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In the era of personalized/precision health care, additional effort is being expended to understand the biology and molecular mechanisms of disease processes. How these mechanisms are affected by individual genetics, environmental exposures, and behavioral choices will encompass an expanding role in the future of optimally preventing and treating diseases. Considering saliva as an important biological fluid for analysis to inform oral disease detection/description continues to expand. This review provides an overview of saliva as a diagnostic fluid and the features of various biomarkers that have been reported. We emphasize the use of salivary biomarkers in periodontitis and transport the reader through extant literature, gaps in knowledge, and a structured approach toward validating and determine the utility of biomarkers in periodontitis. A summation of the findings support the likelihood that a panel of biomarkers including both host molecules and specific microorganisms will be required to most effectively identify risk for early transition to disease, ongoing disease activity, progression, and likelihood of response to standard periodontal therapy. The goals would be to develop predictive algorithms that serve as adjunctive diagnostic tools which provide the clinician and patient important information for making informed clinical decisions., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To examine the influence of acute stress on salivary flow using a validated stressor paradigm., Study Design: This uniform crossover study consisted of 40 healthy adults who underwent the Trier Social Stress Test, consisting of a 5-minute mental arithmetic task (MAT), and a nonstressful task (NST), consisting of a 5-minute free speech task. The order of the tasks was counterbalanced and unstimulated whole saliva (UWS) was measured in 2 groups of 20 participants during each 5-minute task condition, with a 10-minute washout period between tasks. At baseline, mathematical ability was self-reported and psychological distress was measured using the Symptom Checklist-90-Revised. Heart rate (HR) and breathing rate (BR) were recorded during each task., Results: Age, sex, HR, BR, and psychological distress were similar between groups at baseline (P > .05). During the MAT, HR increased significantly and mean UWS flow rate decreased significantly compared with the NST (P < .001)., Conclusions: An acute psychobiological stressor task was associated with a rapid decrease in salivary flow in adults. Thus, stress can contribute to reduced salivary flow and should be considered as a factor during the diagnostic workup of patients who complain of a dry mouth., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Aims: To describe correlations and agreement between salivary and serum B-type natriuretic peptide (BNP), C-reactive protein (CRP), interleukin (IL)-6, and IL-10 and determine which biomarkers predict worse functional class in patients with heart failure (HF)., Methods: Serum and saliva were collected from 75 hospitalized patients with HF (57 ± 12 years, 43% female, New York Heart Association [NYHA] Classes I [4%], II [43%], and III [53%]). Oral inflammation was rated as good, fair, or poor. Spearman's ρ and Bland-Altman were used to determine correlations and agreement of the salivary and serum forms of each biomarker. Logistic regressions were used to determine which biomarkers predicted worse NYHA functional class, controlling for depression, body mass index, smoking, and oral inflammation., Results: Median biomarker concentrations were as follows: BNP (serum 361 pg/ml, saliva 9 pg/ml), CRP (serum 13 ng/ml, saliva 25.6 ng/ml), IL-6 (serum 19.3 pg/ml, saliva 10.5 pg/ml), and IL-10 (serum 64.1 pg/ml, saliva 4.7 pg/ml). There was a moderate-to-strong correlation for serum-salivary CRP, weak correlation for serum-salivary IL-6, and no correlations for serum-salivary BNP and IL-10. The Bland-Altman test showed good salivary-serum agreement for all biomarkers, but as serum concentrations rose, salivary measures underestimated serum levels. Visible oral inflammation was the only predictor of worse NYHA class.

This study investigates the use of saliva, as an emerging diagnostic fluid in conjunction with classification techniques to discern biological heterogeneity in clinically labelled gingivitis and periodontitis subjects (80 subjects; 40/group) A battery of classification techniques were investigated as traditional single classifier systems as well as within a novel selective voting ensemble classification approach (SVA) framework. Unlike traditional single classifiers, SVA is shown to reveal patient-specific variations within disease groups, which may be important for identifying proclivity to disease progression or disease stability. Salivary expression profiles of IL-1ß, IL-6, MMP-8, and MIP-1α from 80 patients were analyzed using four classification algorithms (LDA: Linear Discriminant Analysis [LDA], Quadratic Discriminant Analysis [QDA], Naïve Bayes Classifier [NBC] and Support Vector Machines [SVM]) as traditional single classifiers and within the SVA framework (SVA-LDA, SVA-QDA, SVA-NB and SVA-SVM). Our findings demonstrate that performance measures (sensitivity, specificity and accuracy) of traditional classification as single classifier were comparable to that of the SVA counterparts using clinical labels of the samples as ground truth. However, unlike traditional single classifier approaches, the normalized ensemble vote-counts from SVA revealed varying proclivity of the subjects for each of the disease groups. More importantly, the SVA identified a subset of gingivitis and periodontitis samples that demonstrated a biological proclivity commensurate with the other clinical group. This subset was confirmed across SVA-LDA, SVA-QDA, SVA-NB and SVA-SVM. Heatmap visualization of their ensemble sets revealed lack of consensus between these subsets and the rest of the samples within the respective disease groups indicating the unique nature of the patients in these subsets. While the source of variation is not known, the results presented clearly elucidate the need for novel approaches that accommodate inherent heterogeneity and personalized variations within disease groups in diagnostic characterization. The proposed approach falls within the scope of P4 medicine (predictive, preventive, personalized, and participatory) with the ability to identify unique patient profiles that may predict specific disease trajectories and targeted disease management.

Unlabelled: Generally, clinical parameters are used in dental practice for periodontal disease, yet several drawbacks exist with the clinical standards for addressing the needs of the public at large in determining the current status/progression of the disease, and requiring a significant amount of damage before these parameters can document disease. Therefore, a quick, easy and reliable method of assessing and monitoring periodontal disease should provide important diagnostic information that improves and speeds treatment decisions and moves the field closer to individualized point-of-care diagnostics., Objective: This report provides results for a saliva-based diagnostic approach for periodontal health and disease based upon the abundance of salivary analytes coincident with disease, and the significant progress already made in the identification of discriminatory salivary biomarkers of periodontitis., Methods: We evaluated biomarkers representing various phases of periodontitis initiation and progression (IL-1ß, IL-6, MMP-8, MIP-1α) in whole saliva from 209 subjects categorized with periodontal health, gingivitis, and periodontitis., Results: Evaluation of the salivary analytes demonstrated utility for individual biomarkers to differentiate periodontitis from health. Inclusion of gingivitis patients into the analyses provided a more robust basis to estimate the value of each of these analytes. Various clinical and statistical approaches showed that pairs or panels of the analytes were able to increase the sensitivity and specificity for the identification of disease., Conclusions: Salivary concentrations of IL-1ß, IL-6, MMP-8, MIP-1α alone and in combination are able to distinguish health from gingivitis and periodontitis. The data clearly demonstrated a heterogeneity in response profiles of these analytes that supports the need for refinement of the standard clinical classifications if we are to move toward precision/personalized dentistry for the twenty-first century.

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.

Objectives: Factors influencing xerostomia during intensity-modulated radiation therapy (IMRT) were assessed., Methods: A 6-week study of 32 head and neck cancer (HNC) patients was performed. Subjects completed the Xerostomia Inventory (XI) and provided stimulated saliva (SS) at baseline, week 2 and at end of IMRT. Influence of SS flow rate (SSFR), calcium and mucin 5b (MUC5b) concentrations and radiation dose on xerostomia was determined., Results: HNC subjects experienced mean SSFR decline of 36% by visit 2 (N = 27; P = .012) and 57% by visit 3 (N = 20; P = .0004). Concentrations of calcium and MUC5b increased, but not significantly during IMRT (P > .05). Xerostomia correlated most with decreasing salivary flow rate as determined by Spearman correlations (P < .04) and linear mixed models (P < .0001)., Conclusions: Although IMRT is sparing to the parotid glands, it has an early effect on SSFR and the constituents in saliva in a manner that is associated with the perception of xerostomia., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Salivary biomarker discovery requires identification of analytes with high discriminatory capacity to distinguish disease from health, including day-to-day variations that occur in analyte levels. In this study, seven biomarkers associated with inflammatory and tissue destructive processes of periodontal disease were investigated. In a prospective cohort study design, analyte expression levels were determined in unstimulated whole saliva samples collected on multiple occasions from 30 healthy adults (i.e., orally and systemically) and 50 chronic adult periodontitis patients. Salivary levels of IL-1β, IL-6, MMP-8, and albumin were significantly elevated (5.4 to 12.6X) and levels of IFNα were consistently lower (8.7X) in periodontitis patients compared with the daily variation observed in healthy adults. ROC analyses of IL-1β, IL-6 and MMP-8 yielded areas under the curves of 0.963-0.984 for discriminating periodontitis from health. These results demonstrate that levels of salivary bioanalytes of patients who have periodontitis are uniquely different from normal levels found in healthy subjects, and a panel consisting of IL-1β, MMP-8 and IL-6 shows particular diagnostic potential.

Objective: To determine if salivary biomarkers demonstrate utility for identifying aspects of myocardial necrosis., Methods: Twenty-one patients undergoing alcohol septal ablation (ASA) for treatment of hypertrophic cardiomyopathy provided serum and unstimulated whole saliva at baseline and incremental time points post-ASA. Samples were analyzed for seven biomarkers related to myocardial damage, inflammation, and tissue remodeling using immunosorbent assays. Levels were compared with baseline and levels observed in 97 healthy controls., Results: Biomarkers of myocardial damage and inflammation (ie, troponin I, creatine kinase-MB, myoglobin, C-reactive protein) rose in serum 2- to 812-fold after ASA (P < .01). Significant elevations of 2.0- to 3.5-fold were observed with C-reactive protein and troponin I in saliva (P < .02). Significant correlations between levels in serum and saliva were observed for C-reactive protein, matrix metalloproteinase-9, and myeloperoxidase (P < .001)., Conclusions: Select salivary biomarkers reflect changes that occur during, and subsequent to, myocardial necrosis caused by ASA., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: The aim of this study was to determine the utility of oral fluids for assessment of coronary and cardiovascular (CV) health., Study Design: Twenty-nine patients with preexisting CV disease underwent an invasive cardiac procedure (alcohol septal ablation or percutaneous coronary intervention) and provided unstimulated whole saliva (UWS), sublingual swabs (LS), gingival swabs (GS) and serum at 0, 8, 16, 24, and 48 hours. Concentrations of 13 relevant biomarkers were determined and correlated with levels in serum and the oral fluids., Results: Concentrations of the majority of biomarkers were higher in UWS than in LS and GS. Coronary and CV disease biomarkers in UWS correlated better with serum than with LS and GS based on group status and measures of time effect. Seven biomarkers demonstrated time effect changes consistent with serum biomarkers, including C-reactive protein and troponin I., Conclusions: Changes in serum biomarker profiles are reflected in oral fluids suggesting that oral fluid biomarkers could aid in the assessment of cardiac ischemia/necrosis., (Copyright © 2012 Mosby, Inc. All rights reserved.)

Background: Salivary biomarkers of periodontitis were assessed longitudinally to determine response to therapy., Methods: A 6-month case-controlled study of adults with chronic periodontitis was performed, with 33 participants receiving oral hygiene instructions (OHI) alone and 35 with scaling and root planing (SRP) combined with OHI. Saliva samples collected at week 0, 16 and 28 were analysed for interleukin (IL)-1β, IL-8, macrophage inflammatory protein (MIP)-1α, matrix metalloproteinase-8 (MMP-8), osteoprotegerin (OPG), and tumour necrosis factor-α (TNF)-α. Clinical measures of periodontal disease were recorded at each visit., Results: All parameters of periodontal health improved significantly in both groups by week 16 (p<0.0001) with the SRP group demonstrating greater benefit at week 16 and 28. Baseline OPG and TNF-α levels changed significantly at both follow-up visits (p<0.03), regardless of treatment group. IL-1β and MMP-8 levels decreased significantly from baseline (p<0.04) in the SRP group only. OPG, MMP-8, and MIP-1α were significantly reduced in responders compared with non-responders (p=0.04, 0.01, 0.05, respectively). In receiver-operating characteristic analyses, MMP-8 produced the highest area under the curve (0.7; p=0.01)., Conclusion: Salivary levels of IL-1β, MMP-8, OPG, and MIP-1α reflected disease severity and response to therapy suggesting their potential utility for monitoring periodontal disease status., (© 2011 John Wiley & Sons A/S.)

Aim: To test the hypothesis that rheumatoid arthritis (RA) influenced levels of salivary biomarkers of periodontal disease., Methods: Medical assessments, periodontal examinations and pain ratings were obtained from 35 RA, 35 chronic periodontitis and 35 age- and gender-matched healthy controls in a cross-sectional, case-controlled study. Unstimulated whole saliva samples were analysed for interleukin-1β (IL-1β), matrix metalloproteinase-8 (MMP-8) and tumour necrosis factor-α (TNF-α) concentrations., Results: The arthritis and healthy groups had significantly less oral disease than the periodontitis group (P<0.0001), with the arthritis group having significantly more sites bleeding on probing (BOP) than matched controls (P=0.012). Salivary levels of MMP-8 and IL-1β were significantly elevated in the periodontal disease group (P<0.002), and IL-1β was the only biomarker with significantly higher levels in the arthritis group compared with controls (P=0.002). Arthritis patients receiving anti-TNF-α antibody therapy had significantly lower IL-1β and TNF-α levels compared with arthritis patients not on anti-TNF-α therapy (P=0.016, 0.024) and healthy controls (P<0.001, P=0.011), respectively., Conclusion: RA patients have higher levels of periodontal inflammation than healthy controls, i.e., an increased BOP. Systemic inflammation appears to influence levels of select salivary biomarkers of periodontal disease, and anti-TNF-α antibody-based disease-modifying therapy significantly lowers salivary IL-1β and TNF-α levels in RA., (© 2010 John Wiley & Sons A/S.)

Saliva can be easily obtained in medical and non-medical settings, and contains numerous bio-molecules, including those typically found in serum for disease detection and monitoring. In the past two decades, the achievements of high-throughput approaches afforded by biotechnology and nanotechnology allow for disease-specific salivary biomarker discovery and establishment of rapid, multiplex, and miniaturized analytical assays. These developments have dramatically advanced saliva-based diagnostics. In this review, we discuss the current consensus on development of saliva/oral fluid-based diagnostics and provide a summary of recent research advancements of the Texas-Kentucky Saliva Diagnostics Consortium. In the foreseeable future, current research on saliva based diagnostic methods could revolutionize health care.

Salivary diagnostics is an emerging field that has progressed through several important developments in the past decade, including the publication of the human salivary proteome and the infusion of federal funds to integrate nanotechnologies and microfluidic engineering concepts into developing compact point-of-care devices for rapid analysis of this secretion. In this article, we discuss some of these developments and their relevance to the prognosis, diagnosis and management of periodontitis, as an oral target, and cardiovascular disease, as a systemic example for the potential of these biodiagnostics. Our findings suggest that several biomarkers are associated with distinct biological stages of these diseases and demonstrate promise as practical biomarkers in identifying and managing periodontal disease, and acute myocardial infarction. The majority of these studies have progressed through biomarker discovery, with the identified molecules requiring more robust clinical studies to enable substantive validation for disease diagnosis. It is predicted that with continued advances in this field the use of a combination of biomarkers in multiplex panels is likely to yield accurate screening tools for these diagnoses in the near future.

Objectives: Human herpesviruses, e.g., herpes simplex virus (HSV) type 1, Epstein-Barr virus, and cytomegalovirus, appear in saliva at greater frequency in persons infected with human immunodeficiency virus (HIV) than in healthy individuals. However, it is not known if varicella zoster virus (VZV) and HSV-2 appear simultaneously during HIV infection at greater frequency in saliva in this era of highly active antiretroviral therapy (HAART). The aim of this study was to investigate the prevalence and amounts of VZV and HSV-2 in the saliva of HIV-infected orally asymptomatic patients., Study Design: Quantitative polymerase chain reaction was used to investigate the prevalence, quantity, risk, and correlations of salivary VZV and HSV-2 from 59 HIV-seropositive individuals and 53 healthy control subjects in a case-control cross-sectional study. Seventy-eight percent of the HIV-seropositive patients (46 out of 59) were taking HAART., Results: VZV DNA was detected in the saliva of 5.1% (3 out of 59) of the HIV-positive group and in only 1 healthy control 1.9% (1 out of 53; P = .62). The amount of VZV DNA in the expressors was low, generally <1,100 copies/mL, with no observed difference between the HIV-positive group and the control subjects (P = 1.0). HSV-2 DNA was not detected in either group. In the HIV-infected group, VZV shedding occurred in those on HAART, but was not associated with oral lesions, specific CD4(+) or CD8(+) T-cell levels, or demographic factors., Conclusions: Varicella zoster virus was detected at low prevalence in the saliva of HIV-infected persons, whereas HSV-2 was not detected in the saliva of this cohort. HAART does not appear to diminish the risk for asymptomatic VZV shedding., (Copyright (c) 2010 Mosby, Inc. All rights reserved.)

Background: For adults with chest pain, the electrocardiogram (ECG) and measures of serum biomarkers are used to screen and diagnose myocardial necrosis. These measurements require time that can delay therapy and affect prognosis. Our objective was to investigate the feasibility and utility of saliva as an alternative diagnostic fluid for identifying biomarkers of acute myocardial infarction (AMI)., Methods: We used Luminex and lab-on-a-chip methods to assay 21 proteins in serum and unstimulated whole saliva procured from 41 AMI patients within 48 h of chest pain onset and from 43 apparently healthy controls. Data were analyzed by use of logistic regression and area under curve (AUC) for ROC analysis to evaluate the diagnostic utility of each biomarker, or combinations of biomarkers, in screening for AMI., Results: Both established and novel cardiac biomarkers demonstrated significant differences in concentrations between patients with AMI and controls without AMI. The saliva-based biomarker panel of C-reactive protein, myoglobin, and myeloperoxidase exhibited significant diagnostic capability (AUC = 0.85, P < 0.0001) and in conjunction with ECG yielded strong screening capacity for AMI (AUC = 0.96) comparable to that of the panel (brain natriuretic peptide, troponin-I, creatine kinase-MB, myoglobin; AUC = 0.98) and far exceeded the screening capacity of ECG alone (AUC approximately 0.6). En route to translating these findings to clinical practice, we adapted these unstimulated whole saliva tests to a novel lab-on-a-chip platform for proof-of-principle screens for AMI., Conclusions: Complementary to ECG, saliva-based tests within lab-on-a-chip systems may provide a convenient and rapid screening method for cardiac events in prehospital stages for AMI patients.

Background: Saliva contains a large number of biomolecules, some of which have putative diagnostic usefulness. A potential problem with the use of biomolecules in diagnosis is day-to-day fluctuation due to within-subject variability. This study evaluated the intraindividual variability of six salivary analytes in healthy adults and determined their normal range., Methods: Unstimulated whole saliva (5 ml) was collected every 2 to 3 days on six occasions from 30 subjects in good oral and systemic health. Four of the samples were collected in the clinic, and two were collected by the subject at home. The concentration ranges of interleukin (IL)-1beta, IL-6, matrix metalloproteinase-8, prostaglandin E(2), tumor necrosis factor-alpha, interferon-alpha, and albumin were examined. Descriptive statistics were computed, and a one-way random-effects model was used to quantify within- and between-subject components of variability. Intraclass correlation coefficients (ICCs) were calculated for each subject/analyte combination., Results: Within-subject coefficients of variation for these analytes ranged from 67.6% to 172.1% for the in-clinic samples and from 111.9% to 201.0% for the at-home samples. The ICC for the various analytes ranged from 41% to 61% for the in-clinic samples. The at-home samples exhibited significantly more variability than did those obtained in the clinic under supervision., Conclusions: There was marked within-subject variation in the salivary concentrations of these analytes. With increased interest in salivary diagnostics, the within-subject variability, normal range, and threshold levels for abnormal levels of individual salivary analytes need to be determined if these diagnostics tests are to have clinical usefulness.

Background: Tumor necrosis factor-alpha (TNF-alpha), C-telopeptide pyridinoline cross-links of type I collagen (ICTP), and receptor activator of nuclear factor-kappa B ligand (RANKL) have been associated with bone remodeling and periodontal tissue destruction. This study evaluated the level of these biomarkers in saliva with respect to periodontal disease status., Methods: Levels of TNF-alpha in unstimulated whole saliva of 74 adults (35 subjects with moderate to severe periodontal disease and 39 healthy controls) and salivary levels of RANKL and ICTP of a subset of 21 subjects and 21 matched controls were examined using enzyme immunosorbent assays in a case-control clinical study., Results: Salivary levels of TNF-alpha were detected in all subjects, whereas levels of ICTP and RANKL were detected in only a minority of subjects. Mean salivary levels of TNF-alpha were significantly higher in individuals with periodontal disease (mean: 4.33 pg/ml) than in controls (mean: 2.03 pg/ml; P = 0.02), with a maximum level (27.96 pg/ml) observed in periodontitis. Subjects with salivary TNF-alpha levels above a threshold of 5.75 pg/ml (i.e., two standard deviations above the mean of the controls) had significantly more sites with bleeding on probing, probing depths >/=4 mm, and attachment loss > or =2 mm (P < or =0.01)., Conclusion: Salivary levels of TNF-alpha were elevated in patients who had clinical indicators of periodontitis, suggesting that this biomarker may serve in a panel of salivary biomarkers that could facilitate the screening, diagnosis, and management of periodontal disease.

Salivary secretions contain a variety of molecules that reflect important pathophysiological activities. Quantitative changes of specific salivary biomarkers could have significance in the diagnosis and management of both oral and systemic diseases. Modern point-of-care technologies with enhanced detection capabilities are needed to implement a significant advancement in salivary diagnostics. One such promising technology is the recently described lab-on-a-chip (LOC) assay system, in which assays are performed on chemically sensitized beads populated into etched silicon wafers with embedded fluid handling and optical detection capabilities. Using this LOC system, complex assays can be performed with small sample volumes, short analysis times, and markedly reduced reagent costs. This report describes the use of LOC methodologies to assess the levels of interleukin-1beta (IL-1beta), C-reactive protein (CRP), and matrix metalloproteinase-8 (MMP-8) in whole saliva, and the potential use of these biomarkers for diagnosing and categorizing the severity and extent of periodontitis. This study demonstrates that the results achieved by the LOC approach are in agreement with those acquired with standard enzyme-linked immunosorbent assay (ELISA), with significant IL-1beta and MMP-8 elevations in whole saliva of periodontitis patients. Furthermore, because of the superior detection capacities associated with the LOC approach, unlike those with ELISA, significant differences in CRP levels between periodontitis patients and normal subjects are observed. Finally, principal component analysis (PCA) is performed to yield an efficient method to discriminate between periodontally healthy and unhealthy patients, thus increasing the diagnostic value of these biomarkers for periodontitis when examined with the integrated LOC sensor system.

Human immunodeficiency virus (HIV) infection is associated with an increased risk for human herpesviruses (HHVs) and their related diseases. Methods for limiting the transmission of HHVs require a better understanding of the prevalence and infectiousness of oral HHVs in HIV-infected patients. We performed quantitative PCR to investigate the prevalence, quantity, risk, and correlates of salivary HHVs from 58 HIV-seropositive individuals in a case control study. HHVs were significantly more prevalent in the salivas of HIV-seropositive persons than in those of the controls (odds ratios [ORs], 4.2 to 26.2; P

Background: The authors conducted a study to determine if salivary biomarkers specific for three aspects of periodontitis--inflammation, collagen degradation and bone turnover--correlate with clinica features of periodontal disease., Methods: The relationship between periodontal disease and the levels of interleukin-1 beta (IL-1beta), matrix metalloproteinase (MMP)-8, and osteoprotegerin (OPG) in whole saliva of 57 adults (28 "case" subjects with moderate-to-severe periodontal disease and 29 healthy control subjects) was examined in a case-control trial., Results: Mean levels of IL-1beta and MMP-8 in saliva were significantly higher in case subjects than in controls. Both analytes correlated with periodontal indexes, whereas, after adjustment for confounders, OPG did not. Elevated salivary levels of MMP-8 or IL-1beta (more than two standard deviations above the mean of the controls) significantly increased the risk of periodontal disease (odds ratios in the 11.3-15.4 range). Combined elevated salivary levels of MMP-8 and IL-1beta increased the risk of experiencing periodontal disease 45-fold, and elevations in all three biomarkers correlated with individual clinical parameters indicative of periodontal disease., Conclusion: Salivary levels of MMP-8 and IL-1beta appear to serve as biomarkers of periodontitis., Clinical Implications: Qualitative changes in the composition of salivary biomarkers could have significance in the diagnosis and treatment of periodontal disease.

Human herpesviruses (HHVs) are ubiquitous pathogens that intermittently reactivate from latency. Transmission is believed to be facilitated by their frequent appearance in saliva. This study sought to understand the factors that influence the appearance of these viruses in saliva by examining the prevalence, pattern, and quantity of all eight HHVs in saliva of immunocompetent adults with a history of recurrent oral herpes simplex virus (HSV) infections following dental treatment and antiviral therapy. Valacyclovir or matched placebo was given (2 g twice on the day of treatment and 1 g twice the following day) to 125 patients in a randomized, double-blind controlled trial. Saliva, collected on the day of dental treatment and 3 and 7 days later, was analyzed using real-time quantitative PCR. At all visits, HHVs coinfected saliva. Over the course of the week, the DNAs of HHV-6 and HHV-7 were detected significantly more often (97% to 99% of patients) than Epstein-Barr virus (EBV; 64.8%), HSV-1 (13.0%), HHV-8 (3.2%), cytomegalovirus (2.4%), HSV-2 (0%), and varicella-zoster virus (0%), irrespective of drug treatment (P < 0.002). Mean genome copy numbers were highest for HSV-1 and HHV-6. Dental treatment did not influence asymptomatic viral shedding patterns. However, valacyclovir treatment resulted in significantly fewer patients shedding EBV at both postoperative visits compared with placebo (P < 0.008). These results suggest that HHVs are simultaneously present in the saliva of healthy adults at levels that could facilitate transmission, and valacyclovir therapy decreases the prevalence of EBV in saliva but has little effect on HHV-6 and HHV-7.

In the last decade, saliva has been advocated as a non-invasive alternative to blood as a diagnostic fluid. However, use of saliva has been hindered by the inadequate sensitivity of current methods to detect the lower salivary concentrations of many constituents compared to serum. Furthermore, developments in the areas related to lab-on-a-chip systems for saliva-based point of care diagnostics are complicated by the high viscosity and heterogeneous properties associated with this diagnostic fluid. The biomarker C-reactive protein (CRP) is an acute phase reactant and a well-accepted indicator of inflammation. Numerous clinical studies have established elevated serum CRP as a strong, independent risk factor for the development of cardiovascular disease (CVD). CVD has also been associated with oral infections (i.e. periodontal diseases) and there is evidence that systemic CRP may be a link between the two. Clinical measurements of CRP in serum are currently performed with "high sensitivity" CRP (hsCRP) enzyme-linked immunosorbent assay (ELISA) tests that lack the sensitivity for the detection of this important biomarker in saliva. Because measurement of salivary CRP may represent a novel approach for diagnosing and monitoring chronic inflammatory disease, including CVD and periodontal diseases, the objective of this study was to apply an ultra-sensitive microchip assay system for the measurement of CRP in human saliva. Here, we describe this novel lab-on-a-chip system in its first application for the measurement of CRP in saliva and demonstrate its advantages over the traditional ELISA method. The increased sensitivity of the microchip system (10 pg ml(-1) of CRP with 1000-fold dilution of saliva sample) is attributed to its inherent increased signal to noise ratio, resulting from the higher bead surface area available for antigen/antibody interactions and the high stringency washes associated with this approach. Finally, the microchip assay system was utilized in this study to provide direct experimental evidence that chronic periodontal disease may be associated with higher levels of salivary CRP.

Background and Objective: Biological regulators of periodontal inflammation, collagen degradation, and insulin resistance have not been determined in association with severity of periodontitis and response to periodontal treatment in diabetics. Our objective was to determine whether type 2 diabetes (T2DM) patients with periodontal disease present a distinct salivary biomarker profile compared with T2DM patients without periodontal disease and healthy subjects (without diabetes and periodontitis) pre‐ and post‐nonsurgical therapy. Methods: Clinical parameters of periodontal health and whole unstimulated saliva were collected from 92 participants (31 Not Periodontitis, NP; 32 T2DM without periodontitis, DWoP; and 29 with T2DM with periodontitis, DWP) at baseline. The T2DM groups received scaling and root planning (SRP) and provided saliva at 6‐week follow‐up. Salivary concentrations of interleukin (IL)‐1β, IL‐6, matrix metalloproteinase‐8 (MMP‐8), and resistin were measured by immunoassay. Results: The DWP group had significantly more disease and higher salivary concentrations at baseline for IL‐1β, MMP‐8, and resistin (p's <.01) compared with DWoP and NP. SRP resulted in significant improvement in periodontal parameters for the T2DM groups; however, more disease persisted (p <.001), and IL‐1β, MMP‐8, and resistin concentrations remained significantly higher in the DWP than the DWoP group (p <.01) at 6 weeks post‐treatment. Principal component analysis demonstrated the DWoP group appeared more biologically similar to the NP group than the DWP group. Concentrations of these salivary biomarkers increased with increasing periodontal disease severity (p <.05) in this study population. Conclusion: Salivary concentrations of IL‐1β, MMP‐8, and resistin appear to serve as biomarkers of periodontal status pre‐ and post‐treatment, irrespective of diabetes status. [ABSTRACT FROM AUTHOR]

Background: Leptin and adiponectin, two adipokines involved in glucose and lipid metabolism, have been linked to regulation of growth in early infancy, energy balance, and metabolic disorders in childhood. The aim of this study was to determine if concentrations of leptin and adiponectin could be measured reliably in infants' saliva, to evaluate the degree of agreement with infant serum levels, and to explore their association with infant feeding status. Methods: A total of 34 infants were recruited after birth and followed for 20 weeks. After log-transformation of the values, a Bland-Altman graphical approach was used to summarize the direction of the difference between the serum and saliva values. Repeated measures mixed modeling was used to evaluate differences over time in these outcomes by feeding status. Results: Mean concentration of salivary leptin and adiponectin in infants was 3.7 (SD =.8) ng/mL and 2.9 (SD = 0.7) ng/mL, respectively. The degree of agreement between serum and saliva for log-transformed leptin and adiponectin values were relatively robust, albeit with a non-zero bias between the two methods, given that serum values were greater than corresponding saliva values for both adipokines in all infants. Each of the four repeated measures mixed models (one for each adipokine measure) had a significant main effect; however, the interaction between time and feeding status was not significant in any of the models. Conclusion: This study demonstrated that leptin and adiponectin can be measured in infant saliva, but in some cases leptin concentrations may be more difficult to detect. [ABSTRACT FROM AUTHOR]

Aim: Salivary biomarkers can help in assessment of periodontitis; however, concentrations may be altered in the presence of diabetes. Hence, the ability of salivary biomarkers to discriminate periodontally healthy type II diabetics (T2DM) from T2DM who have periodontitis was examined. Methods: Ninety‐two participants (29 with T2DM with chronic periodontitis, DWP; 32 T2DM without chronic periodontitis, DWoP; and 31 Not Periodontitis, NP) provided saliva and clinical parameters of periodontal health were recorded. Salivary concentrations of interleukin (IL)‐1β, IL‐6, matrix metalloproteinase‐8 (MMP‐8), macrophage inflammatory protein‐1α (MIP‐1α), adiponectin and resistin were measured by immunoassay. Results: Salivary analyte concentrations for IL‐1β, MMP‐8 and resistin correlated with clinical parameters of periodontitis, with MMP‐8 demonstrating the strongest positive correlation with PD ≥5 mm (p < 0.0001). Periodontal health was reflected in salivary analyte concentrations by group, with concentrations of IL‐1β and MMP‐8 showing significant associations with periodontitis (p ≤ 0.04) that increased in concentration from health to DWoP to DWP. Odds ratio (OR) analyses showed that MMP‐8 discriminated periodontitis from NP (OR of 8.12; 95% CI: 1.01–65.33; p = 0.03) and in the presence of T2DM (DWP vs DWoP, OR = 5.09; 95% CI: 1.24–20.92; p = 0.03). Conclusion: Salivary MMP‐8 and IL‐1β discriminate periodontitis in T2DM. [ABSTRACT FROM AUTHOR]

Aim: To investigate the synergistic role of biologic markers from saliva, serum and plaque in modelling periodontitis disease progression. Material and Methods: This longitudinal study evaluated characteristics of disease progression in 114 patients with generalized moderate to severe periodontitis. The primary outcome was detection of sites with progressing attachment loss sites over 6 months in patients who received scaling and root planing or oral hygiene only. The predictive potential of 27 biomarkers in serum, whole saliva and subgingival plaque was evaluated using three classification algorithms (Support Vector Machines; Naïve Bayes Classifier; and Linear Discriminant Analysis) within an ensemble predictive modelling framework. Results: Disease progression occurred in 24.6% of subjects (28/114). Predictive modelling using Naïve Bayes Classifier identified progressors best with sensitivity of ~89%. The use of the three classification algorithms revealed the concerted role of salivary matrix metalloproteinase‐8, serum biomarkers (serum amyloid P, matrix metalloproteinase 1, bactericidal permeability‐increasing protein, isoprostane) along with levels of Porphryomonas gingivalis and Tannerella forsythia in plaque in predicting progressors. Conclusions: Synergistic utility of baseline bacterial and inflammatory biomarkers from saliva, serum and plaque predicted disease progression. [ABSTRACT FROM AUTHOR]

The aim was to determine the effect of chewing gum containing xylitol and freeze-dried blackberry powder on oral bacteria. This was a randomized, controlled, cross-over study (RCT #: NCT05133557). Fifty participants chewed gum over an 8 h period, four times for 20 min at 2-hour intervals, containing 700 mg xylitol (CG) with or without 50 mg blackberry powder (BG), while wearing a stent containing a sterile enamel chip. After a 1 week washout, participants chewed gum from the other group following the same protocol. The primary outcome was the amount of nine oral bacteria in saliva as determined by quantitative PCR. The secondary outcome was bacteria formed on enamel chips. Chewing BG for four twenty-minute intervals reduced mean total bacteria load and the relative abundance of six of the nine bacteria studied in saliva (p < 0.05). In comparison, only four bacteria were reduced in abundance in the CG group. After gum chewing and regardless of group, S. sanguinis and A. naeslundii were the predominant bacteria adherent to enamel, with S. mutans representing < 1 % of the total bacteria on enamel. Bacterial loads in saliva were rapidly, differentially, and significantly reduced after one day of chewing BG. • Blackberry powder enhances antibacterial effects of xylitol-containing chewing gum. • Reduction in bacterial counts occur rapidly in saliva. • Chewing gum rapidly reduced levels of some cariogenic and periodontal pathogens in saliva. [ABSTRACT FROM AUTHOR]

Objective Background Methods Results Conclusion To examine the characteristics of the salivary microbiome in Type 2 diabetes mellitus (T2DM) patients with or without periodontitis.Periodontitis has been identified as clear sequelae of T2DM. This chronic oral disease also impacts the management of the clinical features of diabetes. The oral microbiome characteristics in T2DM with and without periodontitis, as well as the response of this oral microbiome to nonsurgical therapy have not been well described. Knowledge of key oral biological features could help address the observed poorer clinical presentation of T2DM patients.The oral microbiome in saliva of adult cohorts periodontally healthy/non‐diabetic (non‐periodontitis; NP; n = 31), T2DM without periodontitis (DWoP; n = 32), and T2DM with periodontitis (DWP; n = 29) were characterized by microbial molecular analysis using V3–V4 sequencing and Luminex or ELISA techniques for salivary host analytes.Phyla distribution showed DWP with significantly lower levels of Firmicutes and Actinobacteria and higher levels of Fusobacteria and Spirochetes compared to the healthier groups. Approximately 10% of the detected microbial species showed significant differences in frequency and level of colonization among the DWP, DWoP, and NP samples. A subset of bacteria were significantly correlated with clinical disease features, as well as a specific repertoire of salivary analytes, in particular matrix metalloproteinase (MMP)8/MMP9, interleukin‐1ß, B‐cell activating factor, and resistin differed between the groups and were related to specific taxa. Principal component analysis that identified a majority of the DWP subjects microbiome was unique based upon an array of 27 taxa out of up to 255 detected in the saliva samples.T2DM patients with periodontitis show unique oral microbiome and salivary analyte composition compared to diabetics or non‐diabetic persons without periodontitis. Specific members of the oral microbiome relate directly to the clinical disease features and/or salivary biomolecules in T2DM individuals. [ABSTRACT FROM AUTHOR]

Abstract: Previous investigations in our laboratory and others (Chen et al., 1998) have shown that the levels of certain inflammatory, proangiogenic cytokines in saliva and tissue specimens of patients with oral premalignant lesions (OPML) are elevated. We have also shown that these cytokines are elevated in tissue culture of oral squamous cell carcinoma (OSCC) cell lines. The purpose of this pilot study was to determine the level of several inflammatory, NF-κB-dependent cytokines in whole unstimulated saliva (WUS), in subjects with OPML as compared to those with diagnosed OSCC. Subjects (n =13) with OMPL, OSCC (n =13), and age–sex matched controls without oral lesions (C) (n =13) were enrolled. The mean age was 58.7 years. WUS was collected by standard techniques for 5min (Navazesh, 1993). WUS samples were centrifuged and the cytokine analysis was performed on the supernatants by ELISA as previously described by Ondrey et al. (1991). The cytokines analyzed were: TNF-alpha, interleukin-1, interleukin-6, and interleukin-8 (TNF-α, IL-1, IL-6, and IL-8). The results as analyzed by Pairwise t-tests revealed significant differences in the salivary levels of: (1) TNF-α: (mean±S.E.M.: TNF-α–OSSC=28.9±14.6* pcg/ml versus OPML=10.5±7.4* pcg/ml versus controls=3.0±1.0pcg/ml; * p &#60;0.01); (2) IL-1: (IL-1–OSSC=454.4±215.8* pcg/ml versus OPML=255.1±124.8* pcg/ml versus controls=173.2±66.9pcg/ml; * p &#60;0.01); (3) IL-6: (mean±S.E.M.: IL-6–OSSC=88.2±43.2* pcg/ml versus OPML=70.8±24.3* pcg/ml versus controls=1.4±1.0pcg/ml; * p &#60;0.001) and (4) IL-8 in saliva: (mean±S.E.M.: IL-8–OSSC=3154.1±1023.2* pcg/ml versus OPML=1918.2±899.1* pcg/ml versus controls=1580.7±789.0pcg/ml; * p &#60;0.001). There was a significant increase in the levels of all cytokines in the saliva of the OPML as compared to controls, and a significant difference in the cytokines of OSSC saliva compared to the OPML and controls. These results suggest that these proangiogenic, proinflammatory cytokines are elevated in the saliva of patients with OSSC and OPML as compared to controls, which may have diagnostic and/or prognostic significance. [Copyright &y& Elsevier]