Saliva and Serum Levels of Pentraxin‐3 and Interleukin‐1β in Generalized Aggressive or Chronic Periodontitis

Background: Pentraxin‐3 (PTX3) is a multifactorial protein involved in immunity and inflammation, which is rapidly produced and released by several cell types in response to inflammatory signals. The aim of the present study is to evaluate saliva, serum levels of PTX3, interleukin (IL)‐1β in patients with generalized chronic periodontitis (CP) or aggressive periodontitis (AgP), and periodontally healthy individuals. Methods: A total of 94 participants (25 patients with AgP, 25 patients with CP, and 44 periodontally healthy individuals matched with AgP and CP groups) were recruited. Saliva and serum samples were collected. Clinical periodontal measurements were recorded. PTX3, IL‐1β levels in serum, and saliva samples were determined by enzyme‐linked immunosorbent assay. Data were tested statistically using Kruskal‐Wallis, Mann‐Whitney U, and Spearman ρ rank test. Results: Serum and saliva data were similar in CP and AgP groups. Saliva levels of IL‐1β were significantly higher in the AgP and CP groups than controls (P <0.05). Salivary PTX3 levels were similar in the CP and control groups. Significantly higher salivary concentrations of PTX3 were detected in the AgP group than the control group (P <0.05). Saliva PTX3 levels correlated with plaque index and bleeding on probing in the CP group (P <0.05). Serum and saliva PTX3 levels correlated with those of IL‐1β in the AgP group (P <0.05). Conclusions: It may be suggested that PTX3 is related with periodontal tissue inflammation. Its salivary concentrations may have a diagnostic potential. Additional intervention and follow‐up studies coupling PTX3 concentrations with microbiologic analysis would better clarify its role in periodontal diseases.     

Influence of Periodontal Clinical Status on Salivary Levels of Glutathione Reductase

Background: Inadequate antioxidant balance may play a role in the excessive tissue breakdown in periodontitis. Because aggressive periodontitis (AgP) not only differs from chronic periodontitis (CP) in terms of clinical manifestations, this study investigates whether the salivary levels of glutathione reductase (GR) may be linked with periodontal status. Methods: Saliva samples from patients with CP (n = 121), patients with AgP (n = 18), and healthy controls (n = 69) were collected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, and extent and severity of periodontal breakdown. GR salivary levels were analyzed by spectrophotometry. The association among GR concentration and CP or AgP was analyzed individually and adjusted for confounding using multivariate binary logistic regression models. Results: GR levels not only differed significantly between the two periodontitis groups, being significantly greater in patients with AgP, but also were significantly greater than those observed in healthy controls. Synchronously, positive significant correlations between salivary GR concentration and clinical parameters were observed. After binary logistic regression analysis, both GR salivary levels ≥15.38 and ≥24.20 mU/mL were associated independently with CP and AgP, respectively. A significant interaction effect was also detected between increased GR salivary concentration and aging in the CP group. Conclusions: Increased GR salivary concentration may be a strong/independent prognostic indicator of the amount and extent of oxidative stress‐induced periodontal damage in both CP and AgP. Likewise, saliva samples might reflect an interactive effect of GR levels associated with the aging‐related cumulative characteristics of periodontal damage in CP.     

Salivary interleukin-1β levels in patients with chronic periodontitis before and after periodontal phase I therapy and healthy controls: a case-control study

Background: The role of interleukin (IL)‐1β in periodontal disease pathogenesis is well researched. This study aimed to assess and compare the salivary IL‐1β levels in patients with chronic periodontitis before and after periodontal phase I therapy and periodontally healthy controls. Further, relationships between IL‐1β levels and various clinical parameters were explored. Methods: Twenty‐eight patients with moderate‐to‐severe generalized chronic periodontitis and 24 age‐, race‐, and ethnicity‐matched controls participated in this study. Saliva samples were obtained from all patients. The clinical parameters recorded were clinical attachment loss (AL), probing depth, bleeding on probing, periodontal index, and gingival index. Clinical evaluation and sample collection were repeated 1 month after periodontal phase I therapy in patients with periodontitis. IL‐1β levels were assessed using enzyme‐linked immunosorbent assay. Results: Mean IL‐1β levels in patients with periodontitis at baseline (1,312.75 pg/mL) were significantly higher (P <0.0001; eight‐fold) than in controls (161.51 pg/mL). Although treatment in patients with periodontitis resulted in significant reduction in IL‐1β levels (mean: 674.34 pg/mL; P = 0.001), they remained significantly higher (P <0.0001; four‐fold) than control levels. There were significant correlations between IL‐1β levels and all clinical parameters (P <0.01) except percentage sites with clinical AL >2 mm (P >0.05). Conclusions: The data indicate that IL‐1β levels are raised in the saliva of patients with chronic periodontitis, which are reduced after phase I therapy, suggesting a close association between salivary IL‐1β and periodontitis. Additional longitudinal studies are needed to validate salivary IL‐1β as a marker for periodontal disease.     

Saliva and Serum Levels of B‐Cell Activating Factors and Tumor Necrosis Factor‐α in Patients With Periodontitis

Background: B‐lymphocytes play a central and critical role in the adaptive immune response against invading pathogens. This study evaluates saliva and serum levels of APRIL (a proliferation‐inducing ligand), B‐cell activating factor (BAFF), tumor necrosis factor‐α (TNF‐α), interleukin (IL)‐6, and IL‐10 in patients with chronic periodontitis (CP) or aggressive periodontitis (AgP) and periodontally healthy individuals. Methods: Twenty‐five patients with AgP, 20 patients with CP, and 20 periodontally healthy individuals were included. Smoking status was recorded, and all individuals were divided into non‐smokers and smokers. Saliva and serum samples were collected before clinical periodontal measurements. APRIL, BAFF, TNF‐α, IL‐6, and IL‐10 levels in serum and saliva samples were determined by enzyme‐linked immunosorbent assay. Statistical analysis was performed using multivariate analysis of variance and bivariate correlation. Results: Serum and saliva levels of TNF‐α, APRIL, BAFF, IL‐6, and IL‐10 were similar in CP and AgP groups. Serum levels of TNF‐α, APRIL, and BAFF and saliva levels of BAFF were significantly higher in periodontitis groups than healthy controls (P <0.05). Non‐smokers with CP or AgP had lower levels of saliva TNF‐α and APRIL and serum APRIL and IL‐6 than smokers with CP or AgP (P <0.05). Saliva APRIL and serum TNF‐α and IL‐6 levels were significantly higher in healthy smokers than healthy non‐smokers (P <0.05). Clinical periodontal parameters correlated positively with TNF‐family cytokines and negatively with IL‐10 (P <0.05). Conclusions: Within the limits of this study, it may be suggested that elevated salivary and serum TNF‐α, APRIL, and BAFF in patients with periodontitis may contribute to the dominance of B cells in periodontitis lesions. Moreover, higher levels in healthy smokers than non‐smoking counterparts may play a role in detrimental effects of smoking on periodontal tissues.     

Activation of inflammasomes in dendritic cells and macrophages by Mycoplasma salivarium

Interleukin‐1β (IL‐1β) plays crucial roles in the pathogenesis of periodontal disease. It is produced after the processing of pro‐IL‐1β by caspase‐1, which is activated by the inflammasome‐a multiprotein complex comprising nucleotide‐binding domain leucine‐rich repeat‐containing receptor (NLR), the adaptor protein apoptosis‐associated speck‐like protein containing a caspase‐recruitment domain (ASC), and procaspase‐1. Mycoplasma salivarium preferentially inhabits the gingival sulcus and the incidence and number of organisms in the oral cavity increase significantly with the progression of periodontal disease. To initially clarify the association of this organism with periodontal diseases, this study determined whether it induces IL‐1β production by innate immune cells such as dendritic cells or macrophages by using Mycoplasma pneumoniae as a positive control. Both live and heat‐killed M. salivarium and M. pneumoniae cells induced IL‐1β production by XS106 murine dendritic cells as well as pyroptosis. The activities were significantly downregulated by silencing of caspase‐1. Bone‐marrow‐derived macrophage (BMMs) from wild‐type and NLR‐containing protein 3 (NLRP3)‐, ASC‐, and caspase‐1‐deficient mice were examined for IL‐1β production in response to these mycoplasmas. Live M. salivarium and M. pneumoniae cells almost completely lost the ability to induce IL‐1β production by BMMs from ASC‐ and caspase‐1–deficient mice. Their activities toward BMMs from NLRP3‐deficient mice were significantly but not completely attenuated. These results suggest that live M. salivarium and M. pneumoniae cells can activate several types of inflammasomes including the NLRP3 inflammasome. Both M. salivarium and M. pneumoniae cells can activate THP‐1 human monocytic cells to induce IL‐1β production. Hence, the present finding that M. salivarium induces IL‐1β production by dendritic cells and macrophages may suggest the association of this organism with periodontal diseases.     

Alarm anti‐protease trappin‐2 negatively correlates with proinflammatory cytokines in patients with periodontitis

1 Background

Trappin‐2 is a potent biologically active serine protease inhibitor with anti‐inflammatory properties that has also been characterized as an “alarm anti‐protease.” Although the importance of trappin‐2 in several chronic infections has been demonstrated, its potential involvement in periodontitis remains undefined. This study aims to investigate salivary levels of trappin‐2 and interleukin (IL)‐1β in periodontally healthy individuals and patients with gingivitis or generalized chronic periodontitis (CP) or aggressive periodontitis (GAgP).

2 Methods

Whole unstimulated saliva samples were collected from 80 systemically healthy and non‐smoking individuals before full‐mouth periodontal examination. Trappin‐2 and IL‐1β were analyzed by enzyme‐linked immunosorbent assay and reported as nanograms per milligram after calibration for total protein levels.

3 Results

Correlation analysis revealed negative association between trappin‐2 and IL‐1β levels. Trappin‐2 also showed strong negative correlation with clinical periodontal parameters, in contrast to IL‐1β, which showed positive correlation. Trappin‐2 levels were significantly lower in individuals with CP and GAgP, but not gingivitis, compared with healthy individuals. Reduced salivary concentrations of trappin‐2 had high sensitivity and specificity to distinguish health from periodontitis.

4 Conclusions

Trappin‐2 is abundant in the saliva of individuals with healthy periodontium in line with its role as an “anti‐alarm” protease. Decreased salivary trappin‐2 and increased IL‐1β levels in individuals with periodontitis, compared with healthy individuals, may implicate a potential antiprotease/proinflammatory cytokine imbalance, resulting in impaired host protective capacity.     

Salivary and Serum Chromogranin A and α-Amylase in Periodontal Health and Disease

Background: Salivary stress-related biomarkers in connection with periodontal disease have not been extensively studied. In addition to cortisol as a well-known marker of stress loading, chromogranin A (CgA) and α-amylase (AA) are supposed to link the activity of the neuroendocrine system to local and systemic immune functions and to be related to periodontitis. This study aims to determine CgA and AA in saliva and serum in periodontal health and disease to assess their potential relationship to periodontitis. Methods: Patients with aggressive (AgP) (n = 24) and chronic periodontitis (CP) (n = 34) as well as healthy control (CO) (n = 30) individuals participated in this study. CgA and AA were determined in saliva and serum with enzyme-linked immunosorbent assay and an adapted clinical amylase test; salivary cortisol was determined using mass spectrometry. Clinical parameters of periodontal disease were evaluated, and their possible correlations with stress-related biomarkers were assessed. Results: Significantly higher CgA levels were found in the saliva of patients with AgP compared with those in patients with CP and CO individuals (P <0.001). Salivary cortisol levels were higher in the AgP group compared with those in patients with CP (P <0.05). No differences in serum CgA levels and salivary and serum AA activities were found among all groups. A positive correlation was revealed between salivary AA activity or salivary CgA levels and the extent of periodontitis (P <0.05). Conclusion: The results suggest an association of CgA and cortisol levels as well as AA activity in saliva with periodontitis, especially a significant relationship of salivary CgA and cortisol to AgP.     

Activation of NLRP3 inflammasome in macrophages by mycoplasmal lipoproteins and lipopeptides

The NLRP3 inflammasome, an intracellular sensor consisting of the nucleotide‐binding oligomerization domain‐like receptor family, pyrin domain containing 3 (NLRP3), the adaptor protein apoptosis‐associated speck‐like protein containing a caspase‐recruitment domain (ASC), and procaspase‐1, plays critical roles in host defense against microbial pathogens by inducing production of interleukin‐1β (IL‐1β) and IL‐18. Mycoplasma salivarium and Mycoplasma pneumoniae cells activated murine bone marrow‐derived macrophages (BMMs) to induce production of IL‐1α, IL‐1β, and IL‐18. The IL‐1β production‐inducing activities of these mycoplasmas toward BMMs from Toll‐like receptor 2 (TLR2)‐deficient mice were significantly attenuated compared with those from C57BL/6 mice (B6BMMs). This result suggests the possibility that their lipoproteins as TLR2 agonists are involved in the activity. Lipoproteins of M. salivarium and M. pneumoniae (MsLP and MpLP), and the M. salivarium‐derived lipopeptide FSL‐1 induced IL‐1β production by B6BMMs, but not by BMMs from caspase‐1‐, NLRP3‐ or ASC‐deficient mice. The activities of MsLP and MpLP were not downregulated by the proteinase K treatment, suggesting that the active sites are their N‐terminal lipopeptide moieties. B6BMMs internalized the mycoplasmal N‐terminal lipopeptide FSL‐1 at least 30 min after incubation, FSL‐1‐containing endosomes started to fuse with the lysosomes around 2 hours, and then FSL‐1 translocated into the cytosol from LAMP‐1⁺ endosomes. The artificial delivery of FSL‐1 into the cytosol of B6BMMs drastically enhanced the IL‐1β production‐inducing activity. FSL‐1 as well as the representative NLRP3 inflammasome activator nigericin induced the NLRP3/ASC speck, but FSL‐1 located in a compartment different from the NLRP3/ASC speck.     

Influence of smoking on interleukin-1beta level, oxidant status and antioxidant status in gingival crevicular fluid from chronic periodontitis patients before and after periodontal treatment

Toker H, Akp?nar A, Ayd?n H, Poyraz O. Influence of smoking on interleukin‐1beta level, oxidant status and antioxidant status in gingival crevicular fluid from chronic periodontitis patients before and after periodontal treatment. J Periodont Res 2012; 47: 572–577. © 2012 John Wiley & Sons A/S Background and Objective: The aim of this study was to evaluate the impact of smoking on the relationship between interleukin‐1 (IL‐1β) and oxidation in patients with periodontitis and response to nonsurgical periodontal therapy. Material and Methods: Data were obtained from 30 patients with generalized chronic periodontitis (15 smokers and 15 nonsmokers) and from 10 periodontally healthy controls. IL‐1β level, total oxidant status (TOS) and total antioxidant status (TAS) were recorded in gingival crevicular fluid. Probing depth, clinical attachment level, gingival and plaque indices and bleeding on probing were also measured. The gingival crevicular fluid and clinical parameters were recorded at baseline and 6 wk after periodontal treatment. Results: The study showed statistically significant improvement of clinical parameters in both smokers and nonsmokers after periodontal treatment. Moreover, the baseline IL‐1β levels were significantly higher in smokers compared with nonsmokers (p < 0.05). After periodontal treatment, the IL‐1β levels were significantly reduced in both smokers and nonsmokers (p < 0.05). There were no significant differences in TOS and TAS between periodontitis patients and healthy controls at baseline and 6 wk after periodontal treatment. The level of IL‐1β in gingival crevicular fluid was positively correlated with TOS in both smokers and nonsmokers. Conclusions: Periodontal treatment improved the clinical parameters in both smokers and nonsmokers. The results confirm that periodontal therapy has an effect on IL‐1β levels in gingival crevicular fluid, but not on TOS and TAS.     

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.     

Soluble interleukin-1 receptor type II levels in gingival crevicular fluid in aggressive and chronic periodontitis

Background: Interleukin (IL)-1 is closely related to the initiation and progression of periodontal disease. IL-1 levels in the gingival crevicular fluid (GCF) of subjects with periodontitis are higher than those in periodontally healthy controls, and the levels of IL-1 correlate with disease severity. However, soluble IL-1 receptor type II (sIL-1RII), which acts as a decoy receptor for IL-1s, has not been investigated in detail in periodontal disease. The purpose of this study was to measure sIL-1RII levels in the GCF of subjects with chronic or aggressive periodontitis; the correlation between the sIL-1RII levels in GCF and clinical parameters also was examined. Methods: IL-1beta and sIL-1RII were measured in 64 GCF samples collected from 47 subjects with chronic periodontitis (CP) and 17 subjects with aggressive periodontitis (AgP). The clinical characteristics of each site were recorded at the time of GCF sampling. IL-1beta and sIL-1RII were measured by specific non-cross-reactive enzyme-linked immunosorbent assay. Results: The disease severity was comparable in CP and AgP. IL-1beta was detected in 98% of CP GCF samples and 88% of AgP GCF samples. sIL-1RII was detected in 55% of CP GCF samples and 35% of AgP GCF samples. However, the concentrations of IL-beta and sIL-1RII detected in GCF from subjects with CP or AgP were similar. Conclusion: sIL-1RII was detected more often in CP GCF than in AgP GCF, and there was no correlation between GCF sIL-1RII concentration and clinical parameters.     

Proinflammatory and Anti‐Inflammatory Cytokines in Gingival Crevicular Fluid and Serum of Patients With Rheumatoid Arthritis and Patients With Chronic Periodontitis

Background: The aim of this study is to evaluate proinflammatory and anti‐inflammatory cytokine levels in gingival crevicular fluid (GCF) and serum of rheumatoid arthritis (RA) and chronic periodontitis (CP) patients to assess whether cytokine profiles distinguish patients with RA and patients with CP while using healthy patients as background controls. Methods: A total of 49 patients, 17 patients with RA (three males and 14 females; mean age: 47.82 ± 10.74 years), 16 patients with CP (10 males and six females; mean age: 44.00 ± 7.00 years), and 16 controls (eight males and eight females; mean age: 28.06 ± 6.18 years) were enrolled. Patients with RA were under the supervision of rheumatologists; 15 of the patients with RA were being treated with methotrexate–sulfasalazine combined therapy, and two of the patients were being treated with leflunomid therapy. Periodontal parameters (plaque index, gingival index, probing depth, and clinical attachment level) were recorded. Interleukin (IL)‐1β, IL‐4, IL‐10, and tumor necrosis factor‐α (TNF‐α) were determined in GCF and IL‐1β and IL‐10 in serum by enzyme‐linked immunosorbent assay. Results: There were significant differences found among RA, CP, and control groups for all periodontal parameters (P <0.05). The total amount and concentration of GCF IL‐1 β, IL‐4, IL‐10, and TNF‐α were similar in RA and CP patients (P >0.05). Although the total amount and concentration of serum IL‐10 was not significantly different among the groups (P >0.05), serum IL‐1β was significantly lower in the RA group compared to CP patients and controls and was higher in GCF of the RA group compared to the CP group. Conclusions: Although clinical periodontal disease parameters indicated more severe periodontal disease in CP compared to RA patients, immunologic evaluation did not reveal consistent results regarding proinflammatory and anti‐inflammatory cytokine levels. This might be a result of the use of non‐steroidal anti‐inflammatory drugs and rheumatoid agents by patients with RA.     

Level of Interleukin‐35 in Gingival Crevicular Fluid,Saliva, and Plasma in Periodontal Disease and Health

Background: A novel member of the interleukin (IL)‐12 family, IL‐35 is an important inhibitory cytokine released by regulatory T cells. The aim of this study is to evaluate gingival crevicular fluid (GCF), saliva, and plasma levels of IL‐35 in periodontal disease and health. Methods: Samples of GCF, whole saliva, and plasma were obtained from systemically healthy, non‐smoking individuals with gingivitis (n = 20) or chronic periodontitis (CP) (n = 20) and periodontally healthy individuals (n = 20). Full‐mouth clinical periodontal measurements, including probing depth (PD), bleeding on probing, gingival index, and plaque index (PI), were also recorded. Enzyme‐linked immunosorbent assay was used to determine IL‐35 levels in the samples. Data were tested statistically by analysis of variance and Pearson rank correlation test. Results: All clinical parameters were significantly higher in the CP group than the healthy and gingivitis groups (P <0.001). The GCF total amount of IL‐35 was significantly higher in the CP group than the other groups (P = 0.04), whereas the GCF concentration of IL‐35 was significantly higher in the healthy group than the other groups (P = 0.002). There were significant differences among the study groups in terms of salivary IL‐35 level (P <0.001), with the highest level observed in the healthy group and the lowest in the CP group. There was no statistical difference between groups in plasma levels of IL‐35 (P >0.05). There was a positive correlation between GCF total amount of IL‐35 and PD (r = 0.338, P = 0.03) and PI (r = 0.374, P = 0.005) parameters. Conclusions: IL‐35 could have an important role in suppressing periodontal inflammation and maintaining periodontal health. Additional studies are required to evaluate its role in periodontal diseases.     

Hyperglucose Contributes to Periodontitis: Involvement of the NLRP3 Pathway by Engaging the Innate Immunity of Oral Gingival Epithelium

Background: The NLRP3 inflammasome is essentially a family of intracellular innate immune sensors that can respond to bacterial challenge and initiate early host immunity responses. However, the involvement and possible molecular mechanism of the NLRP3 pathway in the context of chronic periodontitis (CP) and diabetes mellitus have yet to be fully elucidated. Methods: Gingival tissues were collected from patients with CP and/or type 2 diabetes mellitus (T2DM), and the expression of NLRP3 and interleukin (IL)‐1β was analyzed by immunohistochemistry. To explore the possible molecular mechanism, human gingival epithelial cells (HGECs) were established in vitro and challenged with lipopolysaccharide (LPS) and/or high glucose. High extracellular K⁺ was applied as an inhibitor of NLRP3. The NLRP3 pathway was analyzed by immunocytochemistry and quantitative polymerase chain reaction. Results: Compared with control individuals, NLRP3 and IL‐1β were significantly upregulated in oral gingival epithelium of patients with CP and/or T2DM (P <0.05). The expression of NLRP3 was significantly upregulated in HGECs when stimulated in vitro by LPS or high glucose (P = 0.00). The simultaneous stimulation of LPS and high glucose contributed to significant upregulation of NLRP3 expression versus LPS or high glucose alone (P = 0.00). Although expression of caspase 1 and IL‐1β protein were increased in HGECs when stimulated by LPS, they were partially inhibited after the NLRP3 was successfully blocked. Conclusion: For patients with T2DM and CP, hyperglycemic status may exacerbate the inflammation state of gingival tissue by activating the NLRP3 pathway, and this abnormal host inflammatory response may contribute to further tissue breakdown.     

Chemerin as a Novel Crevicular Fluid Marker of Patients With Periodontitis and Type 2 Diabetes Mellitus

Background: The objectives of the present study are to: 1) determine whether gingival crevicular fluid (GCF) chemerin is a novel predictive marker for patients with chronic periodontitis (CP) with and without type 2 diabetes mellitus (t2DM); 2) analyze the relationship between chemerin and interleukin (IL)‐6 in periodontally healthy individuals and in patients with CP and with and without t2DM; and 3) evaluate the effect of non‐surgical periodontal therapy on GCF chemerin levels. Methods: Eighty individuals were split into four groups: 20 who were systemically and periodontally healthy (CTRL), 20 with t2DM and periodontally healthy (DM‐CTRL), 20 systemically healthy with CP (CP), and 20 with CP and t2DM (DM‐CP). Individuals with periodontitis were treated with non‐surgical periodontal therapy. GCF sampling procedures and clinical periodontal measures were performed before and 6 weeks after treatment. Enzyme‐linked immunosorbent assay was used to measure chemerin and IL‐6 levels. Results: Greater values for GCF chemerin and IL‐6 levels were found in CP groups than in periodontally healthy groups, in DM‐CP than in CP, and in DM‐CTRL than in CTRL (P <0.008). GCF chemerin and IL‐6 levels decreased following therapy in CP groups (P <0.02). A comprehensive overview of all groups showed a statistically significant positive correlation of chemerin with IL‐6, glycated hemoglobin, sampled‐site clinical attachment level, and gingival index (P <0.05). Conclusions: In this study, periodontitis and t2DM induced aberrant secretion of chemerin, and non‐surgical periodontal therapy influenced the decrease of GCF chemerin levels in patients with CP with and without t2DM. Furthermore, it suggests GCF chemerin levels may be considered a potential proinflammatory marker for diabetes, periodontal disease, and treatment outcomes.     

Total proteoglycan and chondroitin-4-sulfate levels in gingiva of patients with various types of periodontitis

BACKGROUND: The aim of the present study was to investigate the total proteoglycan (PG) and chondroitin-4-sulfate (C4S) levels in gingival tissue samples obtained from patients with aggressive periodontitis (AgP) and chronic periodontitis (CP) before therapy (baseline) and 1 month after completion of non-surgical periodontal therapy. METHODS: Gingival tissue samples were obtained from 10 AgP and 10 CP patients before initiation of treatment (baseline) and 1 month after non-surgical periodontal treatment. The control group comprised 10 systemically and periodontally healthy subjects. Total PG and C4S levels were determined by biochemical techniques. PG levels were analyzed using a modified Bitter and Muir method. C4S assay was carried out using chondroitin sulphate lyase AC and chondroitin-6 sulphate sulphohydrolase enzymes. The results were tested statistically using parametric tests. RESULTS: The clinical periodontal parameters demonstrated significant decreases in the periodontitis groups (P<0.05) after treatment, and there was no significant difference between AgP and CP groups at baseline and after treatment (P>0.05). At baseline, total PG and C4S levels in both of the periodontitis groups were significantly lower than that of the control group (P<0.05). One month after the non-surgical periodontal treatment, total PG levels in the periodontitis groups were comparable to the control group (P>0.05), whereas C4S levels in the AgP group were significantly lower than the other study groups (P<0.05). In the CP group, total PG and C4S levels increased significantly (P = 0.001 and P = 0.006, respectively) after non-surgical periodontal treatment, but they did not increase in the AgP group (P>0.05). CONCLUSION: The significant increases observed in total proteoglycan and chondroitin-4-sulfate levels after non-surgical periodontal treatment in the CP group but not in the AgP group may suggest that healing patterns differ between the two periodontitis types in terms of PG and C4S composition of extracellular matrix.     

Periodontitis and Endothelial Dysfunction: Periodontal Clinical Parameters and Levels of Salivary Markers Interleukin‐1β, Tumor Necrosis Factor‐α, Matrix Metalloproteinase‐2, Tissue Inhibitor of Metalloproteinases‐2 Complex,and Nitric Oxide

Background: Periodontitis has been associated with a greater risk for atherosclerotic cardiovascular disease (ACD). Endothelial dysfunction (ED) is a parameter of early ACD, and its association with periodontitis has rarely been investigated to date. The objective of this study is to evaluate the association between periodontitis and ED by means of periodontal clinical parameters and salivary markers interleukin (IL)‐1β, tumor necrosis factor (TNF)‐α, nitric oxide (NO), and matrix metalloproteinase (MMP)‐2 and tissue inhibitor of metalloproteinases (TIMP)‐2 complex. Methods: Forty‐seven individuals were divided into two groups: 1) 24 individuals with chronic periodontitis (CP); and 2) 23 individuals without CP. Periodontal examinations of bleeding on probing (BOP), probing depth (PD), and clinical attachment level (CAL) were performed. ED was evaluated by means of flow‐mediated dilatation (FMD) of the brachial artery. Salivary concentrations of IL‐1β, TNF‐α, and MMP‐2/TIMP‐2 complex were assessed using enzyme‐linked immunosorbent assay. NO determination was based on the reaction of Griess. Results: Individuals with CP presented higher occurrence of ED than individuals without CP (P = 0.03 after reactive hyperemia; P = 0.05 after sublingual nitrate). A significant association among the production of MMP‐2/TIMP‐2 complex with the presence of CP (P = 0.008) and periodontal parameters PD, CAL, and BOP was identified. Concentration of salivary markers IL‐1β, TNF‐α, and NO was similar in individuals with and without CP. A significant positive correlation between NO and ED was also identified. Conclusions: Periodontitis was positively associated with ED, expressed by a smaller percentage of FMD of the brachial artery and higher salivary levels of MMP‐2/TIMP‐2 complex. Additionally, salivary levels of NO were significantly associated with better functioning of the vascular endothelium.     

Alcohol Consumption and Periodontitis: Quantification of Periodontal Pathogens and Cytokines

Background: There are few studies on periodontal status related to microbiologic and immunologic profiles among individuals not or occasionally using alcohol and those with alcohol dependence. The aim of this study is to determine the effect of alcohol consumption on the levels of subgingival periodontal pathogens and proinflammatory cytokines (interleukin [IL]‐1β and tumor necrosis factor [TNF]‐α) in the gingival fluid among individuals with and without periodontitis. Methods: This observational analytic study includes 88 volunteers allocated in four groups (n = 22): individuals with alcohol dependence and periodontitis (ADP), individuals with alcohol dependence and without periodontitis (ADNP), individuals not or occasionally using alcohol with periodontitis (NAP), and individuals not or occasionally using alcohol without periodontitis (NANP). Levels of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Eikenella corrodens, and Fusobacterium nucleatum were determined by real‐time polymerase chain reaction on the basis of the subgingival biofilm, and IL‐1β and TNF‐α were quantified by enzyme‐linked immunosorbent assay in gingival fluid samples. Results: Individuals with alcohol dependence showed worse periodontal status and higher levels of P. intermedia, E. corrodens, F. nucleatum, and IL‐1β than non‐users. No significant correlations between TNF‐α and bacterial levels were observed. However, in the ADP group, higher levels of E. corrodens were correlated with higher levels of IL‐1β. Conclusion: A negative influence of alcohol consumption was observed on clinical and microbiologic periodontal parameters, as well as a slight influence on immunologic parameters, signaling the need for additional studies.     

不同类型牙周炎患者唾液中8-OHdG含量的比较

目的:检测不同类型牙周炎患者非刺激性全唾液中8-OHdG的含量并评价其与PD,CAL等临床指标的关系。方法:留取10名侵袭性牙周炎,10名慢性牙周炎和10名健康对照非刺激性全唾液,并记录全口PD,CAL,PLI,BI。ELLISA试剂盒检测唾液中8-OHdG含量。结果:8-OHdG含量在3组间存在统计学差异,AgP组:(3.8 ±1.0) μg/L, CP组:(2.37±0.7) μg/L,H组:(1.0±0.6) μg/L。PD,BI,PLI,CAL与8-OHdG含量存在相关关系,慢性与侵袭组中8-OHdG含量与平均CAL, CAL>5mm位点所占百分比,CAL>7 mm位点所占百分比相关。结论:唾液中8-OHdG含量可以反映牙周组织的破坏程度并与牙周炎类型存在相关关系。     

Analysis of YKL‐40 Acute‐Phase Protein and Interleukin‐6 Levels in Periodontal Disease

Background: YKL‐40, a new acute‐phase protein, is shown to be elevated in inflammatory diseases, such as rheumatoid arthritis, type 2 diabetes mellitus, and coronary artery diseases. However, there is no data indicating a relationship between YKL‐40 and periodontal disease. Interleukin‐6 (IL‐6) is the major regulator of acute‐phase protein synthesis and one of the most studied inflammatory markers in periodontal disease. The purpose of the present study is to evaluate YKL‐40 and IL‐6 levels in gingival crevicular fluid (GCF) and serum of patients with periodontal disease and healthy individuals. Methods: Periodontally healthy individuals (n = 15), patients with gingivitis (n = 15), and patients with severe chronic periodontitis (CP) (n = 15) without any systemic disease were included in the study. Clinical measurements were recorded; GCF and blood samples were obtained from each participant. GCF and serum YKL‐40 and IL‐6 levels were analyzed by enzyme‐linked immunosorbent assay. Statistical analysis was performed by parametric and non‐parametric tests. Results: Total amounts of YKL‐40 and IL‐6 in GCF as well as serum YKL‐40 and IL‐6 levels were significantly higher in patients with gingivitis and CP compared with healthy controls (P <0.01). YKL‐40 levels in GCF and serum as well as serum IL‐6 levels were significantly higher in patients with CP compared with patients with gingivitis (P <0.01). Conclusions: YKL‐40 levels in GCF as well as serum YKL‐40 and IL‐6 levels increased from gingivitis to periodontitis. Within the limits of the present study, the YKL‐40 molecule might be a potential novel inflammatory marker of periodontal disease.