of gingival epithelial cells were pre-treated with or without RSV for 24 h under high glucose conditions, and subsequently treated with LPS at 100 ng/mL for 2 h. ELISA was used to detect the secretion of IL-1 beta, IL-6, IL-8 and TNF- alpha; the activation of TLR4 downstream signaling molecules NF-κB p65, p38 MAPK, and STAT3 was determined by Western blot. SPSS17.0 software package was used for statistical analysis.. RSV could reverse the increase of TLR4 level in gingival epithelial cells in high glucose medium.LPS markedly increased the expression and secretion of IL-1β, IL-6, IL-8, and TNF-α in GECs cultured in high glucose medium, which was partly blocked in the presence of RSV. Furthermore, Western blot results showed that RSV significantly suppressed the phosphorylation of TLR4 downstream factors NF-κB p65, p38MAPK, and STAT3.. RSV reduces inflammatory cytokine secretion in gingival epithelial cells, through negative regulation of TLR4 signaling pathway.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Line; Cells, Cultured; Epithelial Cells; Gingiva; Glucose; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; NF-kappa B; Periodontitis; Resveratrol; Signal Transduction; Stilbenes; Toll-Like Receptor 4; Transcription Factor RelA; Tumor Necrosis Factor-alpha

The aim of this study was to clarify the anti-inflammatory effects and its molecular mechanism of glycyrrhizin on LPS-stimulated human gingival fibroblasts (HGFs), which will be of benefit for periodontitis treatment. An MTT assay was performed to assess the effects of glycyrrhizin on cellular viability. The levels of IL-6 and IL-8 were measured by ELISA. The expression of iNOS, COX-2, NF-κB, and LXRα were detected by western blot analysis. The results showed that glycyrrhizin significantly inhibited LPS-induced IL-6 and IL-8 production, as well as COX-2 and iNOS expression. LPS-induced NF-κB activation in HGFs was also inhibited by treatment of glycyrrhizin. Furthermore, glycyrrhizin increased the expression of LXRα in a concentration-dependent manner. In addition, the inhibition of glycyrrhizin on IL-6 and IL-8 production was reversed by LXRα inhibitor GGPP. In conclusion, these results indicated that glycyrrhizin exhibited its anti-inflammatory effects in HGFs by activating LXRα.

Topics: Cell Survival; Cells, Cultured; Cyclooxygenase 2; Cytokines; Fibroblasts; Gene Expression Regulation; Gingiva; Glycyrrhizic Acid; Humans; Interleukin-6; Interleukin-8; Lipopolysaccharides; Liver X Receptors; NF-kappa B; Nitric Oxide Synthase Type II; Periodontitis; Signal Transduction

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

Varying cytokine responses of human gingival epithelial cells (HGECs) by Aggregatibacter actinomycetemcomitans subtypes have been found. Most studies have used reference strains, whereas a few has evaluated the cytokine expression in response to clinical subtypes of this bacterial species. This study aimed to examine whether there was any difference in cytokine responses of HGECs stimulated with cell wall extract (CWE) from A. actinomycetemcomitans subtypes included clinical strains from Thai adult periodontitis, various serotypes and non-serotypeable strains, strains from deep or shallow pockets, and reference serotype strains. Totally 50 clinical strains and 7 reference strains of A. actinomycetemcomitans were analyzed for the expression of IL-1β, IL-6, IL-8, and TNF-α mRNAs in HGECs by real time-PCR, and the IL-8 concentrations in cell-free supernatant measured using ELISA. An in vitro effect of released IL-8 on neutrophil migration was examined using transwell chambers. Result showed that among four cytokines studied, IL-8 mRNA was highly up-regulated by both clinical and reference strains. Serotype f revealed the highest expression compared to other serotypes. The JP2-like leukotoxin promoter gene and non-serotypeable (NS1 and NS2) demonstrated lower IL-8 responses compared to serotypeable strains, and IL-8 responses upon stimulation with clinical strains from deep pockets were also significantly lower than those isolated from shallow pockets (P < 0.01). Our findings suggest that the clinical isolates of A. actinomycetemcomitans associating with deep pockets, JP2-like leukotoxin promoter gene, NS1, and NS2 may interfere neutrophil function via minimal and immunosuppressing IL-8 responses, which may enhance their survival and virulence.

Topics: Aggregatibacter actinomycetemcomitans; Cell Movement; Cell Wall; Cells, Cultured; Exotoxins; Gingiva; Gingival Pocket; Humans; Interleukin-8; Neutrophils; Periodontitis; Promoter Regions, Genetic; RNA, Messenger

Streptococcus gordonii, a Gram-positive oral bacterium, is a life-threatening pathogen that causes infective endocarditis. It is frequently isolated from the periapical lesions of patients with apical periodontitis and has thus been implicated in inflammatory responses. However, little is known about the virulence factors of S. gordonii responsible for the induction of inflammatory responses in the periapical areas. Here, we investigated the role of S. gordonii cell wall-associated virulence factors on interleukin (IL)-8 induction in human periodontal ligament (PDL) cells using ethanol-inactivated wild-type S. gordonii, a lipoteichoic acid (LTA)-deficient mutant (ΔltaS), and a lipoprotein-deficient mutant (Δlgt). Wild-type S. gordonii induced IL-8 expression at both the protein and mRNA levels in human PDL cells in a dose- and time-dependent manner. A transient transfection and reporter gene assay demonstrated that wild-type S. gordonii activated Toll-like receptor 2 (TLR2). Additionally, IL-8 production induced by wild-type S. gordonii was substantially inhibited by anti-TLR2-neutralizing antibodies. Both wild-type S. gordonii and the ΔltaS mutant induced IL-8 production; however, this response was not observed when cells were stimulated with the Δlgt mutant. Interestingly, lipoproteins purified from S. gordonii induced IL-8 production, whereas purified LTA did not. In addition, purified lipoproteins stimulated TLR2 more potently than LTA. Furthermore, S. gordonii-induced IL-8 expression was specifically inhibited by blocking p38 kinase, while lipoprotein-induced IL-8 expression was inhibited by blocking p38 kinase, ERK, or JNK. Of particular note, exogenous addition of purified S. gordonii lipoproteins enhanced Δlgt-induced IL-8 production in human PDL cells to an extent similar to that induced by the wild-type strain. Collectively, these results suggest that lipoproteins are an important component of S. gordonii for the induction of IL-8 production in human PDL cells through TLR2 activation. Therefore, lipoproteins potentially contribute to inflammatory apical periodontitis.

Topics: Bacterial Proteins; Gene Expression Regulation; HEK293 Cells; Humans; Interleukin-8; Lipoproteins; Mutation; p38 Mitogen-Activated Protein Kinases; Periodontal Ligament; Periodontitis; Streptococcus gordonii; Toll-Like Receptor 2

Porphyromonas gingivalis is regarded as a significant contributor in the pathogenesis of periodontitis and certain systemic diseases, including atherosclerosis. P. gingivalis occasionally translocates from periodontal pockets into the circulation, where it adheres to red blood cells (RBCs). This may protect the bacterium from contact with circulating phagocytes without affecting its viability.. In this in vitro study, we investigated whether human peripheral blood neutrophils from 10 subjects with localized aggressive periodontitis (LAgP) and 10 healthy controls release the proinflammatory cytokines interleukin (IL)-6, tumor necrosis factor α (TNF-α), the chemokine (C-X-C motif) ligand 8 (CXCL8; also known as IL-8) and chemokine (C-C motif) ligand 2 (CCL2; also known as monocyte chemotactic protein-1) and intracellular reactive oxygen species (ROS) in response to challenge with P. gingivalis. In addition, the impact of RBC interaction with P. gingivalis was investigated. The actions of resolvin E1 (RvE1), a known regulator of P. gingivalis induced neutrophil responses, on the cytokine and ROS responses elicited by P. gingivalis in cultures of neutrophils were investigated.. Upon stimulation with P. gingivalis, neutrophils from subjects with LAgP and healthy controls released similar quantities of IL-6, TNF-α, CXCL8, CCL2 and intracellular ROS. The presence of RBCs amplified the release of IL-6, TNF-α and CCL2 statistically significant in both groups, but reduced the generation of ROS in the group of healthy controls, and showed a similar tendency in the group of subjects with LAgP. RvE1 had no impact on the production of intracellular ROS, TNF-α, IL-6, CXCL8 and CCL2 by neutrophils from either group, but tended to reduce the generation of ROS in subjects with LAgP in the absence of RBCs.. Our data support that binding to RBCs protects P. gingivalis from ROS and concomitantly enhances neutrophil release of proinflammatory cytokines providing a selective advantage for P. gingivalis growth.

Topics: Adult; Aged; Chemokine CCL2; Eicosapentaenoic Acid; Erythrocytes; Female; Humans; In Vitro Techniques; Interleukin-6; Interleukin-8; Male; Middle Aged; Neutrophils; Periodontitis; Porphyromonas gingivalis; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Periodontitis is a chronic inflammatory disease initiated by bacteria and their virulence factors. Bortezomib (BTZ) is the first proteasome inhibitor for clinical treatment of malignancies. Its anticancer activity is accompanied by an anti-inflammatory effect. However, there are few reports about its anti-inflammatory effect and underlying mechanism in periodontal disease, especially on human periodontal ligament cells (hPDLCs), which are considered a promising cell-based therapy for treating periodontitis.. hPDLCs were treated with lipopolysaccharide (LPS) and pretreated with BTZ. mRNA and protein levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1β, IL-6, and IL-8 were determined. The anti-inflammatory mechanism of BTZ was studied. Further, experimental rat periodontitis was induced with ligature and LPS injection, and simultaneously and locally treated with BTZ (three injections/week). Four weeks after treatment, microcomputed tomography, immunohistochemical, and histopathologic analyses were performed.. Bortezomib administration at safe concentrations (≤1 nM) inhibited production of proinflammatory cytokines in LPS-stimulated hPDLCs via nuclear factor (NF)-kappa B, p38/extracellular signal-regulated kinase, and mitogen-activated protein kinase/activator protein-1 pathways. Moreover, in the LPS and ligature-induced periodontitis rat model, BTZ suppressed expression of TNF-α, IL-1β, IL-6, and IL-8, reduced the ratio of receptor activator of NF-κB ligand/osteoprotegerin, and prevented alveolar bone absorption.. These findings demonstrate the anti-inflammatory activity of BTZ against periodontal inflammatory response and present BTZ as a promising therapy for periodontal disease.

Topics: Adolescent; Animals; Bortezomib; Child; Disease Models, Animal; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Periodontal Ligament; Periodontitis; Proteasome Inhibitors; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

The Wingless/integrase-1 (Wnt) family of protein ligands and their functional antagonists, secreted frizzled-related proteins (sFRPs), regulate various biological processes ranging from embryonic development to immunity and inflammation. Wnt5a and sFRP5 comprise a typical ligand/antagonist pair, and the former molecule was recently detected at the messenger RNA (mRNA) level in human periodontitis. The main objective of this study was to investigate the interrelationship of expression of Wnt5a and sFRP5 in human periodontitis (as compared to health) and to determine their roles in inflammation and bone loss in an animal model. We detected both Wnt5a and sFRP5 mRNA in human gingiva, with Wnt5a dominating in diseased and sFRP5 in healthy tissue. Wnt5a and sFRP5 protein colocalized in the gingival epithelium, suggesting epithelial cell expression, which was confirmed in cultured human gingival epithelial cells (HGECs). The HGEC expression of Wnt5a and sFRP5 was differentially regulated by a proinflammatory stimulus (lipopolysaccharide [LPS] from Porphyromonas gingivalis) in a manner consistent with the clinical observations (i.e., LPS upregulated Wnt5a and downregulated sFRP5). In HGECs, exogenously added Wnt5a enhanced whereas sFRP5 inhibited LPS-induced inflammation, as monitored by interleukin 8 production. Consistent with this, local treatment with sFRP5 in mice subjected to ligature-induced periodontitis inhibited inflammation and bone loss, correlating with decreased numbers of osteoclasts in bone tissue sections. As in humans, mouse periodontitis was associated with high expression of Wnt5a and low expression of sFRP5, although this profile was reversed after treatment with sFRP5. In conclusion, we demonstrated a novel reciprocal relationship between sFRP5 and Wnt5a expression in periodontal health and disease, paving the way to clinical investigation of the possibility of using the Wnt5a/sFRP5 ratio as a periodontitis biomarker. Moreover, we showed that sFRP5 blocks experimental periodontal inflammation and bone loss, suggesting a promising platform for the development of a new host modulation therapy in periodontitis.

Topics: Alveolar Bone Loss; Animals; Biomarkers; Biopsy; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Gingiva; Glycoproteins; Humans; Interleukin-8; Intracellular Signaling Peptides and Proteins; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Periodontitis; Polymerase Chain Reaction; Porphyromonas gingivalis; RNA, Messenger; Wnt-5a Protein

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

Adipokines enhance the synthesis of proinflammatory cytokines and matrix metalloproteinases (MMPs), which play a role in extracellular matrix degeneration. The aim of this study is to determine the levels of some adipokines, proinflammatory cytokines, and MMPs in the saliva of patients with periodontitis and healthy individuals and to evaluate the changes after non-surgical periodontal therapy (NSPT).. Of 32 individuals included in the study, 17 had periodontitis and 15 had healthy gingiva. Saliva samples were obtained from all individuals. In patients with periodontitis, samples were recollected 3 and 6 months after NSPT. Visfatin, chemerin, progranulin, interleukin (IL)-1β, IL-8, MMP-8, and MMP-13 levels were measured using enzyme-linked immunosorbent assay.. In patients with periodontitis, all of the parameters measured in the saliva were higher than those of healthy individuals. At 3 months, visfatin, progranulin, IL-8, and MMP-8 levels were significantly decreased compared with baseline values. The levels of other biochemical parameters, chemerin and IL-1β, were significantly decreased compared with baseline values at 6 months, and the levels became similar to those in healthy individuals. In the periodontitis group, positive correlations were found among visfatin and IL-8 (r = 0.909, P <0.01), MMP-8 (r = 0.702, P = 0.02), and MMP-13 (r = 0.781, P = 0.01); chemerin and IL-8 (r = 0.913, P <0.01), MMP-8 (r = 0.770, P <0.01), and MMP-13 (r = 0.788, P <0.01); and progranulin and IL-8 (r = 0.762, P <0.01), MMP-8 (r = 0.845, P <0.01), and MMP-13 (r = 0.813, P <0.01).. Adipokines may contribute to the breakdown of periodontal tissue in periodontitis by stimulating the expression of proinflammatory cytokines and MMPs.

Topics: Adipokines; Case-Control Studies; Humans; Interleukin-8; Matrix Metalloproteinase 8; Matrix Metalloproteinases; Periodontitis; Saliva

This study aims to evaluate the association of salivary matrix metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) with periodontitis and its screening ability on periodontitis.. We selected 693 participants from the Yanypyeong Cohort: 577 participants with periodontitis and 116 participants without periodontitis. Periodontitis was assessed by dentists using a panoramic radiograph. Salivary MMP-9 and IL-8 were assayed using multiplexed bead immunoassay (Luminex). MMP-9 and IL-8 were categorized into low, medium and high. Age, sex, income, smoking, drinking, exercise, obesity and metabolic syndrome were confounders. Logistic regression analysis was applied to evaluate the association between MMP-9, IL-8 and periodontitis. Receiver operating characteristic curve was applied for sensitivity, specificity and c-statistics.. High MMP-9 and medium IL-8 were associated with periodontitis: adjusted odds ratio were 2.2 [95% confidence interval (CI): 1.3-3.7] for MMP-9 and 1.9 (95% CI: 1.1-3.4) for IL-8. The final screening model using salivary MMP-9 for periodontitis had a sensitivity of 0.46, specificity of 0.77 and c-statistic of 0.63 (p < 0.001).. Our data show that salivary MMP-9 and IL-8 could be potential markers for periodontitis. The screening model for periodontitis could be useful in clinics and home. A future prospective study is indicated for predicting the occurrence of periodontitis.

Topics: Adult; Asian People; Humans; Interleukin-8; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Periodontitis; Prospective Studies

The interaction between human gingival fibroblasts (HGFs) and Porphyromonas gingivalis plays an important role in the development and progression of periodontitis. Porphyromonas gingivalis possesses several virulence factors, including cysteine proteases, the arginine-specific (Rgp) and lysine-specific (Kgp) gingipains. Studying the mechanisms that P. gingivalis, and its derived virulence, use to propagate and interact with host cells will increase the understanding of the development and progression of periodontitis. In this study, we aimed to elucidate how P. gingivalis influences the inflammatory events in HGFs regarding transforming growth factor-β1 (TGF-β1 ), CXCL8, secretory leucocyte protease inhibitor (SLPI), c-Jun and indoleamine 2,3-dioxygenase (IDO). HGFs were inoculated for 6 and 24 h with the wild-type strains ATCC 33277 and W50, two gingipain-mutants of W50 and heat-killed ATCC 33277. The P. gingivalis regulated CXCL8 and TGF-β1 in HGFs, and the kgp mutant gave significantly higher immune response with increased CXCL8 (P < 0.001) and low levels of TGF-β1 . We show that HGFs express and secrete SLPI, which was significantly suppressed by P. gingivalis (P < 0.05). This suggests that by antagonizing SLPI, P. gingivalis contributes to the tissue destruction associated with periodontitis. Furthermore, we found that P. gingivalis inhibits the expression of the antimicrobial IDO, as well as upregulating c-Jun (P < 0.05). In conclusion, P. gingivalis both triggers and suppresses the immune response in HGFs. Consequently, we suggest that the pathogenic effects of P. gingivalis, and especially the activity of the gingipains on the inflammatory and immune response of HGFs, are crucial in periodontitis.

Topics: Adhesins, Bacterial; Cells, Cultured; Cysteine Endopeptidases; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gingipain Cysteine Endopeptidases; Gingiva; Humans; Indoleamine-Pyrrole 2,3,-Dioxygenase; Inflammation; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Proto-Oncogene Proteins c-jun; Real-Time Polymerase Chain Reaction; Secretory Leukocyte Peptidase Inhibitor; Transforming Growth Factor beta

To investigate if there is subspecies specific migration to the placenta by Fusobacterium nucleatum (Fn) and to determine whether experimentally induced periodontitis results in adverse pregnancy outcomes (APO) in mice.. Periodontitis was induced in pregnant mice using an inoculum of Fn and Porphyromonas gingivalis. In parallel, four sub-species of Fn were individually injected into the circulatory system. At day 18 of gestation, the placenta, liver, spleen and blood were harvested and litter size, number of viable fetuses and resorptions, maternal, fetal and placenta weights were recorded. For the direct inoculation group, some mice were allowed to deliver for assessment of length of gestation, litter size, maternal, placental and pup weight. The presence of Fn was assessed by PCR and inflammatory mediators were measured by ELISA or multiplex analysis.. Mice with alveolar bone loss, a marker of periodontitis, demonstrated significantly higher fetal weights (p = 0.015) and fetal/placental weight ratios (p = 0.030). PCR analysis of maternal organs did not identify Fn in any extracted tissues. In mice that received direct injection of Fn subspecies, varying degrees of APO were observed including preterm birth, intrauterine growth restriction, and fetal loss. Haematogenous spread of only Fn subsp. nucleatum to the placenta was confirmed. Litter size was significantly smaller (p = 0.023) and the number of resorptions was higher in inoculated versus control groups. Mice injected with subsp. nucleatum had significantly increased circulating CRP levels (p = 0.020) compared to controls while the mice with induced periodontitis had increased levels of IL-6 (p = 0.047) and IL-8 (p = 0.105).. Periodontitis in mice elevated fetal weight and the fetal weight/placental weight ratio. This study found that subsp. nucleatum migrated haematogenously to the placenta, leading to APO in mice. The study supports the potential role of Fn in the association between periodontitis and APO.

Topics: Alveolar Bone Loss; Animals; C-Reactive Protein; Disease Models, Animal; Female; Fusobacterium nucleatum; Interleukin-6; Interleukin-8; Mice; Mice, Inbred BALB C; Periodontitis; Placenta; Polymerase Chain Reaction; Porphyromonas; Pregnancy; Pregnancy Complications, Infectious; Pregnancy Outcome; Radiography; RNA, Ribosomal, 16S

Imbalance or disruption in the expression of inflammatory mediators contributes greatly to the breakdown of the periodontal supporting tissues. Leptin, through binding to its receptor (obesity-related leptin and leptin receptor [OBR]), has potent effects on immunity and inflammation. However, to date, researchers only indicated a role of leptin in periodontitis. No direct or valid evidence exists about how leptin and its receptor are regulated by local inflammation, what effects they have, and the underlying mechanisms.. Experimental periodontitis was induced by ligation of mandibular second molars in beagle dogs. The expression of leptin, OBR, and interleukin (IL)-1β was examined by immunohistochemistry. Meanwhile, recombinant human IL-1β was used to stimulate human periodontal ligament cells (hPDLCs) in vitro, and mRNA and protein levels of leptin were measured using real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Then, mRNA and protein levels of IL-6 and IL-8 were measured using real-time PCR and ELISA, after stimulation with various concentrations of leptin, knocking down all or only the long form of OBR (OBRb) by small interfering RNA and incubation with multiple intracellular signaling pathway inhibitors, respectively.. Leptin and OBR increased substantially in inflammatory periodontal tissues, which correlated well with the extent of inflammatory infiltration, and was a result of the upregulation in resident cells themselves. A high dose of leptin could induce the expression of mRNA and protein of IL-6 and IL-8 in hPDLCs through binding with OBRb and activating different intracellular signaling pathways.. Upregulated leptin and OBR in periodontitis stimulated proinflammatory cytokine expression in PDL cells to additionally promote local inflammation.

Topics: Alveolar Process; Animals; Cell Culture Techniques; Cells, Cultured; Dental Plaque Index; Dogs; Enzyme Inhibitors; Gene Knockdown Techniques; Gingiva; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Janus Kinase 2; Leptin; MAP Kinase Signaling System; Periodontal Index; Periodontal Ligament; Periodontitis; Phosphoinositide-3 Kinase Inhibitors; Receptors, Leptin; RNA, Small Interfering; Signal Transduction; Up-Regulation

Vitamin D has important roles on control of calcium and phosphate levels in the body. However, the role of vitamin D on the pathogenesis of periodontal disease is still uncertain. Therefore, we examined the effect of the hormonal form of vitamin D, calcitriol, on inflammatory responses of human periodontal ligament cells (HPDLC). We detected vitamin D receptor expression in non-stimulated HPDLC. Calcitriol inhibited interleukin (IL)-6, IL-8, CC chemokine ligand (CCL) 20, CXC chemokine ligand (CXCL) 10, and matrix metalloproteinase (MMP)-3 release from IL-1β-stimulated HPDLC. Tissue inhibitor of metalloproteinase (TIMP)-1 production did not change by calcitriol. Moreover, we found c-jun N-terminal kinase (JNK) phosphorylation and IκB-α degradation in IL-1β-stimulated HPDLC were inhibited by calcitriol, and JNK and nuclear factor (NF)-κB inhibitors could decrease IL-6, IL-8, CCL20, CXCL10, and MMP-3 productions in IL-1β-treated HPDLC. These findings suggest that vitamin D could modulate inflammatory response in periodontal tissues.

Topics: Anti-Inflammatory Agents; Calcitriol; Cells, Cultured; Chemokine CCL20; Chemokine CXCL10; Dose-Response Relationship, Drug; Humans; I-kappa B Proteins; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Matrix Metalloproteinase 3; NF-KappaB Inhibitor alpha; Periodontal Ligament; Periodontitis; Phosphorylation; Proteolysis; Receptors, Calcitriol; Signal Transduction; Time Factors

Periodontitis is the most common chronic inflammatory condition occurring in the human oral cavity, but our knowledge on its contribution to oral cancer is rather limited. To define crosstalk between chronic periodontitis and oral cancer, we investigated whether Porphyromonas gingivalis, a major pathogen of chronic periodontitis, plays a role in oral cancer progression. To mimic chronic irritation by P. gingivalis in the oral cavity, oral squamous cell carcinoma (OSCC) cells were infected with P. gingivalis twice a week for 5 weeks. Repeated infection of oral cancer cells by P. gingivalis resulted in morphological changes of host cancer cells into an elongated shape, along with the decreased expression of epithelial cell markers, suggesting acquisition of an epithelial-to-mesenchymal transition (EMT) phenotype. The prolonged exposure to P. gingivalis also promoted migratory and invasive properties of OSCC cells and provided resistance against a chemotherapeutic agent, all of which are described as cellular characteristics undergoing EMT. Importantly, long-term infection by P. gingivalis induced an increase in the expression level of CD44 and CD133, well-known cancer stem cell markers, and promoted the tumorigenic properties of infected cancer cells compared to non-infected controls. Furthermore, increased invasiveness of P. gingivalis-infected OSCC cells was correlated with enhanced production of matrix metalloproteinase (MMP)-1 and MMP-10 that was stimulated by interleukin-8 (IL-8) release. This is the first report demonstrating that P. gingivalis can increase the aggressiveness of oral cancer cells via epithelial-mesenchymal transition-like changes and the acquisition of stemness, implicating P. gingivalis as a potential bacterial risk modifier.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Matrix Metalloproteinase 1; Mouth Neoplasms; Neoplastic Stem Cells; Periodontitis; Porphyromonas gingivalis

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride (NaF) on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin (IL)-1β, matrix metalloproteinase 9 (MMP9), and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL (receptor activator of nuclear factor κB ligand) and macrophage colony-stimulating factor (M-CSF). The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells (NFAT) c1 was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATc1, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Bacteroidaceae Infections; Bone Density Conservation Agents; Cathepsin K; Interleukin-1beta; Interleukin-6; Interleukin-8; Isoenzymes; Macrophage Colony-Stimulating Factor; Male; Matrix Metalloproteinase 9; Osteoclasts; Periodontitis; Porphyromonas gingivalis; RANK Ligand; Rats; Rats, Sprague-Dawley; Sodium Fluoride; Tartrate-Resistant Acid Phosphatase; Transcription Factors; X-Ray Microtomography

Periodontitis is an inflammatory disease of polymicrobial origin affecting the tissues supporting the tooth. The oral anaerobic bacterium Porphyromonas gingivalis, which is implicated as an important pathogen for chronic periodontitis, triggers a series of host inflammatory responses that promote the destruction of periodontal tissues. Among the virulence factors of P. gingivalis, hemoglobin receptor protein (HbR) is a major protein found in culture supernatants. In this study, we investigated the roles of HbR in the production of inflammatory mediators. We found that HbR induced interleukin-8 (IL-8) production in the human gingival epithelial cell line Ca9-22. p38 mitogen-activated protein kinase (MAPK) and extracellular signal-related kinase 1/2 (Erk1/2) were activated in HbR-stimulated Ca9-22 cells. Inhibitors of p38 MAPK (SB203580) and Erk1/2 (PD98059) blocked HbR-induced IL-8 production. Additionally, HbR stimulated the translocation of NF-κB-p65 to the nucleus, consistent with enhancement of IL-8 expression by activation of the NF-κB pathway. In addition, small interfering RNA (siRNA) targeting activating transcription factor 2 (ATF-2) or cyclic AMP-response element-binding protein (CREB) inhibited HbR-induced IL-8 production. Moreover, pretreatment with SB203580 and PD98059 reduced HbR-induced phosphorylation of CREB and ATF-2, respectively. Combined pretreatment with an inhibitor of NF-κB (BAY11-7082) and SB203580 was more efficient in inhibiting the ability of HbR to induce IL-8 production than pretreatment with either BAY11-7082 or SB203580 alone. Thus, in Ca9-22 cells, the direct activation of p38 MAPK and Erk1/2 by HbR caused the activation of the transcription factors ATF-2, CREB, and NF-κB, thus resulting in the induction of IL-8 production.

Topics: Bacterial Proteins; Bacteroidaceae Infections; Cells, Cultured; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gingiva; Humans; Interleukin-8; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Periodontitis; Porphyromonas gingivalis; Protein Kinase Inhibitors; Receptors, Immunologic; Signal Transduction

Periodontitis is a chronic inflammatory disease induced by bacteria. Exposure of the host to periodontal pathogens and their virulence factors induces a hyporesponsive state to subsequent challenge, which is termed endotoxin tolerance. In this experiment, we studied the cytokine production in THP-1 cells upon single or repeated Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) or Escherichia coli (E. coli) LPS stimulation by ELISA. In addition, the protein expression profiles of Toll-like receptor 2 (TLR2), TLR4, IL-1 receptor-associated kinase 4 (IRAK4) and IRAK-M and the gene expression changes of Toll-interacting protein (Tollip) and suppressor of cytokine-signaling-1 (SOCS1) were explored to identify possible mechanisms for changes in cytokine secretion. After repeated stimulation with P. gingivalis LPS or E. coli LPS, secretions of TNF-α and IL-1β were decreased significantly compared with those following single challenge, while the levels of IL-10 were increased (p < 0.05). Only comparable levels of IL-8 were confirmed in P. gingivalis LPS-tolerized cells (p > 0.05). In addition, severe downregulation of TLR2 was detected in THP-1 cells retreated with P. gingivalis LPS, and the reduction of TLR4 expression was observed in cells restimulated with E. coli LPS (p < 0.05). Precondition with P. gingivalis LPS or E. coli LPS also led to an enhancement of IRAK-M and SOCS1, while maintaining the expressions of IRAK4 and Tollip. This pattern of cytokine production indicates the different effects of endotoxin tolerance triggered by P. gingivalis LPS and E. coli LPS, which might contribute to limiting inflammatory damage. Moreover, TLR2, TLR4, IRAK-M, and SOCS1 might play important roles in developing tolerance.

Topics: Cell Line; Down-Regulation; Endotoxins; Escherichia coli; Humans; Immune Tolerance; Inflammation; Interleukin-1 Receptor-Associated Kinases; Interleukin-10; Interleukin-1beta; Interleukin-8; Intracellular Signaling Peptides and Proteins; Lipopolysaccharides; Periodontitis; Porphyromonas gingivalis; Signal Transduction; Suppressor of Cytokine Signaling 1 Protein; Suppressor of Cytokine Signaling Proteins; Toll-Like Receptor 2; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Periodontitis is a chronic inflammatory disease characterized by destruction of periodontal tissue ultimately leading to bone destruction and has been associated with other inflammatory diseases, such as atherosclerosis. Attachment loss of periodontal tissue is primarily caused by host cell-derived immune responses against subgingival biofilm. The aim of the present study was to determine the cytokine profile in serum, saliva and gingival crevicular fluid (GCF) patients with periodontitis and healthy controls. We show that periodontitis patients exhibit higher numbers of periodontal pathogens and their immune responses are significantly altered. The levels of IL-6 in saliva and GCF were significantly suppressed, and while CXCL8 was not altered in serum, its expression levels were significantly suppressed in saliva and elevated in GCF. The T-cell-derived cytokine IL-2 did not differ between patients and controls in serum and saliva, but there was a significant suppression in GCF of patients. Interestingly, TGF-β1 levels were significantly elevated in serum, saliva and GCF in patients compared to controls. Furthermore, by using cultured gingival fibroblasts stimulated with wild type and proteinase mutant strains of Porphyromonas gingivalis, we show that the suppression of CXCL8 and IL-6, and the induction of TGF-β1 is primarily mediated by the proteolytic activity of lysine-specific proteinases. These results indicate that P. gingivalis is a major contributor to the altered immune responses and the pathology of periodontitis. Furthermore, the ease of sampling and analyzing cytokine expression profiles, including TGF-β1, in saliva and GCF may serve to predict the progression of periodontitis and associated systemic inflammatory diseases.

Topics: Biomarkers; Cells, Cultured; Disease Progression; Female; Fibroblasts; Gingival Crevicular Fluid; Humans; Inflammation; Interleukin-2; Interleukin-6; Interleukin-8; Male; Middle Aged; Peptide Hydrolases; Periodontitis; Porphyromonas gingivalis; Saliva; Transforming Growth Factor beta1

Gingival epithelial cells are part of the first line of host defense against infection. Toll-like receptors (TLRs) serve important immune and nonimmune functions. We investigated how interferon gamma (INF-γ) and interleukin 13 (IL-13) are involved in the TLR4 ligand-induced regulation of interleukin-8 (IL-8) effects on gingival epithelial cells. We used immunohistochemistry to localize TLR4 in ten healthy and ten periodontitis tissue specimens. Gingival epithelial cells then were primed with Th1 cytokine (INF-γ) or Th2 cytokine (IL-13) before stimulation with Escherichia coli-derived lipopolysaccharide (LPS) and enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of IL-8 secretion in cell culture supernatants. Although both healthy and periodontitis gingival tissue samples expressed TLR4, the periodontitis samples showed more intense expression on gingival epithelial cells. Gingival epithelial cell cultures were primed with either INF-γ or IL-13 before stimulation with TLR4 ligand. Supernatants from co-stimulated epithelial cells exhibited IL-8 production in opposite directions, i.e., as one stimulates the release, the other reduces the release. INF-γ significantly increased TLR4 function, whereas IL-13 significantly decreased TLR4 function, i.e., production of IL-8. Pathogen associated molecular pattern-LPS, shared by many different periodonto-pathogenic bacteria, activates the gingival epithelial cells in a TLR-dependent manner. Diminished or increased TLR function in gingival epithelial cells under the influence of different Th cell types may protect or be harmful due to the altered TLR signaling.

Topics: Adult; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Gingiva; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-13; Interleukin-8; Lipopolysaccharides; Male; Middle Aged; Periodontitis; Toll-Like Receptor 4; Young Adult

Neutrophil extracellular traps (NETs) are a recently discovered addition to the defensive armamentarium of neutrophils, assisting in the immune response against rapidly dividing bacteria. Although older adults are more susceptible to such infections, no study has examined whether aging in humans influences NET formation. We report that TNF-α-primed neutrophils generate significantly more NETs than unprimed neutrophils and that lipopolysaccharide (LPS)- and interleukin-8 (IL-8)-induced NET formation exhibits a significant age-related decline. NET formation requires generation of reactive oxygen species (ROS), and this was also reduced in neutrophils from older donors identifying a mechanism for reduced NET formation. Expression of IL-8 receptors (CXCR1 and CXCR2) and the LPS receptor TLR4 was similar on neutrophils from young and old subjects, and neutrophils challenged with phorbol-12-myristate-13-acetate (PMA) showed no age-associated differences in ROS or NET production. Taken together, these data suggest a defect in proximal signalling underlies the age-related decline in NET and ROS generation. TNF-α priming involves signalling through p38 MAP kinase, but activation kinetics were comparable in neutrophils from young and old donors. In a clinical setting, we assessed the capacity of neutrophils from young and older patients with chronic periodontitis to generate NETs in response to PMA and hypochlorous acid (HOCL). Neutrophil extracellular trap generation to HOCL, but not PMA, was lower in older periodontitis patients but not in comparison with age-matched controls. Impaired NET formation is thus a novel defect of innate immunity in older adults but does not appear to contribute to the increased incidence of periodontitis in older adults.

Topics: Adult; Aged; Aging; Case-Control Studies; Chronic Disease; Enzyme Activation; Extracellular Traps; Female; Humans; Immunity, Innate; Interleukin-8; Lipopolysaccharides; Male; Middle Aged; Neutrophils; p38 Mitogen-Activated Protein Kinases; Periodontitis; Reactive Oxygen Species; Receptors, Cell Surface; Tumor Necrosis Factor-alpha

This study aims to evaluate and validate a periodontitis screening model that includes sociodemographic, metabolic syndrome (MetS), and molecular information, including gingival crevicular fluid (GCF), matrix metalloproteinase (MMP), and blood cytokines.. The authors selected 506 participants from the Shiwha-Banwol cohort: 322 participants from the 2005 cohort for deriving the screening model and 184 participants from the 2007 cohort for its validation. Periodontitis was assessed by dentists using the community periodontal index. Interleukin (IL)-6, IL-8, and tumor necrosis factor-α in blood and MMP-8, -9, and -13 in GCF were assayed using enzyme-linked immunosorbent assay. MetS was assessed by physicians using physical examination and blood laboratory data. Information about age, sex, income, smoking, and drinking was obtained by interview. Logistic regression analysis was applied to finalize the best-fitting model and validate the model using sensitivity, specificity, and c-statistics.. The derived model for periodontitis screening had a sensitivity of 0.73, specificity of 0.85, and c-statistic of 0.86 (P <0.001); those of the validated model were 0.64, 0.91, and 0.83 (P <0.001), respectively.. The model that included age, sex, income, smoking, drinking, and blood and GCF biomarkers could be useful in screening for periodontitis. A future prospective study is indicated for evaluating this model's ability to predict the occurrence of periodontitis.

Topics: Adolescent; Adult; Aged; Alcohol Drinking; Biomarkers; Cohort Studies; Early Diagnosis; Female; Gingival Crevicular Fluid; Humans; Income; Interleukin-6; Interleukin-8; Male; Mass Screening; Matrix Metalloproteinase 13; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Metabolic Syndrome; Middle Aged; Periodontal Index; Periodontitis; Republic of Korea; Sensitivity and Specificity; Smoking; Tumor Necrosis Factor-alpha; Young Adult

Porphyromonas gingivalis is a major periodontal pathogen that contains a variety of virulence factors. The antibody titer to P. gingivalis GroEL, a homologue of HSP60, is significantly higher in periodontitis patients than in healthy control subjects, suggesting that P. gingivalis GroEL is a potential stimulator of periodontal disease. However, the specific role of GroEL in periodontal disease remains unclear. Here, we investigated the effect of P. gingivalis GroEL on human periodontal ligament (PDL) cells in vitro, as well as its effect on alveolar bone resorption in rats in vivo. First, we found that stimulation of PDL cells with recombinant GroEL increased the secretion of the bone resorption-associated cytokines interleukin (IL)-6 and IL-8, potentially via NF-κB activation. Furthermore, GroEL could effectively stimulate PDL cell migration, possibly through activation of integrin α1 and α2 mRNA expression as well as cytoskeletal reorganization. Additionally, GroEL may be involved in osteoclastogenesis via receptor activator of nuclear factor κ-B ligand (RANKL) activation and alkaline phosphatase (ALP) mRNA inhibition in PDL cells. Finally, we inoculated GroEL into rat gingiva, and the results of microcomputed tomography (micro-CT) and histomorphometric assays indicated that the administration of GroEL significantly increased inflammation and bone loss. In conclusion, P. gingivalis GroEL may act as a potent virulence factor, contributing to osteoclastogenesis of PDL cells and resulting in periodontal disease with alveolar bone resorption.

Topics: Alveolar Bone Loss; Animals; Bacterial Proteins; Chaperonin 60; Gene Expression Regulation; Host-Pathogen Interactions; Humans; Integrin alpha1; Integrin alpha2; Interleukin-6; Interleukin-8; Male; NF-kappa B; Osteoclasts; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; RANK Ligand; Rats; Rats, Sprague-Dawley; Recombinant Proteins

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

The aim of this study was to evaluate the effects of host modulation therapy on periodontal and biochemical parameters. Sixteen rheumatoid arthritis patients newly scheduled for anti-tumour necrosis factor (TNF) therapy were screened for 30 days. Periodontal parameters (clinical attachment level, probing pocket depth, bleeding on probing, plaque index and gingival index) as well as salivary and gingival crevicular fluid (GCF), interleukin (IL)-1β, IL-8 and monocyte chemoattractant protein-1 (MCP-1) levels of the patients were evaluated at baseline and on the 30th day of therapy. GCF volume, IL-1β and IL-8 levels (p = 0.007, p = 0.017 and p = 0.009, respectively) of the periodontitis patients significantly decreased. Although there was a decrease in all these parameters in healthy patients, it was below statistical significance. Salivary IL-8 and MCP-1 levels significantly decreased in periodontitis patients (p = 0.028 and p = 0.013, respectively), but IL-1β levels remained unchanged. These results suggest that TNF blockers may significantly modify host response in terms of biochemical parameters of the periodontium and may mask significant associations such as those reported between periodontitis and rheumatoid arthritis.

Topics: Adult; Arthritis, Rheumatoid; Chemokine CCL2; Dental Plaque Index; Female; Gingival Crevicular Fluid; Humans; Inflammation; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Periodontal Index; Periodontitis; Periodontium; Saliva; Tumor Necrosis Factor-alpha; Young Adult

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

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

Protease-activated receptor 2 (PAR2) is implicated in the pathogenesis of chronic inflammatory diseases, including periodontitis; it can be activated by gingipain and produced by Porphyromonas gingivalis and by neutrophil protease 3 (P3). PAR2 activation plays a relevant role in inflammatory processes by inducing the release of important inflammatory mediators associated with periodontal breakdown. The effects of periodontal treatment on PAR2 expression and its association with levels of proinflammatory mediators and activating proteases were investigated in chronic periodontitis patients. Positive staining for PAR2 was observed in gingival crevicular fluid cells and was reflective of tissue destruction. Overexpression of PAR2 was positively associated with inflammatory clinical parameters and with the levels of interleukin-6 (IL-6), IL-8, tumor necrosis factor alpha, matrix metalloprotease 2 (MMP-2), MMP-8, hepatocyte growth factor, and vascular endothelial growth factor. Elevated levels of gingipain and P3 and decreased levels of dentilisin and the protease inhibitors secretory leukocyte protease inhibitor and elafin were also associated with PAR2 overexpression. Healthy periodontal sites from individuals with chronic periodontitis showed diminished expression of PAR2 mRNA and the PAR2 protein (P < 0.05). Furthermore, periodontal treatment resulted in decreased PAR2 expression and correlated with decreased expression of inflammatory mediators and activating proteases. We concluded that periodontal treatment resulted in decreased levels of proteases and that proinflammatory mediators are associated with decreased PAR2 expression, suggesting that PAR2 expression is influenced by the presence of periodontal infection and is not a constitutive characteristic favoring periodontal inflammation.

Topics: Adhesins, Bacterial; Adult; Bacterial Proteins; Chymotrypsin; Cysteine Endopeptidases; Elafin; Female; Gingipain Cysteine Endopeptidases; Gingival Crevicular Fluid; Hepatocyte Growth Factor; Humans; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Middle Aged; Myeloblastin; Peptide Hydrolases; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Receptor, PAR-2; RNA, Messenger; Secretory Leukocyte Peptidase Inhibitor; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

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

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

The antigens of the Duffy blood group system (DARC) act as a receptor for the interleukin IL-8. IL-8 plays an important role in the pathogenesis of chronic periodontitis due to its chemotactic properties on neutrophils. The aim of this study was to investigate a possible association of Duffy blood group gene polymorphisms with the -353T>A, -845T>C and -738T>A SNPs of the IL8 gene in chronic periodontitis. One hundred and twenty-four individuals with chronic periodontitis and 187 controls were enrolled. DNA was extracted using the salting-out method. The Duffy genotypes and IL8 gene promoter polymorphisms were investigated by PCR-RFLP. Statistical analyses were conducted using the Chi square test with Yates correction or Fisher's Exact Test, and the possibility of associations were evaluated by odds ratio with a 95% confidence interval. When analyzed separately, for the Duffy blood group system, differences in the genotype and allele frequencies were not observed between all the groups analyzed; and, in nonsmokers, the -845C allele (3.6% vs. 0.4%), -845TC genotype (7.3% vs. 0.7%) and the CTA haplotype (3.6% vs. 0.4%) were positively associated with chronic periodontitis. For the first time to our knowledge, the polymorphisms of erythroid DARC plus IL8 -353T>A SNPs were associated with chronic periodontitis in Brazilian individuals. In Afro-Brazilians patients, the FY*02N.01 with IL8 -353A SNP was associated with protection to chronic periodontitis.

Topics: Brazil; Duffy Blood-Group System; Gene Frequency; Genetic Association Studies; Genotype; Haplotypes; Humans; Interleukin-8; Odds Ratio; Periodontitis; Polymorphism, Genetic; Promoter Regions, Genetic

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

Vitamin D has been known to be closely associated with periodontitis while the exact mechanisms remain unclear. The present study aimed to discover the effects of 1a,25-dihydroxyvitamin D3 (1,25D) on the expressions of interleukin (IL)-6 and IL-8 in human periodontal ligament cells (hPDLCs) stimulated with Porphyromonas gingivalis (P. gingivalis) W83.. Primary cultures of hPDLCs from ten donors were established and the cells of passage four were treated with 1,25D or P. gingivalis individually or 1,25D combined with P. gingivalis. The levels of IL-6 and IL-8 protein in hPDLCs were detected with enzyme-linked immunosorbent assay (ELISA) and the mRNA levels were detected with real-time RT-PCR.. P. gingivalis significantly promoted the protein expressions of IL-6 and IL-8. P. gingivalis at the multiplicity of infection (MOI) 100 exerted the strongest promotion effect on the IL-6 protein expression by 5.83-fold compared with the controls (2482.88±26.53pg/ml versus 425.80±77.25pg/ml, P<0.0005) and the IL-8 protein expression by 12.39-fold (4965.81±1072.55pg/ml versus 400.75±2.27pg/ml, P=0.005) in hPDLCs at 24h. At 48h, 10(-8)mol/L 1,25D had the best inhibition on the IL-8 protein expression in hPDLCs by 2.00-fold compared with the controls (100.76±21.11pg/ml versus 201.75±18.15pg/ml, P<0.0005) and the IL-8 mRNA expression by 2.13-fold (P<0.0005). 10(-8)mol/L 1,25D combined with P. gingivalis (MOI 100) exerted the strongest inhibition effect on the IL-8 protein expression by 1.54-fold compared with P. gingivalis treatment alone (3077.33±210.04pg/ml versus 4738.24±1386.17pg/ml, P=0.018) and the IL-8 mRNA expression by 1.78-fold (P=0.012) in hPDLCs at 12h. 1,25D did not influence the expression of IL-6 in hPDLCs with or without P. gingivalis treatment.. Vitamin D may potentially inhibit the periodontal inflammation induced by P. gingivalis partly by decreasing the IL-8 expression in hPDLCs.

Topics: Adolescent; Analysis of Variance; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-6; Interleukin-8; Male; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Vitamin D; Vitamins; 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

Porphyromonas gingivalis alters cytokine expression in gingival epithelial cells, stimulating inflammatory responses that may lead to periodontal disease. This study explored the effect of Lactobacillus acidophilus on the specific expressions of the interleukins (ILs) IL1B, IL6, and IL8 induced by the pathogen. Human gingival epithelial cells were co-cultured with P. gingivalis, L. acidophilus, or L. acidophilus + P. gingivalis; the control group consisted of the cells alone. Protein and gene expression levels of the ILs were detected using ELISA and qRT-PCR, respectively. The supernatant from the P. gingivalis group held significantly higher protein and mRNA levels of IL1B, IL6, and IL8, compared to the control group. In the mixed bacterial group (L. acidophilus + P. gingivalis), the levels of all three ILs decreased with increasing concentrations of L. acidophilus and were significantly different from the P. gingivalis group. This suggests that in gingival cells, L. acidophilus offsets the P. gingivalis-induced secretion of these ILs in a dose-dependent manner.

Topics: Cells, Cultured; Coculture Techniques; Epithelial Cells; Gingiva; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Interleukins; Lactobacillus acidophilus; Periodontitis; Porphyromonas gingivalis; Probiotics; RNA, Messenger

Fibronectin, an extracellular matrix component, is a substrate for multiple host and bacterial proteinases found in inflamed periodontal sites. In the present study, we investigated the potential contribution of various fibronectin fragments to the inflammatory process of periodontitis. Our results showed that the smaller fragments of fibronectin (30 and 45 kDa) were the most potent inflammatory inducers as they dose-dependently increased the secretion of TNF-α, IL-1β, and IL-8 by human macrophages. The 120-kDa fragment did not induce the secretion of all the cytokines tested, while intact fibronectin only increased IL-8 secretion and to a lesser extent TNF-α secretion. Cytokine secretion was associated with increased amounts of phosphorylated ERK1/2, JNK2, and p38α MAPK in treated macrophages. The combination of fibronectin or fibronectin fragments with Porphyromonas gingivalis lipopolysaccharide had an additive effect, but no synergism appeared to occur. It was also demonstrated that gingival crevicular fluid samples recovered from patients with moderate to severe periodontitis contained more fibronectin fragments than samples obtained from healthy subjects. Finally, both Arg- and Lys-gingipains purified from P. gingivalis were found to modulate fibronectin fragmentation. In conclusion, we showed that specific fibronectin fragments that may be present in diseased periodontal sites may contribute to maintaining and amplifying the inflammatory state and that P. gingivalis gingipains may be involved in the production of these fragments.

Topics: Adhesins, Bacterial; Cells, Cultured; Cysteine Endopeptidases; Extracellular Signal-Regulated MAP Kinases; Fibronectins; Gingipain Cysteine Endopeptidases; Gingival Crevicular Fluid; Humans; Interleukin-1beta; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Macrophages; p38 Mitogen-Activated Protein Kinases; Peptide Fragments; Periodontitis; Phosphorylation; Porphyromonas gingivalis; Tumor Necrosis Factor-alpha

Periodontitis is a multifactorial polymicrobial infection characterized by a destructive inflammatory process. Porphyromonas gingivalis, a Gram-negative anaerobic black-pigmented rod, which produces several virulence factors that stimulate human periodontal ligament cells (HPLCs) to produce various inflammatory mediators, has been implicated as a crucial etiologic agent in the initiation and progression of periodontitis. Since natural polyphenols such as resveratrol have growth-inhibitory effects on some bacterial pathogens and have shown chemo-preventive, anti-inflammatory and antioxidant activity, in the present study we used an HPLC model stimulated with lipopolysaccharide (LPS) of P. gingivalis to simulate the in vivo conditions such as those found in diseased periodontal sites. To determine whether resveratrol interferes with P. gingivalis LPS-activity and reduces the production of pro-inflammatory molecules, we investigated its effect on the cytokines IL-1β, IL-6, IL-8, IL-12 and TNF-α and NO production of HPLCs. The results showed that resveratrol treatment decreased in a dose- and time-dependent manner the NO expression induced by P. gingivalis LPS, correlated to an increased viability of infected HPLCs, and decreased the production of pro-inflammatory cytokines in HPLCs stimulated by P. gingivalis LPS. These results suggest that the ability of resveratrol to determine immunomodulatory effects could provide possible therapeutic applications for the treatment of periodontitis.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cells, Cultured; Cytokines; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Gene Expression; Humans; Inflammation Mediators; Interleukin-12; Interleukin-1beta; Interleukin-8; Lipopolysaccharides; Nitric Oxide; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; Resveratrol; Reverse Transcriptase Polymerase Chain Reaction; Stilbenes; Time Factors; Tumor Necrosis Factor-alpha

Periodontal (gum) disease is one of the main global oral health burdens and severe periodontal disease (periodontitis) is a leading cause of tooth loss in adults globally. It also increases the risk of cardiovascular disease and diabetes mellitus. Porphyromonas gingivalis lipopolysaccharide (LPS) is a key virulent attribute that significantly contributes to periodontal pathogenesis. Baicalin is a flavonoid from Scutellaria radix, an herb commonly used in traditional Chinese medicine for treating inflammatory diseases. The present study examined the modulatory effect of baicalin on P. gingivalis LPS-induced expression of IL-6 and IL-8 in human oral keratinocytes (HOKs). Cells were pre-treated with baicalin (0-80 µM) for 24 h, and subsequently treated with P. gingivalis LPS at 10 µg/ml with or without baicalin for 3 h. IL-6 and IL-8 transcripts and proteins were detected by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The expression of nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) proteins was analyzed by western blot. A panel of genes related to toll-like receptor (TLR) signaling was examined by PCR array. We found that baicalin significantly downregulated P. gingivalis LPS-stimulated expression of IL-6 and IL-8, and inhibited P. gingivalis LPS-activated NF-κB, p38 MAPK and JNK. Furthermore, baicalin markedly downregulated P. gingivalis LPS-induced expression of genes associated with TLR signaling. In conclusion, the present study shows that baicalin may significantly downregulate P. gingivalis LPS-upregulated expression of IL-6 and IL-8 in HOKs via negative regulation of TLR signaling.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Cells, Cultured; Down-Regulation; Flavonoids; Humans; Inflammation; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Keratinocytes; Lipopolysaccharides; Mouth; Periodontitis; Phosphorylation; Porphyromonas gingivalis; Scutellaria; Signal Transduction; Toll-Like Receptors

  To investigate the inflammatory responses of periodontitis patients and healthy patients in a whole-blood model stimulated with Porphyromonas gingivalis..   Whole blood collected from 17 periodontitis patients and six healthy patients was stimulated with Porphyromonas gingivalis cells. The secretion of cytokines and matrix metalloproteinases was quantified by enzyme-linked immunosorbent assay. An analysis of covariance with the ancova model was used to evaluate the significance of differences in secreted host molecules by whole blood from the periodontitis and healthy groups..   Porphyromonas gingivalis induced the secretion of interleukin-1β, interleukin-6, interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1, interferon inducible protein-10 by whole blood from patients in the periodontitis and healthy groups. Matrix metalloproteinase-8 and -9 levels secreted by whole blood also increased following stimulation. No significant differences (P < 0.05) in the amounts of secreted host molecules were observed between periodontitis and healthy patients..   This study suggests that Porphyromonas gingivalis can provoke an inflammatory response and promote the progression of periodontitis by inducing the secretion of high levels of cytokines and matrix metalloproteinases by a mixed leukocyte population. However, the whole-blood model did not reveal any significant differences in the inflammatory response between periodontitis patients (n = 17) and periodontally-healthy patients (n = 6).

Topics: Chemokine CCL2; Chemokine CCL5; Chemokine CXCL10; Disease Progression; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Leukocytes; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Tumor Necrosis Factor-alpha

We have previously reported that human periodontal ligament (hPDL) cells produced many kinds of cytokines as a result of bacterial stimulation, including stimulation with Porphyromonas gingivalis (P. gingivalis). However, the effects of mechanical stress on cytokine production in hPDL cells stimulated by periodontopathogenic bacteria are not clearly understood. In this study, we investigated the effects of mechanical stress on the production of inflammatory cytokines in hPDL cells induced by stimulation with P. gingivalis.. The hPDL cells were exposed to various levels of mechanical stress (1, 6, 10 and 50MPa) and costimulated with mechanical stress and P. gingivalis for 24h. Cytokine mRNA expressions were determined by RT-PCR. Cytokines in the culture supernatant were assessed by ELISA, and morphologic changes in hPDL cells were observed.. The expressions of interleukin (IL)-6, IL-8 and tumor necrosis factor-α mRNA were observed in hPDL cells after exposure to mechanical stress. Moreover, the production of IL-6 and IL-8 increased significantly after exposure to mechanical stress ranging from 1 to 10MPa. The amount of IL-8 in the culture supernatants of hPDL cells costimulated with P. gingivalis and mechanical stress was significantly higher than the expected additive amount. The morphology of hPDL cells did not change after exposure to 6MPa, but these cells were partly detached from the Petri dish after exposure to 50MPa.. These results suggest that local inflammation of the periodontal ligament may be induced mainly by periodontal bacteria, and mechanical stress may promote local inflammation.

Topics: Cells, Cultured; Female; Humans; Hydrostatic Pressure; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Male; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; RNA, Messenger; Stress, Mechanical; Tumor Necrosis Factor-alpha; Young Adult

Current therapies for peri-implantitis apply the same clinical protocols as those used for the treatment of periodontitis; however, outcomes remain unpredictable. We hypothesized that resident fibroblasts of the peri-implantitis stroma and periodontitis stroma differ in their phenotype and response to host immune factors. Fibroblasts are highly heterogeneous and comprise discrete subtypes with the potential of modulating inflammatory activities. The aim of the present study was to characterize the expression of receptors for complement C1q of innate immunity on human peri-implantitis fibroblasts and investigate effects of C1q on the proinflammatory properties of the cells.. Fibroblasts were cultured from gingival tissues exhibiting peri-implantitis and periodontitis, and from healthy gingivae as a control. Expression of C1q receptors for the collagen (cC1qR) and globular domains (gC1qR) of the protein was determined by flow cytofluorometric analysis (FITC) of specific antibodies bound to the surface of the cells. Secretion of C1q-inducible proinflammatory mediators was quantified after 24 h incubation using array-based ELISAs.. The percentage of fibroblasts FITC-positive for cC1qR was 67, 75 and 12% in peri-implantitis, healthy and periodontitis cultures, respectively, whereas the percentage of gC1qR FITC-positive fibroblasts was 5, 3 and 59%, respectively. The C1q interactions with peri-implantitis and healthy fibroblasts increased secretion of the chemokines interleukin-6 and interleukin-8 twofold, and monocyte chemoattractant protein-1 fourfold over baseline values, whereas periodontitis fibroblasts were unresponsive. Complement C1q increased levels of vascular endothelial growth factor sevenfold and transforming growth factor-β1 12-fold over baseline values in peri-implantitis cultures, only.. Peri-implantitis fibroblasts differ from periodontitis fibroblasts in phenotypic expression of cC1qR and function, and from healthy fibroblasts in proinflammatory, angiogenic and fibrogenic function. Peri-implantitis fibroblasts may represent a novel subtype.

Topics: Cells, Cultured; Chemokine CCL2; Complement C1q; Fibroblasts; Fluorescent Antibody Technique; Gingiva; Humans; Immunity, Innate; Immunophenotyping; Inflammation Mediators; Interleukin-6; Interleukin-8; Membrane Glycoproteins; Peri-Implantitis; Periodontitis; Protein Structure, Tertiary; Receptors, Complement; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

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

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

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

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

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

Periodontitis is a bacterial infection of the periodontal tissues. The Gram-negative anaerobic bacterium Porphyromonas gingivalis is considered a major causative agent. One of the virulence factors of P. gingivalis is capsular polysaccharide (CPS). Non-encapsulated strains have been shown to be less virulent in mouse models than encapsulated strains.. To examine the role of the CPS in host-pathogen interactions we constructed an insertional isogenic P. gingivalis knockout in the epimerase-coding gene epsC that is located at the end of the CPS biosynthesis locus. This mutant was subsequently shown to be non-encapsulated. K1 capsule biosynthesis could be restored by in trans expression of an intact epsC gene. We used the epsC mutant, the W83 wild type strain and the complemented mutant to challenge human gingival fibroblasts to examine the immune response by quantification of IL-1beta, IL-6 and IL-8 transcription levels. For each of the cytokines significantly higher expression levels were found when fibroblasts were challenged with the epsC mutant compared to those challenged with the W83 wild type, ranging from two times higher for IL-1beta to five times higher for IL-8.. These experiments provide the first evidence that P. gingivalis CPS acts as an interface between the pathogen and the host that may reduce the host's pro-inflammatory immune response. The higher virulence of encapsulated strains may be caused by this phenomenon which enables the bacteria to evade the immune system.

Topics: Bacterial Capsules; Cells, Cultured; Fibroblasts; Gene Knockout Techniques; Gingiva; Host-Pathogen Interactions; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Mutagenesis, Insertional; Periodontitis; Porphyromonas gingivalis; Virulence

Analysis of peri-implant crevicular fluid (PICF) offers a non-invasive means of studying the host response in peri-implant disease and may provide an early indication of patients at risk for active disease. This study examined the PICF levels of interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8) and macrophage inflammatory protein-1alpha (MIP-1alpha) in patients with non-manifesting inflammation, early and late stages of mucositis. The study group comprised 90 adult healthy volunteers with endosseal titanium implants inserted. Samples were taken from peri-implant sulcus using a filter paper technique. Implant tissues were categorized clinically as healthy, early mucositis or advanced mucositis. Clinical manifestations were determined by: gingival index and bleeding on probing, plaque index and radiographic analyses. Cytokine concentrations were assesed using commercially available enzyme-linked immunosorbent assay kits. Patients from the control group (healthy patients) have significantly lower concentrations of IL-1beta, TNF-alpha, IL-8 and MIP-1alpha in PICF compared with both groups with mucositis. Positive correlation was noted in the control group between IL-1beta and TNF-alpha and between MIP-1alpha and IL-8 in the group with early mucositis. The results suggest that cytokines could be prognostic markers of implant failure.

Topics: Chemokine CCL3; Cytokines; Dental Implantation, Endosseous; Dental Implants; Dental Restoration Failure; Female; Gingival Crevicular Fluid; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Mucositis; Periodontal Index; Periodontitis; Stomatitis; Tumor Necrosis Factor-alpha

Recent epidemiological studies have shown a correlation between periodontitis and hyperlipidemia. We have found high levels of oxidized low-density lipoprotein (OxLDL) in the gingival crevicular fluid of dental patients. In the present study, we tried to examine the possible role of OxLDL in periodontal inflammation in vitro.. Cells of the human gingival epithelial cell line Ca9-22 were cultured in media containing OxLDL, and the amounts of interleukin-8 (IL-8) and prostaglandin E(2) (PGE(2)) produced were measured using ELISAs.. Production of IL-8 by Ca9-22 cells was significantly increased when the cells were treated with OxLDL, but not with native LDL or acetylated LDL. Production of PGE(2) by Ca9-22 cells was enhanced by co-incubation with OxLDL and interleukin-1 beta (IL-1 beta). Scavenger receptor inhibitors, fucoidan and dextran sulfate, inhibited the OxLDL-induced IL-8 and PGE(2) production in the presence of IL-1 beta. The p(38) MAPK inhibitors SB203580 and SB202190 and the ERK inhibitor PD98059 inhibited the OxLDL-induced IL-8 production. Among oxidized lipids and chemically modified LDL, 7-ketocholesterol enhanced IL-8 production.. This is the first report to show that OxLDL enhances IL-8 production in epithelial cells.

Topics: Cell Line, Tumor; Chemokine CCL2; Cholesterol 7-alpha-Hydroxylase; Dextran Sulfate; Dinoprostone; Enzyme Inhibitors; Epithelial Cells; Flavonoids; Fucose; Gingiva; Humans; Imidazoles; Interleukin-1beta; Interleukin-8; Ketocholesterols; Lipoproteins, LDL; Mitogen-Activated Protein Kinases; Oxidation-Reduction; p38 Mitogen-Activated Protein Kinases; Periodontitis; Polysaccharides; Pyridines; Receptors, Scavenger; Sulfuric Acid Esters

Interleukin 8 (IL-8) is a chemokine related to the initiation and amplification of acute and chronic inflammatory processes. Polymorphisms in the IL8 gene have been associated with inflammatory diseases. We investigated whether the -845(T/C) and -738(T/A) single nucleotide polymorphisms (SNPs) in the IL8 gene, as well as the haplotypes they form together with the previously investigated -353(A/T), are associated with susceptibility to chronic periodontitis.. DNA was extracted from buccal epithelial cells of 400 Brazilian individuals (control n=182, periodontitis n=218). SNPs were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Disease associations were analyzed by the chi(2) test, Exact Fisher test and Clump program. Haplotypes were reconstructed using the expectation-maximization algorithm and differences in haplotype distribution between the groups were analyzed to estimate genetic susceptibility for chronic periodontitis development.. When analyzed individually, no SNPs showed different distributions between the control and chronic periodontitis groups. Although, nonsmokers carrying the TTA/CAT (OR=2.35, 95% CI=1.03-5.36) and TAT/CTA (OR=6.05, 95% CI=1.32-27.7) haplotypes were genetically susceptible to chronic periodontitis. The TTT/TAA haplotype was associated with protection against the development of periodontitis (for nonsmokers OR=0.22, 95% CI=0.10-0.46).. Although none of the investigated SNPs in the IL8 gene was individually associated with periodontitis, some haplotypes showed significant association with susceptibility to, or protection against, chronic periodontitis in a Brazilian population.

Topics: Adult; Brazil; Chronic Disease; Female; Haplotypes; Humans; Interleukin-8; Male; Middle Aged; Periodontitis; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide

The periodontal pathogen Porphyromans gingivalis is classified into six groups (types I-V and Ib) based on the genotype of the fimbriae A (fimA) gene. Among genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. The present study was undertaken to develop passive immunotherapy monoclonal antibodies (MAbs) against periodontitis, which are capable of inhibiting virulency and were constructed through the immunization of outer membrane vesicles (OMV) fraction of fimAII strain, TDC60, using mouse hybridoma technology. MAbs that recognized OMV by ELISA assay were identified, and 28 clones were screened by Western blot analysis. After purifying these MAbs using protein G column, the effect of the MAb on IL-8 production from human gingival fibroblasts by OMV was examined. We selected MAb TDC4-33H, which strongly inhibited the IL-8 production with a higher MAb production rate. Since the MAb showed an individual ladder-like profile against OMV by Western blotting, we further examined the reactivity against lipopolysaccharides (LPS) from TDC60, W83 (fimAIV), and ATCC33277 (fimAI). As a result, MAb TDC4-33H recognized all LPSs. Moreover, MAb TDC4-33H significantly inhibited the LPS-stimulated IL-8 production in human gingival fibroblasts. These findings suggest that MAb TDC4-33H reacts with LPS and may be useful for passive immunotherapy through neutralizing IL-8 production in gingival fibroblasts by P. gingivalis LPS.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Fimbriae Proteins; Gingiva; Humans; Hybridomas; Immunotherapy; Interleukin-8; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Periodontitis; Porphyromonas gingivalis

Periodontitis is a destructive disease which is likely to be the result of the activities of different microbial complexes. Recently, sulphate-reducing bacteria (SRB) have been detected in the oral cavity, and they have been found to be common inhabitants of sites showing periodontal destruction. The aim of study was to evaluate the influence of endotoxins of Desulfovibrio desulfuricans bacteria on human gingival fibroblast HGF-1 line.. The immunological response of gingival fibroblasts was evaluated by determination of their IL-6 and IL-8 secretion upon treatment with D. desulfuricans intestinal and type strain LPS, sodium butyrate (NaB) and IL-1beta. The amounts of cytokines were estimated by ELISA immunoassay. The influence of LPS and NaB on fibroblast proliferation was determined using the CyQUANT Cell Proliferation Assay Kit.. No significant growth inhibition of cells exposed to LPS was observed, except for the culture growing in the presence of intestinal strain endotoxin at the highest concentration (100 microg/ml). The secretion of IL-6 and IL-8 by fibroblasts was increased by D. desulfuricans endotoxins. Cells stimulated with proinflammatory cytokine 1L-1beta showed very high levels of both cytokines secretion. The release of IL-6 and IL-8 by cells in response to LPS and 1L-1beta was modulated by butyric acid.. The observed response of gingival fibroblasts to stimulation by endotoxin suggests that D. desulfuricans can be involved in the pathogenesis of periodontitis. Moreover, butyrate present in the oral cavity seems to have immunoregulatory effect on cytokine production by gingival fibroblasts under physiological conditions and during microbe-induced inflammation.

Topics: Butyrates; Cell Line; Cell Proliferation; Desulfovibrio desulfuricans; Fibroblasts; Gingiva; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Periodontitis; Proteins

The aim of this study was to evaluate correlations between levels of cytokines in secreted stimulated saliva in patients with chronic kidney disease (CKD) and hyposalivation.. Seventy patients with clearance <20 mL/min/1.73 m(2) were evaluated; 40 were predialysis, 21 hemodialysis, and 9 peritoneal dialysis, and they were matched with 70 control subjects. Salivary flow rate was measured and submandibular/sublingual saliva collected. Analyses were performed for whole protein content using a protein assay, and levels of tumor necrosis factor (TNF) α, interleukin (IL) 1β, γ-interferon (γ-INF), IL-6, IL-8, IL-10, monocyte chemotactic protein (MCP) 1, and soluble intercellular adhesion molecule (sICAM) 1, by using Luminex technology.. Patients with CKD had lower (P = .03) stimulated salivary secretion rate and higher salivary whole protein concentration (P = .002) than control subjects. Concentrations of IL-8 (P = .03) and MCP-1 (P = .002) were decreased and TNF-α/IL-10 (P = .05) and IL-8/IL10 (P = .03) ratios were decreased in CKD patients. CKD patients with low secretion levels of stimulated saliva expressed decreased levels of TNF-α (P = .04), IL-1β (P = .02), γ-INF (P = .03), IL-6 (P = .003), IL-8 (P = .005), MCP-1 (P = .006), and sICAM-1 (P = .02).. Salivary cytokines and secretion rates are significantly decreased in CKD patients. Further research is necessary to understand operating mechanisms and clinical implications of the down-regulation of inflammatory markers in saliva.

Topics: Case-Control Studies; Chemokine CCL2; Cross-Sectional Studies; Cytokines; Dental Caries; Female; Humans; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Periapical Diseases; Periodontitis; Peritoneal Dialysis; Renal Dialysis; Renal Insufficiency, Chronic; Saliva; Salivary Proteins and Peptides; Secretory Rate; Sublingual Gland; Submandibular Gland; Tumor Necrosis Factor-alpha; Xerostomia

The important inflammatory mediator interleukin-8 (IL-8) is responsible for the migration and activation of neutrophils. The IL8 gene contains a functional single-nucleotide polymorphism (SNP) (rs4073) in its promoter region that may influence the expression of IL-8, and which has been associated with inflammatory diseases. The purpose of this study was to investigate the association of the SNP (rs4073) in the IL8 gene with susceptibility to periodontitis. DNA was extracted from the buccal epithelial cells of 500 individuals (control n = 224 and periodontitis n = 276). Individuals were genotyped for the SNP (rs4073) using sequence-specific primer polymerase chain reaction. Associations between the SNP (rs4073) and subject phenotypes were analyzed using the chi-squared test, followed by univariate and multivariate logistic regression modeling. The genotype distributions in both groups were consistent with Hardy-Weinberg equilibrium. Univariate and multivariate analysis showed that age, skin color, and smoking status were associated with periodontitis. No significant differences were found for sex and frequencies of alleles and genotypes between the control and periodontitis groups in the univariate analysis. These findings were replicated in the multivariate analysis. The SNP (rs4073) in the IL8 gene is not associated with susceptibility to periodontitis in Brazilian individuals, even after controlling for covariates.

Topics: Brazil; Genetic Predisposition to Disease; Interleukin-8; Periodontitis; Polymorphism, Genetic; Polymorphism, Single Nucleotide

A major factor in the pathogenesis of periodontal disease, which is one of the biofilm infectious diseases, is thought to be lipopolysaccharide (LPS), owing to its ability to cause inflammation and promote tissue destruction. Moreover, the elimination of pathogens and their component LPSs is essential for the successful treatment of periodontal disease. Lipopolysaccharide tolerance is a mechanism that prevents excessive and prolonged responses of monocytes and macrophages to LPS. Since persistence of inflammation is necessary for inflammatory cytokine production, cells other than monocytes and macrophages are thought to maintain the production of cytokines in the presence of LPS. In this study, we investigated whether human gingival fibroblasts (HGFs), the most abundant structural cell in periodontal tissue, might be able to maintain inflammatory cytokine production in the presence of LPS bynot displaying LPS tolerance.. Human gingival fibroblasts were pretreated with LPS (from Porphyromonas gingivalis and Escherichia coli) and then treated with LPS, and the amounts of interleukin (IL)-6 and IL-8 in the cell culture supernatants were measured. The expression of negative regulators of LPS signalling (suppressor of cytokine signalling-1, interleukin-1 receptor-associated-kinase M and SH2 domain-containing inositol-5-phosphatase-1) was also examined in LPS-treated HGFs.. Human gingival fibroblasts did not display LPS tolerance but maintained production of IL-6 and IL-8 when pretreated with LPS, followed by secondary LPS treatment. Lipopolysaccharide-treated HGFs did not express negative regulators.. These results demonstrate that HGFs do not show LPS tolerance and suggest that this characteristic of HGFs sustains the inflammatory response in the presence of virulence factors.

Topics: Actins; Cell Line; Cells, Cultured; Escherichia coli; Fibroblasts; Gingiva; Humans; Immune Tolerance; Inositol Polyphosphate 5-Phosphatases; Interleukin-1 Receptor-Associated Kinases; Interleukin-10; Interleukin-6; Interleukin-8; Lipopolysaccharides; Periodontitis; Phosphoric Monoester Hydrolases; Porphyromonas gingivalis; Skin; src Homology Domains; Suppressor of Cytokine Signaling 1 Protein; Suppressor of Cytokine Signaling Proteins; Transforming Growth Factor beta1

Tetracyclines have been extensively used as adjuncts in the treatment of some forms of periodontitis. The aim of this study was to evaluate the capacity of doxycycline to influence the secretion of inflammatory mediators in macrophage and ex vivo human whole blood models stimulated with periodontopathogen lipopolysaccharides (LPS).. Monocyte-derived macrophages were treated with various concentrations of doxycycline prior to being stimulated with Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) LPS. The capacity of doxycycline to mediate the inflammatory response was also tested in an ex vivo whole blood model (whole blood isolated from periodontitis patients and healthy subjects) stimulated with Porphyromonas 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 (ELISA). Changes in phosphorylation state of kinases induced by A. actinomycetemcomitans LPS and doxycycline in the macrophage model were characterized by a multiplex ELISA analysis.. The secretion of IL-1beta and -8 and TNF-alpha by macrophages decreased significantly (P <0.05) when they were pretreated with 2 microM doxycycline, whereas a concentration of 10 microM was required to significantly reduce IL-6 secretion. Pretreatment of macrophages with 10 microM doxycycline prior to A. actinomycetemcomitans LPS stimulation resulted in a marked decrease in the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 (-76%). In the whole blood model, doxycycline, more particularly at 10 microM, was also a potent inhibitor of the proinflammatory cytokine response.. These two models provided clear evidence that some of the clinically proven benefits of doxycycline may be related to its ability to regulate inflammatory mediator release by host cells.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Anti-Inflammatory Agents; Doxycycline; Female; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase 9; p38 Mitogen-Activated Protein Kinases; Periodontitis; Phosphorylation; Porphyromonas gingivalis; Proto-Oncogene Proteins c-akt; Tumor Necrosis Factor-alpha

Periodontitis is an inflammatory disease caused by bacteria. In periodontitis, reactive oxygen species (ROS) are released from inflammatory cells in response to bacteria. Interleukin (IL)-8 is one of pro-inflammatory cytokines. To investigate the role of ROS in pathogenesis of periodontitis, we estimated the effect of H(2)O(2), one of ROS, on the expression of IL-8 in human periodontal ligament (PDL) cells. PDL cells were treated with H(2)O(2). IL-8 expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). The phosphorylation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38) and c-jun NH(2)-terminal kinase (JNK) was estimated by Western blotting. Treatment with H(2)O(2) at concentration of up to 250 microM increased IL-8 mRNA expression and production in a concentration-dependent manner. However, treatment with 500 microM H(2)O(2) did not increase IL-8 production. Catalase, an inhibitor of H(2)O(2), down-regulated the production of IL-8 induced by H(2)O(2). H(2)O(2) increased the phosphorylation of ERK, p38, and JNK. Pretreatment with PD98059 (ERK inhibitor), SB203580 (p38 inhibitor), or SP600125 (JNK inhibitor) decreased the IL-8 production induced by H(2)O(2). These results indicate that H(2)O(2) acts as an inducer of IL-8 secretion via activation of ERK, p38, and JNK in PDL cells. H(2)O(2) deposited in periodontal tissue during inflammation against bacteria may accelerate tissue destruction via induction of IL-8 in PDL cells.

Topics: Cell Survival; Cells, Cultured; Gene Expression; Humans; Hydrogen Peroxide; Interleukin-8; Mitogen-Activated Protein Kinases; Periodontal Ligament; Periodontitis; Phosphorylation; Signal Transduction; Up-Regulation

For clinical applications of biomarkers, there is a need for multiplex assays using high throughput platforms. The objective of this study was to determine the efficacy of Luminex Multianalyte Profiling (xMAP) technology for measurement of salivary proteins and to evaluate whether multiplex assays are as effective as single-plex assays and enzyme-linked immunosorbent assay (ELISA).. The average levels of interleukin-8 (IL-8) from the single-plex assay were 3313.2 +/- 3759.8 pg ml(-1) [oral squamous cell carcinoma (OSCC), n = 20] and 1061.7 +/- 1978.8 pg ml(-1) (control, n = 20). The IL-1beta average levels from the single-plex assay were 945.2 +/- 1134.8 pg ml(-1) (OSCC, n = 20) and 314.2 +/- 444.8 pg ml(-1) (control, n = 20). The average levels of IL-8 from the multiplex assay were 2834.9 +/- 3385.6 pg ml(-1) (OSCC, n = 20) and 947.3 +/- 2036.8 pg ml(-1) (control, n = 20). The IL-1beta average levels from the multiplex assay were 1013.5 +/- 1221.1 pg ml(-1) (OSCC, n = 20) and 376.3 +/- 576.3 pg ml(-1) (control, n = 20). The correlation coefficient between Luminex and ELISA assay for IL-8 (n = 19) and IL-1beta (n = 19) was 0.91 and 0.84, respectively.. Luminex xMAP single-plex and multiplex assays are as effective as ELISA assays for quantification of proteins in saliva. Both IL-8 and IL-1beta were expressed at significantly higher levels in OSCC subjects than in the matched healthy control subjects.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Chromogenic Compounds; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Immunoassay; Interleukin-1beta; Interleukin-8; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Periodontitis; Protein Array Analysis; Saliva; Salivary Proteins and Peptides; Sensitivity and Specificity; Young Adult

The gingival epithelium provides the first line of defense against colonization by periodontal pathogens, both as a physical barrier and by the production of inducible innate immune mediators such as beta-defensins and pro-inflammatory cytokines. The gram-negative bacterium Aggregatibacter actinomycetemcomitans is implicated in the pathogenesis of localized aggressive periodontitis, although the bacterium is found widely in the healthy population. We hypothesized that gingival epithelial cell-derived innate immune mediators triggered in response to A. actinomycetemcomitans infection may play an important role in increased susceptibility to infection.. Primary cultures of human gingival epithelial cells were cultured in the presence of A. actinomycetemcomitans. Total mRNA was examined for the presence of innate immune markers using RT-PCR.. We show here that the mRNA levels of human beta-defensin 2 and interleukin-8 are elevated by live cultures of a clinical isolate of A. actinomycetemcomitans in cultured gingival epithelial cells from healthy individuals, but not by A. actinomycetemcomitans lipopolysaccharide. Cells from a patient with localized aggressive periodontitis, however, did not respond to this bacterial stimulation. In contrast, the pro-inflammatory cytokine interleukin-19 was induced in cells from both localized aggressive periodontitis and healthy subjects. Examination of Toll-like receptors and associated adapter molecules indicated lower levels of Toll-like receptor 2 mRNA in the localized aggressive periodontitis patient-derived cells compared with cells from healthy subjects.. These results suggest that a differential expression of innate immune response genes to A. actinomycetemcomitans in the gingival epithelium could be an underlying factor of susceptibility to localized aggressive periodontitis.

Topics: beta-Defensins; Cell Culture Techniques; Epithelial Cells; Gene Expression Regulation, Bacterial; Genes, MHC Class II; Gingiva; Humans; Interleukin-8; Pasteurellaceae; Periodontitis

Tannerella forsythia is a periodontal pathogen. Recently, we have reported that the cytopathic component of T. forsythia contains two distinct factors. One arrests the cell cycle at the G2 phase and the other, named forsythia detaching factor, detaches adhesion-dependent immortalized human cells. In this study, we investigated the biological function of forsythia detaching factor using human normal fibroblasts.. A recombinant forsythia detaching factor, reported previously, was used. TIG-3 cells, cultured in the absence or presence of forsythia detaching factor, were lysed and the supernatant was analyzed by western blotting with polyclonal forsythia detaching factor antibodies. The cells were subsequently fractionated to isolate the cytoplasmic, mitochondrial and remaining fractions. In order to measure the activity of mitochondria using nicotinamide adenine dinucleotide-linked reductase, the water-soluble tetrazolium method was used. The mitochondrial oxidative membrane potential was estimated by measuring the oxidization-dependent fluorogenic conversion of dihydrotetramethylrosamine using flow cytometry. The concentration of interleukin-8 in the culture supernatant was assayed using a Human IL-8 ELISA kit.. Forsythia detaching factor-treated cells detached from the substratum and aggregated from 3 to 24 h. Then, the detached cells resumed adhesion and proliferated after 48 h. The western blot analysis revealed that most forsythia detaching factor trans-located into the mitochondrial fraction. Forsythia detaching factor suppressed the nicotinamide adenine dinucleotide-linked reductase activity in a dose-dependent manner and consequently increased the mitochondrial oxidative membrane potential. The production of interleukin-8 was reinforced in forsythia detaching factor-treated cells at 72 h through an increase of the mitochondrial oxidative membrane potential.. The forsythia detaching factor might be involved in the virulence of T. forsythia through induction of the pro-inflammatory cytokine interleukin-8.

Topics: Bacterial Proteins; Bacteroides; Cell Adhesion; Cell Extracts; Cells, Cultured; Fibroblasts; Humans; Interleukin-8; Membrane Potential, Mitochondrial; NADH Dehydrogenase; Oxidation-Reduction; Periodontitis; Recombinant Proteins; Virulence Factors

Periodontitis is a chronic inflammatory disease of bacterial etiology, affecting tooth-supporting tissues. The host inflammatory response to periodontopathogens, notably the high and continuous production of cytokines, is considered a major factor causing the local tissue destruction observed in periodontitis. The aim of the present study was to investigate the effect of naringenin, a major flavanone in grapefruits and tomatoes, on the lipopolysaccharide-induced pro-inflammatory cytokine production by host cells, using two different models.. The effect of naringenin was characterized using macrophages stimulated with the lipopolysaccharide of either Aggregatibacter actinomycetemcomitans or Escherichia coli and using whole blood stimulated with A. actinomycetemcomitans lipopolysaccharide, in the presence or absence of naringenin. Lipopolysaccharide-induced interleukin-1 beta, interleukin-6, interleukin-8 and tumor necrosis factor-alpha production by macrophages and whole-blood samples treated with naringenin were evaluated using an enzyme-linked immunosorbent assay. Changes in the phosphorylation states of macrophage kinases induced by A. actinomycetemcomitans lipopolysaccharide and naringenin were characterized by immunoblot screening.. Our results clearly indicated that naringenin is a potent inhibitor of the pro-inflammatory cytokine response induced by lipopolysaccharide in both macrophages and in whole blood. Naringenin markedly inhibited the phosphorylation on serines 63 and 73 of Jun proto-oncogene-encoded AP-1 transcription factor in lipopolysaccharide-stimulated macrophages.. The results from the present study suggest that naringenin holds promise as a therapeutic agent for treating inflammatory diseases such as periodontitis.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Inflammatory Agents; Escherichia coli; Flavanones; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages; p38 Mitogen-Activated Protein Kinases; Periodontitis; Phosphorylation; Phosphotransferases; Protein Serine-Threonine Kinases; Proto-Oncogene Mas; Proto-Oncogene Proteins c-jun; Serine; Transcription Factor AP-1; Tumor Necrosis Factor-alpha; U937 Cells

The periodontal vasculature is profoundly affected during the progression of periodontitis, and several specific bacteria are believed to be involved in this inflammatory disease. Eikenella corrodens is one of the common bacteria detected in periodontitis diseased lesions; however, the function of this organism in periodontitis is not well understood. In this study, we investigated the E. corrodens-induced endothelial cell alteration and inflammation process that leads to leukocyte infiltration in inflamed regions. Soluble products from E. corrodens (EcSP) induced the gene expression and protein production of vascular endothelial growth factor in oral epithelial cells and human umbilical vein endothelial cells (HUVEC). Direct stimulation by EcSP also activated endothelial cell proliferation. Moreover, EcSP induced ERK1/2 (p44/42) and p38 mitogen-activated protein kinase (MAPK) phosphorylation within 10-30 min in HUVEC, as demonstrated by Western blot analysis and up-regulated intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin and interleukin-8 (IL-8) production demonstrated by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. The specific p38 MAPK inhibitor SB203580 reduced the expression of ICAM-1, VCAM-1 and IL-8, whereas the blockade of p44/42 by MAPK kinase (MEK1) inhibitor, PD98059, inhibited only IL-8 expression. Our results indicate that E. corrodens can trigger a cascade of events that induce inflammatory responses in periodontal tissue via the MAPK cascade and may promote chronic periodontitis without bacteria-cell contact.

Topics: Cell Adhesion Molecules; Cell Line; Cell Proliferation; Cells, Cultured; Culture Media, Conditioned; E-Selectin; Eikenella corrodens; Endothelial Cells; Endothelium, Vascular; Enzyme Inhibitors; Epithelial Cells; Flavonoids; Humans; Imidazoles; Intercellular Adhesion Molecule-1; Interleukin-8; KB Cells; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; p38 Mitogen-Activated Protein Kinases; Periodontitis; Phosphorylation; Pyridines; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factor A

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

Treponema denticola, a major pathogen of periodontitis, has also been detected in the lesions of atherosclerosis. The aim of this study was to investigate induction of chemokine production in human umbilical vein endothelial cells (HUVECs) by T. denticola and determine whether those chemokines were degraded by a protease, dentilisin. T. denticola ATCC35405 or dentilisin-deficient mutant K1 were added to HUVECs and levels of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in the culture supernatants were determined by enzyme-linked immunosorbent assay. T. denticola ATCC35405 induced production of IL-8 in a time-dependent manner, with both production of IL-8 and expression of IL-8 mRNA showing higher levels than with exposure to dentilisin-deficient mutant K1. Although exposure to ATCC35405 induced expression of MCP-1 mRNA in the HUVECs, MCP-1 levels were remained similar to that in unstimulated cells. IL-8 and MCP-1 showed partial hydrolysis with exposure to T. denticola ATCC35405, but not with T. denticola K1. These results suggest that T. denticola can evade host defense mechanisms by modulating production of IL-8 and MCP-1, and that this play a role in the development of chronic infections such as periodontitis. The association of T. denticola infection to atherosclerosis was also discussed based on the present study.

Topics: Bacterial Proteins; Chemokine CCL2; Chymotrypsin; Endothelial Cells; Epithelial Cells; Humans; Interleukin-8; Peptide Hydrolases; Periodontitis; Reverse Transcriptase Polymerase Chain Reaction; RNA; Treponema denticola; Umbilical Veins

Neutrophils play a crucial role in the defense of invading bacteria by releasing biologically active molecules. The response of peripheral blood neutrophils was studied in periodontitis-affected patients and in healthy controls towards stimulation to Porphyromonas gingivalis (Pg) and Actinobacillus actinomycetemcomitans (Aa) extracts.. Peripheral venous blood was drawn from 23 adult patients with moderate to advanced chronic periodontitis (probing depth >or=5 mm, attachment loss >or=3 mm), and 30 healthy volunteers. Neutrophil response followed by metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) secretion was assayed by zymography and enzyme-linked immunosorbent assay, respectively, on both whole blood and purified neutrophils. In addition to periodontal pathogen extracts, known stimulating agents were tested, such as Escherichia coli-lipopolysaccharide (LPS), phytohemagglutinin, and zymosan A.. Neutrophil response, expressed as a secretion ratio under stimulated and non-stimulated conditions, measured in whole blood, showed no differences between periodontitis and healthy controls. Instead, in purified neutrophils from patients, MMP-9 exhibited a significantly higher secretion ratio with LPS and Pg (1.5- to 2-fold), whereas IL-8 showed a larger increase in secretion ratio (3- to 7-fold) in the presence of Pg, Aa, LPS, and zymosan A.. Peripheral neutrophils of periodontitis-affected patients are more reactive as suggested by their significantly higher response toward periodontal pathogen extracts and other stimulating agents.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Case-Control Studies; Dental Plaque Index; Female; Humans; Interleukin-8; Male; Matrix Metalloproteinase 9; Neutrophils; Periodontal Index; Periodontitis; Porphyromonas gingivalis

Histamine is an important mediator in immune responses, but it is unclear whether periodontal tissues express histamine receptors and are able to respond to histamine. We hypothesized that histamine, inflammatory cytokines, and bacterial components released in inflamed periodontal tissues may be synergistically involved in periodontitis. The present study showed that human gingival fibroblasts mainly express histamine receptor H1R, and responded to histamine to produce interleukin (IL)-8. Stimulation of gingival fibroblasts with tumor necrosis factor-alpha, IL-1alpha, and lipopolysaccharide markedly induced IL-8 production, and the IL-8 production was synergistically augmented in the presence of or pre-treatment with histamine. Selective inhibitors of mitogen-activated protein kinases (MAPKs), nuclear factor (NF)-kappaB, and phospholipase C (PLC) significantly inhibited the synergistic effect. These results indicate that histamine induces IL-8 production from gingival fibroblasts through H1R, and synergistically augments the inflammatory stimuli by amplification of the MAPK and NF-kappaB through H1R-linked PLC. Abbreviations used: HDC, histidine decarboxylase; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor; HR, histamine receptor; PLC, phospholipase C; MAPK, mitogen-activated protein kinase; NF, nuclear factor; ERK, extracellular signal-related kinase; JNK, c-Jun N-terminal kinase; R, receptor; TLR, Toll-like receptor; alpha-MEM, alpha-minimum essential medium; FCS, fetal calf serum; RT-PCR, reverse-transcriptase polymerase chain-reaction; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; LDH, lactate dehydrogenase.

Topics: Cells, Cultured; Fibroblasts; Gingiva; Histamine; Humans; Interleukin-1alpha; Interleukin-8; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; NF-kappa B; Periodontitis; Receptors, Histamine H1; Recombinant Proteins; Tumor Necrosis Factor-alpha; Type C Phospholipases

Periodontitis is characterized by an accumulation of inflammatory cells in periodontal tissue and subgingival sites. Leukocytes play a major role in the host response to Porphyromonas gingivalis, a major aetiological agent of chronic periodontitis. Secretion of high levels of inflammatory mediators, including cytokines and prostaglandins, by leucocytes is believed to contribute to periodontal tissue destruction. The aim of this study was to investigate the inflammatory response of an ex vivo whole blood model to P. gingivalis stimulation. The production of interleukin-1 beta (IL-1beta), IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, tumour necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), IFN-gamma-inducible protein 10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), Regulated on Activation Normal T cell Expressed and Secreted (RANTES) and prostaglandin E2 (PGE2) were quantified by enzyme-linked immunosorbent assays. P. gingivalis induced the secretion of the pro-inflammatory cytokines IL-1beta, TNF-alpha, IL-6 and IFN-gamma, the chemokines IL-8, RANTES and MCP-1 and the inflammatory mediator PGE2 in an ex vivo human whole blood model. The secretion levels were dependent on the strain and the infectious dose used. While the mediator profiles were comparable between six healthy subjects, a high interindividual variability in the levels of secreted mediators was observed. This study supports the view that P. gingivalis, by inducing high levels of inflammatory mediators from a mixed leucocyte population, can contribute to the progression of periodontitis.

Topics: Analysis of Variance; Bacteriological Techniques; Bacteroidaceae Infections; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Cytokines; Dinoprostone; Enzyme-Linked Immunosorbent Assay; Humans; Interferon-gamma; Interleukin-1; Interleukin-6; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Serum Albumin; Species Specificity; Tumor Necrosis Factor-alpha

Bacteremia frequently occurs after dental treatment. Periodontal inflammation may influence the incidence, magnitude and duration of bacteremia. The presence of circulating oral bacteria or bacterial components may induce cytokine synthesis in blood cells, which may contribute to the development or exacerbation of atherosclerosis. The present study tested the hypothesis that bacteremia occurring after scaling in periodontitis patients results in altered plasma levels of cytokines. Twenty periodontitis patients were subjected to scaling. Blood samples at baseline and at 0.5, 10 and 30 minutes postscaling were examined for bacteremia whereas baseline and eight-hour postscaling blood samples were examined for the levels of IL-1beta, TNF-alpha, IL-6, IL-8, IL-10 and IL-12p70. IL-6 levels were significantly increased eight hours after scaling, while IL-8 was significantly decreased. No systematic changes occurred in the levels of IL-1beta, TNF-alpha, IL-10 and IL-2p70. IL-6 levels may be increased while IL-8 may be decreased due to scaling, which may have implications for general health.

Topics: Adult; Bacteremia; Bacteroidaceae Infections; Dental Plaque Index; Dental Scaling; Female; Follow-Up Studies; Gingival Hemorrhage; Humans; Interleukin-10; Interleukin-12; Interleukin-6; Interleukin-8; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Prevotella; Streptococcal Infections; Streptococcus; Time Factors; Tumor Necrosis Factor-alpha

Chronic periodontitis is correlated with Porphyromonas gingivalis infection. In this study, we found that the expression of secretory leucocyte protease inhibitor (SLPI), an endogenous inhibitor for neutrophil-derived proteases, was reduced in gingival tissues with chronic periodontitis associated with P. gingivalis infection. The addition of vesicles of P. gingivalis decreased the amount of SLPI in the media of primary human gingival keratinocytes compared to untreated cultures. We therefore investigated how arginine-specific gingipains (Rgps) affect the functions of SLPI, because Rgps are the major virulence factors in the vesicles and cleave a wide range of in-host proteins. We found that Rgps digest SLPI in vitro, suppressing the release of SLPI. Rgps proteolysis of SLPI disrupted SLPI functions, which normally suppresses neutrophil elastase and neutralizes pro-inflammatory effects of bacterial cell wall compounds in cultured human gingival fibroblasts. The protease inhibitory action of SLPI was not exerted towards Rgps. These results suggest that Rgps reduce the protective effects of SLPI on neutrophil proteases and bacterial proinflammatory compounds, by which disease in gingival tissue may be accelerated at the sites with P. gingivalis infection.

Topics: Adhesins, Bacterial; Cells, Cultured; Chronic Disease; Cysteine Endopeptidases; Disease Progression; Electrophoresis, Polyacrylamide Gel; Gingipain Cysteine Endopeptidases; Gingiva; Gingival Crevicular Fluid; Humans; Immunoblotting; Interleukin-8; Middle Aged; Periodontitis; Porphyromonas gingivalis; Proteinase Inhibitory Proteins, Secretory; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Secretory Leukocyte Peptidase Inhibitor; Statistics, Nonparametric

Polymorphonuclear neutrophil (PMN) dysfunction is associated with diabetes. We examined the gingival crevicular fluid (GCF) beta-glucuronidase (BG) and interleukin-8 (IL-8) levels of periodontitis patients with and without type 2 diabetes mellitus (DM).. Forty five adults with type 2 DM and 32 adults without DM, both with chronic periodontitis were enrolled. GCF was collected from eight posterior sites in each quadrant, and periodontal parameters were recorded. GCF was assayed for IL-8 by ELISA and BG by a fluorometric assay.. GCF IL-8 was positively correlated with probing depth (PD), and GCF BG but not clinical attachment level (CAL), bleeding on probing (BOP), or plaque index (PI). In contrast, GCF BG was strongly correlated with each of the clinical measures of periodontal disease. Subjects with DM significantly lower levels of both BG (73.0+/-44.8 versus 121.9+/-84.6 pg/sample; p=0.002) and IL-8 (32.1+/-33.1 versus 90.8+/-83.2 pg/sample; p<0.0001) even after adjustments for age, gender, PD, CAL, BOP, and PI. Neither BG nor IL-8 was correlated with HbA1c levels in subjects with DM.. These data suggest that an inadequate local response by PMN, partially explained by an altered chemokine gradient, may contribute to periodontal disease in patients with type 2 DM.

Topics: Adult; Age Factors; Aged; Chronic Disease; Cross-Sectional Studies; Dental Plaque Index; Diabetes Mellitus, Type 2; Female; Gingival Crevicular Fluid; Gingival Hemorrhage; Glucuronidase; Glycated Hemoglobin; Humans; Interleukin-8; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Sex Factors

Host recognition pathways for gram-negative and gram-positive bacteria comprise pattern recognition receptors among which Toll-like receptors (TLRs) play a pivotal role. TLRs share common signaling pathways yet exhibit specificity as well. Periodontal disease is initiated and maintained in the first line by gram-negative but also gram-positive bacterial infection of the gingival sulcus. To date only limited information is available on whether gram-positive and gram-negative bacteria induce different host responses (strength or quality).. To elucidate these differential effects we focused on proinflammatory cytokine releases by assessing ex vivo stimulation of whole blood with heat-killed gram-negative and gram-positive bacteria and thereof derived microbial products associated with distinct TLRs. Tumor necrosis factor-alpha and interleukin-8 release were measured in the supernatants by enzyme-linked immunosorbent assay. In addition, innate immune responses of peritoneal macrophages from mice lacking TLR2 and TLR4 were tested.. We observed that gram-negative and gram-positive species induced distinct patterns of cytokine production. Gram-negative species produced higher amounts of tumor necrosis factor-alpha while gram-positive species released higher amounts of the chemokine interleukin-8. Data from TLR knockout mice and TLR-transfected HEK cells revealed a somehow specific role of TLR4 and TLR2 for the recognition of gram-negative and gram-positive bacteria, respectively, an observation that goes along with the dominant recognition of the respective pathogen associated molecular patterns lipopolysaccharide and lipoteichoic acid.. The results show that gram-negative and gram-positive bacterial species induce different patterns of immunoregulatory activity, which might be the result of activation of different TLRs.

Topics: Animals; Antigens, Bacterial; Cells, Cultured; Gram-Negative Bacteria; Gram-Positive Bacteria; Humans; Immunity, Innate; Interleukin-8; Macrophages, Peritoneal; Mice; Mice, Inbred C3H; Periodontitis; Toll-Like Receptors; Tumor Necrosis Factor-alpha

Tumour necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK), a member of the TNF family, is a multi-functional cytokine that regulates cellular proliferation, angiogenesis, inflammation and apoptosis. In this study, we investigated TWEAK expression in periodontally diseased tissues and the effect of TWEAK on human gingival fibroblasts (HGF). Reverse transcription-polymerase chain reaction (RT-PCR) analysis and immunohistochemistry revealed that TWEAK and the TWEAK receptor, fibroblast growth factor-inducible 14 (Fn14), mRNA and protein were expressed in periodontally diseased tissues. HGF expressed Fn14 and produced interleukin (IL)-8 and vascular endothelial growth factor (VEGF) production upon TWEAK stimulation in a dose-dependent manner. The IL-8 and VEGF production induced by TWEAK was augmented synergistically by simultaneous stimulation with transforming growth factor (TGF)-beta1 or IL-1beta. IL-1beta and TGF-beta1 enhanced Fn14 expression in a dose-dependent manner. Moreover, TWEAK induced intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression on HGF in a dose-dependent manner. The ICAM-1 expression induced by TWEAK was augmented by TGF-beta1. On the other hand, the TWEAK-induced VCAM-1 expression was inhibited by TGF-beta1. Phosphatidylinositol 3-kinase (PI3K) and nuclear factor-kappaB (NF-kappaB) inhibitor inhibit both ICAM-1 and VCAM-1 expression induced by TWEAK. However, mitogen-activated protein kinase (MEK) and c-Jun NH2-terminal kinase (JNK) inhibitor enhanced only VCAM-1 expression on HGF. These results suggest that TWEAK may be involved in the pathophysiology of periodontal disease. Moreover, in combination with IL-1beta or TGF-beta1, TWEAK may be related to the exacerbation of periodontal disease to induce proinflammatory cytokines and adherent molecules by HGF.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Cells, Cultured; Cytokine TWEAK; Dose-Response Relationship, Immunologic; Female; Fibroblasts; Gene Expression; Gingiva; Humans; Intercellular Adhesion Molecule-1; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Periodontitis; Receptors, Tumor Necrosis Factor; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Transforming Growth Factor beta1; Tumor Necrosis Factors; TWEAK Receptor; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factor A

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

Anti-neutrophil cytoplasmic Abs targeting proteinase 3 (PR3) have been detected in relation to a wide range of inflammatory conditions such as periodontitis, and interaction of anti-PR3 Abs with endothelial and epithelial cells provokes cell activation, although the underlying mechanism has been unclear. The present study showed that human oral epithelial cells expressed PR3 mRNA after treatment with proinflammatory cytokines such as IL-1alpha, TNF-alpha, IFN-alpha, IFN-beta, and IFN-gamma. A 29-kDa PR3 was expressed on the cell surface and released into culture supernatants by the cells upon stimulation with these cytokines. The membrane and supernatant fractions of oral epithelial cells exhibited enzymatic activity, which was inhibited by serine proteinase inhibitors, but not by a cysteine proteinase inhibitor or secretory leukocyte protease inhibitor. Addition of anti-PR3 Abs to cytokine-primed oral epithelial cells in culture induced remarkable secretion of IL-8 and monocyte chemoattractant protein 1 and aggregation of PR3 on the cells. RNA interference targeted to protease-activated receptor-2 mRNA and intracellular Ca2+ mobilization assays revealed that anti-PR3 Abs activated the epithelial cells through protease-activated receptor-2, a family of G protein-coupled receptors. The anti-PR3 Ab-mediated cell activation was completely abolished by RNA interference targeted to PR3 mRNA and by inhibition of phospholipase C and NF-kappaB. Immunohistochemistry showed that inflamed oral epithelium actually expresses PR3 protein. These results suggest that oral epithelial cells express functional PR3 in the inflamed sites and respond to anti-PR3 Abs detected in diseased sera, and that these mechanisms may actively participate in the inflammatory process, including periodontitis.

Topics: Adult; Antibodies; Cell Line, Tumor; Cells, Cultured; Chemokine CCL2; Cytokines; Enzyme Induction; Epithelial Cells; Gingiva; HeLa Cells; Humans; Immunohistochemistry; Inflammation Mediators; Interleukin-8; Membrane Proteins; Mouth Mucosa; Myeloblastin; NF-kappa B; Periodontitis; Receptor, PAR-2; RNA, Messenger; Serine Endopeptidases

The aim of this study was to evaluate the relationship between the accumulation of interleukins IL-1beta, TNF-alpha, IL-8, and chemokine RANTES (Regulated upon Activation Normal T-cell Expressed and Secreted) in gingival fluid and periodontal support tissues in patients with periodontitis. A review is also provided of apoptotic processes as events of major importance, highlighting the presence of TUNEL cells and ultrastructural morphologic changes associated with cell apoptosis. There appears to be further evidence to support the important role of inflammation control. Cytokines may be considered as markers of the progression and severity of periodontitis as well as indicators of an appropiate response to treatment. However, further studies are needed to support and characterize this concept.

Topics: Antigens, Bacterial; Apoptosis; Biomarkers; Chemokine CCL5; Cytokines; Gingival Crevicular Fluid; Humans; In Situ Nick-End Labeling; Inflammation Mediators; Interleukin-1; Interleukin-8; Lymphocytes; Neutrophils; Periodontitis; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) is believed to play an important role in the pathogenesis of various forms of periodontitis. In addition, the anti-IL-8 autoantibody has been recently recognized as a potent modulator of IL-8 function. In the current study, the concentrations of IL-8 and its autoantibody in gingival crevicular fluid from patients with chronic generalized periodontitis were compared to those in gingival crevicular fluid from patients with refractory chronic periodontitis. Gingival crevicular fluids were collected from patients treated in a private periodontal clinic. Nine patients who were identified as having chronic generalized periodontitis and four with refractory chronic periodontitis were selected for the study. Patients included in the latter group had undergone supportive periodontal therapy for more than 10 years, and during that time had experienced many episodes of periodontal destruction. The gingival crevicular fluid concentrations of total protein, IL-8, free anti-IL-8 autoantibody and IL-8 bound to the autoantibody (anti-IL-8:IL-8 complexes) were examined. There were no differences in concentration of total protein, but significantly higher levels of IL-8 were detected in patients with chronic generalized periodontitis in comparison to patients with refractory chronic periodontitis (P < 0.05). In addition, anti-IL-8:IL-8 complexes were present in 90% of patients with chronic generalized periodontitis, but in only 50% of patients with refractory chronic periodontitis. The results suggest that elevated concentrations of free and complexed IL-8 can differentiate patients with chronic generalized periodontitis from patients with refractory chronic periodontitis.

Topics: Antigen-Antibody Complex; Autoantibodies; Chronic Disease; Gingival Crevicular Fluid; Humans; Immunoglobulin G; Interleukin-8; Periodontal Index; Periodontitis; Proteins; Recurrence; Statistics, Nonparametric

We previously reported that a capsular polysaccharide (CP) from Actinobacillus actinomycetemcomitans Y4 induces bone resorption in a mouse organ culture system and osteoclast formation in mouse bone marrow cultures. However, the effects of A. actinomycetemcomitans Y4 CP on human gingival fibroblasts (HGF) are still unclear. The present study was undertaken to test the hypothesis that A. actinomycetemcomitans Y4 CP alters the production of inflammatory cytokines, such as interleukin-6 (IL-6) and IL-8 by HGF. When HGF were cultured with various concentrations of Y4 CP for 24 h, IL-6 and IL-8 production decreased in a concentration-dependent manner. Y4 CP (100 microg/ml) suppressed the release of IL-6 from 9.09 +/- 0.08 ng/ml to 0.34 +/- 0.21 ng/ml (P < 0.01) and IL-8 production decreased from 3.76 +/- 0.03 ng/ml to 0.09 +/- 0.01 ng/ml (P < 0.01). Y4 CP suppressed 70-80% of the release of IL-6 and IL-8 from HGF stimulated with Y4 lipopolysaccharide (LPS), too. Interestingly, anti-A. actinomycetemcomitans Y4 CP completely inhibited the effect of A. actinomycetemcomitans Y4 CP on IL-6 and IL-8 production from HGF. These results indicate that Y4 CP inhibits the release of IL-6 and IL-8 from HGF, suggesting that A. actinomycetemcomitans Y4 modulates the inflammatory response in periodontitis. Remarkably, this inhibitory effect was reversed by specific anti-A. actinomycetemcomitans Y4 CP suggesting an important relationship between the organism and the humoral host response.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Analysis of Variance; Antibodies, Monoclonal; Bacterial Capsules; Cell Culture Techniques; Dose-Response Relationship, Drug; Fibroblasts; Gingiva; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Lipopolysaccharides; Periodontitis; Polysaccharides, Bacterial; Statistics as Topic

As some residual inflammation may remain after periodontal therapy, the present pilot study investigated the long-term effect of full-mouth tooth extraction therapy on the responsiveness of peripheral blood monocytes in a case with generalized terminal adult periodontitis.. Before and 3, 9, 20 and 32 months after therapy, venous blood was collected. Total and differential white blood cell counts were determined and whole blood cell cultures (WBCC) were incubated with lipopolysaccharide (LPS) to stimulate the production of inflammatory mediators by monocytes.. After full-mouth tooth extraction, the numbers of total peripheral white blood cells and neutrophils decreased over time. The release of the chemokines interleukin (IL)-8 and macrophage chemoattractant protein (MCP)-1 in the cultures decreased twofold over time, whereas no changes were seen for the other studied cytokines, chemokines and prostaglandin E2.. On the basis of previous studies and the present case, the high production of IL-8 and MCP-1 by monocytes in LPS-stimulated WBCC from periodontitis patients is most likely acquired, as their levels decrease over time when the periodontal infection is controlled. The possible connection between periodontitis and atherosclerosis through IL-8 and MCP-1 is discussed.

Topics: Adult; Arteriosclerosis; Chemokine CCL2; Humans; Interleukin-8; Leukocyte Count; Lipopolysaccharides; Male; Monocytes; Periodontitis; Tooth Extraction

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

It has been suggested that periodontal inflammation may result in an altered immune response. The peripheral immune capacity in periodontitis patients can be investigated using lipopolysaccharide (LPS)-stimulated whole blood cell cultures (WBCC), known to reflect the behavior of monocytes in particular. A previous study in our laboratory revealed that monocytes in the stimulated cultures from periodontitis patients behaved functionally different compared with controls. The present study investigated whether this different response of periodontitis patients' monocytes is intrinsic or acquired.. The release of inflammatory mediators was measured in Escherichia coli LPS-stimulated WBCC from 12 periodontitis patients before and after periodontal therapy. In addition, the total leukocyte and leukocyte differentiation counts were also determined in the patients before and after therapy.. The levels of interleukin (IL)-12p70 in cell culture supernatants increased two times and those of prostaglandin E2 showed a trend towards reduction after therapy, whereas the levels of IL-1beta, IL-6, IL-8, IL-10, IL-12p40 and tumor necrosis factor-alpha did not change. The total number of white blood cells was decreased after periodontal therapy.. After periodontal therapy, the functional phenotype of the peripheral blood monocytes from patients was reconstituted, resembling that of subjects without periodontitis.

Topics: Adult; Dinoprostone; Escherichia coli; Follow-Up Studies; Humans; Inflammation Mediators; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Interleukin-8; Leukocyte Count; Leukocytes; Lipopolysaccharides; Monocytes; Periodontitis; Phenotype; Protein Subunits; Tumor Necrosis Factor-alpha

The ability of different Porphyromonas gingivalis strains (15 clinical isolates and ATCC 33277) to attach to and invade KB cells, in relation to other properties such as release of interleukin (IL)-6 and IL-8, cytotoxicity, proteolytic activity and types of fimbriae genes present, was examined. A hierarchical cluster analysis based on adherence and internalization resulted in four groups. Eight of the 15 clinical isolates belonged to a cluster group whose adherence and internalization were about 10% those of the ATCC strain. A negative correlation between lysine-specific protease activity and adherence was found. In all cases the released concentrations of IL-6 and IL-8 were very low. Only one strain was found to be cytotoxic to KB cells. Principal components analysis demonstrated correlations between adherence, internalization and autoaggregation. Most strains had fimA type I and II, type I being associated with elastase-like activity. The ability of P. gingivalis to invade epithelial cells may be a key factor for maintaining periodontal disease.

Topics: Bacterial Adhesion; Cluster Analysis; Endopeptidases; Fimbriae Proteins; Fimbriae, Bacterial; Genes, Bacterial; Humans; Interleukin-6; Interleukin-8; KB Cells; Periodontitis; Porphyromonas gingivalis; Principal Component Analysis; Species Specificity; Statistics, Nonparametric; Virulence Factors

Adult periodontitis, which is the major cause of adult tooth loss, is commonly characterized by chronic inflammatory disease caused by infection with periodontopathic bacteria such as Porphyromonas gingivalis. Our aims in the present study were to examine the expression of endothelin-1 (ET-1) in cultured HEp-2 epithelial cells after infection with P. gingivalis, and in gingival tissue from adult periodontitis patients. The cell lines were infected with the strains P. gingivalis 33277 and 381 for assessment of bacterial invasion using an antibiotic protection assay, and the expression of ET-1, inflammatory cytokines and cell adhesion molecules was examined by ELISA and reverse transcription-PCR. The expression of ET-1, as well as that of interleukin-1 beta, interleukin-8 and ICAM-1 (intercellular cell adhesion molecule 1), was induced significantly in a time-dependent manner, whereas the expression of MCP-1 (monocyte chemotactic protein-1), RANTES (regulated upon activation, normal T-cell expressed and secreted) and VCAM-1 (vascular cell adhesion molecule 1) was not. Furthermore, in gingival tissues from adult periodontitis patients, we also observed increased expression of ET-1 mRNA compared with tissue from normal healthy donors. These results suggest that infection by periodontopathic bacteria up-regulates the expression of ET-1, together with that of inflammatory cytokines and ICAM-1, in gingival epithelial cells, and that ET-1 expression may be closely involved in the regulation of cytokine responses and cell-cell adhesion in adult periodontitis lesions.

Topics: Adult; Case-Control Studies; Cell Line; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gingiva; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Reverse Transcriptase Polymerase Chain Reaction

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

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

Viable and inactivated Porphyromonas gingivalis dose-dependently induced interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) secretion in human umbilical vein endothelial cells (HUVECs). The inactivated P. gingivalis, in comparison with viable bacteria, tended to enhance the production of both chemokines more strongly. The production of MCP-1 protein began increasing immediately after stimulation by P. gingivalis, and there was a nearly linear increase from 0 to 8 h of incubation, whereas IL-8 production showed a linear increase between 4 and 12 h of incubation. The IL-8 and MCP-1 mRNA expressions in HUVECs as determined by reverse transcriptase-polymerase chain reaction (RT-PCR) or Quantikine mRNA colorimetric quantification kits were found to be enhanced by P. gingivalis. Furthermore, the time courses of IL-8 and MCP-1 mRNA expressions were in accordance with those of protein production. Addition of polymyxin B or boiling did not weaken the stimulatory effect of P. gingivalis, which inhibited the effect of Escherichia coli lipopolysaccharide (E. coli LPS) and tumour necrosis factor-alpha (TNF-alpha), respectively. In contrast, the induction of IL-8 and MCP-1 by P. gingivalis was significantly reduced by anti-CD14 antibody. Our results suggest that some heat-stable component of P. gingivalis, including LPS, may be responsible for the induction of IL-8 and MCP-1 in HUVECs by a CD14-dependent mechanism. These effects might be involved in the accumulation and activation of neutrophils and monocytes at an early stage of the periodontal pathogenesis.

Topics: Bacteroidaceae Infections; Base Sequence; Cells, Cultured; Chemokine CCL2; Endothelium, Vascular; Gene Expression; Hot Temperature; Humans; Interleukin-8; Kinetics; Lipopolysaccharide Receptors; Periodontitis; Polymyxin B; Porphyromonas gingivalis; RNA, Messenger

During the acute inflammatory response in periodontitis, gingival epithelial cells are considered to play important roles in the recruitment of inflammatory cells to the site of infection through the secretion of chemokines. However, little is known about the expression of molecules that are involved in the interaction between the epithelium and neutrophils following bacterial attachment. Earlier work reported that periodontopathogenic Eikenella corrodens strain 1,073 up-regulated the expression and secretion of chemokines such as interleukin-8 (IL-8) from KB cells (a human oral epithelial cell line derived from a human oral epidermoid carcinoma). To elucidate the mechanism of the transmigration of neutrophils through the epithelium, the present study investigated the expression of adhesion molecules on KB cells in response to E. corrodens attachment. Adhesion molecule gene expression was assessed by RT-PCR and adhesion proteins expressed on KB cell surfaces were determined by cell-based ELISA and FACS. In RT-PCR, ICAM-1 mRNA levels were significantly increased within 1 h in response to exposure to E. corrodens and continued to increase over the 12-h period of study. In ELISA, increased surface ICAM-1 expression was paralleled by increased ICAM-1 mRNA levels. Furthermore, the increases in ICAM-1 expression on epithelial cells infected with E. corrodens were observed to be due to the N-acetyl-D-galactosamine (GalNAc) specific bacterial lectin-like substance of E. corrodens (EcLS), which was one of the adhesins of E. corrodens. This is the first study to report that a bacterial lectin-like substance increased the expression of ICAM-1 on gingival epithelial cells.

Topics: Acetylgalactosamine; Bacterial Adhesion; Eikenella corrodens; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Flow Cytometry; Gene Expression Regulation, Bacterial; Gram-Negative Bacterial Infections; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; KB Cells; Lectins; Mouth Mucosa; Neutrophils; Periodontitis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

In other studies, we have found deviant functions in peripheral neutrophils in periodontitis. The aim here was to study (1) the release of cytokines, IL-8 and TNFalpha, from neutrophils in 15 treated periodontitis patients and pair-matched controls as well as (2) the effects of cigarette smoking.. Cytokines released in the incubation medium from un-stimulated and Fcgamma-R-stimulated neutrophils and some acute-phase reactants were measured with ELISA.. Non-smoking patients had trends for lower TNFalpha release compared to non-smoking controls, while corresponding trends were rather similar for Il-8. Smoking had a moderate but inconsistent effect on the release of both cytokines. However, in patients, the ratio between stimulated/un-stimulated release of Il-8 was significantly lowered by smoking (p<0.03). The parameters of inflammation in plasma differed only slightly between patients and controls, indicating that periodontal disease in a quiet phase has a negligible systemic effect with the possible exception for a higher IL-8 level. In contrast, smoking had significant systemic effect on the neutrophil count and IgG levels.. Release of IL-8 and TNF-alpha from peripheral neutrophils and various parameters of inflammation in plasma seem to be affected more by cigarette smoking than periodontal disease.

Topics: Acute-Phase Proteins; Adult; Aged; Antigens, CD; C-Reactive Protein; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin G; Interleukin 1 Receptor Antagonist Protein; Interleukin-6; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Periodontitis; Pilot Projects; Receptors, IgG; Receptors, Interleukin-1; Receptors, Tumor Necrosis Factor; Receptors, Tumor Necrosis Factor, Type I; Sialoglycoproteins; Smoking; Statistics, Nonparametric; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) has been implicated in a wide variety of diseases. Previous studies have demonstrated the expression of IL-8 in periodontal tissues, yet little is known about the exact source(s), mechanisms and factors involved in gingival expression of IL-8. Additionally, nothing is known about the presence and distribution of IL-8 receptors (IL-8R) in gingival cells. Therefore it was hypothesized that, in vivo, periodontal pathogens induce IL-8 expression from gingival keratinocytes (GK) which enhances leukocyte, microvascular endothelial cell (MVEC) and GK migration via specific IL-8 receptors present on these cells. The objective of the present study was to determine the distribution of IL-8 and IL-8R in gingival tissues and cultured human GK in vitro. Standard immunohistochemical and immunocytochemical techniques were utilized in order to localize IL-8 and its receptors CXCR-1 and CXCR-2 in archival gingival specimens (eight periodontitis and four non-inflamed controls) and in cultured gingival keratinocytes. It was demonstrated that, in vivo, IL-8 and IL-8R were present in gingival epithelium, MVEC and leukocytes. In vitro studies verified the above results, by showing expression of IL-8 and IL-8R in cultured gingival keratinocytes. It is concluded that IL-8 and IL-8 receptors are expressed in gingival epithelium both in vivo and in vitro. This new evidence indicates that epithelium plays a critical role in the host defense against invading pathogens and that keratinocytes can actively respond to IL-8 and other host cytokines, via specific receptors.

Topics: Adult; Aged; Antibodies; Cell Movement; Cells, Cultured; Coloring Agents; Endothelium, Vascular; Epithelium; Female; Gingiva; Humans; Immunoenzyme Techniques; Immunohistochemistry; Interleukin-8; Keratinocytes; Leukocytes; Male; Microcirculation; Middle Aged; Muscle, Smooth, Vascular; Periodontitis; Receptors, Interleukin-8A; Receptors, Interleukin-8B

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

Leukocyte migration is essential for immune surveillance of tissues by focusing immune cells to sites of antigenic challenge. The control of leukocyte migration depends on the combined actions of adhesion molecules and a vast array of chemokines and their receptors. The purpose of the present study was to investigate the involvement of Interleukin-8 (IL-8), RANTES, the associated infiltrating cells and expression of CCR5 chemokine receptors in periodontitis; furthermore, the effect of periodontal therapy on these parameters was evaluated. Patients included in the study had moderate to advanced periodontal disease with at least 5-6 teeth with probing depth > 6 mm, attachment loss > or =3 mm and extensive radiographic bone loss. The inflammatory infiltrate was analyzed by immunohistochemistry in gingival biopsies obtained from subjects at the beginning of the study and 2 months after periodontal treatment. Gingival crevicular fluid (GCF) was collected for 30 seconds using periopaper strips, and chemokines were quantified by ELISA. The cellular components of the inflammatory infiltrate included B (CD19) and T (CD3, CD4+ and CD8+) lymphocytes and monocytes/macrophages (CD11c). CCR5 chemokine receptor expressing cells were exclusively found in periodontitis gingiva. IL-8 and RANTES were detected in the periodontitis group, obtaining a total amount of 212.5 pg and 42.0 pg, respectively. However, IL-8 was also detectable in the GCF of the healthy group (total amount of 44.8 pg). Periodontal therapy reduced the cell number in the infiltrate and the levels of IL-8 and RANTES, suggesting a relationship between these chemokines and periodontal status. We propose that the presence of these chemokines and the expression of chemokine receptors may represent a marker of lymphocyte subsets with the ability to migrate to inflammatory sites.

Topics: Adult; Alveolar Bone Loss; Analysis of Variance; B-Lymphocytes; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Movement; Chemokine CCL5; Double-Blind Method; Female; Follow-Up Studies; Gingiva; Gingival Crevicular Fluid; Humans; Immunohistochemistry; Interleukin-8; Leukocytes; Macrophages; Male; Middle Aged; Monocytes; Observer Variation; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Receptors, CCR5; Statistics as Topic; T-Lymphocytes

Impaired polymorphonuclear neutrophil (PMN) functions were generally considered to be related to the onset of generalized aggressive periodontitis (GAgP). However, some research has indicated that the hyperreactivity of PMN seems to be involved in the inflammatory response of GAgP. The present study's main purpose was to provide more evidence about the role of PMN in the pathogenesis of GAgP by surveying PMN infiltration in gingiva and its relationship with the expression of their mediators including intercellular adhesion molecule-1 (ICAM-1), interleukin-8 (IL-8), interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha). The inflammatory response in GAgP was also compared with that in adult periodontitis (AP) and periodontally healthy subjects. Since these PMN mediators were reported to be produced mainly by macrophages, the association between the expression of these PMN mediators and the distribution of macrophages was also investigated.. A total of 25 gingival specimens were obtained from 10 GAgP patients, 10 AP patients, and 5 periodontally healthy subjects. Serial sections were obtained from each specimen, and the following techniques were adopted to investigate the distribution and interrelation of different cells and cytokines. Infiltration of PMN was observed by using hematoxylin and eosin staining. Distribution of the macrophages, identified as CD68+, was shown by using immunohistochemistry. Immunohistochemistry and in situ hybridization were used to detect the expression of ICAM-1, IL-8, IL-1beta, and TNF-alpha in gingival tissues. These techniques were performed in serial sections from each individual specimen.. Large numbers of infiltrating PMNs were observed in gingiva from GAgP. In gingiva from both GAgP and AP, the strongest protein and mRNA expression of IL-8, ICAM-1, IL-1beta, and TNF-alpha were located in pocket epithelium and adjacent connective tissue with large numbers of infiltrating PMNs. In tissues without abundant PMN infiltration, the appearance of positive cells expressing IL-8, ICAM-1, IL-1beta, and TNF-alpha was scattered. CD68+ was distributed sparsely in connective tissue and was hardly seen in pocket epithelium with large numbers of PMN infiltration. The degree of leukocyte infiltration and connective tissue destruction in gingiva from GAgP patients was not distinctly different from that in gingiva from AP. The gingival specimens with heavy PMN infiltration from both GAgP and AP patients presented strong expressions of IL-1beta and TNF-alpha; showed more extensive inflammatory cell infiltration; had severe connective tissue destruction; and presented severe elongation and ulceration of pocket epithelium. In gingiva from healthy subjects, inflammation was minor with visually no PMN, CD68+, or the positive cells of IL-8, ICAM-1, IL-1beta and TNF-alpha expression.. Enhanced accumulation of PMN, which is associated with the upregulation of IL-8, ICAM-1, IL-1beta, and TNF-alpha expression, relates to the severity and activity of GAgP. In addition to macrophages, PMN and/or epithelial cells might also be important sources of IL-8, IL-1beta, and TNF-alpha production in gingiva.

Topics: Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Coloring Agents; Connective Tissue; Epithelium; Female; Fluorescent Dyes; Gingiva; Humans; Immunohistochemistry; In Situ Hybridization; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Macrophages; Male; Neutrophil Infiltration; Neutrophils; Periodontal Pocket; Periodontitis; RNA, Messenger; Tumor Necrosis Factor-alpha; Up-Regulation

Gingival epithelial cells (GEC) are the first cells of the host that encounter the periodontal pathogens. and therefore their role in the initiation of the inflammatory response is critical. We aimed to: 1) characterize the expression of interleukin (IL)- Ialpha and IL-Ibeta in human gingiva and cultured GEC: 2) demonstrate the ability of A. actinomycetemcomitans extracts to upregulate IL-1alpha, IL-1beta and IL-8 expression in GEC in vitro: and 3) characterize the role of IL-1alpha and IL-1beta in the induction of IL-8 expression in GEC in vitro. Ten gingival biopsies (5 inflamed and 5 controls) and cultured GEC were examined for IL-1alpha and IL-Ibeta using immunohistochemical techniques. GEC were also challenged with A. actinomycetemcomitans extracts or IL-1alpha, and secretion of IL-1 and IL-8 was determined by ELISA. In vivo, IL-lalpha and IL-1beta were localized in the gingival epithelium and the infiltrating leukocytes. In vitro, A. actinomycetemcomitans extracts induced a time-dependent expression of IL-1alpha, IL-1beta and IL-8 in GEC. IL-1 inhibitors did not affect A. actinomycetemcomitans-induced IL-8. although they inhibited IL-8 induced by IL-1alpha or IL-1beta. In conclusion, GEC are a major source of IL-1alpha and IL-1beta in the periodontium, which in turn induce additional inflammatory mediators such as IL-8. Therefore GEC can be a potential target for therapeutic intervention in the future.

Topics: Aggregatibacter actinomycetemcomitans; Cell Line, Transformed; Epithelial Cells; Gingiva; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Periodontitis; Up-Regulation

To investigate the rôle of leukocytes in the pathogenesis of Papillon-Lefevre syndrome (PLS).. Peripheral blood polymorphonuclear neutrophils (PMNs), monocytes (MNs) and gingival crevicular fluid (GCF) were obtained from 2 cases of PLS with typical features. The chemotaxis of PMNs and MNs were evaluated using a modified Boyden chamber. The adherence of PMNs was determined by adherence of PMNs to petri dishes. Interleukin-8 (IL-8) in GCF was detected by sandwich ELISA. Elastase activity in GCF was measured with a low molecular weight substrate (S-2484) specific for granulocyte elastase.. PMNs from both patients showed depressed chemotactic response to FMLP and IL-8. Total amounts of IL-8 in GCF from the 2 patients were much higher than those of the normal controls. Elastase activity was not significantly different from that of the controls. The adherence of PMN and the chemotaxis of MN in the 2 patients were normal.. The depressed chemotactic response of PMN leads to decreased recruitment of PMN and/or release of lysozyme from PMN in the diseased gingival tissue, increasing the susceptibility of PLS patients to periodontal infection.

Topics: Adult; Case-Control Studies; Cell Adhesion; Chemotaxis, Leukocyte; Child; Disease Susceptibility; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Leukocyte Elastase; Lymphocyte Activation; Male; Monocytes; N-Formylmethionine Leucyl-Phenylalanine; Neutrophil Infiltration; Neutrophils; Papillon-Lefevre Disease; Periodontitis; Statistics, Nonparametric

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

Our aim was to compare the levels of laminin and interleukin-8 (IL-8) in GCF from inflamed shallow (GP) and deep pockets (PP) in patients with periodontitis to these levels in GCF from inflamed shallow pockets (GG) in subjects with gingivitis alone.. Lactoferrin was used as a marker for the number of neutrophils in the sites sampled. The periodontitis group consisted of 13 subjects, having at least 6 sites with a pocket depth > or = 5 mm. The healthy control group consisted of 12 subjects with no clinical signs of periodontal destruction. GCF was collected with paper strips and the volume was measured immediately after sampling. Laminin, lactoferrin and IL-8 were measured using specific antibodies with an ELISA.. The total amount of laminin was significantly higher in PP than in GG, but no significant difference was seen between GP and PP. The concentration and the total amounts of lactoferrin were similar in the three groups. The ratio between laminin and lactoferrin was higher in the samples from the patient, suggesting that neutrophils in patients are more activated and degrades more of the membrane per cell. The total amounts of IL-8 were very similar in the 3 groups, while the concentration also tended to be higher in the GP.. Our study showed higher amounts of laminin in GCF from patients with periodontitis suggesting the presence of hyperactive neutrophils during the transmigration process through the endothelium/epithelium.

Topics: Adult; Basement Membrane; Chemotaxis, Leukocyte; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Lactoferrin; Laminin; Male; Neutrophil Activation; Periodontitis; Statistics, Nonparametric

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

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

Periodontitis is characterised by tissue destruction caused by reactive oxygen species (ROS) and proteolytic enzymes, which are released by the interaction between bacteria and phagocytes. We estimated the ability of Fusobacterium species to induce release of tissue destructive and proinflammatory mediators from in vitro activated peripheral leukocytes. ROS was measured with the nitroblue tetrazolium (NBT) method, elastase with a specific chromogenic substrate and cytokines, including interleukin 1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), and interleukin 8 (IL-8) with a sandwich ELISA method. Various clinical isolates of unopsonized Fusobacterium species stimulated the neutrophils to an increased NBT- reduction. IL-1beta, TNFalpha, IL-8 and elastase were released in significantly higher levels from neutrophils stimulated by Fusobacterium species. In conclusion, unopsonized Fusobacterium species can induce increased production of oxygen radicals, cytokines and elastase from leukocytes activated in vitro.

Topics: Analysis of Variance; Cells, Cultured; Cytokines; Fusobacterium nucleatum; Humans; Interleukin-1; Interleukin-8; Leukocyte Elastase; Neutrophil Activation; Neutrophils; Periodontitis; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Various cytokines have been identified at sites of chronic inflammation such as periodontitis. Cytokines are synthesized in response to bacteria and their products, inducing and maintaining an inflammatory response in the periodontium. The purpose of the present study was to investigate the involvement of interleukin-1 beta (IL-1 beta), IL-8, and IL-10 and RANTES (regulated on activation, normally T cell expressed and secreted) and the cell populations associated with the immune response in destructive periodontitis, as well as the effect of periodontal therapy on cytokine levels in gingival crevicular fluid (GCF).. Data were obtained from 12 patients with moderate to advanced periodontitis and 6 healthy controls. Patients presenting at least 2 sites with > or =2 mm clinical attachment loss were included in the destructive periodontitis group. After monitoring for 4 months, only 6 patients showed destructive periodontitis and GCF samples and soft tissues biopsies were collected from these patients. GCF samples and biopsies were collected both from active (12 CGF samples and 6 biopsies) and inactive (12 CGF samples and 6 biopsies) sites. The comparison with healthy controls was carried out by collecting GCF samples from 6 healthy volunteers (12 samples) and biopsies during the surgical removal of wisdom teeth. In periodontal patients, clinical data and GCF samples were obtained prior to periodontal treatment (72 samples) and 2 months after periodontal therapy (72 samples). GCF was collected using a paper strip; eluted and enzyme-linked immunoabsorbent assays (ELISA) were performed to determine cytokine levels. The inflammatory infiltrate was analyzed by immunohistochemistry of gingival biopsy samples with monoclonal antibodies against CD3, CD8, CD4, CD11c, and CD19 antigens.. Cellular components of the inflammatory infiltrate include B and T lymphocytes and monocyte/macrophages. Active sites contained a higher number of B lymphocytes and macrophages. IL-8 and IL-1 beta and RANTES in GCF were detected in the majority of sites from periodontal patients (100%, 94% and 87%, respectively); IL-10 was found in only 43%. IL-8 was the only cytokine detected in the GCF (75%) of the control group. Moreover, IL-1 beta levels were significantly higher in active sites versus inactive sites (P <0.05). IL-8 and IL-10 and RANTES were increased in active sites; however, differences were not significant (P>0.05). A positive correlation between the IL-8 and RANTES (r = 0.677, P<0.05) was observed in periodontitis patients. Periodontal therapy reduced the total amount of IL-1 beta, IL-8, and IL-10 and RANTES. Data showed a weak correlation between the clinical parameters and the total amount of cytokines in periodontitis.. These data suggest that the amount of crevicular IL-1 beta, IL-8, and IL-10 and RANTES is associated with periodontal status. Removal of the bacterial plaque reduces the antigenic stimuli and consequently could modulate the chemokines present in GCF. We propose that the dynamic interactions between cytokines, their production rates, and their quantity could represent factors controlling the induction, perpetuation, and collapse of the cytokine network present in the periodontal disease.

Topics: Adult; Chemokine CCL5; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Humans; Immunohistochemistry; Interleukin-1; Interleukin-10; Interleukin-8; Male; Middle Aged; Periodontitis

We hypothesized that bacterial viability and strain characteristics of Porphyromonas gingivalis could affect the induction of pro-inflammatory mediator secretion by human gingival fibroblast cultures. Both killed and viable P. gingivalis elicited production of prostaglandin E2, interleukin-1 beta (IL-1 beta), IL-6 and IL-8, although killed P. gingivalis induced generally higher levels, particularly IL-6 and IL-8, compared with the viable bacteria. P. gingivalis strains, which exhibited wild-type levels of trypsin-like protease activity, stimulated human gingival fibroblasts to secrete increased levels of prostaglandin E2 and IL-1 beta, although minimal levels of IL-6 and IL-8 were noted in supernatants from the gingival fibroblast cells. P. gingivalis strains BEI and NG4B19, which have either decreased or undetectable levels of trypsin-like protease, respectively, induced significantly greater IL-6 and IL-8 levels in gingival fibroblast cultures compared with the other strains. The ability of antibody to P. gingivalis to alter human gingival fibroblast production of pro-inflammatory mediators was tested using nonhuman primate antisera. Both immune and nonimmune sera altered the P. gingivalis-generated pattern of mediators from the gingival fibroblasts. We conclude that: (i) viable and killed P. gingivalis were capable of inducing various pro-inflammatory cytokines from human gingival fibroblasts; (ii) strain differences in cytokine induction were noted, and the expression of a trypsin-like protease activity was related to decreased extracellular levels of IL-6 and IL-8; and (iii) the presence of serum, particularly with specific antibody to P. gingivalis, significantly altered human gingival fibroblast cytokine production compared with P. gingivalis alone.

Topics: Animals; Antibodies, Bacterial; Bacterial Proteins; Cell Line; Cytokines; Dinoprostone; Endopeptidases; Female; Fibroblasts; Gingiva; Humans; Inflammation Mediators; Interleukin-1; Interleukin-6; Interleukin-8; Leukotrienes; Macaca fascicularis; Periodontitis; Porphyromonas gingivalis; Species Specificity

The aim of this study was to investigate the relation between local expression of IL-8 and the localization of neutrophilic granulocytes, using CD16 as a marker of neutrophils. We also investigated the correlation between IL-8 and epithelial proliferation using proliferating cell nuclear antigen (PCNA) as a marker of proliferation. The distribution of IL-8, CD16 and PCNA/cyclin was determined by immunocytochemical techniques. We used cryostat-cut sections from gingival biopsies harvested from 5 subjects with and 5 subjects without periodontitis. Our histological examination demonstrated that the localization of neutrophilic granulocytes in gingival tissue from patients with periodontitis did not correlate with the expression of IL-8. In all tissue sections from patients and controls, the inflammatory cells accumulated near the pocket epithelium and only a few leukocytes deviated from this pattern. In the patient group, keratinocytes not belonging to the pocket or junctional epithelium expressed IL-8 without any evidence of a chemoattractant effect on neutrophils. The marker of proliferation, PCNA/cyclin, was expressed in keratinocytes in the basal cell layer. The expression was less pronounced in the control group. Our finding that IL-8 was expressed in proliferating cells suggests that IL-8 may have a role in keratinocyte proliferation.

Topics: Adult; Aged; Biomarkers; Case-Control Studies; Cell Division; Female; Gingiva; Humans; Immunoenzyme Techniques; Interleukin-8; Keratinocytes; Male; Middle Aged; Neutrophil Infiltration; Periodontitis; Proliferating Cell Nuclear Antigen; Receptors, IgG

Periodontitis is the inflammatory response in periodontal tissues elicited by bacterial colonization in periodontal pockets. In this response, pocket epithelial cells are the first cells to come into contact with bacteria. To elucidate this mechanism, we determined the adherence of the periodontopathic bacterium Eikenella corrodens 1073, which has a GalNAc-sensitive lectin-like adhesin (EcLS), to a human oral epithelial carcinoma cell line (KB) and the induction of proinflammatory cytokine production in the cells following exposure to this bacterium in vitro. In the adherence assay, EcLS played a role as the adhesin of this bacterium in adherence to KB cells. In a reverse transcriptase PCR, significant interleukin-8 (IL-8) and IL-6 mRNA levels were induced in response to exposure to this bacterium. In an enzyme-linked immunosorbent assay after an 8-h bacterial exposure, the IL-8 and IL-6 protein levels were 13.5- and 8.3-fold higher than those in the nonexposed controls, respectively. These protein responses were time dependent. Interestingly, when E. corrodens was separated from KB cells by cell culture inserts, a slight stimulation of the IL-6 and IL-8 mRNA and secreted protein levels was seen. These results imply that the direct contact of E. corrodens 1073 with oral epithelial cells is not necessarily required for the stimulation of IL-6 and IL-8 secretion. We suggest that E. corrodens induces the epithelial cells to secrete proinflammatory cytokines which serve as an early signaling system to host immune and inflammatory cells in underlying connective tissues.

Topics: Adhesins, Bacterial; Bacterial Adhesion; Eikenella corrodens; Epithelial Cells; Gram-Negative Bacterial Infections; Humans; Interleukin-6; Interleukin-8; KB Cells; Kinetics; Lectins; Mouth Mucosa; Periodontitis; RNA, Messenger; Time Factors

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

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

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

Polymorphonuclear neutrophils (PMN) are the most abundant immune cells in inflammatory gingival sites of patients with early onset periodontitis, localized juvenile periodontitis, and rapidly progressive periodontitis (RPP). In the latter, the large number of PMN in connective tissue may explain the marked gingival destruction. Because interleukin-8 (IL-8) is a potent PMN chemoattractant, we evaluated circulating levels and gingival mRNA expression of IL-8. We found high IL-8 plasma levels as well as strong IL-8 mRNA expression in both epithelial and connective gingival cells from patients with RPP. Moreover, the gingival PMN themselves contained IL-8 mRNA, suggesting an autoamplification of PMN recruitment and activation in the gingiva. We also measured the expression of adhesion molecules at the PMN surface as well as the oxidative burst in whole blood from 14 patients with RPP, using flow cytometry to avoid irrelevant stimulations and to analyze single cells. In RPP patients, resting PMN showed reduced L-selectin, Lewis x, and sialyl Lewis x antigen expression as well as increased H2O2 production. These modifications of PMN adhesion molecule expression, together with their increased basal oxidative burst and excessive IL-8 production, may contribute to the noxious inflammatory reaction, which may in turn be autopotentiated by PMN production of IL-8. In addition, PMN showed a lack of increased response (H2O2 production) to formyl peptides after ex vivo priming with IL-8, possibly owing to IL-8 desensitization that may be involved in the increased susceptibility of RPP patients to infection. After appropriate treatment of RPP, the reduction in inflammation was associated with a return to control levels of both plasma IL-8 and PMN functions, suggesting that these features are linked.

Topics: Adult; Cell Adhesion Molecules; Cytokines; Female; Humans; Hydrogen Peroxide; Interleukin-8; Male; Middle Aged; Neutrophils; Oxidants; Periodontitis; RNA, Messenger

Periodontitis is a chronic inflammation of the supporting structures of the dentition which constitutes one of the most common causes of adult tooth loss. While certain microorganisms have been associated with the onset of the disease process, the exact pathogenetic mechanisms underlying periodontal destruction are still poorly understood. We have tested the hypothesis that gingival fibroblasts from diseased sites contribute to pathogenesis by possessing a secretory phenotype characterized by an exuberant secretion of inflammatory mediators and cytokines. Of the cytokines and mediators tested, fibroblast IL-1beta and prostaglandin E2 (PGE2) secretion was not different between health and disease. However, we have shown that fibroblasts from periodontal lesions produce in vitro greater amounts of IL-6 and IL-8 constitutively than healthy controls. When fibroblasts were stimulated with a panel of endogenous or exogenous response modifiers, the magnitude of cytokine and mediator stimulation above constitutive levels did not differ between health and disease. A strong positive correlation was identified between IL-6 or IL-8 constitutive secretion levels in vitro and the in situ expression of these cytokines within the connective tissues from where these cells originated, indicating that the in vitro phenotype mirrors their in vivo function. Furthermore, we present evidence which indicates that increased cytokine secretion by fibroblasts in disease is due to an elevated proportion of subpopulations with higher cytokine secretory capacity. Finally, we demonstrated that cultures from diseased sites are composed of cells with higher levels of constitutive CD40 expression, which may contribute to the increased IL-6 and IL-8 secretory phenotype.

Topics: Adult; Aged; CD40 Antigens; Cell Lineage; Cells, Cultured; Chronic Disease; Connective Tissue; Dinoprostone; Fibroblasts; Fluorescent Antibody Technique, Indirect; Gingiva; Humans; Inflammation Mediators; Interleukin-1; Interleukin-6; Interleukin-8; Middle Aged; Periodontitis; Phenotype; Vimentin

The aim of this study is to identify the nature of the IL-8 inhibitor(s) in gingival crevicular fluid (GCF).. Part 1: Fourteen GCF samples were collected from 13 adult periodontitis (AP) patients, and 9 samples were taken from 8 healthy subjects. Each GCF sample was divided into two aliquots. A serine protease-specific inhibitor, PMSF, was added to one aliquot, and PBS was added to the other aliquot as control. ELISA was used to measure IL-8 level in the samples. Part 2: Forty-one GCF samples were collected from 15 AP patients and indirect ELISA was performed to detect the anti-IL-8 IgG antibody.. Part 1: IL-8 level in the PMSF samples was significantly greater than that in the control group (3.01 +/- 5.79 mg/L vs 0.05 +/- 0.15 mg/L, respectively P < .001). Part 2: The mean value of anti-IL-8 IgG in GCF was greater than that of negative control + 3 x SD.. A serine protease that can "cleave" IL-8 exists in GCF. Gingival crevicular fluid from AP sites contains an autoantibody against IL-8.

Topics: Adult; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Humans; Immunoglobulin G; Interleukin-8; Linear Models; Male; Middle Aged; Periodontitis; Recombinant Proteins; Serine Endopeptidases

It has been reported that 48%-85% of rapidly progressive periodontitis (RPP) are associated with a defect in polymorphonuclear neutrophil (PMN) chemotactic function. However, some reports failed to detect any defect in PMN chemotaxis in RPP patients. Further, the influence of race on neutrophil function has been suggested. The purpose of the present study was to determine the PMN chemotactic function in Chinese RPP patients.. Neutrophils were obtained from heparinized peripheral blood of 16 RPP and 14 periodontally healthy control subjects matched for age and sex. Cells were isolated by Percoll discontinuous density gradient centrifugation. Chemotaxis was evaluated using modified Boyden chamber. (N-formyl-methionyl-leucyl-phenylalanine, FMLP) (10(-8) mol/L) and IL-8 (100 micrograms/L) which is a more specific neutrophil chemokines were used in this study. The results were determined by staining the filters followed by visual counting under microscopy at 600 x. Data was analyzed using the Students' t test and the value of RPP was compared with that of control subjects(> mean +/- 2s or < mean +/- 2s).. The results showed all RPP patients had normal PMN chemotactic behavior.

Topics: Adult; Chemotaxis, Leukocyte; Female; Humans; Interleukin-8; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Periodontitis

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

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

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

Chemotactic activity in gingival crevicular fluids (GCF) from 39 cases of patients with periodontitis and 10 cases of healthy donors was determined via Boyden method.. Chemotactic activity in GCF from patients was much higher than that from healthy donors. Chemotactic activity in GCF was not relative to pocket depth and attachment loss, but markely correlative to gingival index and bleeding index.. The results indicates that leukocyte chemokine activity would not reflect the degree of periodontal damage, but might be a marker for periodontal inflammatory reaction and disease activity. It is likely that interleukin-8 might be a major component among the chemokines in GCF.

Topics: Adult; Aged; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Male; Middle Aged; Periodontal Index; Periodontitis

The role of interleuking-8 (IL-8), a neutrophil-attracting and-activating cytokine, was investigated in gingival crevicular fluid (GCF). ELISA was used to detect the levels of IL-8 in GCF collected from 54 adult periodontitis (AP) patients (105 teeth) and 24 healthy subjects (54 teeth). The results showed that 1. The role of IL-8 was dependent upon the concentration of IL-8 in GCF. IL-8, in the low concentration (< or = 30 micrograms/L), was positively correlated with bleeding index (r = 0.36, P < 0.01). While in the high concentration (> 30 micrograms/L), was negatively correlated with bleeding index and probing depth (r = -0.54 and r = -0.65 respectively, P < 0.01). In the majority of periodontitis sites (91%), the concentration of IL-8 in GCF were lower than 30 microliters/L. IL-8 most likely acted as pro-inflammatory factor in these teeth. 2. IL-8 was a two-way regulator of inflammation, pro-inflammation and anti-inflammation. The threshold of IL-8 between inducing and suppressing inflammation was approximately 30 micrograms/L. Within the confined range, IL-8 concentration greater than 30 micrograms/L could be inflammation suppressive, while a less-than-30 micrograms/L concentration of IL-8 might become inflammation inducing.

Topics: Adult; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Male; Middle Aged; Periodontal Index; Periodontitis

In order to confirm the existence of Interleukin-8 (IL-8) inhibitor and its biological activity in gingival crevicular fluid (GCF), this study examined the GCF taken from 7 adult periodontitis (AP) patients. In neutralization test of IL-8, the results showed that the mean level of IL-8 was less than 1 ng/ml, which had been added into the GCF before ELESAs was performed to measure the amount of IL-8 in GCF. The mean level of IL-8 in the GCF of AP group was significantly lower than that of healthy group (P < 0.001). In biological activity test of IL-8 inhibitor, using pooled GCF taken from 8 AP patients (23 teeth), the results showed that the GCF (without recombinant human IL-8, rhIL-8) caused more white blood cell (WBC) migration than blank control group (physiological saline) did. When the amount of rhIL-8 increased in GCF from 0.1 microgram to 1 microgram, the WBC count increased by 18.6% which was less than the increase rate (49.1%) in control group with same dose of IL-8. In the saline group containing rhIL-8, the WBC chemotactic response appeared as an inverted "V"-shape curve. All these data suggested that 1. Certain kinds of IL-8 inhibitor exist in GCF which can "cleave" IL-8. 2. The level of IL-8 inhibitor(s) increases significantly in the GCF from periodontitis sites. 3. The GCF of adult periodontitis patient has strong chemotactic effects on WBC. IL-8 inhibitor(s) in GCF can slightly suppress the chemotactic effect induced by IL-8. When assessing the role of IL-8 in pathophysiology, the high and low dose of IL-8 might have different sense.

Topics: Adult; Chemotaxis, Leukocyte; Gingival Crevicular Fluid; Humans; Interleukin-8; Periodontitis; Recombinant Proteins

Bacteria can indirectly affect the course of periodontal diseases by activating host cells to produce and release inflammatory mediators and cytokines. These mediators and cytokines manifest potent proinflammatory and catabolic activity and may play key roles in local amplification of the immune response as well as in periodontal tissue breakdown. This study tested the effect of Actinobacillus actinomycetemcomitans (Aa) and Campylobacter rectus (Cr) challenge on PGE2, IL-1 beta, IL-6 and IL-8 production by human gingival fibroblasts (HGF). Contact-inhibited HGF were prepared and formalin-killed bacterial cells (Aa JP2, ATCC 29523 & 33384 and Cr ATCC 33238) at 10(6)-10(9) were added to the HGF. Culture supernatants were collected at varying time intervals and analyzed for cytokine and mediator content. All concentrations of Aa JP2 and Cr ATCC 33238 suppressed IL-1 beta production up to approximately 50% during the initial 3-12-h period. No bacterial concentration tested was able to increase IL-1 beta production above the maximum basal levels. Both bacterial species stimulated production of IL-6 and IL-8. Aa JP2 did not affect PGE2 levels significantly, whereas Cr ATCC 33238 was stimulatory only at the highest concentration tested (10(9)). There were no significant differences among the three Aa strains with respect to IL-1 beta production. However, Aa ATCC 29523 and ATCC 33384 were less capable of stimulating IL-6 secretion and more efficient in stimulating IL-8 production than Aa JP2. In general, Cr was the most potent enhancer of cytokine and mediator production by HGF. In conclusion, Aa and Cr are capable of amplifying the local immune response and promoting periodontal tissue inflammation by stimulating HGF to secrete mainly IL-6 and IL-8.

Topics: Aggregatibacter actinomycetemcomitans; Campylobacter; Cells, Cultured; Cytokines; Dinoprostone; Fibroblasts; Gingiva; Humans; Immunization; Inflammation Mediators; Interleukin-1; Interleukin-6; Interleukin-8; Periodontitis; Periodontium; Serotyping

Within the in vivo environment human gingival fibroblasts (HGF) may be challenged with bacteria or bacterial products. This interaction may result in the release of cytokines which are directly or indirectly involved in connective tissue and bone catabolism, such as interleukin (IL)-1 beta, IL-6, and IL-8. Our investigation has tested the hypothesis that HGF from Actinobacillus actinomycetemcomitans (Aa)-infected patients with rapidly destructive forms of periodontitis, such as localized juvenile periodontitis (LJP), respond to Aa challenge with an exaggerated secretion of IL-1 beta, IL-6, and IL-8. We have compared the in vitro profiles of these cytokines by Aa-challenged HGF obtained from 2 healthy subjects, 2 Aa-infected, slowly progressing adult periodontitis (AP) patients and 2 LJP patients. HGF were challenged throughout a 48-hour period with formalinized whole bacterial cells, and culture supernatants were analyzed for cytokine content using RIA. No differences were noted in the IL-1 beta secretion levels among the different HGF cultures. Although basal (unchallenged) IL-6 and IL-8 production was similar in all HGF cultures, HGF from the two LJP patients responded to Aa challenge with a more rapid IL-6 and a more pronounced IL-8 secretion than healthy or AP HGF. We also tested the ability of human serum antibodies against Aa to moderate the Aa-elicited HGF cytokine secretion by adding human serum, with normal or elevated antibody content. Both sera appeared to have an upregulating effect on IL-6 and IL-8 secretion. Depletion of 95% of the anti-Aa antibody from serum by absorption did not affect its activity. Based on the response of HGF from two LJP patients, we propose that Aa-induced pathology in LJP may be modulated by stimulation of rapid and/or exaggerated secretion of cytokines with potential catabolic effects, although studies with a larger group of LJP patients are needed to further test this hypothesis. Furthermore, serum antibodies against this microorganism do not appear to have a neutralizing effect in cytokine-eliciting HGF-Aa interactions.

Topics: Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Antibodies, Bacterial; Cells, Cultured; Female; Fibroblasts; Gingiva; Humans; Immunoglobulin G; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Male; Middle Aged; Periodontitis

Cytokines play an important role in the pathology associated with chronic inflammatory diseases. We measured the total amounts [picograms (pg)] and concentrations.(pg/microliter) of interleukin-1 alpha (IL-1 alpha), interleukin-8 (IL-8) and interferon-alpha (IFN-alpha) in 20 s gingival crevicular fluid (GCF) samples obtained from 2 diseased and 2 healthy sites in 20 subjects with periodontitis, and from 2 healthy sites in 20 subjects without disease. Both the mean amount and concentration of IL-1 alpha were significantly higher (p < 0.001) in diseased sites compared to healthy sites in subjects with disease. The results for IL-8 and IFN-alpha differed depending on the method of reporting. Whereas the amount of IL-8 was significantly higher (p < 0.01) in diseased sites, the mean concentration of IL-8 was lower compared to healthy sites. The mean amount of IFN-alpha was similar in health and disease; however, the concentration of IFN-alpha was significantly lower in diseased sites (p < 0.001) corresponding to the significant increase in crevicular fluid volume (p < 0.001). There were no significant differences in the amount or concentrations of the 3 cytokines between healthy sites from subjects with disease and healthy sites from healthy controls. The total amounts of both IFN-alpha and IL-8 were correlated between healthy and diseased sites in subjects. These data suggest that, while the disease status of a site is the major determinant of the levels of these cytokines locally, subjects with high levels of IL-8 and IFN-alpha in healthy sites also tend to have high levels of these cytokines in diseased sites. Finally, both the concentrations and total amounts of IL-8 and IFN-alpha were significantly correlated in diseased sites, suggesting that levels of these two cytokines rise or fall in tandem. The combination of decreased IL-8 and decreased IFN-alpha concentrations at diseased sites may reflect the reduced anti-bacterial host defense activity at that site.

Topics: Adult; Aged; Case-Control Studies; Chi-Square Distribution; Female; Gingival Crevicular Fluid; Humans; Interferon-alpha; Interleukin-1; Interleukin-8; Linear Models; Male; Middle Aged; Periodontal Index; Periodontitis; Sensitivity and Specificity; Statistics, Nonparametric

Recent in vitro findings indicate that cytokines represent an important pathway of connective tissue destruction in human periodontitis. The biological effects of interleukin-1 beta (IL-1 beta) and interleukin-8 (IL-8) are relevant in this regard, and the objective of this study was to compare the levels of these molecules in gingival crevicular fluids (GCF) from patients with adult periodontitis (experimental group) and from individuals with clinically healthy gingiva (control group). GCF was collected for 30 seconds using a periopaper strip and the volume of the sample determined. Following elution of the fluid, assays for IL-1 beta and IL-8 were carried out by ELISA. The concentrations (ng/ml) of cytokines were calculated in the original volume of GCF on each strip. The total amounts (pg/site) of cytokines were expressed as the concentrations multiplied by volumes of GCF: The total amounts of IL-1 beta and IL-8 of the experimental group were significantly higher than the control group. The total amounts of both cytokines were markedly reduced following phase 1 periodontal treatment. The clinical parameters were positively related to the total amounts of IL-1 beta and IL-8. IL-1 beta concentrations and total amounts were also positively related to IL-8 suggesting that the GCF IL-8 levels are influenced by local IL-1 beta activities. These data indicate that the total amounts of IL-1 beta and IL-8 exhibited dynamic changes upon severity of periodontal disease. The levels of IL-1 beta and IL-8 in GCF are valuable in detecting the inflammation of periodontal tissue.

Topics: Adult; Dental Plaque Index; Enzyme-Linked Immunosorbent Assay; Female; Gingiva; Gingival Crevicular Fluid; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Tooth Mobility

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

Gingival crevicular fluid (GCF) IL-8 and IL-1 beta levels were determined by sandwich enzyme-linked immunosorbent assays. Associations between IL-8 and IL-1 beta GCF levels, and between these cytokines and patient estrogen status were evaluated. IL-8 and IL-1 beta were detected more frequently and in higher amounts/30 s GCF sample in estrogen-deficient patients than in estrogen-sufficient patients. IL-8 and IL-1 beta GCF levels were significantly correlated. These findings suggest that GCF IL-8 levels are associated with patient estrogen status and local IL-1 beta concentrations.

Topics: Adult; Age Factors; Chi-Square Distribution; Enzyme-Linked Immunosorbent Assay; Estrogens; Female; Gingival Crevicular Fluid; Humans; Interleukin-1; Interleukin-8; Linear Models; Menopause; Middle Aged; Periodontal Attachment Loss; Periodontitis

Topics: Base Sequence; Collagenases; Cytokines; DNA Primers; Extracellular Matrix; Gene Expression Regulation, Enzymologic; Humans; Interleukin-8; Matrix Metalloproteinase 3; Matrix Metalloproteinase 8; Metalloendopeptidases; Molecular Sequence Data; Neutrophils; Periodontitis; Polymerase Chain Reaction; RNA, Messenger

Cells expressing messenger RNA (mRNA) for inflammatory cytokines [interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha), IL-6, IL-8] were demonstrated by in situ hybridization in human inflamed gingiva. When this technique was used in conjunction with immunohistochemistry, IL-1 alpha and/or beta and IL-8 messages were observed predominantly on macrophages infiltrating the gingiva. TNF-alpha messages were abundant on macrophages and T cells. In contrast, the IL-6 mRNA were more widely distributed on many types of cells such as macrophages, T cells, fibroblasts, endothelial cells and B cells. This study clearly identified the cells which express mRNA for inflammatory cytokines in inflamed gingiva and suggested an involvement of cytokine network in the generation of human periodontitis.

Topics: Adult; Gingiva; Humans; Immunoenzyme Techniques; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Nucleic Acid Hybridization; Periodontitis; RNA, Messenger; Tumor Necrosis Factor-alpha

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