Gingival Crevicular Fluid IL-1beta Research Articles

Cytokine-based Predictive Models to Estimate the Probability of Chronic Periodontitis: Development of Diagnostic Nomograms

Although a distinct cytokine profile has been described in the gingival crevicular fluid (GCF) of patients with chronic periodontitis, there is no evidence of GCF cytokine-based predictive models being used to diagnose the disease. Our objectives were: to obtain GCF cytokine-based predictive models; and develop nomograms derived from them. A sample of 150 participants was recruited: 75 periodontally healthy controls and 75 subjects affected by chronic periodontitis. Sixteen mediators were measured in GCF using the Luminex 100™ instrument: GMCSF, IFNgamma, IL1alpha, IL1beta, IL2, IL3, IL4, IL5, IL6, IL10, IL12p40, IL12p70, IL13, IL17A, IL17F and TNFalpha. Cytokine-based models were obtained using multivariate binary logistic regression. Models were selected for their ability to predict chronic periodontitis, considering the different role of the cytokines involved in the inflammatory process. The outstanding predictive accuracy of the resulting smoking-adjusted models showed that IL1alpha, IL1beta and IL17A in GCF are very good biomarkers for distinguishing patients with chronic periodontitis from periodontally healthy individuals. The predictive ability of these pro-inflammatory cytokines was increased by incorporating IFN gamma and IL10. The nomograms revealed the amount of periodontitis-associated imbalances between these cytokines with pro-inflammatory and anti-inflammatory effects in terms of a particular probability of having chronic periodontitis.

Interleukin‐1β levels in gingival crevicular fluid and serum under naturally occurring and experimentally induced gingivitis

To evaluate the interleukin-1 beta (IL-1 beta) levels in gingival crevicular fluid (GCF) and serum in either naturally occurring (N-O) or experimentally induced (E-I) plaque-associated gingivitis. Thirty-seven periodontally healthy subjects were evaluated in real life conditions (N-O gingivitis) as well as after 21 days of experimental gingivitis trial (E-I gingivitis). During the experimental gingivitis trial, in one maxillary quadrant (test quadrant), gingival inflammation was induced by oral hygiene abstention, while in the contralateral (control) quadrant, oral hygiene was routinely continued. IL-1 beta concentrations in N-O and E-I gingivitis were investigated for IL-1B(+3954) and IL-1B(-511) gene polymorphisms. (i) GCF IL-1 beta concentrations in E-I gingivitis were significantly higher compared with N-O gingivitis; (ii) an intra-individual correlation between GCF concentrations of IL-1 beta detected in N-O and E-I gingivitis was observed in control quadrants, but not in test quadrants; (iii) IL-1 beta concentration in GCF was associated with IL-1B(+3954) genotype only at test quadrants; (iv) IL-1 beta was detectable in serum only at low levels in a limited number of subjects, without difference between gingivitis conditions. Aspects of the bacterial challenge to the gingival tissues, such as the amount of plaque deposits and plaque accumulation rate, appear to affect the IL-1 beta levels in GCF in subjects with a specific IL-1B genotype.

Gingival changes during pregnancy: I. Influence of hormonal variations on clinical and immunological parameters

To test whether exacerbated gingival inflammation in pregnancy is associated with increased salivary hormone levels and changes in gingival crevicular fluid (GCF) interleukin-1beta (IL-1beta) and prostaglandin-E2 (PGE2) levels. In this cohort study, 48 pregnant women without periodontitis were evaluated in the first, second, and third trimesters and at 3 months postpartum. Twenty-eight non-periodontitis non-pregnant women were evaluated twice, with a 6-month interval. Plaque and gingival indices (PlI, GI), salivary progesterone and estradiol and GCF IL-1beta and PGE2 levels were determined. anova for repeated measures or Friedman's test were used for intragroup analyses. Inter-group comparisons were analysed with t-test or Mann-Whitney U-test. Correlations were evaluated with Pearson's and Spearman's test. Pregnant women showed an increase in GI (p<0.05) despite maintaining low PlI values. No changes in IL-1beta and PGE2 levels were observed during pregnancy. No significant correlation was found between the GI increase and salivary hormone levels. GI (p<0.05) and IL-1beta levels (p<0.001) were lower in non-pregnant than in pregnant women. This study confirms the presence of an exacerbated gingival inflammation during pregnancy, but this phenomenon could not be associated with an increase in progesterone or estradiol or with changes in PGE2 or IL-1beta.

IL‐1β, TNF‐α, total antioxidative status and microbiological findings in chronic periodontitis treated with fluorescence‐controlled Er:YAG laser radiation

Although some studies have attempted to elucidate the possible advantages of the use of Er:YAG laser radiation as a coadjuvant of scaling and root planing (SRP), the results have often been contradictory. A new possibility to improve the results of the laser therapy is the control of the laser radiation by a feedback system that selectively detects subgingival calculus. This study compared the effects of fluorescence-controlled Er:YAG radiation after SRP with SRP alone on the treatment of chronic periodontitis. Thirty patients with chronic periodontitis were randomly distributed into two groups of 15 patients to receive: SRP or SRP followed by fluorescence controlled Er:YAG laser (SRP+ERL). Clinical parameters including probing pocket depth (PPD), bleeding on probing (BOP), and plaque index (PI) were recorded and samples of gingival crevicular fluid (GCF) and subgingival plaque were taken at baseline, 4 and 8 weeks postoperatively. The laser therapy was performed 1 day after the SRP procedure. The GCF samples were analyzed for interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and total antioxidative status (TAS). Subgingival plaque was examined by culture for 10 bacteria. During the observation periods (4 and 8 weeks postoperatively), no statistically significant differences were detected for clinical, microbiological variables and TAS of GCF between SRP and SRP+ERL groups. However, the amount of IL-1beta and TNF-alpha in GCF was increased after SRP procedure in contrast to a slight decrease of their levels after SRP+ERL treatment, where in addition the process of recolonization seems to be more delayed. SRP+ERL allowed a decrease of the levels of proinflammatory cytokines and prevented a fast process of recolonization.

Local and systemic biomarkers in gingival crevicular fluid increase odds of periodontitis

To determine the independent and combined associations of interleukin-1beta (IL-1beta) and C-reactive protein (CRP) in gingival crevicular fluid (GCF) on periodontitis case status in the Australian population. GCF was collected from 939 subjects selected from the 2004-2006 Australian National Survey of Adult Oral Health: 430 cases had examiner-diagnosed periodontitis, and 509 controls did not. IL-1beta and CRP in GCF were detected by enzyme-linked immunosorbent assays. Odds ratios (OR) and 95% confidence intervals (CIs) were calculated in bivariate and stratified analysis and fully adjusted ORs were estimated using multivariate logistic regression. Greater odds of having periodontitis was associated with higher amounts of IL-1beta (OR=2.4, 95% CI=1.7-3.4 for highest tertile of IL-1beta relative to lowest tertile) and CRP (OR=1.9, 95% CI=1.5-2.5 for detectable CRP relative to undetectable CRP). In stratified analysis, there was no significant interaction between biomarkers (p=0.68). In the multivariate analyses that controlled for conventional periodontal risk factors, these relationships remained (IL-1beta OR=1.8, 95% CI=1.1-2.6; CRP OR=1.7, 95% CI=1.3-2.3). Elevated odds of clinical periodontitis was associated independently with each biomarker. This suggests that people with elevated biomarkers indicative of either local (IL-1beta) or systemic (CRP) inflammation are more likely to suffer from periodontal disease.

Levels of Serum Interleukin (IL)‐6 and Gingival Crevicular Fluid of IL‐1β and Prostaglandin E2 Among Non‐Smoking Subjects With Gingivitis and Type 2 Diabetes

The goal of this study was to assess whether non-smoking patients with type 2 diabetes present with increased levels of local and systemic proinflammatory mediators and, if so, whether such an increase is associated with enhanced clinical gingival inflammation compared to non-smoking patients without diabetes. We used a cross-sectional database consisting of 725 self-reported lifelong non-smokers aged 53 to 74 years. Gingival crevicular fluid (GCF) levels of interleukin (IL)-1beta and prostaglandin E(2) (PGE(2)) and serum levels of IL-6 were measured using enzyme-linked immunosorbent assay. No participant had probing depth >3 mm. Participants with bleeding on probing (BOP) in <10% of sites were classified as healthy, whereas those with BOP in >or=10% of sites were defined as having biofilm-gingival interface (BGI) gingivitis. Approximately 53% (n = 385) and 11% (n = 80) of the sample had BGI gingivitis and type 2 diabetes, respectively. The mean age-adjusted level of GCF IL-1beta was significantly elevated in the diabetic group compared to the non-diabetic group (P = 0.048), but serum IL-6 (P = 0.14) and GCF PGE(2) were not (P = 0.98). The mean GCF IL-1beta and PGE(2) levels were significantly elevated in subjects with BGI gingivitis (136.2 +/- 112.9 ng/ml and 277.2 +/- 187.2 ng/ml, respectively) compared to subjects with gingival health (95.9 +/- 82.9 ng/ml and 205.7 +/- 149.6 ng/ml, respectively), regardless of diabetic status (P <0.001 for both). However, serum IL-6 was elevated in subjects with BGI gingivitis compared to subjects with gingival health only among subjects with diabetes (2.9 +/- 3.2 pg/ml versus 1.5 +/- 1.4 pg/ml; P = 0.008). With the exception of serum IL-6 in subjects without diabetes, an increase in the levels of proinflammatory mediators was associated with increased odds of having BGI gingivitis. The associations were stronger in the diabetic group. Type 2 diabetes may increase the host inflammatory response to oral biofilm, which, in turn, may exacerbate preconditions associated with gingivitis in susceptible individuals. Furthermore, systemic inflammation, as demonstrated by the increased level of serum IL-6, is associated with BGI gingivitis among non-smoking patients with diabetes.

The effect of adjunctive chlorhexidine mouthrinse on clinical parameters and gingival crevicular fluid cytokine levels in untreated plaque-associated gingivitis

To examine the effectiveness of chlorhexidine mouthrinse (CHX) in addition to daily plaque control on gingival inflammation. Fifty gingivitis patients were randomized to CHX or placebo groups. In addition to proper plaque control, CHX group rinsed with CHX, while placebo group rinsed with placebo mouthrinse for 4 weeks. Gingival crevicular fluid (GCF) samples were collected and clinical parameters including plaque index (PI), papillary bleeding index (PBI), calculus index and probing depth (PD) were recorded at baseline and repeated at 4 week. GCF IL-1alpha, IL-1beta, IL-1Ra, and IL-8 levels were determined by ELISA. Whole mouth clinical parameters were significantly improved in both groups at 4 weeks. CHX group showed greater reduction in the mean PI scores than placebo at 4 weeks (p < 0.05). GCF IL-8 levels of anterior sites significantly reduced in CHX and placebo group at 4 weeks (p < 0.05). GCF IL-1alpha, IL-1beta, IL-1Ra levels remained unchanged at 4 weeks in both groups. GCF cytokine levels of CHX group were similar to those of placebo at 4 weeks. Within the limitations of this study, CHX mouthrinse as adjuncts to daily plaque control could be useful in management of plaque-associated gingivitis, although ineffective on GCF cytokine levels.

Daytime variations of interleukin‐1β in gingival crevicular fluid

Interleukin-1beta (IL-1beta) is an important parameter in periodontal research because of its role in inflammation and bone resorption. One measure used to assess local IL-1beta concentrations is analysis of its levels in gingival crevicular fluid (GCF). While studies on serum IL-1beta concentrations indicate a circadian rhythm of this parameter, nothing is known about daytime variations of IL-1beta in GCF. The present study thus aimed to analyse such variations. Daytime variations of GCF-IL-1beta between 08:00 and 22:00 h were assessed, with a time resolution of 2 h, in 28 periodontally healthy subjects. The data showed a significant variation throughout the day, with the lowest concentrations and total amounts in the morning and the highest in the evening. The effect sizes of comparisons between morning and evening samples were medium to high and corresponded in magnitude to those reported in other published research comparing healthy sites and those affected by periodontitis. The smallest daytime variations were found to occur between 12:00 h and 18:00 h. It is concluded that daytime variations in GCF-IL-1beta are large enough to be able to mimic or mask differences caused by clinical factors.

Periodontal Disease at the Biofilm–Gingival Interface

A molecular epidemiologic study provided the opportunity to characterize the biology of the biofilm-gingival interface (BGI) in 6,768 community-dwelling subjects. Disease classifications and multivariable models were developed using clinical, microbial, inflammatory, and host-response data. The purpose was to identify new clinical categories that represented distinct biologic phenotypes based upon DNA checkerboard analyses of eight plaque bacteria, serum immunoglobulin G (IgG) titers to 17 bacteria, and the gingival crevicular fluid (GCF) levels of 16 inflammatory mediators. Five BGI clinical conditions were defined using probing depths (PDs) and bleeding on probing (BOP) scores. Subjects with all PDs < or = 3 mm were grouped as BGI-healthy (14.3% of sample) or BGI-gingivitis (BGI-G, 15.1%). Subjects with one or more PDs > or = 4 mm [deep lesion (DL)] were divided into low BOP (18.0%), moderate BOP (BGI-DL/MB, 39.7%), and severe BOP (BGI-DL/SB, 12.9%). Subjects with BGI-G had increased levels of Campylobacter rectus-specific serum IgG levels (P = 0.01), and those with BGI-DL/SB had increased IgG levels to Porphyromonas gingivalis (P < 0.0003) and C. rectus (P < 0.01). BGI-DL/SB subjects had an excessive GCF interleukin (IL)-1beta and prostaglandin E2 response and an enhanced chronic inflammatory response with significant increases in GCF IL-6 and monocyte chemotactic peptide-1. Within BGI-DL/SB subjects, more severe pocketing and BOP were associated with higher levels of GCF IL-1beta, not higher microbial counts or plaque scores. New BGI classifications create categories with distinct biologic phenotypes. The increased titers of C. rectus IgG among 68.5% of the BGI-G subjects and elevated P. gingivalis titers among BGI-DL/MB and BGI-DL/SB subjects (63.8% and 75.7%, respectively) are strongly supportive of the microbial specificity of pathogenesis for BGI categories.

Periodontal disease increases the risk of severe pre‐eclampsia among pregnant women

To evaluate the possible link between the severity of periodontal disease and pre-eclampsia and to correlate this link to clinical periodontal parameters and interleukin (IL)-1beta, tumour necrosis factor-alpha (TNF-alpha), and prostaglandins (PGE(2)) levels in both gingival crevicular fluid (GCF) and serum. Fifty-nine pregnant women (20 mild pre-eclampsia, 18 severe pre-eclampsia, and 21 healthy pregnant women) were included in the study. Dental and periodontal recordings as well as GCF and blood samples were obtained within 48 h preceding delivery. The results of multivariate logistic regression showed a highly significant association between mild to severe pre-eclampsia and severe periodontal disease (p<0.001). After adjusting for potential confounders (smoking, body weight, socioeconomic status, education level, and age), severe pre-eclamptic women were 3.78 (1.77-12.74) times more likely to present severe periodontal disease than normotensive pregnant women. This odds ratio (OR) was 2.43 (1.13-8.19) for mild pre-eclamptic women. IL-1beta, TNF-alpha, and PGE(2) levels in both serum and GCF were also significantly higher in the pre-eclamptic groups than the normotensive women. These results indicate that the presence and severity of periodontal disease seems to increase the risk for not only the occurrence but also the severity of pre-eclampsia in pregnant women.

Gingival crevicular fluid interleukin‐1β, prostaglandin E2 and periodontal status in a community population

Interleukin-1 beta (IL-1beta) and prostaglandin E(2) (PGE(2)) are key inflammatory mediators involved in periodontal disease. The purposes of this molecular cross-sectional epidemiological study were to investigate relationships in a community sample between mean concentrations of IL-1beta and PGE(2) in gingival crevicular fluid (GCF) and (1) clinical periodontal signs and (2) risk factors of host inflammatory response and/or periodontal disease. The sample comprised 6277 community-dwelling adults aged 52-74 years enrolled in the Atherosclerosis Risk in Communities (ARIC) study. IL-1beta and PGE(2) concentrations were measured using enzyme-linked immunosorbent assay. Person-level summary variables were computed for maximum pocket depth (MaxPD), maximum clinical attachment level (MaxCAL) and presence/absence of bleeding on probing (BOP). Mean GCF IL-1beta and PGE(2) concentrations were dependent variables in multiple linear regression models with periodontal measures and covariates as explanatory variables. Both GCF IL-1beta and PGE(2) were positively related to MaxPD and BOP in multiple regression models (p<0.01). Increased levels of IL-1beta and PGE(2) were associated with body mass index >or=30 kg/m(2). Higher levels of GCF IL-1beta and PGE(2) were significantly associated with clinical signs of periodontal disease and independently related to patient-based anthropomorphic measures, behaviours and exposures in community-dwelling adults.

Effect of meloxicam on gingival crevicular fluid IL-1beta and IL1 receptor antagonist levels in subjects with chronic periodontitis, and its effects on clinical parameters

The aim of the present study was to determine the effects of meloxicam after initial periodontal treatment on interleukin-1beta (IL-1beta) and IL-1 receptor antagonist (IL-1ra) in gingival crevicular fluid (GCF) and clinical parameters in the chronic periodontitis patients. Data were obtained from 30 patients with chronic periodontitis. Fifteen chronic periodontitis patients received 7.5 mg meloxicam, and 15 patients received placebo tablets in a 1x1 regimen for 1 month. All subjects were nonsmokers and had not received any periodontal therapy. The plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) were recorded. The GCF was collected using a paper strip: eluted and enzyme-linked immunoabsorbent assays (ELISAs) were performed to determine the cytokine levels. The clinical data and GCF samples were obtained after periodontal therapy and 1 month after periodontal therapy. The PI, GI, PD, and GCF IL-1ra decreased significantly (p<0.05) in meloxicam group at first month when comparing the initial levels. While decrease of the PI was statistically significant in control group (p<0.05), statistically significant changes were not determined in the other clinical parameters and GCF cytokine levels (p>0.05). There were no significant differences between two groups in any of the investigated parameters. Our observations did not reveal any influence of meloxicam on levels of IL-1beta and IL-1ra in chronic periodontitis. Additional clinical studies are advisable to determine whether COX-2 selective drugs alter periodontal disease outcome with greater safety.

Chronic Oral Inflammation and the Progression of Periodontal Pathology in the Third Molar Region

To assess the association between risk markers of chronic oral inflammation and changes over time in periodontal probing depth (PD) in the third molar region, the distal of a second molar, or around a third molar. The data from these analyses are part of a study of subjects enrolled with 4 asymptomatic third molars with adjacent second molars in an institutional review board-approved longitudinal trial. Full-mouth periodontal probing was conducted at enrollment and follow-up. Enrollment levels of periodontal pathogens and gingival crevicular fluid inflammatory mediators were assayed as indicators of the degree of oral inflammation. Subjects were categorized as those who had at least a 2 mm change in periodontal PD between baseline and follow-up in the third molar region and those who did not. The relationship between aggregated subject baseline PD, levels of periodontal pathogens, and gingival crevicular fluid IL-1 beta, and the proportion of subjects with changes in PD >or=2 mm versus those with PD <2 mm were compared with Cochran-Mantel-Haenzsel statistics. Level of significance was set at 0.05. Risk assessment models for a change in PD >or=2 mm were developed using logistic regression analysis. Twenty-four percent of 254 subjects exhibited a change in PD from baseline to follow-up of >or=2 mm in the third molar region. Of these, 95% had a baseline PD of >or=4 mm. Both high (>or=10(5)) "orange" and "red" complex bacteria and PD of >or=4 mm detected at enrollment were significantly associated with a change in PD >or=2 mm. Odds of a change in PD >or=2 mm were increased if baseline pathogen levels were >or=10(5) or a PD of >or=4 mm was detected at enrollment. Our findings are consistent with chronic oral inflammation leading to a progression of periodontal disease in the third molar region.

Gingival Crevicular Fluid Levels of Interleukin‐1β and Glycemic Control in Patients With Chronic Periodontitis and Type 2 Diabetes

Patients with diabetes have increased incidence and severity of periodontal disease not accounted for by differences in the subgingival microbial infection. Poor glycemic control has been consistently associated with periodontal disease severity. Also, recent evidence suggests that hyperglycemia may induce inflammatory cytokine production. Few studies, however, have examined local biochemical measures of periodontal inflammation in patients with type 2 diabetes. The aim of this study was to determine whether glycemic control was related to gingival crevicular fluid (GCF) levels of interleukin-1beta (IL-1beta). GCF samples were collected from 45 patients with type 2 diabetes and untreated chronic periodontitis. Plaque index (PI), bleeding on probing (BOP), probing depth (PD), and attachment level (AL) were recorded at six sites per tooth. IL-1beta levels were determined from individual GCF samples by enzyme-linked immunoabsorbent assay (ELISA). Individual site and mean patient values were calculated. Glycated hemoglobin (HbA1c) levels were measured from anticoagulated whole blood using an automated affinity chromatography system. Serum glucose was also determined. Clinical periodontal measures (PD, AL, BOP) and measures of glycemic control (HbA1c, random glucose) were significantly correlated with GCF IL-1beta. Patients with greater than 8% HbA1c had significantly higher mean GCF IL-1beta levels than patients with less than 8% HbA1c. In a multivariate model adjusting for age, gender, PD, AL, BOP, and PI, HbA1c and random glucose were independent predictors of high GCF IL-1beta. Poor glycemic control is associated with elevated GCF IL-1beta. These data are consistent with the hypothesis that hyperglycemia contributes to an heightened inflammatory response, and suggests a mechanism to account for the association between poor glycemic control and periodontal destruction.

The investigation of glutathione peroxidase, lactoferrin, myeloperoxidase and interleukin-1beta in gingival crevicular fluid: implications for oxidative stress in human periodontal diseases.

Human periodontal diseases are inflammatory disorders that are the result of complex interactions between periodontopathogens and the host's immune response. Two important and interrelated factors are involved in the pathophysiological progression of periodontal diseases, i.e. the activation of immune system and the production of oxygen radicals and their related metabolites. Increased production of oxygen radicals may contribute to oxidative stress, which is reported to be involved in many diseases, including periodontal diseases. The objective of this study was to investigate glutathione peroxidase, lactoferrin and myeloperoxidase, which play an essential role in free radical production and defenses, and the proinflammatory cytokine interleukin-1beta (IL-1beta), which is important in the regulation of immunological and inflammatory reactions in human periodontal diseases. Gingival crevicular fluid (GCF) samples were collected from 27 subjects, 19 periodontitis patients and eight healthy controls, ranging in ages from 24 to 62 years. Clinical parameters were recorded. GCF glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta were analyzed by enzyme-linked immunosorbent assays (ELISA). The periodontitis sites exhibited significantly greater total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta than healthy sites. Total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta was positively correlated with plaque index, gingival index, probing depth and probing attachment level (p < 0.05). The imbalance between the levels of myeloperoxidase/IL-1beta and glutathione peroxidase/lactoferrin could result in tissue damage of reactive oxygen species (ROS) in periodontitis which is initiated and perpetuated by the chronic insults of periodontopathogens.

Bone resorbing activity and cytokine levels in gingival crevicular fluid before and after treatment of periodontal disease.

The aim of the present study was to investigate bone resorption activity (BRA), interleukin-1 alpha (IL-1 alpha), IL-1 beta and interleukin-1 receptor antagonist (IL-1ra) in gingival crevicular fluid (GCF) in sites with no signs of periodontal disease and in sites with horizontal or angular loss of periodontal bone. These assessments were performed before and after periodontal treatment. GCFs were collected from 10 individuals with filter strips from two healthy sites and four sites with deep pathological periodontal pockets, two of which showed horizontal bone loss and two with angular bone loss. All diseased pockets were treated with flap surgery and systemic Doxyferm. Twelve months later GCF was collected again and treatment outcome evaluated. BRA in GCFs was assessed in a bone organ culture system by following the release of (45)Ca from neonatal mouse calvariae. The amounts of IL-1 alpha, IL-1 beta and IL-1ra in GCFs were quantified by enzyme-linked immunosorbent assay (ELISA). Treatment resulted in reduction of pocket depths with 3.5+/-0.5 mm in sites with angular bone loss and 2.8+/-0.3 mm in sites with horizontal bone loss. Initially, BRA, IL-1 alpha, IL-1 beta and IL-1ra were significantly higher in GCFs from diseased sites compared with healthy sites. No differences in BRA and cytokine levels were seen between GCFs from pockets with horizontal and angular bone losses. The levels of IL-1 alpha, IL-1 beta and IL-1ra were significantly reduced after treatment of diseased pockets. Pocket depths were significantly correlated to BRA only in pre-treatment sites with angular bone loss. BRA was correlated to Il-1 alpha, IL-1 beta, but not to IL-1ra, in diseased sites with angular bone loss, before and after treatment. The reductions of BRA in the individual sites, seen after treatment, were not correlated to the reductions of Il-1 alpha, IL-1 beta or IL-1ra. These data show that BRA and cytokine levels are increased in GCFs from sites with periodontal disease and that periodontal treatment results in reduction of the cytokines. Our findings further indicate that IL-1 alpha and IL-1 beta play important roles for the BRA present in GCFs, but that other factors also contribute to this activity.

Effects of scaling and root planing on the amounts of interleukin-1 and interleukin-1 receptor antagonist and the mRNA expression of interleukin-1beta in gingival crevicular fluid and gingival tissues.

The purpose of this study was to evaluate the relationship between the clinical changes after non-surgical periodontal therapy and interleukin 1 (IL-1) in gingival crevicular fluid (GCF) and gingival tissues from patients with chronic periodontitis. The inflammatory responses mediated by IL-1 play an important role in periodontal tissue destruction. Although numerous studies have attempted to elucidate the dynamic movement involved in chronic periodontitis, the results have often conflicted. Such discrepancies may have been due to the inability to determine clinical disease activity. Seven patients with chronic periodontitis were examined. The severity of periodontal inflammation was expressed using clinical parameters before and after a scaling and root planing (SRP) procedure. The amounts and concentrations of IL-1alpha, IL-1beta and IL-1 receptor antagonist in GCF were measured by enzyme-linked immunosorbent assay (ELISA) and IL-1 activity index was calculated. A needle biopsy in matching gingival tissues was also performed before and after the SRP procedure. The localization and mRNA expression of IL-1beta were determined using histological methods. Clinical parameters improved slightly after the SRP procedure. Only the probing pocket depth (PPD) was reduced significantly (p < 0.05). However, the amount of IL-1beta in GCF was slightly increased. The localization and mRNA expression of IL-1beta could still be observed after the SRP procedure. Therefore, none of the clinical parameters showed a high sensitivity or specificity for evaluating subgingival inflammation. These observations suggest that IL-1 is effective for evaluating in detail the state of subgingival inflammation.

Impact of increased occlusal contact, interleukin-1 genotype, and periodontitis severity on gingival crevicular fluid IL-1beta levels.

The study of occlusal therapy in humans with periodontitis is problematic due to potential irreversible bone loss in control subjects. The hypothesis of this pilot investigation was that increased interocclusal contact initiated by short-term occlusal splint disuse would increase tooth mobility and the bone-resorptive cytokine interleukin (IL)-1beta in gingival crevicular fluid (GCF), comparable to IL-1-positive genotype and increased periodontitis severity. Nineteen non-smoking chronic periodontitis patients using nocturnal occlusal splints and undergoing periodontal maintenance in a private practice were evaluated at five time points: 24 hours after continuous splint use; 1, 2, and 3 days after no occlusal splint use; and 14 days after resumption of customary nighttime splint use. Subjects were evaluated to confirm that the plaque index and gingival index were < or = 1.0, and to categorize past periodontitis (moderate or severe) and IL-1 genotype (1A +4845 plus IL-1B +3954). Test sites on two anterior teeth vulnerable to occlusal trauma were sampled for mobility, GCF IL-1beta, and IL-1 receptor antagonist (ra). Tooth mobility remained low during the 3-day period when patients were not wearing their occlusal appliance. GCF IL-1beta decreased after not wearing the appliances (P = 0.016), especially at 48 hours. At this time, genotype-positive subjects had higher levels of GCF IL-1beta/IL-1ra than genotype-negative subjects (P = 0.045), and patients who had experienced severe periodontitis had higher IL-1beta levels than moderate periodontitis subjects (P = 0.004). These findings suggest that short-term discontinuation of occlusal splint therapy in non-smoking periodontitis patients undergoing periodontal maintenance does not result in potential signs of early occlusal trauma (increasing mobility or GCF IL-1beta). Longer-term studies may be needed to determine appropriate therapy applications for periodontitis-susceptible patients with definable occlusal discrepancies and/or parafunction.

GCF IL-1beta profiles in periodontal disease.

Studies suggest a genetic influence on levels of interleukin-1beta (IL-1beta) in gingival crevicular fluid (GCF). Levels of IL-1beta in GCF, however, are also dependent upon the clinical parameters at the site of collection, including probing depth (PD) and level of attachment (AL). To examine this issue, IL-1beta in GCF was evaluated from patients with varying degrees of periodontal disease. The influence of both the status of the patient and the probing depth at the sampled sites were considered in the analysis. GCF IL-1beta was determined by ELISA at 6-8 molar sites from 29 non-smoking adults with mild, moderate, or severe periodontal disease at baseline, 2 weeks, and 24 weeks following scaling and root planing. For later analysis, patients were dichotomized on the basis of disease severity (mild/moderate vs severe). Sampled sites were classified at baseline by PD as, shallow (<4 mm), intermediate (4-6 mm), or deep (>6 mm). PD and AL were each strongly correlated with IL-1beta levels at baseline. However, patients with severe disease had higher levels of IL-1beta in each PD category than those with mild/moderate disease. As compared to patients with mild/moderate disease, IL-1beta levels in shallow sites from patients with severe disease was elevated nearly 2 fold (p<0.001). IL-1beta levels were reduced in all groups at 2 weeks and were still significantly reduced in patients with mild/moderate disease at 24 weeks. At 24 weeks IL-1beta returned to near baseline levels in patients with severe disease. While PD and AL are each associated with increased GCF IL-1beta, patients with severe disease show higher IL-1beta GCF levels in shallow sites, suggesting that high GCF IL-1beta expression is in part a host trait, and not strictly a function of clinical parameters.

Levels of Interleukin‐1β, ‐8, and ‐10 and RANTES in Gingival Crevicular Fluid and Cell Populations in Adult Periodontitis Patients and the Effect of Periodontal Treatment

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.