Localized adhesion molecule expression and circulating LFA-3 levels in adult and early onset forms of periodontitis

Because of their importance in mediating cellular interactions in chronic inflammatory diseases, this study has examined the expression of a number of adhesion molecules in adult (n=11), generalized early onset (n=5) and localized early onset (n=2) forms of periodontitis. In comparison with immunostaining profiles of cryostat sections of healthy gingival tissue (n=7), the beta 1 integrins VLA-1, VLA-2 and VLA-4 were found to be up-regulated in periodontitis, with VLA-6 being markedly elevated. Although only small differences were observed in ICAM-1 and LFA-3 expression in the gingival epithelium, there was particularly notable up-regulation of these adhesion molecules within the inflammatory infiltrates of the diseased tissues. However, there were no statistically significant differences between the serum levels of a soluble form of LFA-3 in periodontitis patients (n=47) compared with healthy control subjects (n=40), although the generalized early onset and adult periodontitis groups exhibited wider ranges of circulating LFA-3. These findings show that there is localized modulation of adhesion molecule expression in the chronic inflammatory periodontal diseases studied, but that the levels of LFA-3 in the circulation nevertheless remain unaffected.     

Salivary malondialdehyde levels in clinically healthy and periodontal diseased individuals

Background: Lipid peroxidation (LPO) has been implicated in the pathogenesis of several pathologic disorders, including periodontal disease. Malondialdehyde (MDA) is one of many low molecular weight end products of LPO. Objective: This study was conducted to evaluate salivary MDA levels in generalized chronic periodontitis (GCP) subjects. Materials and methods: The MDA levels were measured in the saliva of 104 subjects, aged 18–65 years. Three groups with different degrees of severity of GCP were established: 30 early (group 1), 30 moderate (group 2) and 14 severe (group 3). Thirty individuals (aged 25–29 years) with clinically healthy periodontium were served as control. Unstimulated whole saliva samples from study subjects were collected, centrifuged at 3000 g for 15 min and were then stored at ?70°C until analysed. The MDA level was determined with 2‐thiobarbituric acid by a colorimetric method at 532 nm. Results: A significant increase in the MDA level existed in the samples obtained from the three groups of patients compared to the control subjects. Conclusion: Increased MDA levels are with closely associated with the severity and patients status of periodontal disease that has not been previously reported. The detection of salivary MDA level may provide additional advantages in elucidating the pathogenesis of periodontal disease.     

Crevicular fluid endothelin-1 levels in periodontal health and disease

Background and Objective:  Endothelin-1 is a 21-amino-acid peptide with multifunctional regulation. Initial research indicated that endothelin-1 levels in the gingival crevicular fluid from patients with chronic periodontitis were higher than those in the gingival crevicular fluid from healthy subjects. The aim of the present study was to assess the relationship between the clinical parameters and the concentrations of endothelin-1 within the gingival crevicular fluid from inflamed gingiva and periodontitis sites and, subsequently, after the treatment of periodontitis sites.
Material and Methods:  A total of 60 subjects were divided into three groups – healthy (group I), gingivitis (group II) and chronic periodontitis (group III) – based on gingival index, pocket probing depth and clinical attachment loss. A fourth group consisted of 20 subjects from group III, 6–8 wk after treatment (i.e. scaling and root planing). Gingival crevicular fluid samples collected from each patient were quantified for endothelin-1 using an enzymatic immunometric assay.
Results:  Endothelin-1 was not detected in any sample from any of the study groups.
Conclusion:  The results showed that all the gingival crevicular fluid samples were negative for the endothelin-1 molecule. Therefore, endothelin-1 cannot be considered as a potential biomarker of periodontal disease progression.     

GCF IL-1beta profiles in periodontal disease

OBJECTIVES: 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. MATERIAL AND METHODS: 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). RESULTS: 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. CONCLUSION: 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.     

Estimation of interleukin-1beta levels in the gingival crevicular fluid in health and in inflammatory periodontal disease

Initial research indicated that the levels of interleukin-1beta (IL-1beta) are higher in sites of inflammation than in healthy sites. However, subsequent studies suggest heterogenous responses and indicate the quantitative levels of IL-1beta to be the characteristic of an individual rather than simply being the reflection of the inflammatory status of the tissues. This study has been designed to find out the relationship between IL-1beta levels in the gingival crevicular fluid and the inflammatory status of the periodontal tissues in the Indian population. Sixty patients were selected for the study. They were categorized in to three groups based on their periodontal tissue status as group I (clinically healthy gingiva with no loss of attachment), group II (gingivitis with no attachment loss) and group III (gingivitis with attachment loss). Microcapillary pipettes were used to collect gingival crevicular fluid samples from one site in each person and the samples were analysed for IL-1beta using a commercially available ELISA kit. The concentration of IL-1beta in the gingival crevicular fluid of patients in group III is statistically higher (P < 0.0001) than that in group II and the concentration of IL-1beta in groups II and III is statistically at much higher levels (P < 0.0001) than in the group I subjects. However, there is a significant overlap in the values obtained in groups II and III and the values in both the groups range over a wide spectrum. The composite values obtained within the groups and the overlapping values in groups II and III could indicate the role of genetic polymorphism in determining the quantity of IL-1beta produced and also the contributory role of other cytokines that share similar biologic activity.     

Salivary biomarkers of existing periodontal disease: a cross-sectional study

BACKGROUND: The authors conducted a study to determine if salivary biomarkers specific for three aspects of periodontitis--inflammation, collagen degradation and bone turnover--correlate with clinica features of periodontal disease. METHODS: The relationship between periodontal disease and the levels of interleukin-1 beta (IL-1beta), matrix metalloproteinase (MMP)-8, and osteoprotegerin (OPG) in whole saliva of 57 adults (28 "case" subjects with moderate-to-severe periodontal disease and 29 healthy control subjects) was examined in a case-control trial. RESULTS: Mean levels of IL-1beta and MMP-8 in saliva were significantly higher in case subjects than in controls. Both analytes correlated with periodontal indexes, whereas, after adjustment for confounders, OPG did not. Elevated salivary levels of MMP-8 or IL-1beta (more than two standard deviations above the mean of the controls) significantly increased the risk of periodontal disease (odds ratios in the 11.3-15.4 range). Combined elevated salivary levels of MMP-8 and IL-1beta increased the risk of experiencing periodontal disease 45-fold, and elevations in all three biomarkers correlated with individual clinical parameters indicative of periodontal disease. CONCLUSION: Salivary levels of MMP-8 and IL-1beta appear to serve as biomarkers of periodontitis. CLINICAL IMPLICATIONS: Qualitative changes in the composition of salivary biomarkers could have significance in the diagnosis and treatment of periodontal disease.     

Salivary infectious agents and periodontal disease status

Saygun I, Nizam N, Keskiner I, Bal V, Kubar A, Aç?kel C, Serdar M, Slots J. Salivary infectious agents and periodontal disease status. J Periodont Res 2011; 46: 235–239. © 2011 John Wiley & Sons A/S Background and Objectives: The potential of salivary microorganisms to diagnose periodontal disease and to guide periodontal treatment is a research topic of current interest. This study aimed to determine whether the salivary counts of periodontopathic microbes correlated with the periodontal pocket counts of the same infectious agents, and whether the salivary counts of the test infectious agents could distinguish among individuals with periodontal health and various types of periodontal disease. Material and Methods: The study included 150 systemically healthy adults, of whom 37 were periodontally healthy, 31 had gingivitis, 46 had chronic periodontitis and 36 had aggressive periodontitis. Each study subject contributed microbial samples from the two deepest periodontal pockets of the dentition and from whole saliva. Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Epstein–Barr virus were identified using the TaqMan real‐time PCR methodology. Statistical analysis was performed using the Mann–Whitney U‐test and the receiver operating characteristic statistics. Results: C. rectus, F. nucleatum, P. gingivalis, P. intermedia and T. forsythia occurred with significantly higher copy‐counts in salivary samples from patients with gingivitis, chronic periodontitis and aggressive periodontitis than from periodontally healthy individuals. A. actinomycetemcomitans only showed higher salivary copy‐counts in subjects with aggressive periodontitis compared with subjects with healthy periodontium, and the salivary copy‐counts of Epstein–Barr virus did not reveal any significant difference among the four subject groups studied. The diagnostic sensitivity for periodontitis was 89.19 for P. gingivalis and for T. forsythia and 86.49 for P. intermedia, with specificities ranging from 83.78 to 94.59. The optimal copy‐counts per mL saliva for identifying periodontitis were 40,000 for P. gingivalis, 700,000 for T. forsythia and 910,000 for P. intermedia. Conclusion: Salivary copy‐counts of P. gingivalis, T. forsythia and P. intermedia appear to have the potential to identify the presence of periodontitis, whereas the salivary level of the other test infectious agents may possess little or no diagnostic utility. Longitudinal studies are warranted to determine the ability of salivary copy‐counts of major periodontopathic bacteria to predict future periodontal breakdown.     

Salivary interleukin-1β concentration and the presence of multiple pathogens in periodontitis

Aim: This study aimed to find salivary enzymes and/or cytokines that would reflect periodontitis, alone or in combination with salivary microbial markers.
Material and Methods: The salivary concentrations of elastase, lactate dehydrogenase, interleukin-1 β (IL-1 β ), interleukin-6, and tumour necrosis factor- α , and the presence of five periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia , and Treponema denticola , were analysed from salivary specimens of 165 subjects, a subpopulation of Health 2000 Health Examination Survey in Finland; 84 of the subjects had probing pocket depth (PPD) of 4 mm at 14 or more teeth (the advanced periodontitis group), while 81 subjects had no teeth with PPD of 4 mm (the control group). All subjects had at least 20 teeth and no systemic diseases.
Results: Among the salivary cytokines and enzymes tested, IL-1 β was the only biomarker associated with periodontitis. An association was also found with the presence of multiple periodontal pathogens. Salivary IL-1 β and the presence of multiple periodontal pathogens were associated with periodontitis at the same magnitude, when they were in the logistic regression model individually or together.
Conclusion: We suggest that salivary IL-1 β and the presence of multiple periodontal pathogens in saliva should be studied more thoroughly as markers of periodontitis.     

Correlation between salivary IL-1beta levels and periodontal clinical status

OBJECTIVE: To assess the concentration of the proinflammatory cytokine IL-1beta in saliva of periodontally diseased and healthy patients and their relationship with the periodontal status. DESIGN: Unstimulated whole saliva samples from patients with chronic periodontitis (n=30), aggressive periodontitis (n=18) and healthy controls (n=18) were obtained for the study. The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss and the extent/severity of periodontal breakdown. The levels of IL-1beta were measured in saliva samples with a high sensitivity enzyme-linked immunosorbent assay (ELISA). RESULTS: Although no significant difference (P=0.624) was found for salivary IL-1beta levels between periodontitis groups, they were significantly greater (P<0.01) than those detected for healthy controls. Furthermore, Spearman correlation analysis showed statistically significant correlations (P<0.01) between data from salivary IL-1beta levels and clinical measurements. CONCLUSION: The findings of the present study reemphasize the importance of whole saliva as sampling method in terms of immunological purposes in periodontal disease and suggest that the elevated IL-1beta concentration may be one of the host-response components associated to the clinical manifestations of periodontal disease.     

Salivary cytokine levels in subjects with chronic periodontitis and in periodontally healthy individuals: a cross-sectional study

Background and Objective: Saliva has been proposed as a noninvasive diagnostic fluid that could be used in the diagnosis of oral and systemic diseases. The levels of salivary biomarkers, such as cytokines, could potentially be used as a surrogate to distinguish periodontally healthy individuals from subjects with periodontitis. Therefore, the goal of the present investigation was to determine if the levels of 10 different cytokines in saliva differed between a group of periodontally healthy individuals and a group of subjects with periodontitis. Correlations between the concentrations of these 10 cytokines and clinical parameters of periodontal disease were also examined. Material and Methods: In this cross‐sectional study, 74 subjects with chronic periodontitis and 44 periodontally healthy individuals were periodontally examined and had the levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐1β, interleukin‐2, interleukin‐4, interleukin‐5, interleukin‐6, interleukin‐8, interleukin‐10, interferon‐γ and tumor necrosis factor‐α measured in whole saliva using a multiplexed bead immunoassay (Luminex). Significance of statistical differences in the levels of salivary cytokines between groups was determined using nonparametric analysis of covariance, adjusting for age and smoking status. The Spearman rank correlation coefficient was used to explore associations between the mean levels of salivary cytokines and mean clinical parameters. Results: There were no statistically significant differences between groups for any of the cytokines. There were weak, statistically significant positive associations between salivary interleukin‐8 and pocket depth (r_s = 0.2, p < 0.05) and bleeding on probing (r_s = 0.2, p < 0.05), and weak negative correlations between salivary interleukin‐10 and attachment level (r_s = ?0.2, p < 0.05) and bleeding on probing (r_s = ?0.3, p < 0.001). Conclusion: Mean salivary levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐1β, interleukin‐2, interleukin‐4, interleukin‐5, interleukin‐6, interleukin‐8, interleukin‐10, interferon‐γ and tumor necrosis factor‐α could not discriminate between periodontal health and disease.     

Gingival fluid IL-1βin postmenopausal females on supportive periodontal therapy

Abstract. Posterior interproximal alveolar bone in 59 women, within 5 years after menopause, was assessed at baseline and after 2 years of supportive periodontal therapy (history of moderate/advanced periodontitis) using digitized image analysis. Baseline lumbar spine bone mineral density, smoking status, and yearly serum estradiol (E₂) levels also were obtained to group subjects. An additional 16 non-periodontitis postmenopausal women were followed 2 years for clinical and estrogen status, 2-min GCF IL–1β levels averaged from 2 baseline periodontal pockets (in periodontitis subjects) and 2 non-periodontitis sites (in non-penodontitis and periodontitis subjects) were determined with an enzyme immunoassay, A progressive and stable site were also monitored every 6 months for GCF IL–1β in 15 patients. Results after 2 years indicated that 17 subjects had no posterior interproximal sites losing ≥0.4 mm of alveolar crest bone height, while 13 subjects had ≥3 such sites. Using analysis of variance, none of the above clinical groupings resulted in a significant difference in mean baseline or longitudinal GCF IL–1≥ levels, However, when subjects who lost alveolar crest bone height were considered. E₂-sufficient subjects had significantly depressed baseline GCF IL-β (in past-periodontitis sites) compared to E₂;-deficient patients (9.1 ± 2,1 versus 31,7±10.2 pg/2-min sample. p <0.05), suggesting E₂ influences gingival IL-lβ production in progressive periodontitis patients.     

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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。     

Effect of periodontal treatment on IL-1beta, IL-1ra, and IL-10 levels in gingival crevicular fluid in patients with aggressive periodontitis

Aim: The aim of this study was to examine the effect of phase I periodontal treatment on the levels of interleukin (IL)‐1β, IL‐1ra, and IL‐10 in gingival crevicular fluid (GCF) in patients with generalized aggressive periodontitis (G‐AgP). Material and Methods: Data were obtained from 15 patients with aggressive periodontitis and 15 healthy controls. GCF was collected from at least four pre‐selected sites (one shallow, at least two moderate, or at least one deep pockets) in patients with G‐AgP. In the healthy group, GCF samples were collected from one site. The cytokine levels were determined by an enzyme‐linked immunosorbent assay. Probing depth, clinical attachment level (CAL), gingival and plaque indices, and bleeding on probing were measured. The GCF sampling and clinical measurements were recorded at baseline and 6 weeks later after periodontal treatment. Results: IL‐1β levels were significantly higher at the moderate and deep pocket sites compared with the shallow sites (p<0.05). After periodontal therapy, IL‐1β levels were significantly reduced in the moderate and deep pocket sites (p<0.05). IL‐1ra levels at baseline of the moderate and deep pocket sites were significantly lower than the control sites (p<0.05). IL‐10 levels were similar in all pockets and did not change after periodontal therapy. Conclusions: The periodontal treatment improves the clinical parameters in G‐AgP, and this improvement is evident in deep pocket sites for pocket depth and CAL values. These results confirm that IL‐1β is effective for evaluating the periodontal inflammation and can thus be used as a laboratory tool for assessing the activity of periodontal disease.     

Protease inhibitor levels in periodontal health and disease

Kretschmar S, Yin L, Roberts F, London R, Flemmig TT, Arushanov D, Kaiyala K, Chung WO. Protease inhibitor levels in periodontal health and disease. J Periodont Res 2012; 47: 228–235. © 2011 John Wiley & Sons A/S Background and Objective: Our previous study showed that protease inhibitors were attenuated by the periodontal pathogen Porphyromonas gingivalis in cultured gingival epithelial cells. We hypothesize that fewer protease inhibitors would be present in more advanced periodontal disease sites, where the level of P. gingivalis may be high. The goal of this study was to investigate the relationship between the protease inhibitor [secretory leukocyte protease inhibitor (SLPI), elastase‐specific inhibitor (ELAFIN) and squamous cell carcinoma antigen (SCCA)] levels in gingival crevicular fluid and the number of P. gingivalis micro‐organisms in subgingival plaque. Material and Methods: Plaque samples from subjects without (n = 18) and with moderate to advanced periodontitis (n = 41) were used to quantify P. gingivalis using real‐time PCR. Protease inhibitor levels in the gingival crevicular fluid of all the subjects were determined by ELISA. Results: P. gingivalis was detected in 68.3% of patients with periodontitis, while 16.7% of subjects without periodontitis had a detectable level of P. gingivalis. Patients with periodontitis and P. gingivalis in their plaque exhibited lower SLPI and ELAFIN levels (p < 0.001) compared with control subjects without periodontitis. Secretory leukocyte protease inhibitor was also reduced (p < 0.05) in gingival crevicular fluid of periodontitis patients without a detectable level of P. gingivalis. Periodontitis patients with high vs. low levels of P. gingivalis exhibited reciprocal mean levels of SLPI and ELAFIN concentrations. Conclusion: The reduced concentrations of SLPI and ELAFIN may contribute to the loss of host protective capacity and increase susceptibility to breakdown from chronic infection. The work of this investigation may aid in finding diagnostic and prognostic markers in periodontal health and disease and may also help in finding pharmacological targets directed against periodontal inflammation.     

Smoking and GCF levels of IL-1beta and IL-1ra in periodontal disease

AIMS: GCF levels of the cytokine IL-1beta and its receptor antagonist IL-1ra were analyzed with respect to smoking in patients with moderate to severe periodontal disease. The study population included 22 smokers and 18 non-smokers in the age range 32-86 years. Concomitantly, the GCF levels of IgA, IgG, albumin and total protein were analyzed. METHOD: Samples of GCF were obtained from 2 diseased sites in each patient by means of an aspiration method. IL-1beta, IL-1ra, IgA and IgG were determined with immunoelectrophoresis. Total protein was determined by the BCA method. RESULTS: The clinical characteristics in terms of probing depth and frequency of diseased sites and supragingival plaque did not differ between smokers and non-smokers. Gingival bleeding, however, was significantly depressed in smokers. IL-1beta was detected in GCF of 95% of both smokers and non-smokers and IL-1ra in all patients. The GCF level of IL-1ra was approximately 1,000-fold that of IL-1beta. The GCF levels of IL-1beta and IL-1ra were high in comparison with those of TNF-alpha and IL-6 determined by the same method in our earlier studies. CONCLUSION: Our observations did not reveal any influence of smoking on the levels of IL-1beta and IL-1ra in GCF.     

Interleukin-17 and interleukin-18 levels in saliva and plasma of patients with chronic periodontitis

Özçaka Ö, Nalbantsoy A, Buduneli N. Interleukin‐17 and interleukin‐18 levels in saliva and plasma of patients with chronic periodontitis. J Periodont Res 2011; 46: 592–598. © 2011 John Wiley & Sons A/S Background and Objective: This study was planned to investigate whether patients with chronic periodontitis exhibit different salivary and/or plasma concentrations of interleukin (IL)‐17 and IL‐18 compared with clinically healthy subjects. Material and Methods: Whole saliva and blood samples, together with full‐mouth clinical periodontal recordings, were obtained from 22 otherwise healthy untreated nonsmokers with chronic periodontitis and from 21 systemically and periodontally healthy control subjects. The concentrations of IL‐17 and IL‐18 in saliva and plasma were determined using ELISAs. Results: The healthy control group exhibited significantly lower values in all clinical periodontal measurements (p < 0.001). The salivary concentration of IL‐17 was significantly lower, and that of IL‐18 significantly higher, in patients from the chronic periodontitis group compared with healthy control subjects (p = 0.025 and p = 0.009, respectively). Plasma IL‐17 and IL‐18 concentrations were similar in the two study groups (p > 0.05). Conclusion: Within the limits of the present study, it may be suggested that an elevated salivary IL‐18 level in untreated nonsmoker chronic periodontitis patients has the potential to be a biomarker for periodontal tissue destruction.