Tissues Of Chronic Periodontitis Patients Research Articles

Role of ubiquitin-specific protease 5 in the inflammatory response of chronic periodontitis.

The systemic inflammatory response caused by chronic periodontitis is a risk factor for multiple diseases. Ubiquitin-specific protease 5 (USP5) is a kind of deubiquitinase which mainly responsible for dissociating unanchored polyubiquitin. However, the functions of USP5 in chronic periodontitis have not been reported. Chronic periodontitis patients were recruited, and their periodontal samples were collected. The levels of USP5, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in gingival crevicular fluid were evaluated by ELISA. The expression of USP5, TNF-α, IL-6, and IL-1β in human periodontal ligament stem cells (PDLSCs) was estimated by qRT-PCR assay. The activation of STAT3 signaling was examined by Western blot. USP5 was upregulated in the gingival crevicular fluid and gingival tissues of chronic periodontitis patients. USP5 expression was positively correlated with the expression of proinflammatory factors. USP5 knockdown and deubiquitinase inhibitor inhibited LPS-induced inflammatory responses in PDLSCs. Suppressing USP5 inhibited STAT3 signaling in PDLSCs. Suppression deubiquitinase USP5 inhibits the inflammatory response of chronic periodontitis by suppressing STAT3 signaling.

Expression of NLRP3 and P2X7 transcripts in gingival tissues of chronic periodontitis patients and its correlation with P. gingivalis load and periodontal parameters

BackgroundInflammasomes are pivotal regulators of the host innate defense in periodontal tissues. Porphyromonas gingivalis modulates NLRP3 inflammasome complex and P2X7 receptor for survival in tissues through its virulence factors. AimTo estimate the expression of NLRP3 and P2X7 in gingival tissues of chronic periodontitis patients and to assess its correlation with periodontal clinical parameters and P. gingivalis load in subgingival plaque samples. Materials and methodsInterproximal gingival tissue and subgingival plaque samples (collected from same site) from systemically healthy chronic periodontitis patients (n = 22) and healthy controls (n = 22) were collected and stored in −80 °C. NLRP3 and P2X7 gene expression in gingival tissues and load of P. gingivalis in sub gingival plaque were assessed using real time Polymerase Chain Reaction. ResultsThe expression of NLRP3 in chronic periodontitis was marginally higher than healthy controls though the difference was statistically insignificant. P2X7 expression was higher in healthy controls and down regulated in chronic periodontitis patients (p = 0.04). A statistically significant correlation was observed between the probing pocket depth and expression of P2X7 gene in chronic periodontitis (p = 0.04). Correlation between P. gingivalis load and expression of the genes in tissues was either moderate (P2X7 control group) or negligible (both test and control of NLRP3). ConclusionA marginal increase in expression of NLRP3 and statistically significant decrease in P2X7 expression in chronic periodontitis patients was present. A statistically significant correlation was observed between P2X7 and P. gingivalis load in control group and periodontal parameters in test group. The study confirms involvement of genes in periodontal pathogenesis.

Leptin regulates OPG and RANKL expression in Gingival Fibroblasts and Tissues of Chronic Periodontitis Patients.

Objective: Chronic periodontitis is a bone-destructive disease affecting periodontal support structures. Although leptin has a protective effect against periodontitis, the underlying mechanism remains unclear. Therefore, this study aimed to investigate the possible role of leptin by examining its relationship with OPG and RANKL in human gingival tissues obtained from patients with chronic periodontitis.Method: Twenty-two patients with chronic periodontitis were enrolled (10 with moderate periodontitis and 12 with severe periodontitis) in the experimental group, and 12 healthy individuals were enrolled in the control group. Gingival tissue samples were collected, and the protein levels and localization of leptin, OPG, and RANKL were studied using immunohistochemistry (IHC). The staining intensities of leptin, OPG, and RANKL were correlated with the periodontal clinical index. Moreover, real-time quantitative PCR (RT-qPCR) was used to determine OPG and RANKL mRNA levels in gingival fibroblasts stimulated with gradient concentrations of leptin protein in vitro.Result: Leptin, OPG, and RANKL were located in the cytoplasm of gingival epithelial cells and the connective tissue. Leptin was widely and significantly expressed in the control group, whereas it was lightly stained in the severe group. RANKL was lightly stained in the control group, whereas it was widely and significantly expressed in the severe group. The control and the moderate groups had similar OPG levels, which were significantly higher than that in the severe group. Leptin was positively correlated with OPG(r = 0.905, p < 0.01) and negatively correlated with RANKL (r = -0.635, p < 0.01). In vitro low concentrations of leptin led to an increased OPG/RANKL mRNA ratio, whereas the opposite effect was observed at high concentrations.Conclusion: Leptin can regulate OPG and RANKL expression in gingival fibroblasts and may thus play a role in the development of chronic periodontitis by modulating the OPG/RANKL ratio.

Clinical and Microbiological Effects of Weekly Supragingival Irrigation with Aerosolized 0.5% Hydrogen Peroxide and Formation of Cavitation Bubbles in Gingival Tissues after This Irrigation: A Six-Month Randomized Clinical Trial.

Introduction The study investigated the effect of weekly supragingival irrigation with aerosolized 0.5% hydrogen peroxide (H2O2) solution as a maintenance periodontal therapy on clinical and microbiological parameters in patients with chronic periodontitis. The other purpose was to investigate whether cavitation bubbles can penetrate not only into periodontitis-damaged tissues but also into ex vivo porcine healthy periodontal tissues. Materials and Methods The study included 35 systemically healthy patients with chronic periodontitis (CP). After nonsurgical periodontal debridement (NSPD), all patients were randomized into two groups: the Control group (NSDP alone, n = 18) and the Test group (NSDP plus supragingival irrigation, n = 17). Clinical (Approximal Plaque Index (API), Bleeding Index (BI), and Modified Gingival Index (MGI)) and microbiological (Polymerase Chain Reaction technology (using a micro-IDent® kit)) measurements were performed at the initial time point, 3 months, and 6 months after NSPD. The impact of supragingival irrigation on diseased gingival tissues of CP patients (n = 5) and on ex vivo porcine healthy gingival tissue samples (n = 3) was evaluated to estimate morphological changes in healthy and diseased gingival tissues. Results Morphological data revealed that supragingival irrigation caused the formation of cavitation bubbles in diseased gingival tissue of CP patients and in healthy porcine gingival tissues. The decrease in API, BI, and MGI scores after 6 months in the Test group significantly (p ≤ 0.01, p ≤ 0.05, and p ≤ 0.01, respectively) exceeded that in the Control group. Test group patients demonstrated a decrease in periodontal sites showing Pocket Probing Depth > 4 mm and, after 6 months, a statistically significant decrease in the proportion of periopathogenic bacteria. Conclusion The effectiveness of mechanical periodontal treatment combined with weekly supragingival irrigation with aerosolized 0.5% H2O2 solution on clinical and microbiological parameters of periodontal tissues of periodontitis patients is reliably higher than that of mechanical periodontal debridement alone. It has been found that cavitation bubbles as a result of irrigation with the aerosolized 0.5% hydrogen peroxide solution can form not only in periodontal tissues of periodontitis patients but also in ex vivo porcine healthy gingival tissues.

Comparative estimation of sulfiredoxin levels between chronic periodontitis and healthy patients - A case-control study.

Growing evidence suggests that oxidative stress forms a key component in the etiopathogenesis of periodontitis. Studies have shown potential antioxidants responsible for combating the pro-oxidants which stress the periodontium. But, peroxiredoxin-sulfiredoxin system is the least explored in periodontal disease. A case-control study was conducted on 30 participants who fulfilled the inclusion criteria from the Department of Periodontics, Saveetha Dental College and Hospital, Chennai, India. The patients were divided into two groups: 1) Group A- healthy controls (n=18), 2) Group B- patients with generalized chronic periodontitis (n=17). Following clinical examination, gingival tissue samples were procured from both the groups and subjected to protein quantification by Lowry method. The samples with adequate protein concentration (n=30) from the two groups were further analyzed by enzyme-linked immunosorbent assay for estimation of sulfiredoxin levels. Sulfiredoxin levels were significantly higher in the gingival tissues of chronic periodontitis patients (171.20±16.97ng/mL) than in healthy controls (131.20±22.87) with P<0.001. Also, the levels of sulfiredoxin in gingival tissue of periodontitis patients positively correlated with site-specific probing depth (r=0.67; P=0.007) and clinical attachment level (r=0.55; P=0.035). The present study was a novel attempt to estimate the levels of sulfiredoxin which was significantly elevated in the diseased sites of patients with chronic periodontitis. Future studies are required to probe the role of sulfiredoxin in the etiopathogenesis of periodontal disease.

Leukocyte receptor expression in chronic periodontitis.

Microbial recognition in the periodontium through specific leukocyte receptors gives rise to the response which in susceptible individuals can lead to periodontal diseases. The aim of this study was to explore the expression of leukocyte receptors in the gingival tissues of chronic periodontitis patients and to analyse differences between diseased and control sites (sites with probing pocket depth <4mm). Thirty-seven chronic periodontitis patients were included in the study. Gingival biopsies were harvested from diseased and control sites and processed by flow cytometry for the determination of the expression of 16 leukocyte receptors (CD4, CD8, CD11b, CD14, CD16, CD19, CD25, CD28, CD49d, CD49e, CD62, CD71, CD80, CCR7, Ly6G and HLA-DR). Expression of all studied receptors was higher in test compared with control sites (p<0.005). Sampled sites with less bleeding on probing exhibited higher expression of CD16 and CD14 receptors (p=0.020 and 0.011, respectively). This study points towards considerable differences in the expression of leukocyte receptors between diseased and control sites in the same periodontal patients.

Gingival, plasma and salivary levels of melatonin in periodontally healthy individuals and chronic periodontitis patients: a pilot study.

Periodontal disease is an inflammatory condition affecting tooth supporting structures in which dysregulated immune response and oxidative stress mediate tissue destruction. Melatonin, the pineal gland hormone is a regulator of circadian rhythm, an antioxidant and an immunomodulator. Previous studies have shown lowered melatonin levels in saliva, plasma and gingival crevicular fluid (GCF) of patients with periodontal disease. Till date no study has assessed the melatonin levels in gingival tissues. Five healthy individuals and 15 chronic periodontitis patients were recruited for this pilot study. 5ml of whole saliva, 2 ml peripheral blood and gingival tissue samples were obtained from each individual at 8.00 am in fasting state. Melatonin assay was performed with a commercially available ELISA kit. Statistical analysis was done to assess the difference in mean melatonin levels among the groups. No statistically significant difference was found in mean melatonin levels between healthy individuals and chronic periodontitis patients in saliva (p=.266) and plasma (p=.933) samples, whereas in gingival tissue samples (p=.015), the melatonin levels were significantly lowered in chronic periodontitis patients compared to healthy individuals. This study demonstrates the presence of melatonin in gingival tissue. Furthermore, melatonin levels are lowered in gingival tissues of chronic periodontitis patients.

Protease-activated receptors expression in gingiva in periodontal health and disease

ObjectiveProtease-activated receptors (PARs) are a unique class of receptors which are implicated in mediating inflammation, pain and other functions. The aim of this study was to elucidate the role of PARs in the pathogenesis of chronic periodontitis by differential expression analysis of PARs in the gingival tissues of chronic periodontitis patients compared with those of healthy control individuals. DesignGingival tissue specimens were collected from chronic periodontitis patients (n=20) and control individuals (n=20). The expression of PAR-1, -2, -3 and -4 was determined in these tissues by immunohistochemistry and differential expression between the two groups was investigated by quantitative real-time reverse transcription-polymerase chain reaction analysis. ResultsPAR-1, -2, -3 and -4 were expressed in all gingival tissues. A significant overexpression of PAR-3 was detected in chronic periodontitis-affected tissues compared to healthy gingival tissues. However, expression of PAR-2 was decreased in periodontal lesions. ConclusionsOur study shows that PAR-1, -2, -3 and -4 are expressed in both healthy and inflamed gingival tissues. Furthermore, PAR-2 and PAR-3 may contribute to the inflammatory responses associated with chronic periodontitis.

Effect of Non-Surgical Periodontal Therapy on Superoxide Dismutase Levels in Gingival Tissues of Chronic Periodontitis Patients: A Clinical and Spectophotometric Analysis

BACKGROUND: Superoxide dismutase (SOD), an antioxidant acting against superoxide (oxygen radical, O2.-), it is released in inflammatory pathways and causes connective tissue breakdown. Increased SOD activity in inflamed gingiva may indicate increased O2.-radical generation by neutrophils and other inflammatory cells at the diseased site. The aim of the study was to evaluate the effects of non-surgical periodontal therapy (NSPT) on SOD levels in gingival tissues of chronic periodontitis patients.METHODS: Forty subjects: 20 periodontally healthy (Control) and 20 chronic periodontitis (Test); age range 24–55 years were recruited. Gingival tissue samples were collected by excising the inner lining of the periodontal pocket at baseline (prior to non-surgical periodontal therapy) and 2 months post therapy. In controls, tissue samples were obtained immediately after tooth extraction scheduled for orthodontic reasons. Clinical parameters included probing depth, clinical attachment level, gingival index, bleeding index, plaque index. SOD activities were assessed spectrophotometrically at baseline and 2 months post NSPT, results were analysed statistically.RESULTS: At baseline, patients with chronic periodontitis had higher mean SOD activity (2.73 ± 1.36) than the control subjects (1.12 ± 1.13) withp= 0.00003 (p&lt; 0.05). At 2 months post NSPT median SOD level (1.00) had come close to median SOD value of control group (0.85);p= 0.99 (p&gt; 0.05). The resolution of inflammation with successful NSPT resulted in decreased SOD levels as in control group. Clinical parameters in patients with chronic periodontitis showed a significant improvement 2 months post NSPT (p&lt; 0.05).CONCLUSION: Non-surgical periodontal therapy significantly improves the clinical parameters and restores previously increased SOD levels to normal in chronic periodontitis patients.

Analysis of Expression and Localization of TLR-2 by Immunofluorescent Technique in Healthy and Inflammed Oral Tissues

Toll-like receptors (TLRs) are an important component of immune system. Among them, TLR-2 plays a dominant role in the oral tissues in initiating inflammation in chronic periodontitis. Not many studies have been done on quantitative expression of TLR-2 by using immunofluorescent techniques (IFT) in oral tissues. In this study, the expression and localisation of TLR-2 were detected in gingival tissues of chronic periodontitis patients and healthy individuals. Immuno Fluorescent Technique (IFT) was used for the expression and localization of TLR-2 in gingival tissue samples from 25 chronic periodontitis patients and from 25 healthy controls. Haematoxylin and Eosin staining was also done for all the samples to determine the histological characteristics of the gingival tissue samples. Both healthy and periodontitis gingival tissues expressed TLR-2. We found that the expression level of TLR-2 was higher in all the periodontitis patients than in healthy individuals. We also found out that the expression of TLR-2 was higher in the epithelial cells than in the connective tissue cells. These data suggest a definite involvement of TLR-2 in initiating an inflammatory response in periodontal tissues. More studies are required to define the mechanisms and expression levels of TLR-2 in oral health and diseases.

Effect of non-surgical periodontal therapy on superoxide dismutase levels in gingival tissues of chronic periodontitis patients: a clinical and spectophotometric analysis.

BACKGROUND: Superoxide dismutase (SOD), an antioxidant acting against superoxide (oxygen radical, O2.-), it is released in inflammatory pathways and causes connective tissue breakdown. Increased SOD activity in inflamed gingiva may indicate increased O2.- radical generation by neutrophils and other inflammatory cells at the diseased site. The aim of the study was to evaluate the effects of non-surgical periodontal therapy (NSPT) on SOD levels in gingival tissues of chronic periodontitis patients.METHODS: Forty subjects: 20 periodontally healthy (Control) and 20 chronic periodontitis (Test); age range 24–55 years were recruited. Gingival tissue samples were collected by excising the inner lining of the periodontal pocket at baseline (prior to non-surgical periodontal therapy) and 2 months post therapy. In controls, tissue samples were obtained immediately after tooth extraction scheduled for orthodontic reasons. Clinical parameters included probing depth, clinical attachment level, gingival index, bleeding index, plaque index. SOD activities were assessed spectrophotometrically at baseline and 2 months post NSPT, results were analysed statistically.RESULTS: At baseline, patients with chronic periodontitis had higher mean SOD activity (2.73 ± 1.36) than the control subjects (1.12 ± 1.13) with p = 0.00003 (p < 0.05). At 2 months post NSPT median SOD level (1.00) had come close to median SOD value of control group (0.85); p = 0.99 (p > 0.05). The resolution of inflammation with successful NSPT resulted in decreased SOD levels as in control group. Clinical parameters in patients with chronic periodontitis showed a significant improvement 2 months post NSPT (p < 0.05).CONCLUSION: Non-surgical periodontal therapy significantly improves the clinical parameters and restores previously increased SOD levels to normal in chronic periodontitis patients.

Interleukin‐21 Expression and Its Association With Proinflammatory Cytokines in Untreated Chronic Periodontitis Patients

Interleukin-21 (IL-21) controls the differentiation of T-helper Th17 cells and induces the production of IL-17 in this T-cell subtype. The aim of this study is to determine the relative expression of IL-21 in gingival tissues of chronic periodontitis patients and correlate/associate this expression with proinflammatory cytokines and clinical parameters of disease. Samples of gingival biopsies were collected from chronic periodontitis patients (n = 10) and controls (n = 8). The mRNA expressions of IL-21, IL-1β, IL-6, IL-17, IL-23, IL-10, and transforming growth factor-β1 (TGF-β1) were quantified using real-time reverse transcription-polymerase chain reaction. IL-21 levels were compared between chronic periodontitis and healthy gingival tissues and correlated with cytokine and clinical parameters of tissue destruction. A significant overexpression of IL-21, IL-1β, IL-6, IL-17, and IL-23p19 was detected in periodontal disease-affected tissues compared to healthy gingival tissues. IL-10 and TGF-β1 were, however, downregulated in periodontal lesions. IL-21 yielded significant positive correlations with probing depth, clinical attachment level, IL-1β, and IL-6. In addition, IL-21 was negatively correlated with IL-10 and TGF-β1. IL-21 was overexpressed in chronic periodontitis gingival tissues and correlated with clinical parameters of periodontal destruction and with proinflammatory cytokines. Therefore, IL-21 might play a role in the tissue destruction that characterizes chronic periodontal disease.

Syndecan-1 immunohistochemical expression in gingival tissues of chronic periodontitis patients correlated with various putative factors

Limited information is available on the expression and distribution of syndecan-1 within human gingival tissues/cells and on putative factors that might affect its expression. Therefore, the objective of the present study was to determine immunohistochemically the expression and distribution of syndecan-1 in the gingival tissues of patients with chronic periodontitis and to examine the correlation of syndecan-1 expression with various putative factors (environmental, patient/systemic and local factors). Gingival specimens were surgically excised from the area of the junctional/pocket epithelium (study group 1, including 30 chronic periodontitis patients) or the gingival oral epithelium (study group 2, comprising another 30 chronic periodontitis patients), adjacent to teeth with poor prognosis. Standard two-step immunohistochemistry and semi-quantitative evaluation of immunohistochemical staining were used to determine syndecan-1 expression. Statistical analyses on the impact of various putative factors were performed. In the junctional/pocket epithelium or the oral epithelium, syndecan-1 expression was weak to moderate in the suprabasal and basal epithelial cells and absent to weak in the internal basal lamina, external basal lamina and gingival connective tissue matrix. Syndecan-1 expression in the junctional/pocket epithelium was statistically significantly stronger than in the oral epithelium in inflammatory cells within the underlying gingival connective tissue (primarily plasma cells and lymphocytes) and in scattered fibroblast-like cells. Syndecan-1 expression in the junctional/pocket epithelium or the oral epithelium can exhibit a significant positive correlation with the severity/degree of histologically evaluated local gingival inflammation, but in general is not significantly correlated with age, smoking, full-mouth and local clinical (probing pocket depth and clinical attachment level) and radiographical parameters (radiographical bone loss) of periodontal status.

High Expression Levels of Receptor Activator of Nuclear Factor‐Kappa B Ligand Associated With Human Chronic Periodontitis Are Mainly Secreted by CD4+ T Lymphocytes

Chronic periodontitis is an infectious disease characterized by alveolar bone destruction and teeth loss. Receptor activator of nuclear factor-kappa B ligand (RANKL) is an osteoclastogenic cytokine, a central regulatory factor in the osteoclast's lifespan, and a participant in physiological and pathological bone resorption. Gingival T cells synthesize RANKL, contributing to molecular local imbalance that entails the alveolar bone resorption seen in periodontitis. Our study was aimed at associating the levels of RANKL with the CD4(+) T-cell activity present in gingival tissues of chronic periodontitis patients. Gingival biopsies were obtained from 33 chronic periodontitis patients and 20 healthy controls. Specimens were either formalin fixed and paraffin embedded for real-time reverse transcription-polymerase chain reaction (RT-PCR) and histologic analysis or tissue digestion processed for cell culture and flow-cytometry analysis. RANKL mRNA and protein levels were determined by quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) in gingival-cell culture supernatants. Gingival leukocytes were quantified by flow cytometry. RANKL and CD4 immunoreactivity were analyzed by flow cytometry and confocal microscopy. RANKL mRNA levels were higher in patients with periodontitis than in healthy subjects, and spontaneous and lipopolysaccharide (LPS)- and phytohemagglutinin (PHA)-stimulated RANKL synthesis were higher also in patients than controls. CD4(+) T lymphocytes were the predominant infiltrate cell subset present in gingival tissues of periodontitis patients. Furthermore, an association between RANKL and CD4(+) T cells was determined by double-staining flow cytometry and confocal microscopy. Taken together, these data demonstrate that gingival CD4(+) T cells are the main cells responsible for higher levels of RANKL observed in human chronic periodontitis patients.