Salivary biomarkers in association with periodontal parameters and the periodontitis risk haplotype.
ABSTRACT: Genetic factors play a role in periodontitis. Here we examined whether the risk haplotype of MHC class III region BAT1-NFKBIL1-LTA and lymphotoxin-? polymorphisms associate with salivary biomarkers of periodontal disease. A total of 455 individuals with detailed clinical and radiographic periodontal health data were included in the study. A 610?K genotyping chip and a Sequenom platform were used in genotyping analyses. Phospholipid transfer protein activity, concentrations of lymphotoxin-?, IL-8 and myeloperoxidase, and a cumulative risk score (combining Porphyromonas gingivalis, IL-1? and matrix metalloproteinase-8) were examined in saliva samples. Elevated IL-8 and myeloperoxidase concentrations and cumulative risk scores associated with advanced tooth loss, deepened periodontal pockets and signs of periodontal inflammation. In multiple logistic regression models adjusted for periodontal parameters and risk factors, myeloperoxidase concentration (odds ratio (OR); 1.37, P?=?0.007) associated with increased odds for having the risk haplotype and lymphotoxin-? concentration with its genetic variants rs2857708, rs2009658 and rs2844482. In conclusion, salivary levels of IL-8, myeloperoxidase and cumulative risk scores associate with periodontal inflammation and tissue destruction, while those of myeloperoxidase and lymphotoxin-? associate with genetic factors as well.
Project description:Periodontitis is a chronic inflammatory disease with a multifactorial etiology. We investigated whether human major histocompatibility complex (MHC) polymorphisms (6p21.3) are associated with periodontal parameters. Parogene 1 population samples (n = 169) were analyzed with 13,245 single nucleotide polymorphisms (SNPs) of the MHC region. Eighteen selected SNPs (P ? 0.001) were replicated in Parogene 2 population samples (n = 339) and the Health 2000 Survey (n = 1,420). All subjects had a detailed clinical and radiographic oral health examination. Serum lymphotoxin-? (LTA) concentrations were measured in the Parogene populations, and the protein was detected in inflamed periodontal tissue. In the Parogene 1 population, 10 SNPs were associated with periodontal parameters. The strongest associations emerged from the parameters bleeding on probing (BOP) and a probing pocket depth (PPD) of ?6 mm with the genes BAT1, NFKBIL1, and LTA. Six SNPs, rs11796, rs3130059, rs2239527, rs2071591, rs909253, and rs1041981 (r(2), ?0.92), constituted a risk haplotype. In the Parogene 1 population, the haplotype had the strongest association with the parameter BOP, a PPD of ?6 mm, and severe periodontitis with odds ratios (95% confidence intervals) of 2.63 (2.21 to 3.20), 2.90 (2.37 to 3.52), and 3.10 (1.63 to 5.98), respectively. These results were replicated in the other two populations. High serum LTA concentrations in the Parogene population were associated with the periodontitis risk alleles of the LTA SNPs (rs909253 and rs1041981) of the haplotype. In addition, the protein was expressed in inflamed gingival connective tissue. We identified a novel BAT1-NFKBIL1-LTA haplotype as a significant contributor to the risk of periodontitis. The genetic polymorphisms in the MHC class III region may be functionally important in periodontitis susceptibility.
Project description:Several studies have addressed cytokine gene polymorphisms and their possible associations with periodontitis. We examined the association between salivary anti- and pro-inflammatory mediator polymorphisms and initial periodontitis in Finnish adolescents, taking into account the effect of smoking. Salivary samples of 93 clinically examined adolescents from Eastern Finland were analyzed. Their oral health and smoking habits were recorded. Periodontal probing depth (PPD), and bleeding on probing (BOP) at four sites per tooth, root calculus (RC), and visible plaque index (VPI) were recorded from the index teeth. Salivary MMP-8 median values were assessed. The sites with ?4 mm PD were categorized as follows: PPD1 = one or more ?4 mm pocket, PPD2 = two or more ?4 mm pockets, and PPD3 = three or more ?4 mm pockets. Genomic DNA was extracted from 300 ?l of the saliva samples by genomic QIAamp® DNA Blood Mini Kit and genotyped for polymorphisms. Genetic variants for genotyping were selected from the following genes of interest: S100A8, FCGR2A, FCGR2B, IL10, MMP8, MMP3, MMP13, VDR, TLR4, MMP2, MPO, ELANE, IL1A, IL1B, IL1RN, CD28, MMP9, DDX39B, NFKBIL1, LTA, TNF, SOD2, IL6, TLR4, TIMP1, and SYN1. After false discovery rate control (FDR), polymorphisms in MMP3 (rs679620, rs520540, rs639752), CD28 (rs3116496), and VDR (rs2228570) associated (FDR q < 0.05) with deepened periodontal pockets. Smoking did not affect the results. Genetic polymorphisms of pro-inflammatory mediators MMP3, CD28, and VDR seem to link to initial periodontitis.
Project description:Background:Periodontitis is associated with increased concentration of inflammatory markers and saliva has been proposed as a non-invasive diagnostic fluid in oral and systemic diseases. The levels of salivary biomarkers, such as cytokines, could potentially be used to distinguish periodontal healthy individuals from subjects with periodontal disease. The purpose of this study was to characterize the salivary levels of two inflammatory biomarkers associated with periodontitis, interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-?), in order to assess whether these cytokines salivary levels could potentially be used to complement periodontitis pregnant women diagnose. Methods:Forty-four pregnant women were distributed into three groups, according to their periodontal status: healthy, mild/moderate periodontitis and severe periodontitis. Unstimulated saliva was collected and analysis of TNF-? and IL-6 salivary levels were performed with Immulite®. Results:Women with periodontitis exhibited significantly higher levels (p = 0.001) of salivary IL-6 and TNF-? compared with the healthy group: 25.1 (±11.2) pg/mL vs. 16.3 (±5.0) pg/mL and 29.7 (±17.2) pg/mL vs. 16.2 (±7.6) pg/mL, approximately 1.5 and 1.8 times more, respectively. Additionally, cytokines were significantly increased (p < 0.05) in severe periodontitis compared to periodontal healthy pregnant women. Conclusions:These results revealed that IL-6 and TNF-? salivary biomarkers provide high discriminatory capacity for distinguishing periodontal disease from periodontal health in pregnant women.
Project description:AIMS:Higher intakes of fruits and vegetables, and vitamin C are associated with improved periodontal healing post-scaling and root planing (SRP). This study determined if this association was sustained at 3-4 years post-SRP, and if flavonoid intake is associated with periodontal health. Whether reduced probing depth (PD) is sustained and whether PD is correlated with salivary IL-1?, IL-6 and CRP at 3-4 years post-SRP were also studied. MATERIALS AND METHODS:Clinical periodontal outcomes, dietary intakes and salivary markers of inflammation were measured in patients (n = 43, 23 females, 37-93 years) who had undergone SRP 3-4 years earlier and had been part of a periodontal maintenance programme. RESULTS:Flavonoid intake was inversely associated with PD (p = .042) and salivary IL-1? concentration (p = .015) after adjustment for multiple confounders. When changes in PD were considered, the association of flavonoid intake with reduced PD became borderline significant (p = .051) but persisted for IL-1? (p = .018). PD at 3-4 years and 2-4 months post-SRP was similar. There was a positive correlation between PD and salivary IL-1? (p = .005) but not with salivary CRP and IL-6. CONCLUSION:Higher flavonoid intake is associated with lower IL-1?. Also, regular supportive periodontal therapy maintained the improved PD at 3-4 years post-SRP regardless of smoking status.
Project description:OBJECTIVE:This study aimed to determine serum and salivary levels of neutrophil gelatinase-associated lipocalin (NGAL) and evaluate NGAL correlation with key anti-interleukin 10 (IL-10) and pro-inflammatory (IL-1?) cytokines in different severities of periodontal diseases. We also calculated the systemic inflammation using the periodontal inflamed surface area (PISA) to evaluate its correlation with NGAL in the study groups. METHODOLOGY:Eighty systemically healthy and non-smoking individuals were separated into four groups of 20: clinically healthy (Group 1), gingivitis (Group 2), stage I generalized periodontitis (Group 3, Grade A), and stage III generalized periodontitis (Group 4, Grade A). Sociodemographic characteristics and periodontal parameters were recorded, and PISA was calculated. The serum and salivary levels of interleukin (IL)-1?, IL-10, and NGAL were determined using the enzyme-linked immunosorbent assay (ELISA). RESULTS:We observed a significant increase in serum and salivary NGAL levels from healthy to periodontitis groups (p=0.000). Group 2 presented significantly higher serum and salivary IL-10 levels and salivary IL-1? levels than Group 3 (p=0.000). Serum and salivary parameters (IL-1?, IL-10, and NGAL levels) were strongly positively correlated to periodontal parameters and PISA values (p=0.000). Groups 2 and 3 showed overlapping PISA values. CONCLUSION:The overlapping PISA values found in Groups 2 and 3 suggest that gingivitis might progress to a systemic inflammatory burden somewhat comparable to stage I periodontitis. This finding is supported by the higher serum and salivary cytokines/mediators levels in the gingivitis group than in stage I periodontitis group. Serum and salivary NGAL levels increased proportionally to disease severity and PISA. NGAL seems to play a role in the pathogenesis of periodontal disease, within the limitation of our study.
Project description:The purpose of this study was to determine the impact of nonsurgical periodontal therapy considering the salivary stress-related hormone and cytokine levels in the gingival crevicular fluid (GCF) on pregnant and nonpregnant women.Thirty non-pregnant (control group) and 30 pregnant women (test group) that met the study inclusion criteria were chosen. Only participants with gingivitis were included. Clinical data and samples of GCF and saliva were collected at baseline and after periodontal therapy. The levels of interleukin-1 beta (?-1?) and IL-10, and concentration of salivary chromogranin A (CgA) hormone were analyzed by enzyme-linked immunosorbent assay (ELISA). The repeated measures analysis of variance was used for intragroup and intergroup analyses.A major decrease in the gingival inflammation was observed in both groups after periodontal therapy (p<0.05). Periodontal treatment decreased the level of IL-1? in GCF (p<0.05) in control group, but no statistical difference was determined for GCF IL-1? in the test group. However, after periodontal therapy, the CgA hormone concentration was reduced in both groups (p<0.05). However, there was no difference in salivary CgA concentration, GCF IL-10 levels, and perceived stress scale (PSS)-10 between the groups (p>0.05).Within the limitations of this study, periodontal therapy significantly improved the periodontal status and stress level. In addition, the severity of the gingival inflammation during pregnancy was related to stress. However, further studies will be needed to substantiate these early findings.
Project description:OBJECTIVE:Interleukin-14?-transgenic (IL-14?TG) mice develop an autoimmune exocrinopathy with characteristics similar to Sjögren's syndrome, including sialadenitis and hyposalivation. The P2Y2 receptor (P2Y2 R) for extracellular ATP and UTP is upregulated during salivary gland inflammation (i.e., sialadenitis) where it regulates numerous inflammatory responses. This study investigated the role of P2Y2 Rs in autoimmune sialadenitis in the IL-14?TG mouse model of Sjögren's syndrome. MATERIALS AND METHODS:IL-14?TG mice were bred with P2Y2 R-/- mice to generate IL-14?TG × P2Y2 R-/- mice. P2Y2 R expression, lymphocytic focus scores, B- and T-cell accumulation, and lymphotoxin-? expression were evaluated in the submandibular glands (SMG) along with carbachol-stimulated saliva secretion in IL-14?TG, IL-14?TG × P2Y2 R-/- , and C57BL/6 control mice at 9 and 12 months of age. RESULTS:Genetic ablation of P2Y2 Rs in IL-14?TG mice significantly reduced B and T lymphocyte infiltration of SMGs. However, reduced sialadenitis did not restore saliva secretion in IL-14?TG × P2Y2 R-/- mice. Decreased sialadenitis in IL-14?TG × P2Y2 R-/- mice correlated with decreased lymphotoxin-? levels, a critical proinflammatory cytokine associated with autoimmune pathology in IL-14?TG mice. CONCLUSIONS:The results of this study suggest that P2Y2 Rs contribute to the development of salivary gland inflammation in IL-14?TG mice and may also contribute to autoimmune sialadenitis in humans.
Project description:OBJECTIVE:Interleukin (IL)-17A and IL-18 have been proposed to play important roles in periodontitis and type 2 diabetes mellitus (DM), but human data are conflicting. The present study aimed to investigate the roles of IL-17A and IL-18 in periodontitis and DM by measuring salivary and serum levels, respectively. MATERIALS AND METHODS:A total of 49 participants with type 2 DM and 25 control subjects without type 2 DM were recruited. A periodontal screening and recording (PSR) index (0, 1-2, 3, and 4) was used to classify whether these subjects had periodontitis. Salivary and serum IL-17A and IL-18 levels were measured by enzyme-linked immunosorbent assay. Multiple linear regression analyses were used to evaluate the associations between these cytokines and clinical parameters. RESULTS:Salivary IL-17A levels were not significantly different between patients with DM and controls, however, the levels were significantly higher in controls with periodontitis than those without periodontitis (p = 0.031). Salivary IL-17A levels were significantly associated with the PSR index (? = 0.369, p = 0.011). Multiple linear regression analyses revealed the association of salivary IL-18 levels and fasting plasma glucose (? = 0.270, p = 0.022) whereas serum IL-18 levels were associated with HbA1C (? = 0.293, p = 0.017). No correlation between salivary and serum levels of IL-17A and IL-18 was found. CONCLUSION:Salivary IL-17A was strongly associated with periodontitis, whereas salivary IL-18 was associated with FPG and serum IL-18 was associated with HbA1C. These results suggest the role of these cytokines in periodontal inflammation and DM.
Project description:OBJECTIVE: Interleukin-17 (IL-17) is a cytokine that induces neutrophil recruitment and the release of inflammatory mediators in several inflammatory conditions; nevertheless, the involvement of IL-17 gene polymorphisms in chronic periodontitis (CP) has not been addressed yet. Our aim was to evaluate the association between periodontal status and the polymorphisms IL-17A G197A and IL-17F C7488T in subjects with CP along with their impact on levels of inflammatory mediators. MATERIAL AND METHODS: Genomic DNA was obtained from 30 CP patients and 30 healthy controls (HCs). IL-17A G197A and IL-17F C7488T polymorphisms were determined using PCR-RFLP. Serum and periodontal tissues were collected and processed for ELISA, myeloperoxidase (MPO), and/or microscopic analysis. RESULTS: The frequencies of genotypes in the CP group were significantly different from those of HC. Odds ratio indicated that increased risks for CP were associated with the -197A allele, not with the -7488T allele. In addition, the -197A allele was correlated with worse clinical parameters, higher MPO activity, and increased expression of inflammatory mediators (IL-17A and IL-8) than the other genotypes. CONCLUSIONS: These results indicate that the IL-17A -197A allele is associated with increased risk for CP, likely because this genotype relates to the enhanced inflammation in periodontal tissues.
Project description:Periodontal disease (PD) is an inflammatory condition of the tissues supporting the teeth, which is widespread among the adult population. Evidence shows a relationship between PD and vitamin D levels, which is involved in the regulation of bone metabolism, mineral homeostasis, and inflammatory response. This study aimed to perform a simultaneous evaluation of inflammatory mediators and vitamin D levels in saliva in periodontopathic patients to better understand their role in periodontal disease. In this observational study, clinical periodontal parameter examination was performed for each patient. Moreover, the saliva levels of 25(OH)D3, TGF?, IL-35, IL-17A, and MMP9 were evaluated using an ELISA assay. An increase in TGF?, IL-35, MMP9, and IL-17A salivary levels and a reduction in 25(OH)D3 levels were observed in periodontopathic patients with respect to the healthy controls. The present study revealed significant positive correlation between cytokines and highly negative correlation between 25(OH)D3 and salivary cytokine levels. Further studies are needed to better understand if salivary cytokines and vitamin D evaluation may represent a new approach for detection and prevention of progressive diseases, such as PD.