interleukin-8 and Aggressive-Periodontitis

This study aims to determine the effects of Er,Cr:YSGG and diode laser treatments on IL-1β, IL-8, and TNF-α levels in patients with generalized aggressive periodontitis.. Twenty-six generalized aggressive periodontitis patients were enrolled in the study. We performed three treatment models: "scaling and root planning (SRP-only)," "SRP + Er,Cr:YSGG laser," and "SRP + diode laser." Each experimental quadrant was randomly allocated to the control group or the test group. The IL-1β, IL-8, and TNF-α levels were analyzed with an enzyme-linked immune-sorbent assay.. When the baseline and post-treatment IL-1β, IL-8, and TNF-α levels were compared, the most significant difference was observed in the SRP + Er,Cr:YSGG group and the least difference was observed in the SRP-only group.. The use of Er,Cr:YSGG laser as an addition to the conventional mechanical periodontal treatment was found to be more successful than the diode laser + SRP use in aggressive periodontitis treatment.

Topics: Aggressive Periodontitis; Humans; Interleukin-8; Lasers, Semiconductor; Lasers, Solid-State; Tumor Necrosis Factor-alpha

The association between herpetic/bacterial co-infection and periodontal diseases has been reported. However, how interactions between herpesviruses and periodontal bacteria dampen periodontal inflammation is still unclear. This study determined effects of co-infection with oral bacteria, including Streptococcus sanguinis, Fusobacterium nucleatum or Aggregatibacter actinomycetemcomitans, in herpes simplex virus type 1 (HSV-1)-infected oral epithelial cells.. Cell viability was determined by detection the activity of mitochondrial dehydrogenase. Viral production was measured using the plaque assay. Levels of bacterial and viral DNA were determined by real-time polymerase chain reaction. Secretion of interleukin (IL)-6 and IL-8 was measured using the enzyme-linked immunosorbent assay.. Viability was not further reduced by bacterial co-infection in HSV-1-infected cells. Co-infection with HSV-1 and S. sanguinis or F. nucleatum reduced the viral yield whereas co-infection with HSV-1 and A. actinomycetemcomitans significantly enhanced the viral yield in oral epithelial cells. The enhancing effect of A. actinomycetemcomitans was not affected by bacterial heat-inactivation. Co-infection with HSV-1/A. actinomycetemcomitans increased intracellular levels of both viral and bacterial DNA. Secretion of IL-6 and IL-8 stimulated by A. actinomycetemcomitans infection was partly reduced by co-infection with HSV-1 in oral epithelial cells.. In contrast to S. sanguinis and F. nucleatum, A. actinomycetemcomitans enhanced the yield of HSV-1. Either HSV-1 or A. actinomycetemcomitans may be benefited from co-infection, in aspects of increases in production of viral and bacterial DNA as well as reductions in cytokine secretion. These findings echoed with previous clinical studies showing co-infection of HSV and A. actinomycetemcomitans in patients with aggressive periodontitis.

Topics: Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Coinfection; DNA, Bacterial; Epithelial Cells; Herpesvirus 1, Human; Humans; Interleukin-6; Interleukin-8

The aim of the study is to assess the long-term effect of active periodontal therapy on serum inflammatory parameters in patients with aggressive (AgP) and chronic (ChP) periodontitis in a non-randomised clinical study.. Twenty-five ChP and 17 AgP were examined clinically prior to (baseline), 12 weeks and 60 months after subgingival debridement of all pockets within 2 days. Systemic antibiotics were prescribed if Aggregatibacter actinomycetemcomitans was detected (10 AgP, 8 ChP), flap surgery was rendered if required. Neutrophil elastase (NE), C-reactive protein (CRP), lipopolysaccharide binding protein, interleukin 6, 8, and leukocyte counts were assessed at baseline, 12 weeks and 60 months.. Clinical parameters improved significantly in both groups from 12 weeks to 60 months. Eleven AgP and 18 ChP patients received surgical treatment after the 12 weeks examination. Only 3 patients in each group attended ≥ 2 supportive maintenance visits per year. NE and CRP were significantly higher in AgP than ChP at baseline and 60 months (p < 0.01). For leukocyte counts in ChP, significant changes were observed (baseline: 6.11 ± 1.44 nl. Despite comprehensive periodontal treatment, AgP patients exhibit higher NE and CRP levels than ChP patients up to 5 years after therapy.. Systemic inflammatory burden in AgP patients is higher than in ChP patients even 5 years after periodontal treatment.

Topics: Acute-Phase Proteins; Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Anti-Bacterial Agents; Biomarkers; C-Reactive Protein; Carrier Proteins; Chronic Periodontitis; Debridement; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Male; Membrane Glycoproteins; Surgical Flaps

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

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

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

Calprotectin, a heterodimer of S100A8 and S100A9 subunits, is associated with inflammatory disorders such as rheumatoid arthritis and cystic fibrosis. Although calprotectin levels are increased significantly in the gingival crevicular fluid (GCF) of periodontitis patients, its effects on periodontal ligament cells (PDLCs) remain largely unknown. The aim of this study was to evaluate calprotectin levels in the GCF of generalized aggressive periodontitis (AgP) patients and to investigate the effects of recombinant human calprotectin (rhS100A8/A9) and its subunits (rhS100A8 and rhS100A9) in PDLCs. Both the concentration and amount of crevicular calprotectin were significantly higher in the AgP group compared with healthy controls. In addition, the GCF calprotectin levels were correlated positively with clinical periodontal parameters including bleeding index, probing depth, and clinical attachment loss. rhS100A8/A9 promoted cell apoptosis, whereas rhS100A8 and rhS100A9 individually exerted little effect on apoptosis in PDLCs. rhS100A9 and rhS100A8/A9 increased the activation of nuclear factor-κB (NF-κB) by promoting the nuclear translocation of p65 in PDLCs, subsequently inducing expression of the pro-inflammatory cytokines IL-6, IL-8, TNFα, and COX2. Treatment with an NF-κB inhibitor partially reversed the rhS100A9- and rhS100A8/A9-induced upregulation of the pro-inflammatory cytokines. rhS100A9, and not rhS100A8, was mainly responsible for the pro-inflammatory role of calprotectin. Collectively, our results suggest that calprotectin promotes apoptosis and the inflammatory response in PDLCs via rhS100A9. These findings might help identify novel treatments for periodontitis.

Topics: Adult; Aggressive Periodontitis; Apoptosis; Calgranulin A; Calgranulin B; Case-Control Studies; Connective Tissue Cells; Cyclooxygenase 2; Female; Gene Expression; Gingival Crevicular Fluid; Humans; Interleukin-6; Interleukin-8; Leukocyte L1 Antigen Complex; Male; NF-kappa B; Periodontal Ligament; Primary Cell Culture; Proline; Recombinant Proteins; Thiocarbamates; Tumor Necrosis Factor-alpha

Assessment of the effect of non-surgical periodontal therapy (SRP) on serum inflammatory parameters in patients with untreated aggressive (AgP) and chronic (ChP) periodontitis.. Overall, 31 ChP and 29 AgP were examined clinically prior to and 12 weeks after SRP (subgingival scaling of all pockets within 2 days) with systemic antibiotics for patients positive for Aggregatibacter actinomycetemcomitans (14 AgP, 9 ChP). Blood was sampled prior to, one day, 6, and 12 weeks after the first SRP visit. Serum elastase, C-reactive protein (CRP), lipopolysaccharide-binding protein (LBP), interleukin (IL) 6, 8, and leukocyte counts were assessed.. At baseline, serum elastase, CRP, and LBP were significantly (p < 0.01) higher in AgP than ChP. Serum elastase, CRP, LBP, and IL-6 were significantly (p < 0.001) elevated one day after scaling in both groups. Both groups showed significant clinical improvement (p < 0.001). A significant difference was observed regarding change of serum elastase 12 weeks after SRP between AgP and ChP (p = 0.015). Multiple regression analysis revealed AgP, African origin, and bleeding on probing to be associated with more pronounced elastase reduction. CRP reduction was associated with African origin, systemic antibiotics, and baseline probing pocket depth.. SRP results in serum elastase reduction in AgP but not in ChP.

Topics: Acute-Phase Proteins; Adolescent; Adult; Aggressive Periodontitis; Analysis of Variance; Anti-Bacterial Agents; Asian People; Black People; C-Reactive Protein; Carrier Proteins; Chronic Periodontitis; Dental Scaling; Female; Humans; Interleukin-6; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Linear Models; Male; Membrane Glycoproteins; Periodontal Index; Statistics, Nonparametric; White People; Young Adult

Periodontitis is an inflammatory disease characterized by connective tissue loss and alveolar bone destruction. Interleukin-8 (IL8) is important in the regulation of the immune response. The aim of this study was to analyze four polymorphisms in the IL8 gene in relation to chronic (CP) and aggressive (AgP) periodontitis.. A total of 492 unrelated subjects were included in this case-control association study. Genomic DNA of 278 patients with CP, 58 patients with AgP, and 156 controls were genotyped, using the 5' nuclease TaqMan assay, for IL8 (rs4073, rs2227307, rs2227306, and rs2227532) gene polymorphisms. Subgingival bacterial colonization was investigated by the DNA-microarray detection kit in a subgroup of subjects (N = 247).. Allele and genotype frequencies of all investigated IL8 polymorphisms were not significantly different between the subjects with CP and/or AgP and controls (P > 0.05). Nevertheless, the A(-251)/T(+396)/T(+781) and T(-251)/G(+396)/C(+781) haplotypes were significantly less frequent in patients with CP (2.0% versus 5.1%, P < 0.02, OR = 0.34, 95% CI: 0.15-0.78, resp., 2.0% versus 4.5%, P < 0.05, OR = 0.41, 95% CI: 0.18-0.97) than in controls.. Although none of the investigated SNPs in the IL8 gene was individually associated with periodontitis, some haplotypes can be protective against CP in the Czech population.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Case-Control Studies; Haplotypes; Humans; Interleukin-8; Middle Aged; Polymorphism, Single Nucleotide

Interleukin (IL)-8 is an important chemokine for regulation of the inflammatory response. A single nucleotide polymorphism (SNP) reference sequence (rs) 4073 in the IL8 gene has been shown to regulate IL-8 levels after stimulation with lipopolysaccharide. This study investigates the transmission pattern of the IL8 rs4073 risk allele A and its association with susceptibility to aggressive periodontitis (AgP) in families and in a case-control cohort of unrelated individuals from a Brazilian population.. Genotyping was performed by standard polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) in 13 nuclear families and 184 unrelated subjects. Statistical analysis was performed using the transmission disequilibrium test (TDT) for the family dataset and Chi-square test and multivariate logistic regression modelling for the case-control dataset.. TDT analyses did not detect evidence of over transmission of IL8 rs4073 alleles in affected and unaffected family members (allele T: 52%; allele A: 48%; p=0.2252). How expected, analyses of cases and unrelated controls showed a significant and inverse association of age with AgP; however, a lack of association between genotypes, ethnic groups and generalized AgP was observed.. The SNP (rs4073) was not associated with AgP in unrelated individuals and there is no evidence of over transmission of the alleles in families with AgP, from Brazilian individuals.

Topics: Aggressive Periodontitis; Alleles; Brazil; Case-Control Studies; Female; Genotype; Humans; Interleukin-8; Male; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide

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

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

Papillon-Lefèvre syndrome (PLS) is characterised by aggressively progressive periodontitis combined with palmo-plantar hyperkeratosis. It is caused by "loss of function" mutations in the cathepsin C gene. The hypothesis behind this study is that PLS patients' polymorphonuclear leukocytes (PMNs) produce more proinflammatory cytokines to compensate for their reduced capacity to neutralize leukotoxin and to eliminate Aggregatibacter actinomycetemcomitans. Production of more interleukin (IL)-8 would result in the attraction of more PMNs. The aim of this study was to evaluate the cytokine profile in PLS patients' blood cultures. Blood was sampled from eight PLS patients (one female) from six families (antiinfective therapy completed: six; edentulous: two) with confirmed cathepsin C mutations and deficient enzyme activity. Nine healthy males served as controls. Whole blood cultures were stimulated with highly pure lipopolysaccharide (LPS) from Escherichia coli R515 and IL-1β plus tumor necrosis factor (TNF)-α. Thereafter, release of IL-1β (stimulation: LPS and LPS plus adenosine triphosphate), IL-6, IL-8, interferon-inducible protein (IP)-10, and interferon (IFN)-γ (stimulation: LPS, IL-1β/TNFα) were detected by ELISA. Medians of cytokine release were, with the exception of IP-10, slightly higher for PLS than for controls' cultures. None of these differences reached statistical significance. Increased production of IL-1β, IL-6, IL-8, IP-10, or IFNγ as a significant means to compensate for diminished activity and stability of polymorphonuclear leukocyte-derived proteases could not be confirmed in this study. Cytokine profiles in blood cultures may not be used to identify PLS patients.

Topics: Adenosine Triphosphate; Adolescent; Adult; Aggressive Periodontitis; Biomarkers; Cathepsin C; Chemokine CXCL10; Child; Cytokines; Escherichia coli; Female; Humans; Inflammation Mediators; Interferon-gamma; Interleukin-1beta; Interleukin-6; Interleukin-8; Leukocytes; Lipopolysaccharides; Male; Mutation; Neutrophils; Papillon-Lefevre Disease; Tumor Necrosis Factor-alpha; Young Adult

The inflammatory mediators, serum elastase and C-reactive protein (CRP), are associated with an increased risk for coronary heart disease. Thus, the aim of this study is to compare systemic inflammatory mediators in periodontally healthy controls (C), patients with untreated aggressive (AgP) and chronic (ChP) periodontitis. C [periodontal pocket probing depth (PPD)  <3.6 or <5 mm without bleeding (BOP), BOP < 10%], ChP (PDD ≥ 3.6 mm and probing attachment loss ≥5 mm at >30% of sites; age >35 years), and AgP (clinically healthy; PDD ≥ 3.6 mm at >30% of sites, bone loss ≥50% at ≥2 teeth; age ≤35 years) were examined clinically, and the body mass index was assessed. Blood was sampled for assessment of serum levels of elastase, CRP, lipopolysaccharide binding protein (LBP), interleukin (IL) 6, 8, and leukocyte counts. Thirty C, 31 ChP, and 29 AgP were analyzed. Elastase, CRP, LBP, and IL-6 levels were elevated in AgP compared to C (p < 0.013), whereas leukocyte counts and IL-8 were similar. Multiple regression analysis identified AgP (p < 0.001) and education level (p < 0.001) to explain 47% of the variation of elastase. AgP (p = 0.003), African origin (p = 0.006), female sex (p = 0.002), and BMI (p < 0.001) explained 39% of the variation of CRP. Serum elastase and CRP are significantly elevated in AgP compared to C. AgP patients exhibit a stronger systemic inflammatory burden than C patients.

Topics: Acute-Phase Proteins; Adult; Aggressive Periodontitis; Alveolar Bone Loss; Black People; Body Mass Index; C-Reactive Protein; Carrier Proteins; Chronic Periodontitis; Dental Plaque Index; Educational Status; Female; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Lipopolysaccharides; Male; Membrane Glycoproteins; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Sex Factors

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

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

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

Studies evaluating the methylation status of cytokine genes may have relevance for inflammatory diseases in which the expression of some cytokines is altered, such as periodontitis. This study observes the DNA methylation status in the interleukin-8 (IL8) gene promoter in cells of the oral epithelium of subjects affected by generalized aggressive periodontitis (AgP) and compares it to those of control subjects.. Genomic DNA from epithelial oral cells of 37 generalized AgP patients and 37 controls were purified and modified by sodium bisulphite. Modified DNA was submitted by methylation-specific polymerase chain reaction, electrophoresed on 10% polyacrylamide gels, and stained.. Subjects who presented generalized AgP have a higher frequency of hypomethylation of the IL8 gene promoter in oral epithelium cells than that of controls (86.5% in the generalized AgP group versus 62% in the control group; P = 0.016; chi(2) test).. A marked hypomethylated status is found in the oral epithelial cells of subjects presenting with generalized AgP, compared to controls, in the promoter region of the IL8 gene. This hypomethylated status may reflect a generalized condition of oral epithelial cells, including gingival epithelium, because gingival epithelial cells were also collected during mouthwash use.

Topics: Adult; Aggressive Periodontitis; Case-Control Studies; Chi-Square Distribution; DNA; DNA Methylation; Female; Humans; Interleukin-8; Male; Polymerase Chain Reaction; Promoter Regions, Genetic; Sulfites; Young Adult

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

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

In view of the reports that polymorphonuclear leukocytes (PMN) of patients with localized aggressive periodontitis (LAP) exhibit hyper-responsiveness to stimulation, it has been suggested that such abnormalities could lead to PMN-mediated tissue damage during inflammation. To determine whether these abnormalities include signal transduction, we compared cytoplasmic calcium concentration (Delta[Ca2+](i)) and cytoplasmic pH (DeltapH(i)) changes, early stimulus responses to chemotactic agents, of LAP versus control (C)-PMN and explored whether these could be modulated by sensitizing cytokines or calcium channel-blocking agents. PMN responses of LAP patients were compared with age- and gender-matched controls. Delta[Ca2+](i) and DeltapH(i) were measured fluorimetrically using 1H-indole-6-carboxylic acid, 2-[4-[bis[2-[(acetyloxy)methoxy]-2-oxoethyl]amino]-3-[2-[2-[bis[2-[(acetyloxy)methoxy]-2-oxoethyl]amino]-5-methylphenoxy]ethoxy]phenyl]-1 and 2',7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein as respective probes. Not only was the maximal calcium response to chemoattractants higher in LAP-PMN, but also their subsequent intracellular calcium redistribution was significantly slower. The slower calcium redistribution of LAP-PMN, but not their higher maximal calcium response, was successfully mimicked in C-PMN treated with Nifedipine or 1-[b-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole-HCl, both known to be inhibitors of membrane-associated calcium influx, but this redistribution was not affected when inhibitors of other calcium influx mechanisms, Diltiazem or Verapamil, were used. Taken together, our findings indicate that certain early stimulus responses are aberrant in LAP-PMN, that internal redistribution of cytoplasmic-free calcium is compromised, and, additionally, that a membrane-associated Ca2+ transport defect may be present.

Topics: Aggressive Periodontitis; Calcium; Cytokines; Cytoplasm; Diltiazem; Dose-Response Relationship, Drug; Humans; Hydrogen-Ion Concentration; Imidazoles; Interleukin-8; Intracellular Fluid; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Nifedipine; Substance P; Time Factors; Verapamil

The point of this study was to examine the presence or absence of cytokine-positive cells by means of immunohistochemical methods in the samples of inflamed gingival tissues obtained from an 11-year-old girl with Papillon-Lefevre syndrome (PLS). Interleukin-8 (IL-8)-positive cells were found to be present. In addition, IL-1alpha-and IL-1beta-positive cells were detected. No dysfunction in the phagocytosis and the bacterial killing of peripheral blood polymorphonuclear neutrophils (PMNs) was observed in this patient. Our findings suggest that these cytokines may be members responsible for modulating the process of rapidly progressive periodontitis for patient with PLS.

Topics: Aggressive Periodontitis; Blood Bactericidal Activity; Child; Female; Gingiva; Humans; Immunohistochemistry; In Vitro Techniques; Interleukin-1; Interleukin-8; Neutrophils; Papillon-Lefevre Disease; Phagocytosis

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

Localized juvenile periodontitis (LJP) is an early-onset periodontal disease characterized by progressive bone loss involving the permanent first molar and incisor teeth. Approximately 70% to 75% of LJP patients have impaired neutrophil chemotaxis towards a number of chemoattractants including N-formyl-methionyl-leucyl-phenyl-alanine, complement fragment C5a, leukotriene B4, and interleukin 8 (IL-8). The aim of the present study was to observe the role of IL-8 in the pathogenesis of LJP. Fourteen individuals who were systemically and periodontally healthy and 24 systemically healthy individuals diagnosed with LJP (based on the results of clinical periodontal assessments and radiographic examination) were recruited for this study. Gingival crevicular fluid (GCF) samples were obtained from anterior teeth in each subject before treatment. After evaluation of GCF amount from paper strips, enzyme-linked immunoabsorbent assay was employed to determine the amount of IL-8 in GCF. The amount and concentration of IL-8 measured was 894.5 +/- 435 pg, and 445.3 +/- 468 pg/microl for the experimental group and 747.3 +/- 543 pg and 684.7 +/- 548 pg/microl, for the control group. The correlation among the levels of cytokine and clinical parameters was assessed. It was observed that the concentration of IL-8 demonstrated a negative correlation with gingival index in the LJP group. In addition, no significant correlation was found among the total amount and concentration of IL-8, GCF volume, and clinical parameters in the control group. IL-8 is thought to enhance host defense mechanisms against gram-negative bacteria, thus providing protection against periodontal infections. Our data demonstrate that, when both the total amount and concentration of IL-8 are taken into consideration, no significant difference between LJP and healthy subjects is shown. This may indicate a less active IL-8 production compared with healthy subjects in spite of the dense Gram bacterial stimulation in LJP.

Topics: Adult; Aggressive Periodontitis; Case-Control Studies; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Male; Periodontal Index; Smoking; Statistics, Nonparametric

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