Salivary Biomarkers Associated With Gingivitis and Response to Therapy

Background: Salivary biomarkers are potentially important for determining the presence, risk, and progression of periodontal disease. However, clinical translation of biomarker technology from lab to chairside requires studies that identify biomarkers associated with the transitional phase between health and periodontal disease (i.e., gingivitis). Methods: Eighty participants (40 with gingivitis, 40 healthy) provided saliva at baseline and 7 to 30 days later. An additional sample was collected from gingivitis participants 10 to 30 days after dental prophylaxis. Clinical parameters of gingival disease were recorded at baseline and the final visit. Salivary concentrations of interleukin (IL)‐1β, IL‐6, matrix metalloproteinase (MMP)‐8, macrophage inflammatory protein (MIP)‐1α, and prostaglandin E₂ (PGE₂) were measured. Results: Clinical features of health and gingivitis were stable at both baseline visits. Participants with gingivitis demonstrated significantly higher bleeding on probing (BOP), plaque index (PI), and gingival index (GI) (P ≤0.002) and a significant drop in BOP, PI, and GI post‐treatment (P ≤0.001). Concentrations of MIP‐1α and PGE₂ were significantly higher (2.8 times) in the gingivitis group than the healthy group (P ≤0.02). After dental prophylaxis, mean biomarker concentrations did not decrease significantly from baseline in the gingivitis group, although concentrations of IL‐1β, IL‐6, and MMP‐8 approached healthy levels, whereas MIP‐1α and PGE₂ concentrations remained significantly higher than in the healthy group (P ≤0.04). Odds ratio analyses showed that PGE₂ concentrations, alone and in combination with MIP‐1α, readily discriminated gingivitis from health. Conclusions: Salivary PGE₂ and MIP‐1α discriminate gingivitis from health, and patients with gingivitis who return to clinical health continue to produce inflammatory mediators for weeks after dental prophylaxis.     

Salivary cytokine levels in subjects with chronic periodontitis and in periodontally healthy individuals: a cross-sectional study

Background and Objective: Saliva has been proposed as a noninvasive diagnostic fluid that could be used in the diagnosis of oral and systemic diseases. The levels of salivary biomarkers, such as cytokines, could potentially be used as a surrogate to distinguish periodontally healthy individuals from subjects with periodontitis. Therefore, the goal of the present investigation was to determine if the levels of 10 different cytokines in saliva differed between a group of periodontally healthy individuals and a group of subjects with periodontitis. Correlations between the concentrations of these 10 cytokines and clinical parameters of periodontal disease were also examined. Material and Methods: In this cross‐sectional study, 74 subjects with chronic periodontitis and 44 periodontally healthy individuals were periodontally examined and had the levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐1β, interleukin‐2, interleukin‐4, interleukin‐5, interleukin‐6, interleukin‐8, interleukin‐10, interferon‐γ and tumor necrosis factor‐α measured in whole saliva using a multiplexed bead immunoassay (Luminex). Significance of statistical differences in the levels of salivary cytokines between groups was determined using nonparametric analysis of covariance, adjusting for age and smoking status. The Spearman rank correlation coefficient was used to explore associations between the mean levels of salivary cytokines and mean clinical parameters. Results: There were no statistically significant differences between groups for any of the cytokines. There were weak, statistically significant positive associations between salivary interleukin‐8 and pocket depth (r_s = 0.2, p < 0.05) and bleeding on probing (r_s = 0.2, p < 0.05), and weak negative correlations between salivary interleukin‐10 and attachment level (r_s = ?0.2, p < 0.05) and bleeding on probing (r_s = ?0.3, p < 0.001). Conclusion: Mean salivary levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐1β, interleukin‐2, interleukin‐4, interleukin‐5, interleukin‐6, interleukin‐8, interleukin‐10, interferon‐γ and tumor necrosis factor‐α could not discriminate between periodontal health and disease.     

Salivary Visfatin Concentrations in Patients With Chronic Periodontitis

Background: Visfatin, also known as pre‐B‐cell colony‐enhancing factor, is secreted from a variety of cells and is thought to have some proinflammatory and immunomodulating effects. It is indicated that serum/plasma levels of visfatin increase in a number of inflammatory disorders. The present study aims to evaluate salivary concentrations of visfatin in patients with chronic periodontitis (CP). Methods: Twenty patients with CP and 20 periodontally healthy individuals were enrolled in this study. For each patient, the values of clinical parameters, such as bleeding index, plaque index, probing depth, and clinical attachment level (CAL), were recorded. Whole saliva samples were collected, and concentrations of visfatin were evaluated using standard enzyme‐linked immunosorbent assay. Statistical analysis was performed using statistical software. Results: Visfatin was detectable in all samples. Salivary visfatin concentrations were significantly higher in the periodontitis group. In addition, there was a positive significant relationship between salivary visfatin concentrations and CAL in the periodontitis group. However, no significant association was observed between salivary visfatin levels and other periodontal parameters or body mass index. Conclusion: These results indicate that there is a relationship between salivary visfatin and CP; however, further studies are needed to confirm this finding.     

Evaluation of Salivary Procalcitonin Levels in Different Periodontal Diseases

Background: The present study aims to investigate the levels of salivary procalcitonin (ProCT) in patients with different periodontal diseases. Methods: Seventy‐two non‐smokers are included in this study: 21 individuals with chronic periodontitis (CP), 14 individuals with generalized aggressive periodontitis (GAgP), 18 individuals with gingivitis (G), and 19 periodontally healthy (H) participants. Clinical periodontal parameters, including probing depth (PD), clinical attachment level (CAL), plaque index, and gingival index (GI), were assessed in all participants. Saliva samples were collected and examined for evaluating ProCT levels. Results: It was found that the median (interquartile range) salivary ProCT level was lowest in the H group: 0.00 (0.09) ng/mL; followed by the G group: 0.09 (0.11) ng/mL; the CP group: 0.15 (0.29) ng/mL; and highest in the GAgP group 0.28 (0.68) ng/mL. These differences were statistically significant between the H group and the other groups (P <0.05). There were positive correlations between the mean salivary ProCT level and GI, CAL, and PD. Conclusion: According to the present results, ProCT might play a role during periodontal inflammation, and an elevated salivary ProCT level is suggested as a potential biomarker for periodontal diseases.     

Salivary osteocalcin levels are decreased in smoker chronic periodontitis patients

Oral Diseases (2011) 17 , 200–205 Objectives: This study was planned to investigate whether smoker chronic periodontitis patients exhibit different salivary concentrations of C‐telopeptide pyridinoline cross‐links of type I collagen (ICTP) and osteocalcin (OC) compared to the non‐smoker counterparts. Methods: Whole saliva samples, full‐mouth clinical periodontal recordings were obtained from 33 otherwise healthy chronic periodontitis patients and 36 systemically, periodontally healthy control subjects. Chronic periodontitis patients and healthy control subjects were divided into smoker and non‐smoker groups according to their self reports. Salivary ICTP, OC levels were determined by Enzyme‐linked Immunoassays. Results: Healthy control groups exhibited significantly lower values in all clinical periodontal measurements (P < 0.001). Smoker periodontitis patients revealed similar clinical periodontal index values with non‐smoker counterparts (P > 0.05). Chronic periodontitis patients exhibited significantly higher salivary OC levels than healthy controls (P < 0.05). Smoker periodontitis patients revealed lower salivary OC levels than non‐smoker counterparts (P < 0.001). Log ICTP levels in non‐smoker chronic periodontitis patients were higher than non‐smoker controls (P < 0.05). Smoker healthy control group revealed higher log ICTP levels than non‐smoker counterparts (P < 0.001). Conclusions: Within the limits of this study, it may be suggested that suppression of salivary osteocalcin level by smoking may at least partly explain the deleterious effects of smoking on periodontal status.     

Salivary malondialdehyde levels in clinically healthy and periodontal diseased individuals

Background: Lipid peroxidation (LPO) has been implicated in the pathogenesis of several pathologic disorders, including periodontal disease. Malondialdehyde (MDA) is one of many low molecular weight end products of LPO. Objective: This study was conducted to evaluate salivary MDA levels in generalized chronic periodontitis (GCP) subjects. Materials and methods: The MDA levels were measured in the saliva of 104 subjects, aged 18–65 years. Three groups with different degrees of severity of GCP were established: 30 early (group 1), 30 moderate (group 2) and 14 severe (group 3). Thirty individuals (aged 25–29 years) with clinically healthy periodontium were served as control. Unstimulated whole saliva samples from study subjects were collected, centrifuged at 3000 g for 15 min and were then stored at ?70°C until analysed. The MDA level was determined with 2‐thiobarbituric acid by a colorimetric method at 532 nm. Results: A significant increase in the MDA level existed in the samples obtained from the three groups of patients compared to the control subjects. Conclusion: Increased MDA levels are with closely associated with the severity and patients status of periodontal disease that has not been previously reported. The detection of salivary MDA level may provide additional advantages in elucidating the pathogenesis of periodontal disease.     

Salivary biomarkers: Relationship between oxidative stress and alveolar bone loss in chronic periodontitis

Objectives. Oxidative stress is implicated in the pathogenesis of many systemic and oral diseases such as periodontal disease. The main aim of this study is to explore a possible association between salivary markers of OS and alveolar bone loss. Materials and methods. The study included 20 patients with chronic periodontitis and 20 controls. Salivary OS biomarkers 8-hidroxy-desoxguanosine (8-HOdG), malondialdehyde (MDA), uric acid, total antioxidant capacity (TAC) and glutathione peroxidase (GPx) were evaluated. Bone loss markers such as C-terminal telopeptide of type I collagen (CTX I), matrix metalloproteinases-8 (MMP-8), osteocalcin and 25-hydroxy vitamin D₃ (25- OH D) were detected in this study. The methods included general biochemical tests and ELISA. Results. Salivary 8-OHdG, MDA levels were significantly higher in the chronic periodontitis group compared with controls (p < 0.05). Salivary activities for uric acid, TAC and GPx were significantly decreased in patients with chronic periodontitis vs controls (p < 0.05). Salivary levels for CTX I, MMP-8, 25-OH D and Osteocalcin were significantly higher in the chronic periodontitis group compared to the controls (p < 0.05). A significant positive correlation was observed between salivary levels of MDA and CTX I. Significant negative correlations between uric acid and CTX I and between MMP-8 and uric acid have been found. Significant positive correlations were observed between CTX I, MMP-8, 25-OH D, osteocalcin and clinical parameters of periodontal disease. Conclusions. Important oxidative stress associated with alveolar bone loss biomarkers can be detected in saliva of patients with periodontal disease.     

Salivary human beta-defensins and cathelicidin levels in relation to periodontitis and type 2 diabetes mellitus

Abstract

Objective: Type 2 diabetes mellitus (T2DM) is a well-defined risk factor of periodontitis and it can affect expression of human beta-defensins (hBDs) and cathelicidin (LL-37) as well. The aim of the present study was to evaluate the impact of periodontitis and T2DM on salivary concentrations of these antimicrobial peptides.

Material and methods: Unstimulated saliva samples, together with full-mouth periodontal recordings were collected from 92 individuals with periodontitis (63 with T2DM and 21 smokers) and 86 periodontally healthy controls (58 with T2DM and 21 smokers). Salivary hBD-1, -2, -3, LL-37, and advanced glycalization end products (AGE) concentrations were measured by enzyme-linked immunosorbent assay.

Results: Among the periodontitis patients, T2DM group demonstrated lower levels of hBD-1 (p?=?.006), hBD-2 (p?<?.001) and hBD-3 (p?<?.001), and higher levels of LL-37 (p?<?.001) compared to systemically healthy controls. When only periodontally healthy controls were included into the analysis, higher hBD-1 (p?=?.002) and LL-37 (p?<?.001) levels were found in T2DM patients in comparison to systemically healthy controls. Salivary LL-37 levels were associated with HbA1c and periodontitis, while hBD-2, hBD-3 and levels associated only with HbA1c.

Conclusion: In the limits of this study, hyperglycaemia can be proposed as a regulator of salivary hBD and cathelicidin levels. Periodontitis, on the other hand, affects only salivary LL-37 levels.     

Salivary interleukin-1β concentration and the presence of multiple pathogens in periodontitis

Aim: This study aimed to find salivary enzymes and/or cytokines that would reflect periodontitis, alone or in combination with salivary microbial markers.
Material and Methods: The salivary concentrations of elastase, lactate dehydrogenase, interleukin-1 β (IL-1 β ), interleukin-6, and tumour necrosis factor- α , and the presence of five periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia , and Treponema denticola , were analysed from salivary specimens of 165 subjects, a subpopulation of Health 2000 Health Examination Survey in Finland; 84 of the subjects had probing pocket depth (PPD) of 4 mm at 14 or more teeth (the advanced periodontitis group), while 81 subjects had no teeth with PPD of 4 mm (the control group). All subjects had at least 20 teeth and no systemic diseases.
Results: Among the salivary cytokines and enzymes tested, IL-1 β was the only biomarker associated with periodontitis. An association was also found with the presence of multiple periodontal pathogens. Salivary IL-1 β and the presence of multiple periodontal pathogens were associated with periodontitis at the same magnitude, when they were in the logistic regression model individually or together.
Conclusion: We suggest that salivary IL-1 β and the presence of multiple periodontal pathogens in saliva should be studied more thoroughly as markers of periodontitis.     

Salivary Stress Markers,Stress, and Periodontitis: A Pilot Study

Background: Some cross‐sectional and longitudinal studies attempted to link periodontitis with stress. To our knowledge, only one small study was published on the mechanism by which stress may influence periodontal diseases, suggesting behavioral and physiologic mechanisms and investigating the role of inflammation as a potential mediator. The present study is planned to explore the associations among periodontal disease, psychologic factors, and salivary markers of stress, psychoneuroimmunologic variables, and health behaviors. Methods: One hundred periodontitis patients were selected, and participants provided information on general health, chronic stress, and demographics. Stress markers (choromogranin A, cortisol, α‐amylase, and β‐endorphin) were measured from saliva. A dentist assessed the presence of dental plaque on lingual and buccal surfaces, the gingival index, and the number of remaining teeth with periodontal disease. Results: Stress and salivary stress markers were significantly correlated with clinical parameters of periodontal disease (ranging from 0.19 to 0.59; P <0.001). Neglecting to brush teeth during stress was associated with missing teeth. After adjusting for stress variables, salivary cortisol and β‐endorphin were significantly associated with tooth loss and periodontal clinical parameters (P <0.001; R² = 0.59). Conclusions: This study suggests that stress might be associated with periodontal disease through physiologic and behavioral mechanisms. In making diagnoses of psychiatric patients, the association between salivary stress markers and periodontal disease needs to be included. Further exploration of relationships between periodontitis and stress is warranted.     

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 blood group antigens and microbial flora

To cite this article:
Int J Dent Hygiene 9 , 2011; 117–121
DOI: 10.1111/j.1601‐5037.2010.00451.x
Tabasum ST, Nayak RP. Salivary blood group antigens and microbial flora. Abstract: Objective: Secretor is an individual with ability to produce blood group reactive substances in the exocrine glands and to secrete these substances in the body fluids such as saliva. In saliva the blood reactive antigens are found primarily on mucins which exhibit microheterogenicity and different subtypes of these molecules. This study was done to find out if the salivary blood group antigens have any role in the adherence of certain selected microorganism in the oral cavity. Methods: Unstimulated whole saliva and sub gingival plaque samples were collected from subjects with clinically healthy gingival, chronic gingivitis and chronic periodontitis with probing pocket depth ≥4 mm and attachment loss ≥1 mm. Secretor status was determined by using Haemagglutination Inhibition Assay. Unstimulated whole saliva and sub gingival plaque samples were collected to culture and isolate the selected microorganisms. The clinical scores, secretor status and the presence or absence of selected microorganisms were compared within the groups using Chi‐square test and students unpaired t‐test. Results: The numbers of secretors were more in the healthy group (22.2%) and non secretors were more in the chronic periodontitis group (22.2%). The clinical scores were higher in the in non secretors compared to the secretors in all the three groups. P intermedia and P gingivalis were prevalent among non secretors in chronic gingivitis group. (P = 0.075 and P = 0.032) and chronic periodontitis group (P = 0.068 and P = 0.009).     

Salivary lactoferrin and low-Mr mucin MG2 in Actinobacillus actinomycetemcomitans-associated periodontitis

Concentrations and output of lactoferrin and of low-Mr mucin MG2 were determined in saliva of subjects suffering from Actinobacillus actinomycetemcomitans-associated periodontal disease and healthy subjects. Periodontal patients were clinically examined and a microbiological sample was taken from the deepest bleeding pockets in each quadrant. The number of viable A. actinomycetemcomitans was determined in the sampled sites of each patient. The MG2 output in the diseased subjects (13.6 microg protein/min) was decreased at least by a factor three compared to periodontal healthy subjects (44.3 microg protein/min). On the other hand, output of lactoferrin was not significantly different in healthy (9.5 microg/min) and diseased subjects (7.6 microg/min). Western analyses demonstrated a higher iron-saturation of lactoferrin in diseased subjects in comparison with the healthy subjects. Lactoferrin degrading enzymes, probably derived from microbial sources, could be detected in saliva of the periodontally diseased subjects, but not in saliva of healthy subjects. The combination of iron-saturation and degradation of lactoferrin suggests that anti-microbial properties of lactoferrin are diminished in periodontitis patients. Moreover, the low concentration of mucin MG2 suggests a decline in mucin defence and consequently a higher susceptibility for oral infection. A negative correlation (r= -0.4, p < 0.05) between the number of subgingival A. actinomycetemcomitans and lactoferrin in saliva suggested that low concentrations of lactoferrin favour the growth of the bacterium. These data indicate that a decline in the salivary defence system might increase the risk for oral infection by A. actinomycetemcomitans.     

Salivary and Serum Markers Related to Innate Immunity in Generalized Aggressive Periodontitis

Background: Host inflammatory and immune responses play an important role in aggressive periodontitis (AgP). Thus, this study aims to evaluate levels of the innate immunity‐related markers calprotectin, colony‐stimulating factor (CSF)‐1, macrophage migration inhibitory factor (MIF), monokine induced by interferon‐γ (MIG), and matrix metalloproteinase (MMP)‐8 in serum and saliva from patients with generalized AgP and those with gingivitis or a healthy periodontium. Methods: This study enrolled 40 individuals (17 males and 23 females; mean age 33.30 ± 9.31 years), 15 with generalized AgP, 15 with gingivitis, and 10 who were periodontally healthy. Full‐mouth periodontal examinations were performed, and serum and saliva were collected. Levels of calprotectin, CSF‐1, MIF, MIG, and MMP‐8 were measured using enzyme‐linked immunosorbent assays. Results: In serum, mean levels of calprotectin were 2.06‐fold higher in patients with AgP than in healthy patients (P = 0.01). Serum levels of MMP‐8 were significantly elevated in patients with AgP compared with both healthy patients and those with gingivitis, by 2.60‐fold and 2.77‐fold, respectively (P = 0.03 and P = 0.009, respectively). In saliva, levels of MMP‐8 were 5.66‐fold higher in patients with AgP than in healthy patients (P = 0.02). CSF‐1, MIF, and MIG levels in both serum and saliva did not differ significantly among the groups. Conclusions: Serum levels of calprotectin and MMP‐8 are elevated in patients with AgP. MMP‐8 levels are also increased in saliva from patients with AgP. These results support involvement of innate immune response in the pathogenesis of AgP.     

Salivary and Serum Leptin Concentrations in Patients With Chronic Periodontitis

Background: The discovery of leptin has led to the elucidation of a robust physiologic system that not only maintains fat stores but is also an integral part of the host defense mechanism. However, leptin concentrations in the saliva of patients with chronic periodontitis (CP) has not been explored despite the potential role of salivary biomarkers in determining the presence, risk, and progression of periodontal disease. Methods: Eighty‐four participants (44 with generalized severe CP and 40 without periodontitis) were enrolled. For each patient, the values of periodontal parameters were recorded, such as plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment loss (AL), and percentage of sites with bleeding on probing (BOP) and clinical AL ≥5 mm. Saliva and serum samples were collected to estimate the leptin concentrations using enzyme‐linked immunosorbent assay kits. Statistical analysis was performed using software. Results: Participants with CP demonstrated significantly higher BOP, PI, GI, and percentage of sites with clinical AL >5 mm (P <0.05). Leptin was detectable in all the clinical samples. Salivary leptin concentrations in patients with CP were significantly lower than in healthy volunteers (6,200.61 ± 2,322.11 versus 8,799.60 ± 901.70 pg/mL), whereas serum leptin concentrations were significantly higher in patients with CP than in healthy volunteers (11,600.00 ± 1,705.01 versus 7,616.62 ± 1,169.83 pg/mL). In addition, the results reflected a significant negative correlation of salivary leptin and a positive correlation of serum leptin with PD (P <0.05). Conclusion: The results suggest that leptin concentrations in saliva and serum are significantly altered in CP and relate closely to current disease activity; however, further studies are needed to confirm the findings.     

Salivary IgA responses to bacteria in dental plaque as related to periodontal and HIV infection status

Levels of total IgA and specific IgA reactive with Streptococcus mutans, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Fusobacterium nucleatum were measured by ELISA in parotid saliva from HIV⁺ and HIV⁻ persons with healthy gingiva (HG), chronic gingivitis, chronic marginal periodontitis (CMP), or neerotizing ulcerative periodontitis (NUP). When the HIV+ group was compared with the HIV⁻ group regardless of periodontal status, total salivary IgA concentration was higher in HTV⁺ patients, but no such difference was observed for total IgA output. HIV⁺ CMP displayed higher total IgA concentration as compared with HIV⁻ CMP. No significant differences in specific IgA outputs and ratios were detected between HTV⁺ and HIV⁻ subgroups with similar periodontal status. HIV⁻ NUP displayed increased specific IgA output towards S. mutans and increased specific IgA ratio values towards S. mutans, P. gingivalis and P. nigrescens as compared with HIV⁺ CMP, and increased specific IgA ratio values towards S. mutans and P. nigrescens as compared with HIV⁺ HG. No such differences were observed between the HIV⁻ subgroups. In sum. salivary IgA responses to bacteria in dental plaque seem not to be related to chronic periodontal disease and HIV infection, but are possibly influenced by acute periodontal infection.