The gingival crevicular fluid Interleukin-1β and tumour necrosis factor-α levels in patients with rapidly progressive periodontitis

Cytokines are believed to play an important role in the pathogenesis of periodontal diseases. In the present study, gingival crevicular fluid (GCF) levels of two important cytokines, interleukin 1-/3 (IL-1β) and tumour necrosis factor-α (TNF-α) and, in addition, serum IL-1β levels, were determined in patients with severe and rapid periodontal breakdown by use of ELISA. While IL-1β was detected in all of the GCF samples studied, TNF-α could only be detected in about half the samples. The mean GCF IL-1β level was 38.45 ± 13.99 pg/mL, and the mean TNF-α level was 3.20 ± 1.39 pg/mL, respectively. The GCF IL-1β levels also presented a strong positive correlation with the mean pocket depths. Although weak, both of the cytokines also presented correlations with the presence of bleeding on probing. Additionally GCF samples contained increased IL-1β levels when compared with the serum samples suggesting local production mechanisms. The findings of the present study suggest that these cytokines may be involved in the pathogenesis of periodontal diseases (IL-1β being more significant), and also may help in defining the active phase of periodontal breakdown.     

Association between body weight and periodontal infection

Background: Besides being a risk factor for cardiovascular diseases, certain cancers and type II diabetes, obesity has been suggested to be a risk factor for periodontitis. A number of epidemiological studies have studied the association between obesity and periodontitis, but the results have been partly inconclusive. The aim of this study was to examine the association of body weight with periodontal infection. Material and Methods: The association between body weight and periodontal infection was examined using a nationally representative Health 2000 Health Examination Survey. The study was based on a subpopulation of dentate non‐diabetic subjects aged 30–49 (n=2841). Periodontal infection was measured by the number of teeth with periodontal pockets of 4 mm or deeper and 6 mm or deeper. Body weight was measured using body mass index (BMI). Results: We detected a weak exposure–response association of BMI with teeth with deepened periodontal pockets after controlling for smoking habits by restricting the sample to subjects who have never smoked and for other potential confounders by including them in the multivariate models. Conclusions: The results showed an association between body weight and periodontal infection among the non‐diabetic, non‐smoking population aged 30–49. Additional research is needed to determine the nature of this association.     

Association between serum lipid levels and periodontal infection

Objective: The aim of the study was to investigate the association between serum lipids and periodontal infection and the role of serum lipids in the association between body weight and periodontal infection. Material and Methods: The Health 2000 Health Examination Survey, which included 8028 subjects aged 30 or older living in continental Finland. This study was based on a subpopulation of dentate, non‐diabetic subjects who had never smoked and were aged under 50 years (n=1297). Periodontal infection was defined as the presence of teeth with deepened periodontal pockets. Serum levels of triglycerides, high‐density lipoprotein (HDL)‐cholesterol and low‐density lipoprotein (LDL)‐cholesterol were analysed enzymatically. Results: We found no consistent association between serum lipid levels and periodontal infection among normoweight subjects. There was an association of high serum triglycerides and low HDL with periodontal infection among obese subjects. The association between body mass index and periodontal infection was not essentially affected by serum lipids. Conclusion: In this study population serum lipid levels were not associated with periodontal infection among normoweight subjects. Obese subjects with a high serum triglyceride level and/or a low HDL‐cholesterol level could be at higher risk of periodontal infection. Our results suggest that the association between body weight and periodontal infection was mainly mediated through a mechanism other than serum lipids.     

Salivary interleukin-6 and tumor necrosis factor-alpha in patients with burning mouth syndrome

Burning mouth syndrome (BMS) is characterized by burning symptoms on the clinically healthy oral mucosa. To date, etiology of BMS is still unknown. We hypothesized that maybe inflammation which is not clinically apparent might lead to burning symptoms which would then result in altered cytokine profile. In the 28 female patients with BMS (age range 48-80 years, mean 64.05 years) and 28 female controls (age range 40-75 years, mean 63.82 years) by use of enzyme-linked immunosorbent assay, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) levels were determined. Statistical analysis included use of independent sample t-test and P < 0.05 was considered as significant. Our results show no significant differences between patients and controls regarding salivary IL-6 and TNF-alpha.     

Oral and skin keratinocytes are stimulated to secrete monocyte chemoattractant protein-1 by tumour necrosis factor-α and interferon-γ

Monocyte chemoattractant protein-1 (MCP-1) is a member of the CC subfamily of chemokines. It is chemoattractant for monocytes, activated memory T cells and dendritic cells. We studied MCP-1 mRNA and protein production in cultured human oral keratinocytes (OK) and skin keratinocytes (SK). MCP-1 production was undetectable in resting keratinocytes. However, stimulation with tumour necrosis factors (TNF-alpha) or interferon-gamma (IFN-gamma) induced MCP-1 mRNA and protein synthesis in both cell types. Together, TNF-alpha and IFN-gamma acted synergistically to increase MCP-1 production. In each case MCP-1 production was greater for SK than OK. The kinetics of IFN-gamma-induced MCP-1 production were similar for both cell types, peaking around 72 h. In contrast, TNF-alpha induced a more rapid increase in MCP-1 production by SK than OK, peak production occurring after 24 and 72 h, respectively. IL-1alpha and IL-4 did not induce MCP-1 production. However, in combination with IFN-gamma, they induced a small extra increase in MCP-1 production in SK but not OK.     

Lipopolysaccharide from Actinobacillus actinomycetemcomitans stimulates production of interleukin-1beta, tumor necrosis factor-alpha, interleukin-6 and interleukin-1 receptor antagonist in human whole blood

Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) is supposed to be an important etiological agent in localized juvenile periodontitis (LJP). We have studied the effect of lipopolysaccharide (LPS) extracted from these periodontopathogenic bacteria on synthesis of the proinflammatory cytokines, interleukin-1beta(IL-1beta), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and the anti-inflammatory cytokine IL-1 receptor antagonist (IL-1ra) in human whole blood. LPS from A. actinomycetemcomitans in concentrations > or =1 ng/ml induced a significant production of all these proinflammatory cytokines, whereas LPS from Escherichia coli (E. coli), strain 026:B6 had to be added in concentrations > or =1 microg/ml to obtain a similar effect. Similarly, LPS from A. actinomycetemcomitans > or =0.1 ng/ml resulted in production of IL-1ra, while LPS from E. coli 026:B6 had to be added at > or =10 ng/ml to obtain similar effects. It has been suggested that the ratio between production of proinflammatory and anti-inflammatory cytokines may influence the outcome of periodontal diseases. Other in vitro and in vivo studies have, however, indicated that very large excesses (100-1000 times) of IL-1ra compared to IL-1beta are required to shift the IL-1ra:IL-1beta ratio in favor of an inhibition of IL-1 bioactivity. In our ex vivo system, we found that stimulation with extremely low doses of A. actinomycetemcomitans LPS (0.1-1 ng/ml) resulted in IL-1ra production solely, without concomitant production of IL-1beta, the excess of IL-1ra over IL-1beta peaking at 1 ng/ml, which accordingly should suggest that LPS from A. actinomycetemcomitans primarily has proinflammatory effects.     

Polymorphisms in the CD14 and IL-6 genes associated with periodontal disease

AIM: To compare the frequencies of cytokine and receptor molecule genotypes in patients with chronic periodontitis with the corresponding frequencies in a reference population and to study the relationship between periodontal disease severity and polymorphisms in the studied genes. SUBJECTS AND METHODS: CD14, IL-6, TNF-alpha, IL-10, IL-1alpha, IL-1beta, and TLR-4 polymorphisms of 51 periodontitis patients were studied using polymerase chain reaction. The genotype frequencies in the periodontitis patients and a reference population (n=178) were compared. Probing pocket depth (PD), periodontal attachment level (AL), and alveolar bone level (BL) were related to the genotypes. RESULTS: No statistically significant differences could be found between the frequencies of the cytokine genotypes in the periodontitis patients and in the reference group. The extent of periodontal disease was higher in subjects with the T-containing genotype of CD14(-260) and the GG genotype of IL-6(-174) when compared with the extent in the rest of the group. Subjects carrying the composite genotype of the above two were most severely affected by periodontal disease. CONCLUSION: According to the present results, an evident association exists between the carriage of the T-containing genotype of CD14(-260) and the GG genotype of IL-6(-174) and the extent periodontal disease.     

The effect of periodontal therapy on serum TNF-α and HbA1c levels in type 2 diabetic patients

Background:  To determine the effect of non-surgical periodontal therapy on serum TNF-α and HbA1c levels in poorly and well-controlled type 2 diabetic patients.
Methods:  In total, 45 patients were enrolled in the study; 30 patients with type 2 diabetes mellitus with periodontitis (15 with poorly controlled diabetes, HbA1c ≥ 7%, group 1A and 15 with well-controlled diabetes, HbA1c < 7%, group 1B) and 15 patients that were systemically healthy with periodontitis (group 2). The plaque index, gingival index, probing depth, clinical attachment loss, gingival bleeding index, HbA1c value, and circulating TNF-α concentration were measured at baseline and three months after the non-surgical periodontal therapy.
Results:  All periodontal parameters and serum TNF-α levels were significantly decreased three months after the non-surgical periodontal therapy compared to the baseline values in all groups. The HbA1c values were significantly decreased only in well-controlled diabetic patients. We found no significant differences in the periodontal parameters or TNF-α levels at baseline and after three months between the two groups.
Conclusions:  Although non-surgical periodontal therapy eliminates local/systemic infection and inflammation via decreases in TNF-α, it is insufficient for significantly reducing HbA1c levels without strict glycaemic control in poorly controlled diabetic patients in a short time period.     

Role of IL-6, TNF-A and LT-A variants in the modulation of the clinical expression of plaque-induced gingivitis

AIM: The purpose of the present study was to assess the association of interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-A) and lymphotoxin alpha (LT-A) gene polymorphisms with the clinical parameters of gingivitis in a large experimental gingivitis trial and with each of two subgroups, "high responder" (HR, n=24) and "low responder" (LR, n=24), with distinct susceptibility to gingivitis. MATERIAL AND METHODS: Ninety-six systemically and periodontally healthy non-smokers, 46 males (mean age: 23.9+/-1.7) and 50 females (mean age: 23.3+/-1.6), were included in a randomized split-mouth localized 21-day experimental gingivitis trial. Plaque index, gingival index, gingival crevicular fluid volume and angulated bleeding score were recorded. HR and LR subgroups were characterized by substantially different severities of gingival inflammation despite a similar plaque accumulation rate. All subjects were genetically characterized for IL-6(-174), IL-6(-597), TNF-A(-308) and LT-A(+252) polymorphisms. RESULTS: None of the variants analysed, either as single polymorphisms or as a combined genotype, was associated with the clinical parameters in the overall population. For the polymorphisms studied, genotypic distributions in HR and LR subjects were not significantly different. CONCLUSIONS: The present results suggest an absence of association between IL-6, TNF-A and LT-A polymorphisms and subject-based clinical behaviour of the gingiva in response to de novo plaque accumulation.     

interleukin-1β and tumor necrosis factor-α stimulate synergistically the expression of monocyte chemoattractant protein-1 in fibroblastic cells derived from human periodontal ligament

The present study demonstrates that interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) induce and synergistically stimulate monocyte chemoattractant protein-1 (MCP-1) expression in fibroblasts from human periodontal ligament. IL-1β and TNF-α induced in a dose-dependent manner the expression of the MCP-1 gene in the fibroblasts from the human periodontal ligament. However, such an inducing effect was not observed with IL-6 and interferon-γ. The peak expression of the MCP-1 gene by IL-1β or TNF-α was observed at 3 h after initiation of their treatment. Furthermore, IL-1β in combination with TNF-α synergistically stimulated the MCP-1 gene expression in the cells. We also observed significant chemotactic activity for human monocytes in conditioned medium of fibroblasts from the human periodontal ligament treated with both cytokines. The stimulated chemotactic activity induced by these cytokines depended on both dose and treatment time. The chemotactic activity in conditioned medium of IL-1β-treated fibroblasts from the human periodontal ligament was neutralized by antiserum specific for MCP-1 protein. The MCP-1 gene product in conditioned medium of IL-1β-treated fibroblasts from the human periodontal ligament was shown to have a molecular mass of 11,000 Da by immunoprecipitation with the specific antiserum, and IL-1β also stimulated synergistically MCP-1 protein expression in combination with TNF-α. These results suggest that IL-1β and TNF-α may contribute to the infiltration of monocytes into inflammatory sites of periodontal ligament tissues via the MCP-1 gene product produced by fibroblasts from the human periodontal ligament.     

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目的通过观察分析中重度牙周炎患者血清中对氧磷酶-1（PON1）活性及白细胞介素-6（IL-6）在牙周基础治疗前后的变化,探讨PON1和IL-6与牙周病之间的相关性以及两者在牙周病病情变化中的作用。方法2012年5月至2013年2月在辽宁省人民医院体检中心体检的牙周健康人群30名（牙周健康组）及口腔科就诊的中重度牙周炎患者39例（牙周炎组）,对所有受试者进行基础治疗,检查治疗前后PON1、IL-6以及临床牙周检查指标。结果牙周健康组与牙周炎组在基础治疗前比较：牙周炎症指标[出血指数（BI）、牙周探诊深度（PD）]及血清IL-6差异均有统计学意义（P〈0．05）,血清中PON1活性差异无统计学意义（P〈0．05）。牙周健康组在牙周基础治疗前后各项指标差异均无统计学意义（P〉0．05）。牙周炎组在牙周基础治疗前后比较：牙周炎症指标BI、PD、血清PON1及IL-6与治疗前比较差异均有统计学意义（均P〈0．05）。结论牙周健康人群和中重度牙周炎患者血清PON1活性无差异;牙周基础治疗可改善中重度牙周病患者牙周健康状况,提高血清中PON1活性,降低IL-6水平;中重度牙周炎患者在牙周基础治疗前后血清中PON1活性和IL-6水平的变化存在负相关性。     

Involvement of tumor necrosis factor-α and interleukin-8 in antigen-induced arthritis of the rabbit temporomandibular joint

BACKGROUND: In temporomandibular joint (TMJ) arthritis, knowledge is limited about the source of the inflammatory mediators. The aim of this study was to investigate the immunohistochemical role of tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) in the development of the antigen-induced arthritis of the rabbit TMJ. METHODS: Unilateral TMJ arthritis was induced in 28 adult rabbits. From 6 h to 6 weeks after induction of arthritis, the topology of TNF-alpha and IL-8 was observed. RESULTS: Positive reaction for TNF-alpha of synovial cells was observed within 3 days after induction and at 3 weeks after induction. TNF-alpha positive vascular endothelial cells and chondrocytes were identified throughout the observation period. IL-8 was detected only during the acute stage. CONCLUSIONS: The cytokines TNF-alpha and IL-8 were observed in specific cells depending on the stage. TNF-alpha was particularly related with angiogenesis and cartilage destruction and IL-8 was involved in the acute stage of inflammation.     

Levamisole can modulate the serum tumor necrosis factor-α level in patients with recurrent aphthous ulcerations

BACKGROUND: Recurrent aphthous ulcerations (RAU) are common oral inflammatory lesions. Tumor necrosis factor (TNF)-alpha is an important inflammatory mediator and a critical cytokine for adequate host defense. Our previous studies have shown that 14-43% and 59-63% of patients in the ulcerative stage of major, minor or herpetiform RAU have significantly higher than normal serum levels of interleukin (IL)-6 and IL-8, respectively. In this study, we examined whether RAU patients in the ulcerative stage had a significantly higher than normal serum level of TNF-alpha and assessed whether treatment with levamisole can modulate serum TNF-alpha levels in RAU patients. METHODS: This study used a solid phase, two-site sequential chemiluminescent immunometric assay to determine the baseline serum levels of TNF-alpha in 146 patients with RAU, nine patients with traumatic ulcers (TU), and 54 normal control subjects. Fifty-five RAU patients with serum TNF-alpha levels higher than 5.0 pg/ml were treated with levamisole for 0.5-4 months and their serum TNF-alpha levels were measured after treatment. RESULTS: We found that 29% (42 of 146) RAU patients as well as 39% (24 of 61) major type, 20% (14 of 69) minor type, and 25% (four of 16) herpetiform type RAU patients had a serum level of TNF-alpha greater than the upper normal limit of 7.4 pg/ml. The mean serum level of TNF-alpha in patients with RAU (9.1 +/- 1.0 pg/ml, P < 0.001), major type RAU (11.6 +/- 1.9 pg/ml, P < 0.001), minor type RAU (6.9 +/- 0.9 pg/ml, P < 0.005), or herpetiform type RAU (9.6 +/- 2.7 pg/ml, P < 0.001) was higher than that (3.8 +/- 0.2 pg/ml) in normal control subjects. The mean serum TNF-alpha level was significantly higher in patients with major type RAU than in patients with minor type RAU (P < 0.05) and was significantly higher in major type RAU patients in the exacerbation stage than in the post-exacerbation stage (P < 0.05). In 55 RAU patients with serum TNF-alpha levels higher than 5.0 pg/ml, treatment with levamisole for a period of 0.5-4 months could significantly reduce the serum TNF-alpha level from 16.4 +/- 1.9 to 5.8 +/- 0.6 pg/ml (P < 0.001). CONCLUSIONS: We conclude that a significantly higher than normal serum level of TNF-alpha can be detected in 20-39% of patients in the ulcerative stage of major, minor or herpetiform RAU. The serum TNF-alpha level may be associated with the severity and the stage of RAU. Levamisole can modulate serum TNF-alpha levels in RAU patients.     

Exposure measurement in the association between periodontal disease and prematurity/low birth weight

AIM: To compare the use of different definitions for exposure measurement in cases of association between periodontal disease (PD) and prematurity and/or low birth weight (PLBW). MATERIAL AND METHODS: A database from a previous case-control study was used to compare four different definitions for periodontitis: at least one site with probing depth > or =4 mm (1); at least one site with clinical attachment loss (CAL)> or =3 mm (2); at least four teeth with one or more sites presenting probing depth > or =4 mm, with CAL> or =3 mm at the same site (3); and at least four teeth with one or more sites with probing depth > or =4 mm, with CAL> or =3 mm at the same site and presence of bleeding on probing (4). The PD frequency, diagnostic values and adjusted association measurements were calculated. RESULTS: PD frequency ranged from 33.1% to 94.7%. Odds ratio(adjusted) varied slightly according to the exposure measurement used. CONCLUSIONS: The association between PD and PLBW weight was consistent, except for exposure measurement 1, i.e. using at least one site with CAL> or =3 mm for periodontitis diagnosis, while the magnitude of this varied according to the definition established.     

The extent of periodontal disease and the IL-6 -174 genotype as determinants of serum IL-6 level

AIM: To study the extent of periodontal disease and the IL-6(-174) genotype as determinants of serum and mouthwash IL-6 concentration in subjects with moderate to severe periodontal disease. MATERIAL AND METHODS: Fifty-two generally healthy subjects volunteered to participate. Probing pocket depth (PD) and periodontal attachment level (AL) were clinically examined and alveolar bone level (BL) was measured on orthopantomographs. IL-6 concentrations in mouthwash, collected by rinsing with 3 ml saline for 30 s and in serum, obtained by venipuncture, were measured using ELISA. IL-6(-174) polymorphism was studied using a polymerase chain reaction. RESULTS: Eleven subjects carried the GG genotype, and 41 subjects, carried the CG/CC genotype. The mean (+/- SD) concentration of IL-6 in serum was 1.6 (+/- 1.5) pg/ml and, 2.8 (+/- 5.04) pg/ml in mouthwash. The serum concentration of IL-6 was higher in subjects with the GG genotype than with the CG/CC genotype. In regression analyses the percentages of sites with PD> or =6 mm, AL> or =6 mm and BL> or =8 mm, the IL-6(-174) genotype, body mass index and gender associated significantly with serum IL-6 concentration. CONCLUSIONS: The extent of moderate to severe periodontal disease and the IL-6(-174) genotype contribute significantly to serum IL-6 concentration.     

Clinical expression of TNF-α in smoking-associated periodontal disease

Abstract. The level of TNF-α in gingival crevicular fluid (GCF) was analyzed with respect to smoking in patients with untreated moderate to severe periodontal disease including 30 current smokers, 19 former smokers and 29 non-smokers, in the age range 31–79 years, Concomitantly the occurrence of the periopathogens Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg) and Prevotella intermedia (Pi) and the GCF levels of albumin, IgA and IgG were analyzed. With regard to clinical characteristics, there were no statistically significant differences between smoking groups. The occurrence of patients positive for the periopathogens Aa, Pg and Pi was 28.2%, 41.0% and 91.0%, respectively. There were no statistically significant differences between smoking groups with regard to occurrence or relative frequency of these periopathogens. An exception was a significantly lower occurrence of Aa in former smokers as compared to non-smokers. The chief novelty of the study was the observation of a clearly increased level of TNF-α in GCF associated with smoking. Both current and former smokers exhibited significantly higher levels of TNF-α in comparison to non-smokers, whereas the levels of albumin, IgA and IgG were the same irrespective of smoking. In conclusion, the present observations in patients with moderate to severe periodontal disease suggest that smoking is associated with elevated GCF levels of the cytokine TNF-α.