Detection of Treponema denticola in saliva obtained from patients with various periodontal conditions

The aim of the study was to determine the prevalence of Treponema denticola in saliva of periodontally diseased and healthy patients and its relationship with the periodontal status. A 16S rRNA-based polymerase chain reaction detection method was used to determine the prevalence of T. denticola in whole saliva samples from patients with chronic periodontitis (CP, n = 37), aggressive periodontitis (AgP, n = 24), and healthy subjects (n = 28). The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss, and extent of periodontal breakdown. Risk factors were assessed individually and adjusted for confounding using a binary logistic regression model. The results showed that the prevalence of T. denticola in CP patients was significantly higher than those in healthy and AgP subjects (P < 0.05). Odds ratio analysis revealed a positive association for CP group/T. denticola-positive and smoking/T. denticola-positive subjects. Furthermore, all clinical measurements were significantly greater (P < 0.05) for T. denticola-positive subjects compared to T. denticola-negative subjects. After binary logistic regression analysis, both T. denticola and smoking were independently and strongly associated with development of CP. It was concluded that when used in conjunction with an optimized clinical examination protocol, this assay may offer a rapid, useful, and cost-effective tool for monitoring the presence of T. denticola in noninvasive clinical samples from both healthy and diseased patients and correlating it with the amount and extent of periodontal breakdown.     

Salivary levels of antibacterial peptide (LL-37/hCAP-18) and cotinine in patients with chronic periodontitis

Background: The purpose of this study is to examine the relationship between salivary LL‐37 levels and clinical severity in patients with chronic periodontitis (CP). The presence/absence of four periodontopathic bacteria and salivary cotinine levels were also examined to assess the impact of these factors on LL‐37 production. Methods: Unstimulated salivary samples were collected from 69 patients with CP. Salivary concentrations of LL‐37 and cotinine were measured by enzyme‐linked immunosorbent assay. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola in saliva were detected by polymerase chain reaction. Periodontal examination included determination of probing depth (PD), clinical attachment level, bleeding on probing, and plaque control record. Results: Mean salivary LL‐37 concentration was 225.0 ± 227.2 ng/mL, and a high prevalence of periodontopathic bacteria was observed. The stepwise ordinal logistic regression model showed that high salivary LL‐37 levels were significantly associated with the presence of T. denticola and higher percentage of teeth with PD ≥5 mm. In addition, higher salivary cotinine levels (≥8 ng/mL) were negatively associated with salivary LL‐37 levels. Conclusions: Salivary LL‐37 level was positively correlated with severe periodontal destruction, and production was apparently associated with periodontopathic bacterial infection. The negative correlations between salivary LL‐37 and cotinine levels also suggest that smoking or long‐term exposure to environmental tobacco smoke can lead to lower LL‐37 levels in the oral cavity and increased risk of periodontitis.     

Microbial Analysis of Subgingival Plaque Samples Compared to That of Whole Saliva in Patients With Periodontitis

Background: The detection of special bacterial species in patients with periodontitis is considered to be useful for clinical diagnosis and treatment. The collection of subgingival plaque samples is the common way for the determination of periodontopathic bacteria. However, recently, salivary analysis has been discussed as an advantageous future diagnostic method for periodontitis because it offers simple quantitative sampling and the possibility to assess various bacteria. The aim of this cross‐sectional study is to investigate whether there is a correlation between the results of different bacterial species in saliva and subgingival plaque samples from individuals with aggressive periodontitis (AgP) and chronic periodontitis (CP). Methods: Whole saliva and subgingival plaque samples from the deepest pocket of each quadrant were collected from 43 patients with CP and 33 patients with AgP. Twenty different bacterial species from both samplings were determined by the 16S ribosomal RNA‐based polymerase chain reaction with microarray technique. Results: All bacterial species were detected in salivary and subgingival plaque samples. For Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, as well as Actinomyces viscosus, Campylobacter rectus/showae, Prevotella intermedia, Parvimonas micra, Eubacterium nodatum, and Campylobacter gracilis, a significant positive correlation between salivary and subgingival plaque samples was detected in patients with both types of periodontitis. There were no significant differences in bacteria in salivary and subgingival plaque samples between AgP and CP. Conclusion: Salivary analysis might be discussed as a potential alternative to subgingival plaque sampling for microbiologic analysis in both AgP and CP.     

The Influences of Periodontal Status and Periodontal Pathogen Quantity on Salivary 8‐Hydroxydeoxyguanosine and Interleukin‐17 Levels

Background: Periodontitis is a biofilm‐initiated disease that is characterized by elevated inflammatory status. 8‐Hydroxydeoxyguanosine (8‐OHdG) and interleukin (IL)‐17 are highly associated with inflammation and bone resorption and therefore are regarded as potential biomarkers for periodontitis. In this study, the associations between salivary 8‐OHdG and IL‐17 levels and clinical and microbial parameters before and after non‐surgical treatment are investigated. Methods: Forty‐five patients with chronic periodontitis (CP) and 47 periodontally healthy volunteers were recruited for the study. Clinical parameters, including the probing depth (PD), clinical attachment level (CAL), sulcular bleeding index, and simplified oral hygiene index (OHI‐S), were examined for each participant. Microbial parameters including the quantities of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola in the subgingival plaque and saliva were determined by real‐time polymerase chain reaction at baseline and 1 and 3 months after the non‐surgical treatment. Salivary 8‐OHdG and IL‐17 levels were detected by enzyme‐linked immunosorbent assays. Results: Compared with healthy volunteers, CP group patients had significantly higher salivary 8‐OHdG and IL‐17 levels at baseline. Baseline salivary 8‐OHdG and IL‐17 levels were positively correlated with all clinical parameters as well as the quantities of T. forsythia and T. denticola. After non‐surgical treatment, baseline levels of salivary 8‐OHdG and IL‐17 were reduced significantly at both the 1‐ and 3‐month follow‐ups. The hierarchical linear model revealed that variations in the PD, CAL, and OHI‐S had significant positive effects on variation in the salivary 8‐OHdG level. However, variations in the PD; quantity of T. forsythia in the subgingival plaque; and quantities of P. gingivalis, T. forsythia, and T. denticola in saliva were associated significantly with variation in the salivary IL‐17 levels. Conclusions: There was a strong association between salivary 8‐OHdG and IL‐17 levels and periodontitis. Variation in the salivary 8‐OHdG level was correlated with variations in the clinical parameters, whereas variation in the IL‐17 level was correlated with variation in the microbial parameters.     

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.     

Influence of Periodontal Status and Periodontopathogens on Levels of Oral Human β‐Defensin‐2 in Saliva

Background: Expression patterns of human β‐defensin‐2 (HBD‐2) mRNA or HBD‐2 protein concentration and periodontal diseases have been a focus of scientific research. This study compares the salivary levels of HBD‐2 protein concentration of healthy patients and patients with gingivitis and chronic periodontitis (CP) and correlates these levels with the presence of periodontopathogens. Methods: A total of 89 patients were enrolled in this study: 31 periodontally healthy, 27 with gingivitis, and 31 with CP. Plaque and gingival indices, probing depth, and clinical attachment level were measured. The presence of Campylobacter rectus, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Prevotella intermedia was evaluated qualitatively by conventional polymerase chain reaction. HBD‐2 quantification in saliva was performed using an immune enzymatic assay. Frequency of periodontopathogens and HBD‐2 protein concentration was assessed. Association between HBD‐2 protein concentration (≥100 pg/mL) and the simultaneous presence of one to two, three to four, or five to six periodontopathogens was tested. Results: Although periodontally healthy individuals and patients with gingivitis showed similar HBD‐2 levels, the CP group displayed an increased level of HBD‐2. P. gingivalis, P. intermedia, T. forsythia, and T. denticola were more prevalent in CP; however, their mere presence was not related to the increased levels of HBD‐2 (Pearson correlation and multinomial logistic regression model). Conclusions: Salivary HBD‐2 protein concentration was higher in patients with CP compared with healthy individuals or patients with gingivitis. These different protein concentrations were not related to the frequency of periodontopathogens. Clinical inflammatory profile had a higher impact on salivary HBD‐2 levels than bacteria.     

Soluble Triggering Receptor Expressed on Myeloid Cells 1 (sTREM‐1) in Gingival Crevicular Fluid: Association With Clinical and Microbiologic Parameters

Background: Soluble triggering receptor expressed on myeloid cells 1 (sTREM‐1) belongs to the immunoglobulin superfamily and is involved in amplification of the inflammatory response to bacterial infection. This cross‐sectional study aims to investigate the levels of sTREM‐1 in gingival crevicular fluid (GCF) of individuals without periodontitis and with chronic periodontitis (CP) or generalized aggressive periodontitis (GAgP) and their association with the levels of key periodontal pathogens in subgingival plaque. Methods: GCF and subgingival plaque samples were obtained from healthy sites of participants without periodontitis (n = 20) and periodontitis sites of patients with CP (n = 22) and GAgP (n = 20). sTREM‐1 levels in GCF were measured by enzyme‐linked immunosorbent assay. Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans levels in subgingival plaque were analyzed by quantitative real‐time polymerase chain reaction. Results: sTREM‐1 levels in GCF were higher in CP and GAgP than healthy sites by 3.6‐ and 4.4‐fold, respectively, with no significant differences between the two forms of periodontitis. Moreover, sTREM‐1 levels in GCF were positively correlated with site‐specific clinical periodontal parameters and levels of P. gingivalis, T. denticola, and T. forsythia, but not A. actinomycetemcomitans, in subgingival plaque. Conclusion: Increased GCF levels of sTREM‐1 at diseased sites and their positive correlation with clinical and microbiologic parameters strengthen the association of this inflammatory marker with periodontitis.     

Association study between salivary levels of interferon (IFN)-gamma,interleukin (IL)-17, IL-21, and IL-22 with chronic periodontitis

Objective

To investigate if the salivary levels of IL-17, IL-21, IL-22, and its ratio regarding salivary IFN-γ may be linked with the periodontal clinical status.

Design

One hundred and five chronic periodontitis (CP) subjects and 44 healthy controls (HC) were recruited. Periodontal status was assessed based on full-mouth clinical periodontal measurements. Cytokine salivary levels were analyzed by ELISA. The association between the analytes with CP was analyzed using a binary logistic regression model.

Results

A statistically significant increase in salivary levels of IFN-γ and IFN-γ/IL-22 ratio in CP group could be detected, but there was no significant domination of any Th17 cytokine that could be of predictive value for health/disease status. Univariate and binary logistic regression analyses revealed a strong and independent association of IFN-γ salivary levels and IFN-γ/IL-22 ratio with disease status. An interaction effect of ageing on IFN-γ levels also could be noted.

Conclusion

While salivary levels of IFN-γ and IFN-γ/IL-22 ratio may act as strong/independent indicators of the amount and extent of periodontal breakdown, the low detection frequency of Th17 cytokines in saliva samples make these determinations useless for the detection of disease presence and/or its severity.     

Prognostic Value of 8‐Hydroxy‐2′‐Deoxyguanosine and Human Neutrophil Elastase/α1‐Proteinase Inhibitor Complex as Salivary Biomarkers of Oxidative Stress in Chronic Periodontitis

Background: 8‐Hydroxy‐2′‐deoxyguanosine (8‐OHdG) and human neutrophil elastase/α1‐proteinase inhibitor (HNE/α₁‐PI) complex have been regarded as reliable biomarkers of oxidative stress in inflammatory conditions. This study investigates whether the salivary levels of these two analytes may be linked with periodontal health status. Methods: One hundred ten patients with chronic periodontitis (CP) and 50 healthy controls were selected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, and extent and severity of periodontal breakdown. 8‐OHdG and HNE/α₁‐PI salivary levels were analyzed by enzyme‐linked immunosorbent assay. The association of these analytes with CP was analyzed individually and adjusted for confounding factors using a multivariate binary logistic regression model. Results: Significantly higher levels of both markers were detected in the CP group in comparison to controls. Weak‐to‐moderate positive significant correlations between salivary biomarkers and clinical parameters were observed. After binary logistic regression analysis, salivary levels of 8‐OHdG >17.35 ng/mL and HNE/α1‐PI complex >158.28 ng/mL were independently associated with disease status. Interaction effects among candidate prognostic variables were also noted. Conclusions: Increased salivary levels of 8‐OHdG and HNE/α₁‐PI complex may be strong, independent prognostic indicators of the amount and extent of oxidative stress–induced periodontal breakdown. In addition, unstimulated whole saliva samples might reflect a synergistic biologic interactive effect of HNE/α₁‐PI associated with the aging and smoking cumulative characteristics of periodontal damage.     

Comparative Analysis of Calcium‐Binding Myeloid‐Related Protein‐8/14 in Saliva and Serum of Patients With Periodontitis and Healthy Individuals

Background: This study aims to investigate calcium‐binding myeloid‐related protein (MRP)‐8/14 in the saliva and serum of individuals with periodontitis and periodontally healthy individuals for the assessment of its role in the pathogenesis and clinical diagnosis of periodontitis. Methods: This cross‐sectional study includes 56 patients with periodontitis and 44 periodontally healthy individuals. Saliva and serum were collected for the detection of MRP‐8/14 and calcium levels. Periodontopathic bacteria were determined by polymerase chain reaction in saliva. Correlations between salivary and serum MRP‐8/14 levels and clinical parameters, bacteria, and calcium were analyzed with Pearson correlation in a multiple regression model. MRP‐8/14 levels were documented with receiver operating characteristic (ROC) curves. Results: Compared with healthy individuals, MRP‐8/14 levels were significantly higher in both the saliva and serum of patients with periodontitis, but calcium was increased only in saliva. A high diagnostic potential of salivary MRP‐8/14 was detected for periodontitis (ROC = 0.86). Salivary MRP‐8/14 levels correlated significantly with the presence of the periodontopathogen Treponema denticola, as well as with the clinical parameters of periodontitis. Conclusion: MRP‐8/14 in saliva might be a potential diagnostic parameter for periodontal disease.     

Acute myocardial infarction elevates serine protease activity in saliva of patients with periodontitis

Mäntylä P, Buduneli E, Emingil G, Tervahartiala T, Pussinen PJ, Bar?? N, Ak?ll? A, Atilla G, Sorsa T. Acute myocardial infarction elevates serine protease activity in saliva of patients with periodontitis. J Periodont Res 2012; 47: 345–353. © 2011 John Wiley & Sons A/S Background and Objective: There are indications that acute myocardial infarction (AMI) may have an effect on the oral environment, which is reflected in the expression of salivary and gingival proteinases. According to our knowledge, no studies have been carried out to investigate the effect of AMI on the activities of two major tissue‐destructive serine protease and microbial effectors, elastase and cathepsin G, produced by oral fluid polymorphonuclear granulocytes (PMN). Therefore, we compared the activities of elastase and cathepsin G in saliva from patients with AMI and from systemically healthy subjects (non‐AMI) with similar periodontal conditions. Material and Methods: A total of 92 patients (47 AMI and 28 non‐AMI patients with gingivitis or periodontitis, and 17 systemically and periodontally healthy subjects as a control group) were recruited. Clinical periodontal measurements were recorded, and stimulated whole‐saliva samples were collected. The patients with AMI were clinically examined within 3–4 d after admission to the coronary care unit. The activities of saliva neutrophil elastase and cathepsin G were measured after collection, at specific time‐points during incubation (from baseline to 23 h) by specific synthetic peptide substrate assays. Results: The saliva of patients with AMI and periodontitis had a significant trend for the highest elastase activities among the study groups. Elastase and cathepsin G activities correlated significantly with each other in the AMI periodontitis group (r = 0.8, p < 0.01). In a logistic regression analysis, the level of salivary elastase activity associated significantly with periodontitis. Conclusion: AMI may be reflected in PMN serine protease elastase activity in saliva, despite its strong association with periodontitis.     

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.     

Elevated levels of salivary lactoferrin,a marker for chronic periodontitis?

Glimvall P, Wickström C, Jansson H. Elevated levels of salivary lactoferrin, a marker for chronic periodontitis? J Periodont Res 2012; 47: 655–660. © 2012 John Wiley & Sons A/S Background and Objective: Whole saliva is a complex mixture of fluids essential for the well‐being of the oral hard and soft tissues. Saliva contains numerous antimicrobial proteins that help protect the oral ecosystem from infectious agents. Chronic periodontitis is an infectious chronic inflammatory condition that affects the tooth‐supporting structures and leads to their destruction. The aim of the present study was to investigate differences in concentrations of salivary lactoferrin in subjects with and without periodontal disease and correlate these values with clinical variables associated with periodontal disease. Material and Methods: Stimulated whole saliva was collected from 17 subjects with chronic periodontitis and 17 periodontally healthy control subjects. Data relating to bleeding on probing, probing pocket depth and horizontal bone loss were registered. Concentrations of lactoferrin, lysozyme and IgA in stimulated whole saliva were quantified using ELISA. Results: Subjects with chronic periodontits showed higher concentrations of lactoferrin in stimulated whole saliva compared with periodontally healthy control subjects (p < 0.05). Salivary concentrations of lactoferrin were positively correlated with bleeding on probing (p < 0.001) and the number of sites with probing pocket depth ≥ 6 mm (p < 0.001). Conclusion: Lactoferrin is raised in stimulated whole saliva in subjects with chronic periodontitis and is correlated with probing pocket depth ≥ 6 mm.     

The Sociodemographic Characteristics,Periodontal Health Status,and Subgingival Microbiota of Patients With Chronic Periodontitis and Type 2 Diabetes Mellitus: A Case‐Control Study in a Chinese Population

Background: In China, chronic periodontitis (CP) is common in patients with type 2 diabetes mellitus (T2DM). The purpose of this study is to identify the sociodemographic characteristics associated with such patients and to assess the periodontal health status and subgingival microbiota of patients with CP and T2DM (T2DMCP) in the Chinese population. Methods: A total of 150 patients with T2DMCP and 306 patients with CP without any systemic disease completed questionnaires, underwent clinical periodontal examinations and participated in diabetes‐related parameter examinations. Subgingival plaques were obtained to determine the prevalence and amounts of selected oral bacterial species using polymerase chain reaction (PCR) and real‐time PCR, respectively. Results: The income level and mean body mass index (BMI) of the patients with T2DMCP were significantly higher than those of the patients with CP. Additionally, the patients with T2DMCP were more likely to be urban residents, and they had significantly more severe periodontitis than did the patients with CP. In the patients with T2DMCP, the prevalence and amounts of Treponema denticola and Tannerella forsythia were significantly higher than those in the patients with CP. Finally, compared with the patients with CP, the patients with T2DMCP had a significantly lower prevalence and amount of Prevotella intermedia. Conclusions: Compared with the patients with CP, the patients with T2DMCP were more likely to be urban residents and generally had higher incomes, higher mean BMI, and poorer periodontal health status. Higher levels of T. denticola and T. forsythia and lower levels of P. intermedia were identified in the subgingival plaque of the patients with T2DMCP.     

Trefoil factors in saliva and gingival tissues of patients with chronic periodontitis

Background: Trefoil factors (TFFs) are secreted molecules that are involved in cytoprotection against tissue damage and the immune response. TFFs have been detected in saliva and oral tissues, but their clinical significance has never been investigated in patients with chronic periodontitis. The objective of this study is to determine whether TFF expression in saliva and gingival tissues is associated with periodontal pathology. Methods: Saliva and gingival tissue samples were collected from 25 non‐periodontitis individuals and 25 patients with chronic periodontitis (CP). Enzyme‐linked immunosorbent assay and immunohistochemical methods were used to evaluate the expression of TFFs (TFF1, TFF2, and TFF3) in saliva and gingival tissues, respectively. Periodontopathic bacteria were quantified by real‐time polymerase chain reaction. Results: Reduced salivary TFF1 and TFF3 concentrations were observed in patients with CP (P = 0.003 and P <0.001, respectively). Decreased TFF3 expression in gingival tissues of patients with CP was demonstrated (P = 0.041). Levels of salivary TFF3 concentrations were negatively correlated with periodontal pathology and number of Porphyromonas gingivalis and Tannerella forsythia (formerly known as Bacteroides forsythus). Conclusions: Altered expression of TFFs in saliva and gingival tissues was detected in patients with CP. The results suggest that TFF3 may be involved in the pathogenesis of periodontal disease.     

TaqMan real‐time polymerase chain reaction assay for the correlation of Treponema denticola numbers with the severity of periodontal disease

Treponema denticola has been implicated in periodontitis, and the presence of this organism in periodontal pockets has been investigated. However, qualitative analysis is insufficient for the clinical evaluation of periodontal treatments, and quantification of T. denticola populations is essential for monitoring therapeutic efficacy. Therefore, we developed a quantitative method for T. denticola that uses the TaqMan real‐time polymerase chain reaction assay. Using this system, we evaluated the relative and absolute numbers of this organism in saliva and subgingival plaque. Furthermore, we analyzed the relationship between the numbers of T. denticola and pocket depth, and found a significant positive correlation (P < 0.0001) between these parameters. This report demonstrates the broad potential of real‐time polymerase chain reaction applications in periodontology.     

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.     

Chronic Periodontal Disease,Periodontal Pathogen Colonization,and Increased Risk of Precancerous Gastric Lesions

Background: This study assesses the association between periodontal pathogen colonization and the potential risk of developing precancerous lesions of gastric cancer (PLGC) in a clinical setting. Methods: Included were 35 newly diagnosed patients with PLGC and 70 age‐matched individuals without PLGC. A full‐mouth intraoral examination was performed to assess periodontal conditions. Stimulated whole saliva and pooled plaque samples were collected to evaluate colonization by Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans and to characterize oral microbial diversity in saliva and dental plaque. Results: Compared with the control group, patients with PLGC experienced higher prevalence of bleeding on probing (31.5% versus 22.4%; P <0.05), higher levels of T. denticola (P <0.01) and A. actinomycetemcomitans (P <0.01), and less bacterial diversity in their saliva (P <0.01). The final multivariate logistic regression model consisting of all key sociodemographic characteristics, oral health behavioral factors, and periodontal assessments revealed that elevated colonization with periodontal pathogens, specifically T. forsythia, T. denticola, and A. actinomycetemcomitans, decreased bacterial diversity in dental plaque, and not flossing teeth regularly was a significant predictor of increased risk of PLGC (P = 0.022). Conclusion: Findings of the present study provide new evidence suggesting that periodontal pathogen burdens and bacterial diversity in the oral cavity are important factors contributing to a potentially increased risk of developing precancerous gastric lesions.     

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.     

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.