Understanding the role of salivary proteomic markers in detecting chronic periodontitis in adults with obesity—A systematic review

Adult periodontitis is a chronic destructive disease characterized by an interaction between gram-negative bacteria and the host inflammatory response. Microbial substances such as lipopolysaccharide can activate host cells, e.g., macrophages, fibroblasts and keratinocytes, to secrete proinflammatory cytokines including tumor necrosis factor alpha and interleukin 1 beta (IL-1 beta). This study examined the hypothesis that periodontitis tissue contains increased levels of cytokines that promote osseous and connective tissue destruction. To test this hypothesis, diseased and healthy gingival biopsies were examined for differences in the expression of cytokine mRNA for the pro-inflammatory cytokines tumor necrosis factor alpha and IL-1 beta and the anti-inflammatory cytokine IL-1ra using quantitative reverse transcriptase polymerase chain reaction and in situ hybridization methods. The levels of tumor necrosis factor alpha and IL-1ra mRNA were shown to be significantly higher in diseased than healthy tissues. Additionally, a significantly correlated expression of IL-1 beta and IL-1ra mRNA was seen in all tissue examined. Analysis of tissue sections by immunohistochemical and in situ hybridization techniques revealed a mononuclear cell infiltrate that consisted of a higher average number of cells staining positive for tumor necrosis factor alpha mRNA, CD14, and CD3 in the diseased than healthy tissues. Although both diseased and healthy tissues expressed IL-1 beta and IL-1ra mRNA in the epithelium, the diseased tissue biopsies expressed more IL-1 beta and IL-1ra mRNA in the connective tissue. These results implicate the potential involvement of both the pro- and anti-inflammatory cytokines in the regulation of the chronic inflammatory disease adult periodontitis.

To investigate whether sub-antimicrobial dose doxycycline (SDD) therapy for 120 d in chronic adult periodontitis patients had significant effects on gingival crevicular fluid (GCF) matrix metalloproteinase-8 (MMP-8) levels, and on gingival tissue MMP-9, tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) and interleukin-6 (IL-6) levels. Tetracycline can significantly inhibit MMP activity in GCF and in gingival tissue, even in much lower dosage then a traditional antimicrobial dosage used in conventional therapy. Sub-antimicrobial dose doxycycline (SDD) therapy has been shown to reduce periodontal disease activity to control MMP and pro-inflammatory cytokines. A total of 32 patients with incipient to moderate (probing pocket depth approximately 4-7 mm) chronic adult periodontitis were included in the study. Subjects were randomly assigned to two groups. After scaling and root planning (SRP), the SRP + SDD group received SDD, 20 mg bid, whereas the SRP + placebo group received placebo, 20 mg bid. In the follow-up, efficacy measures included the change in probing pocket depth (PD), clinical attachment level (CAL), bleeding on probing (BOP) and gingival crevicular fluid MMP-8 levels, gingival tissue MMP-9, TIMP-1 and IL-6 levels from baseline to 120 d. After 120 d, PD and CAL improved significantly in the SRP + SDD group. Initial MMP-8 levels for the SRP + SDD group and the SRP + placebo group were 407.13 +/- 114.45 ng/ml and 378.71 +/- 189.39 ng/ml, respectively, with no statistical difference between the two groups. MMP-8 levels for the SRP + SDD group and the SRP + placebo group were: 235.35 +/- 134.58 ng/ml and 364.04 +/- 219.27 ng/ml at 30 d; 157.50 +/- 95.95 ng/ml and 236.60 +/- 186.16 ng/ml at 60 d; 102.70 +/- 67.64 ng/ml and 208.56 +/- 124.54 ng/ml at 90 d; and 63.77 +/- 53.33 ng/ml and 229.13 +/- 168.09 ng/ml at 120 d, respectively. The amount of decrease in MMP-8 levels for the SRP + SDD group was statistically significant compared to that for the SRP + placebo group, especially apparent at 120 d (p < 0.05). TIMP-1 levels in both groups increased from the baseline to 120 d with statistical significance (p-value < 0.05), but there was no significant difference between the two groups. Changes in MMP-9 and IL-6 levels were not statistically significant. Adjunctive SDD therapy can improve the clinical parameters and this clinical improvement is reflected by controlled level of MMP-8 in chronic adult periodontitis after the therapy.

American Academy of Periodontology Task Force Report on the Update to the 1999 Classification of Periodontal Diseases and Conditions.

Periodontal diseases, including chronic gingivitis, chronic periodontitis, and aggressive periodontitis, are driven by complex immune responses, with mast cells (MCs) playing a pivotal role through the release of mediators during degranulation. Although MC involvement in chronic periodontal conditions has been documented, its role in aggressive periodontitis and the relationship between MC degranulation and inflammation intensity remain poorly understood. This study aimed to quantify MCs and assess their degranulation in gingival tissues across healthy gingiva, chronic gingivitis, chronic periodontitis, and aggressive periodontitis using histochemical staining to elucidate their role in periodontal inflammation and disease progression. This cross-sectional study included 120 systemically healthy subjects recruited from the Periodontology Outpatient Department. The groups included healthy periodontium (n = 30), chronic gingivitis (n = 30), chronic periodontitis (n = 30), and aggressive periodontitis (n = 30) groups. Gingival biopsies were obtained, fixed, sectioned, and stained with toluidine blue to identify intact and degranulated MCs in juxta-epithelial (Z1) and deep connective tissue (Z2) zones. MC counts were performed under 40x magnification, with examiner calibration to ensure reliability (intraclass correlation(ICC) ≥ 0.85). Nonparametric tests (Kruskal-Wallis with Dunn's test) were used to analyze MC density and degranulation, with a significance level of p < 0.05. MC counts were highest in chronic periodontitis (7.10 ± 2.50 in Z1, 6.70 ± 2.10 in Z2) and aggressive periodontitis (6.10 ± 1.70 in Z1, 6.50 ± 2.00 in Z2), followed by chronic gingivitis and healthy gingiva (p < 0.00001). Degranulated MCs were significantly elevated in diseased groups, particularly chronic periodontitis (4.69 ± 2.09 in Z1, 4.51 ± 1.89 in Z2) compared to healthy controls (p = 0.0001). Intact MCs showed less variation, with significant differences between the healthy and aggressive periodontitis groups (p = 0.045). Increased MC degranulation was associated with periodontal disease severity, particularly in chronic and aggressive periodontitis, suggesting a key role of MC degranulation in inflammatory progression. These findings highlight the potential of MCs as therapeutic targets in periodontal management.

Bacteria involved in chronic periodontal disease induce inflammatory cytokines. These cytokines can enter the brain through systemic circulation and cause depression. In this study, we investigated the association between depressed mood and chronic periodontitis in older adults in Korea. This study used data from the Life-changing Period Health Checkup (2007-2008), performed only at 66 years of age, which is available from the National Health Insurance Service-Senior Cohort Database. The depressed mood status was evaluated by three questions in the mental health examination. The definition of chronic periodontitis was determined based on the diagnosis and treatment codes for chronic periodontitis. Based on the three questions, we investigated the association between depressive mood and chronic periodontal disease by dividing the study population into 9622 depressed mood subjects (DMS) and 9091 non-depressed mood subjects (NDMS). Multivariable-adjusted logistic regression analysis was performed, and odds ratios (ORs) with 95% confidence intervals (95% CIs) were presented. Among the NDMS, 5390 patients with chronic periodontitis were identified, and among the DMS, 5964 patients with chronic periodontitis were identified. The crude OR (95% CI) for chronic periodontitis in DMS compared to NDMS was 1.12 (1.06-1.19). The adjusted OR (95% CI) for chronic periodontitis was 1.12 (1.06-1.19). As a result of confirming the effect of sex and comorbidity on the association between depressed mood and chronic periodontitis, a significant association was observed for women at 1.15 (1.07-1.25). In addition, the adjusted OR (95% CI) for depressive mood and chronic periodontitis was 1.15 (1.07-1.25) for patients with hypertension, 1.13 (1.03-1.25) for patients with diabetes, 1.12 (1.02-1.22) for patients with dyslipidemia, and 1.18 (1.04-1.34) for patients with heart disease. This study confirmed the relationship between depressed mood and chronic periodontitis in older adults. Therefore, education to strengthen the emotional management of older adults, especially with respect to depression, would play an auxiliary role in preventing and treating periodontitis.

Background and ObjectiveGenetic markers associated with disease are often non-functional and generally tag one or more functional “causative” variants in linkage disequilibrium. Markers may not show tight linkage to the causative variants across multiple ethnicities due to evolutionary divergence, and therefore may not be informative across different population groups. Validated markers of disease suggest causative variants exist in the gene and, if the causative variants can be identified, it is reasonable to hypothesize that such variants will be informative across diverse populations. The aim of this study was to test that hypothesis using functional Interleukin-1 (IL-1) gene variations across multiple ethnic populations to replace the non-functional markers originally associated with chronic adult periodontitis in Caucasians.Material and MethodsAdult chronic periodontitis cases and controls from four ethnic groups (Caucasians, African Americans, Hispanics and Asians) were recruited in the USA, Chile and China. Genotypes of IL1B gene single nucleotide polymorphisms (SNPs), including three functional SNPs (rs16944, rs1143623, rs4848306) in the promoter and one intronic SNP (rs1143633), were determined using a single base extension method or TaqMan 5′ nuclease assay. Logistic regression and other statistical analyses were used to examine the association between moderate to severe periodontitis and IL1B gene variations, including SNPs, haplotypes and composite genotypes. Genotype patterns associated with disease in the discovery study were then evaluated in independent validation studies.ResultsSignificant associations were identified in the discovery study, consisting of Caucasians and African Americans, between moderate to severe adult chronic periodontitis and functional variations in the IL1B gene, including a pattern of four IL1B SNPs (OR = 1.87, p < 0.0001). The association between the disease and this IL1B composite genotype pattern was validated in two additional studies consisting of Hispanics (OR = 1.95, p = 0.04) or Asians (OR = 3.27, p = 0.01). A meta-analysis of the three populations supported the association between the IL-1 genotype pattern and moderate to severe periodontitis (OR 1.95; p < 0.001). Our analysis also demonstrated that IL1B gene variations had added value to conventional risk factors in predicting chronic periodontitis.ConclusionThis study validated the influence of IL-1 genetic factors on the severity of chronic periodontitis in four different ethnicities.

Chronic inflammation induced by bacteria often leads to host-mediated destruction of tissues adjacent to the sites of microbial insult. The chronic inflammatory process of adult periodontitis results in the destruction of supporting osseous and connective tissues of the teeth. We hypothesized that virulence factors of periodontal pathogens such as lipopolysaccharide stimulate inflammatory cytokine expression by mononuclear cells of the host which contribute to disease development. In this study, to elucidate the role of these cytokines in chronic adult periodontitis, we tested whether the prevalence of mRNA for inflammatory cytokines generally associated with mononuclear phagocytes was higher in diseased than in healthy gingival tissue. Gingival mononuclear cells or whole gingival biopsies from 32 adult periodontitis patients and five healthy individuals used as controls were evaluated for inflammatory cytokine mRNA expression by reverse-transcription polymerase chain-reaction (RT-PCR) procedures. The cytokines assessed included IL-1 alpha, IL-1 beta, IL-1ra, IL-6, IL-8, IL-12, IL-13, TNF-alpha, TGF-beta, and IFN-gamma. The monocyte/macrophage lipopolysaccharide (LPS) receptor CD14 was also assessed. Results showed that TNF-alpha mRNA was present significantly more frequently in diseased than in healthy biopsies, whereas IL-1 alpha, IL-1 beta, and IL-1ra mRNA were found in most (from 80 to 100%) healthy tissues. Message for CD14 was present in both healthy and diseased tissue samples (100%). This study provides evidence for a major role of TNF-alpha in chronic adult periodontitis. Moreover, our results suggest that the mononuclear cells derived from periodontal tissues have the capacity to respond to components of periodontal pathogens and express both pro- and anti-inflammatory cytokines in these tissues.

A number of studies have suggested a bidirectional relationship of periodontitis with rheumatoid arthritis (RA) and type 2 diabetes mellitus (T2DM). However, the genetic factors that underlie these relationships have not been elucidated. We conducted a multicenter case-control study that included 185 patients with RA and chronic periodontitis (CP), 149 patients with T2DM and CP, 251 patients with CP, and 130 systemically and periodontally healthy controls from a cohort of Japanese adults to assess the shared genetic risk factors for RA and CP as well as for T2DM and CP. A total of 17 candidate single nucleotide polymorphisms (SNPs) associated with RA, T2DM, and CP were genotyped. Multiple logistic regression analyses revealed that the KCNQ1 rs2237892 was significantly associated with comorbidity of RA and CP (P=0.005) after adjustment for age, sex, and smoking status. The carriers of the T allele among patients with RA and CP showed significantly higher disease activity scores including 28 joints using C-reactive protein values than the non-carriers (P=0.02), although the age, female percentage, and smoking status were comparable. Other SNPs were not associated with comorbidity of RA and CP, T2DM and CP, or susceptibility to CP. The results of the present pilot study suggest for the first time that the KCNQ1 rs2237892 may constitute a shared genetic risk factor for RA and CP, but not for T2DM and CP in Japanese adults.

Background: Tagetes lucida Cav (T. lucida) is an herbaceous plant from the family Asteraceae, native to Mexico, Guatemala, and other Mesoamerican countries. Their common names are: (i) Spanish names: “hierbanís”, “jericón”, “pericón vomol”, “Santa María”, “cuchrucumín”, “falso hipericón”, “hierba añil”, “pericón”, “periquillo”; (ii) indigenous names: “ichka”, “cuahuyauhtli”, “yahuhtli”, “naná uarhi”, and (iii) English names: “sweet-scented marigold”, “sweet mace”, and “sweet marigold”., Its chemical composition of coumarins, estragole, quercetin, and flavonoids has antioxidant, anti-inflammatory, and antimicrobial effects. This study aimed to evaluate the effect of a mouthwash made with T. lucida for control of chronic periodontitis (CP) in older adults. Methods: We carried out a quasi-experimental study with a convenience sample of 60 older adults with CP. The sample was divided into a placebo group (PG) n = 30, administered with a mouthwash made with alcohol diluted with purified water (10%), and the experimental group (EG) n = 30 treated with a mouthwash made with T. lucida extract in 10% alcohol; both groups were administered three times a day for three months. Results: We found a significant improvement in the EG group compared with the PG group in: probing pocket depth (baseline, 5.2 ± 0.81 vs. post-treatment, 3.1 ± 0.43, p &lt; 0.05), the clinical attachment loss (baseline, 6.6 ± 1.9 vs. post-treatment, 2.1 ± 1.5, p &lt; 0.05), lipoperoxides (baseline, 0.056 ± 0.03 vs. post-treatment, 0.034 ± 0.02 µmol/L, p &lt; 0.05), and chronic inflammation markers (IL-1, IL-8, and TNF-α) (p &lt; 0.05). Conclusions: Our findings suggest that mouthwash made with T. lucida extract, administered three times a day, clinically improved chronic periodontitis in older adults, associated to a significant decrease in lipoperoxides and proinflammatory markers.

Gingival fibroblasts from patients with chronic adult periodontitis are known to produce cytokines in response to changing levels of bacterial lipopolysaccharide (LPS). Cytokine production is one of numerous cell processes that involve calcium dependent enzymes. It is possible that inflammation may induce changes in the amount of the Ca++-pump protein in gingival fibroblasts which could alter Ca++-dependent activities in these cells including the production and release of cytokines. The purpose of this study was to determine if differences exist in the amount of Ca++-pump protein in the gingival fibroblasts of periodontitis patients relative to control individuals without periodontal disease. Fibroblast explants from healthy tissue and from inflamed tissue from patients with chronic adult periodontitis, grown in culture, were analyzed for quantitative differences in the amount of Ca++-pump protein. Fibroblasts from chronic adult periodontitis patients exhibited significantly lower levels of Ca++-pump protein than fibroblasts from healthy subjects (p = 0.0015). However, fibroblasts from chronic adult periodontitis patients, when activated with LPS, did not exhibit significant differences in the amounts of Ca++-pump protein as compared to untreated controls (p = 0.2177). Similarly, cells from healthy subjects did not show significant reduction in Ca++-pump protein following activation with LPS (p = 0.1732). Our results suggest that plasma membrane Ca++-pump is significantly reduced in fibroblasts derived from patients with chronic periodontitis. However, factors other than LPS may be involved in the down-regulation of Ca+ -pump protein.

It is well known that interactions between microbial dental plaque and the host immune system play a major role in the etiopathogenesis of periodontal disease. The aim of the present study was to analyze the phenotypic properties of gingival T lymphocytes and subsets in patients with chronic inflammatory adult periodontitis (AP) showing various degrees of inflammation and to relate the results to the immunopathogenesis of AP. Gingival biopsies were obtained from patients aged between 26 and 52 yr who were grouped according to gingival index scores (GI) of 1, 2, and 3. Using immunohistochemical techniques, T cells (CD2+), T-helper cells (CD4+) and T-suppressor cells (CD8+) were identified in three well-defined areas of the biopsy samples. Moreover, peripheral blood was collected from the same patients, and relative counts of B cells (CD19+), HLA-DR+ cells and IL-2R+ cells as well as CD3+, CD4+, CD8+ cells were determined using three color flow cytometry. While the blood results were found to be within the normal ranges, the relative counts of CD4+ cells showed statistically significant decreases as the GI score increased. Similarly, the CD4+/CD8+ ratio also decreased. Moreover, gingival T lymphocyte and subset counts appeared to be related to the severity of gingival inflammation. Particularly, CD4+ cells showed a significant increase with the GI score. Furthermore, the CD4+/CD8+ ratio beneath the pocket epithelium was apparently correlated with increasing GI score (p < 0.05). The cytotoxic effect of CD8+ cells seems to be more prominent at the local level while the suppressor effect is more active systematically. This means that the price of systemic protection appears to be local destruction.

Blood vessel features in periodontal pocket soft tissues may be significant in the pathogenesis and progression of chronic periodontitis. The aim of this study was to make a quantitative histological assessment of the vasculature in soft tissue biopsies from patients with chronic adult periodontitis and patients with healthy periodontal tissues. We have also investigated changes in tissue morphology at both the histological and ultrastructural level. Twelve interdental biopsies were obtained, 6 from chronic adult periodontitis patients and 6 from healthy volunteers. The specimens were sliced, fixed in 3% glutaraldehyde, postfixed in 1% buffered osmium tetroxide, dehydrated, and embeded in araldite. One micron semithin sections were differentially stained with a dichromatic technique. The number of blood vessels (BV) for sub-epithelial, superficial and deep connective tissue layers were then assessed. Only in the sub-epithelial connective tissue layer was there a significant increase in the number of blood vessels (95% Confidence interval [CI]) in the chronic adult periodontitis specimens when compared to healthy specimens. The results of this study seem to indicate that a dichromatic staining technique facilitates the identification and quantification of blood vessels in epoxy resin embedded specimens at light microscope level, and that there is an increase in the number of blood vessels in the chronic adult periodontitis lesions.

Introduction: Triggering receptor expressed on myeloid cells-1 (TREM-1) was reported to be up-regulated in various inflammatory diseases as well as in periodontal disease. The important role of E-cadherin against bacterial invasion has already been established in the junctional epithelium. This study aimed to evaluate the potential value of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) and E-cadherin as salivary diagnostic markers in patients with progressive chronic periodontitis. Material and method: A total of 79 subjects 41 females and 38 males, divided into 4 groups: healthy group (HG), early generalized chronic periodontitis group (EGP), moderate generalized chronic periodontitis group (MGP) and advanced generalized chronic periodontitis group (AGP), were included in the study. Enzymelinked immunosorbent assay (ELISA) test was used for quantification of sTREM-1 and E-cadherin in the saliva samples. Clinical and periodontal parameters were recorded and statistical analysis, using SPSS version 25.0, was performed. Results: Elevated salivary sTREM-1 levels were evident in EGP, MGP and AGP compared to the HG. Statistically significant differences in sTREM-1 concentrations were found between HG and AGP. Salivary E-cadherin was found to be upregulated during the progression of periodontal disease and statistically significant differences were found between HG and MPG. Conclusion: Salivary sTREM-1 may be considered a new biomarker in chronic periodontitis progression. E-cadherin, although detected, does not seem to have diagnostic value at salivary level in chronic adult periodontitis.

Periodontology 2000Volume 53, Issue 1 p. 28-44 Epidemiologic patterns of chronic and aggressive periodontitis Ryan T. Demmer, Ryan T. DemmerSearch for more papers by this authorPanos N. Papapanou, Panos N. PapapanouSearch for more papers by this author Ryan T. Demmer, Ryan T. DemmerSearch for more papers by this authorPanos N. Papapanou, Panos N. PapapanouSearch for more papers by this author First published: 12 April 2010 https://doi.org/10.1111/j.1600-0757.2009.00326.xCitations: 177Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume53, Issue1June 2010Pages 28-44 RelatedInformation

Cytokines, such as interleukin-10, and related genetic polymorphisms, have been implicated in the pathogenesis of chronic periodontitis. The aim of this study was to investigate a possible correlation between chronic periodontitis and genetic polymorphisms coding for two interleukin-10 related chemokines [interleukin-24 and regulated on activation, normal T cells expressed and secreted (RANTES)] as well as a RANTES receptor [CC chemokine receptor 5 (CCR5)]. A single-blind, two-centre, case-controlled study was carried out with test patients from the Clinic of Periodontics, Göteborg University, and from the Department of Periodontology, Glasgow University, and control subjects from the undergraduate clinics of both schools. Blood samples were collected from 106 patients (56 women and 50 men, mean age 51.7 yr) with generalized, severe chronic periodontitis and from 69 periodontally healthy subjects (37 women and 32 men, mean age 53.3 yr). The polymerase chain reaction (PCR) was used to identify the genetic coding for interleukin-24, RANTES and CCR5. Genotype and allele frequencies were compared between the test and control groups using Fischer's exact test at the 5% level of significance. There were no statistically significant differences between patients with chronic periodontitis and control subjects, regarding genotype distribution or allele frequency, irrespective of smoking status, in the combined Glasgow and Gothenburg cohort or in the specific location cohorts. The allele frequencies for healthy and control subjects for RANTES gave a p-value of 0.80 (allele G was 58.8% in healthy subjects and and 54.4% in subjects with periodontitis), for interleukin-24 the p-value was 0.90 (allele T was 56.2% in healthy subjects and and 54.9% in subjects with periodontitis) and for CCR5 the p-value was 0.90 (the wild-type allele was 85% in healthy subjects and and 82.7% in subjects with periodontitis). The interleukin-24, RANTES and CCR5 polymorphisms investigated are not associated with chronic periodontitis.