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Developing affordable, rapid, and accurate biosensors is essential for SARS-CoV-2 surveillance and early detection. We created a bio-inspired peptide, using the SAGAPEP AI platform, for COVID-19 salivary diagnostics via a portable electrochemical device coupled to Machine Learning algorithms. SAGAPEP enabled molecular docking simulations against the SARS-CoV-2 Spike protein's RBD, leading to the synthesis of Bio-Inspired Artificial Intelligence Peptide 1 (BIAI1). Molecular docking was used to confirm interactions between BIAI1 and SARS-CoV-2, and BIAI1 was functionalized on rhodamine-modified electrodes. Cyclic voltammetry (CV) using a [Fe(CN)6]3−/4 solution detected virus levels in saliva samples with and without SARS-CoV-2. Support vector machine (SVM)-based machine learning analyzed electrochemical data, enhancing sensitivity and specificity. Molecular docking revealed stable hydrogen bonds and electrostatic interactions with RBD, showing an average affinity of −250 kcal/mol. Our biosensor achieved 100% sensitivity, 80% specificity, and 90% accuracy for 1.8 × 10⁴ focus-forming units in infected saliva. Validation with COVID-19-positive and -negative samples using a neural network showed 90% sensitivity, specificity, and accuracy. This BIAI1-based electrochemical biosensor, integrated with machine learning, demonstrates a promising non-invasive, portable solution for COVID-19 screening and detection in saliva. [ABSTRACT FROM AUTHOR]

Background: The purpose was to determine the perception of the use of salivary diagnostics amongst dental students and faculty and assess if this perception changed after personally experiencing salivary diagnostic testing. Materials and Methods: Participants completed a pretest survey on salivary diagnostic testing perception. Participants supplied a saliva sample and samples were analyzed for detection of pathogens associated with periodontal disease. Test results were distributed to respective participants with a standardized verbal explanation. Participants completed a posttest survey. A paired sample t-test was conducted to compare the differences in perception between pre- and post-survey results. Results: A total of 50 participants completed the study; among the participants, 92% were dental students and 8% were dental faculty. The distribution of familiarity with the concept of salivary diagnostics indicated that most participants were unfamiliar with it. Perception towards salivary diagnostics increased after first-hand experience with salivary diagnostic testing. The overall satisfaction on the experience of salivary diagnostics was positive. Furthermore, linear regression analysis showed that age had a significant negative effect on overall satisfaction, p = 0.008 Conclusions: The overall satisfaction on the experience of salivary diagnostics as a screening tool in dentistry was positive. Furthermore, dental students and faculty's perceptions of salivary diagnostics changed after first-hand experience with salivary diagnostic testing. Practical Implications: Exposure to personal experiences with salivary diagnostics significantly improved the perception on salivary diagnostics, indicating the need for increased curricular emphasis on this topic at dental schools. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to characterize the inflammatory profile of systemically healthy children’s saliva and its association with clinical diagnoses of caries and gingival inflammation. Materials and methods: Unstimulated saliva was collected from 100 children before clinical dental examinations. The saliva samples were analyzed for total protein and specific inflammatory cytokines (IL-10, IL-8, IL-6, and TNFα) with Bradford and ELISA assays, respectively. Salivary bacteria were quantified using a quantitative real-time polymerase chain assay. The salivary values were then correlated with age, DMFT index, plaque index (PI), and gingival index (GI). Results: The mean age of the cohort was 8.08 ± 0.23 years with 49% females, the mean DMF of the cohort was 2.64 ± 0.31, the mean GI was 0.51 ± 0.06, and the mean PI was 1.33 ± 0.07. Significant correlations were found between PI with DMFT and GI. Children with DMFT > 2 had significantly higher levels of IL-8 compared with children with DMFT ≤ 2. IL-6 and TNFα were significantly higher among children with PI > 1 than among children with PI ≤ 1. Conclusions: Salivary cytokine were found to be associate with clinical parameters as DMFT and PI, thus may be a potential tool that reflects dental health status. Clinical relevance: The presence of salivary cytokines in children may reflect evaluation of dental caries and oral inflammation. [ABSTRACT FROM AUTHOR]

Simple Summary: This research explores non-invasive methods for detecting Human Papillomavirus (HPV) in patients with oral squamous cell carcinoma (OSCC). The study compares two self-collection techniques, oral rinse vs. salivary sponge, to determine their effectiveness in identifying HPV DNA. Results from 26 patients indicated that the salivary sponge method was more accurate and sensitive compared to the oral rinse. This finding suggests that a salivary sponge may be a superior option, especially for patients with functional limitations caused by OSCC. Our research could lead to improved non-invasive diagnostic tools for HPV in clinical settings. Background/Objectives: Human Papillomavirus (HPV) is a significant etiological factor in the development of oropharyngeal carcinogenesis. The detection of HPV in oral squamous cell carcinoma (OSCC) could be also crucial for diagnosis, prognosis, and treatment planning. This study compares the efficacy and accuracy of two non-invasive sampling methods, oral rinse, and oral sponge, in detecting HPV DNA in patients with OSCC. Methods: Twenty-six patients with histologically confirmed OSCCs were recruited (M/F = 15/11; mean age 68.6). From each patient, two self-collected oral specimens, in the form of an oral rinse and a salivary sponge (i.e., LolliSponge), were collected, and subsequently processed, utilizing INNO-LiPA HPV Genotyping Extra II for HPV DNA detection; Results: Oral sponge detection showed high specificity (100%), sensitivity (85.7%), and accuracy (96.2%) compared to the oral rinse sampling, also demonstrating an area AUC for its diagnostic performance significantly greater than 0.5 (0.93 vs. 0.5, p < 0.0001). Conclusions: This study supports that oral sponge sampling offers valuable non-invasive alternatives for HPV detection in patients with OSCC, with the potentiality to facilitate saliva sampling in patients that may exhibit functional deficit due to OSCC. Further research is recommended to validate these findings in larger cohorts and to explore the integration of these methods into routine clinical practice for the management of HPV-related OSCC. [ABSTRACT FROM AUTHOR]

IntroductionPeriodontitis affects a significant portion of the global population and is associated with systemic health issues. Salivary biomarkers such as salivary matrix metalloproteinase-8 (MMP-8) and its activated form (aMMP-8) have been studied for their roles in tissue degradation and inflammation in periodontitis. This meta-analysis investigates the association between salivary MMP-8 and aMMP-8 levels and periodontitis.MethodsA systematic literature search was conducted utilizing PubMed, Embase, Web of Science, and Cochrane Library databases up to October 2023, yielding 35 studies that quantified MMP-8 or aMMP-8 in saliva from patients with periodontitis and healthy controls. Data were extracted, and standardized mean differences (SMD) with 95% confidence intervals (CI) were calculated. Heterogeneity was assessed, and subgroup analyses were performed based on saliva collection techniques. Meta-regression analysis evaluated the impact of publication year on heterogeneity.ResultsThe meta-analysis included 35 studies. Pooled results indicated significantly higher levels of MMP-8 and aMMP-8 in periodontitis cases compared to healthy controls (SMD: 2.71, 95% CI: 1.04–4.38, p = 0.002) with substantial heterogeneity (I2 = 94.5%). No significant difference was found between MMP-8 and aMMP-8 (p = 0.445). Subgroup analyses by saliva collection technique did not reduce heterogeneity significantly. Meta-regression showed that publication year did not impact heterogeneity. Small-study effects and publication bias were present, suggesting caution in interpreting the results.DiscussionThe findings support the potential of MMP-8 and aMMP-8 as biomarkers for periodontitis, although substantial heterogeneity and methodological differences among studies pose challenges. Standardized protocols and larger sample sizes are necessary to enhance the reliability of these biomarkers in clinical practice. Despite limitations, salivary diagnostics hold promise for non-invasive, early detection and monitoring of periodontitis.ConclusionSalivary MMP-8 and aMMP-8 levels are significantly associated with periodontitis, highlighting their potential as diagnostic biomarkers. However, methodological improvements and standardization are essential for their clinical application. Collaborative efforts and advancements in salivary diagnostics are crucial for improving periodontitis management and patient outcomes.

Amelogenesis imperfecta (AI) is a genetic disease characterized by poor formation of tooth enamel. AI occurs due to mutations, especially in AMEL, ENAM, KLK4, MMP20, and FAM83H, associated with changes in matrix proteins, matrix proteases, cell-matrix adhesion proteins, and transport proteins of enamel. Due to the wide variety of phenotypes, the diagnosis of AI is complex, requiring a genetic test to characterize it better. Thus, there is a demand for developing low-cost, noninvasive, and accurate platforms for AI diagnostics. This case-control pilot study aimed to test salivary vibrational modes obtained in attenuated total reflection fourier-transformed infrared (ATR-FTIR) together with machine learning algorithms: linear discriminant analysis (LDA), random forest, and support vector machine (SVM) could be used to discriminate AI from control subjects due to changes in salivary components. The best-performing SVM algorithm discriminates AI better than matched-control subjects with a sensitivity of 100%, specificity of 79%, and accuracy of 88%. The five main vibrational modes with higher feature importance in the Shapley Additive Explanations (SHAP) were 1010 cm−1, 1013 cm−1, 1002 cm−1, 1004 cm−1, and 1011 cm−1 in these best-performing SVM algorithms, suggesting these vibrational modes as a pre-validated salivary infrared spectral area as a potential biomarker for AI screening. In summary, ATR-FTIR spectroscopy and machine learning algorithms can be used on saliva samples to discriminate AI and are further explored as a screening tool. [ABSTRACT FROM AUTHOR]

Context: Oral cancer is highly prevalent in India. Lack of awareness and delay in diagnosis and treatment of patients with oral cancer leads to high mortality and poor survival of patients. Salivary endothelin-1 is proposed as a prospective biomarker for oral squamous cell carcinoma. Aims: Aim of the study was to evaluate salivary level of endothelin-1 in oral cancer and precancer as a biomarker. Settings and Design: We planned a case control study to evaluate salivary level of Endothelin-1 in oral cancer and precancer as a biomarker. Materials and Methods: A total of 72 subjects were taken in study out of which 24 cases were of histopathologically confirmed premalignat oral lesion (oral leukoplakia and oral submucous fibrosis), 24 cases were of histopathologically confirmed oral squamous cell carcinoma, and 24 cases of healthy age and gender matched controls without any addiction to tobacco in any form from a tertiary care hospital were taken. Saliva was collected from all following standard guidelines and estimation of salivary endothelin-1 was done by ELISA. Statistical Analysis Used: SPSS software version 15. Results: Salivary endothelin-1 values of controls ranged between 0.09 and 1.88 pg/ml while that of premalignant cases ranged between 1.16 and 16.135 pg/ml and of SCC cases ranged between 2.567 and 22.98 pg/ml. Conclusions: Salivary endothelin-1 is raised in oral squamous cell carcinoma compared to premalignant and controls therefore, shows capability to differentiate between premalignant lesion and oral cancer. So, it could be used as a biomarker for early diagnosis. [ABSTRACT FROM AUTHOR]

Background The purpose was to determine the perception of the use of salivary diagnostics amongst dental students and faculty and assess if this perception changed after personally experiencing salivary diagnostic testing.Materials and Methods Participants completed a pretest survey on salivary diagnostic testing perception. Participants supplied a saliva sample and samples were analyzed for detection of pathogens associated with periodontal disease. Test results were distributed to respective participants with a standardized verbal explanation. Participants completed a posttest survey. A paired sample t-test was conducted to compare the differences in perception between pre- and post-survey results.Results A total of 50 participants completed the study; among the participants, 92% were dental students and 8% were dental faculty. The distribution of familiarity with the concept of salivary diagnostics indicated that most participants were unfamiliar with it. Perception towards salivary diagnostics increased after first-hand experience with salivary diagnostic testing. The overall satisfaction on the experience of salivary diagnostics was positive. Furthermore, linear regression analysis showed that age had a significant negative effect on overall satisfaction, p = 0.008Conclusions The overall satisfaction on the experience of salivary diagnostics as a screening tool in dentistry was positive. Furthermore, dental students and faculty’s perceptions of salivary diagnostics changed after first-hand experience with salivary diagnostic testing.Practical Implications Exposure to personal experiences with salivary diagnostics significantly improved the perception on salivary diagnostics, indicating the need for increased curricular emphasis on this topic at dental schools.

The activity of dental caries, combined with its multifactorial etiology, alters salivary molecule composition. The present systematic review was developed to answer the following question: “Are salivary biomarkers reliable for diagnosis of dental caries?”. Following the “Preferred Reporting Item for Systematic Reviews and Meta-analysis” (PRISMA) guidelines, the review was conducted using multiple database research (Medline, Web of Science, and Scopus). Studies performed on healthy subjects with and without dental caries and providing detailed information concerning the clinical diagnosis of caries (Decayed, Missing, Filled Teeth-DMFT and International Caries Detection and Assessment System-ICDAS criteria) were included. The quality assessment was performed following a modified version of the Joanna Briggs Institute Prevalence Critical Appraisal Checklist. The protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO, ID: CRD42022304505). Sixteen papers were included in the review. All studies reported statistically significant differences in the concentration of salivary molecules between subjects with and without caries (p < 0.05). Proteins were the most investigated molecules, in particular alpha-amylase and mucins. Some studies present a risk of bias, such as identifying confounding factors and clearly defining the source population. Nevertheless, the 16 papers were judged to be of moderate to high quality. There is evidence that some salivary compounds studied in this review could play an important diagnostic role for dental caries, such as salivary mucins, glycoproteins (sCD14), interleukins (IL-2RA, 4,-13), urease, carbonic anhydrase VI, and urea.

Developing affordable, rapid, and accurate biosensors is essential for SARS-CoV-2 surveillance and early detection. We created a bio-inspired peptide, using the SAGAPEP AI platform, for COVID-19 salivary diagnostics via a portable electrochemical device coupled to Machine Learning algorithms. SAGAPEP enabled molecular docking simulations against the SARS-CoV-2 Spike protein’s RBD, leading to the synthesis of Bio-Inspired Artificial Intelligence Peptide 1 (BIAI1). Molecular docking was used to confirm interactions between BIAI1 and SARS-CoV-2, and BIAI1 was functionalized on rhodamine-modified electrodes. Cyclic voltammetry (CV) using a [Fe(CN)6]3−/4 solution detected virus levels in saliva samples with and without SARS-CoV-2. Support vector machine (SVM)-based machine learning analyzed electrochemical data, enhancing sensitivity and specificity. Molecular docking revealed stable hydrogen bonds and electrostatic interactions with RBD, showing an average affinity of −250 kcal/mol. Our biosensor achieved 100% sensitivity, 80% specificity, and 90% accuracy for 1.8 × 10⁴ focus-forming units in infected saliva. Validation with COVID-19-positive and -negative samples using a neural network showed 90% sensitivity, specificity, and accuracy. This BIAI1-based electrochemical biosensor, integrated with machine learning, demonstrates a promising non-invasive, portable solution for COVID-19 screening and detection in saliva.

Salivary pH is one of the crucial biomarkers used for non-invasive diagnosis of intraoral diseases, as well as general health conditions. However, standard pH sensors are usually too bulky, expensive, and impractical for routine use outside laboratory settings. Herein, a miniature hydrogel sensor, which enables quick and simple colorimetric detection of pH level, is shown. The sensor structure was manufactured from non-toxic hydrogel ink and patterned in the form of a matrix with 5 mm × 5 mm × 1 mm individual sensing pads using a 3D printing technique (bioplotting). The authors' ink composition, which contains sodium alginate, polyvinylpyrrolidone, and bromothymol blue indicator, enables repeatable and stable color response to different pH levels. The developed analysis software with an easy-to-use graphical user interface extracts the R(ed), G(reen), and B(lue) components of the color image of the hydrogel pads, and evaluates the pH value in a second. A calibration curve used for the analysis was obtained in a pH range of 3.5 to 9.0 using a laboratory pH meter as a reference. Validation of the sensor was performed on samples of artificial saliva for medical use and its mixtures with beverages of different pH values (lemon juice, coffee, black and green tea, bottled and tap water), and correct responses to acidic and alkaline solutions were observed. The matrix of square sensing pads used in this study provided multiple parallel responses for parametric tests, but the applied 3D printing method and ink composition enable easy adjustment of the shape of the sensing layer to other desired patterns and sizes. Additional mechanical tests of the hydrogel layers confirmed the relatively high quality and durability of the sensor structure. The solution presented here, comprising 3D printed hydrogel sensor pads, simple colorimetric detection, and graphical software for signal processing, opens the way to development of miniature and biocompatible diagnostic devices in the form of flexible, wearable, or intraoral sensors for prospective application in personalized medicine and point-of-care diagnosis. [ABSTRACT FROM AUTHOR]

Periodontal diseases, including gingivitis and periodontitis, are among the most prevalent diseases in humans. Gingivitis is the mildest form of periodontal disease, characterized by inflammation of the gingiva caused by the accumulation of dental plaque. Salivary diagnostics are becoming increasingly popular due to the variation in saliva composition in response to pathological processes. We used a metabolomics approach to investigate whether a specific saliva metabolic composition could indicate preclinical stage of gingivitis. 1H-NMR spectroscopy was used to obtain the salivary metabolite profiles of 20 healthy subjects. Univariate/multivariate statistical analysis evaluated the whole saliva metabolite composition, and the Full-Mouth Bleeding Score (FMBS) was employed as a classification parameter. Identifying a signature of specific salivary metabolites could distinguish the subjects with high FMBS scores but still within the normal range. This set of metabolites may be due to the enzymatic activities of oral bacteria and be associated with the early stages of gingival inflammation. Although this analysis is to be considered exploratory, it seems feasible to establish an FMBS threshold that distinguishes between the absence and presence of early inflammatory alterations at the salivary level. [ABSTRACT FROM AUTHOR]

Primary Sjögren syndrome (pSS) is a chronic autoimmune disease mainly affecting salivary and lacrimal glands. The current pSS biomarkers, serum autoantibodies, are negative in many pSS patients diagnosed with histopathology changes, indicating the need of novel biomarkers. The current therapies of pSS are merely short–term symptomatic relief and can't provide effective long–term remedy. Extracellular vehicles (EVs) are nano–sized lipid bilayer–delimited particles spontaneously released by almost all types of cells and carrying various bioactive molecules to mediate inter–cellular communications. Recent studies found that EVs from salivary gland epithelial cells and immune cells play essential roles in pSS pathogenesis. Correspondingly, EVs and their cargos in plasma and saliva are promising candidate biomarkers for pSS diagnosis. Moreover, EVs from mesenchymal stem cells have shown promises to improve pSS treatment by modulating immune responses. This review summarizes recent findings in roles of EVs in pSS pathogenesis, diagnosis, and treatment of pSS, as well as related challenges and future research directions.

Aim: Enzymatic antioxidants are the primary line of defense against oxidative and nitrosative stress. However, their involvement in the progression of periodontitis is still not well understood. The study aimed to determine the activity of enzymatic antioxidants in the gingival crevicular fluid (GCF) and saliva of patients with periodontitis. Materials and methods: The study group of 56 patients with periodontitis (stage III and IV) and 28 healthy controls were involved. The modified plaque index, probing depth, the clinical attachment level, the percentage of sites with bleeding on probing, papilla bleeding index, and maximum value of tooth mobility (Periotest®) were tested. Saliva (stimulated and non‐stimulated) and GCF were collected from the participants, and activity of peroxidase, catalase, superoxide dismutase, and glutathione reductase were determined colorimetrically. Results: Lower activity of peroxidase (p < 0.0001), catalase (p < 0.0001), superoxide dismutase (p = 0.0188), and glutathione reductase (p < 0.0001) was noted in non‐stimulated saliva of patients with periodontitis compared to healthy subjects. Peroxidase (p < 0.0001), catalase (p < 0.0001) and superoxide dismutase (p < 0.0001) showed lower activity in stimulated saliva of patients with periodontitis compared to healthy subjects. The peroxidase (p < 0.0029), catalase (p < 0.0001), and glutathione reductase (p = 0.0028) activity in GCF of stage III + IV were significantly higher compared to healthy controls. Superoxide dismutase (p < 0.0001) showed lower activity in GCF of patients with periodontitis. Conclusions: The demonstrated decrease in activity of all analyzed enzymatic antioxidants in non‐stimulated saliva may result from long‐lasting periodontitis and exhaustion of the safeguard mechanism against reactive oxygen species. [ABSTRACT FROM AUTHOR]

ABSTRACTBackground The oral cavity is the most important link between the human body and the external environment and is well-equipped to protect the body against invading pathogens. Emerging data on the bidirectional relationship between oral health and the severity of COVID-19 symptoms has prompted us to reexamine our role as the guardians of the oral cavity - the true gateway to the body. Recent studies have shown that the oral microbiome is a complex ecological system and the interactions between its residents and the host cells influence human health and disease. This narrative review elaborates on the oral-systemic connection from an oral microbiome standpoint and discusses the future role of oral health professionals.Methods A PubMed literature search using the keywords “Oral Microbiome and Diseases” and “Oral microbiome and Health” for a range of ten years between 2012-2022 was completed. Meta-Analyses, Reviews, Randomized Control Trials, Clinical Trials, and books were included in the search.Results “Oral microbiome and Diseases” yielded 6392 results and “Oral microbiome and Health” yielded 4287 results.Conclusion Oral health professionals need to be trained on the oral-systemic connection periodically to appreciate the recent advances in this field.Practical Implications A deeper understanding of the impact of existing treatment modalities on the oral microbiome could expand the role of oral health professionals in preventing and maintaining patients' overall health. Implementing point-of-care salivary diagnostics to detect salivary biomarkers for oral and non-oral diseases and conditions may be a significant leap toward this goal.

Introduction: Saliva is a clinicaly important biologic fluid. The present study aims at determining the role of saliva in detection of pregnancy using home-based pregnancy detection kits to provide an alternative biofluid that is more user friendly, acceptable, noninvasive, rapid and easy for home use. Materials and method: The study was conducted among 100 women visiting a gynecology clinic for confirmation of pregnancy with a history of a missed menstrual cycle not more than 4 weeks. The test will be performed using pregnancy detection kit with 20 mIU specification for estimation of salivary hCG. Routine tests was prescribed to the patients such as laboratory based urine hCG and/ultrasound examination was used for confirmation of pregnancy status & correlation with the saliva based result. Results: About 84% participants with a confirmed pregnancy showed a positive result, of which 62% showed a dark band & a light band was noted in 22% participants. A negative results was noted in 16% participants & were positive for urine test result. Conclusion: The result of this study showed that traditionally used biofluid can be replaced with more user friendly & acceptable biofluids. i.e, saliva as a biomarker for detection of pregnancy. In this study urinary hCG based kit was used but it opens new avenues towards development of new Salivary based diagnostic kits for detection of pregnancy. INTRODUCTION: The evolution of salivary diagnosis has reached a level towards the goal of using saliva as a powerful fluid for early detection of pregnancy and also it is used for first line diagnosis for life threatening diseases such as cancer, metabolic disorders, infections 1. The collection of saliva is simple, painless, cheap and safe procedure both for patient and medical staff. An additional advantages of saliva is the fact that it may be retrieved several times a day, which make repeat analysis much easier. Furthermore, saliva has very high durability. Although 94-99% of saliva content is water, saliva also contains numerous cellular elements and many organic and inorganic substances, including most biological markers present in blood and urine that may be used in the early detection and monitoring of many dental and general disease. [ABSTRACT FROM AUTHOR]

Pain is one of the most common complaints leading to a pediatric emergency department visit and is associated with various painful procedures, leading to increased anxiety and stress. Assessing and treating pain in children can be challenging, so it is crucial to investigate new methods for pain diagnosis. The review aims to summarize the literature on non-invasive salivary biomarkers, such as proteins and hormones, for pain assessment in urgent pediatric care settings. Eligible studies were those that included novel protein and hormone biomarkers in acute pain diagnostics and were not older than 10 years. Chronic pain studies were excluded. Further, articles were divided into two groups: studies in adults and studies in children (<18 years). The following characteristics were extracted and summarized: study author, enrollment date, study location, patient age, study type, number of cases and groups, as well as tested biomarkers. Salivary biomarkers, such as cortisol, salivary α-amylase, and immunoglobulins, among others, could be appropriate for children as saliva collection is painless. However, hormonal levels can differ among children in different developmental stages and with various health conditions, with no predetermined levels of saliva. Thus, further exploration of biomarkers in pain diagnostics is still necessary. [ABSTRACT FROM AUTHOR]

Human saliva is an ideal body fluid for developing non-invasive diagnostics. Saliva contains naturally-occurring nanoparticles with unique structural and biochemical characteristics. The salivary exosome, a nanoscale extracellular vesicle, has been identified as a highly informative nanovesicle with clinically-relevant information. Salivary exosomes have brought forth a pathway and mechanism by which cancer-derived biomarkers can be shuttled through the systemic circulation into the oral cavity. Despite such clinical potential, routine and reliable analyses of exosomes remain challenging due to their small sizes. Characterization of individual exosome nanostructures provides critical data for understanding their pathophysiological condition and diagnostic potential. In this review, we summarize a current array of discovered salivary biomarkers and nanostructural properties of salivary exosomes associated with specific cancers. In addition, we describe a novel electrochemical sensing technology, EFIRM (electric field-induced release and measurement), that advances saliva liquid biopsy, covering the current landscape of point-of-care saliva testing.

Head and neck cancer is the sixth most common cancer worldwide. It remains one of the leading causes of death, and its early detection is crucial. Liquid biopsy has emerged as a promising tool for detecting and monitoring the disease status of patients with early and advanced cancers. Circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and exosomal miRNAs have received enormous attention because of their apparent clinical implications. Analyses of these circulating biomarkers have paved the way for novel therapeutic approaches and precision medicine. A growing number of reports have implicated the use of circulating biomarkers for detection, treatment planning, response monitoring, and prognosis assessment. Although these new biomarkers can provide a wide range of possible clinical applications, no validated circulating biomarkers have yet been integrated into clinical practice for head and neck cancer. In this review, we summarize the current knowledge of circulating biomarkers in this field, focusing on their feasibility, limitations, and key areas of clinical applications. We also highlight recent advances in salivary diagnostics and their potential application in head and neck cancer.

Salivary biomarkers are increasingly being used as an alternative to diagnose and monitor the progression of various diseases due to their ease of use, on site application, non-invasiveness, and most likely improved patient compliance. Here, we highlight the role of salivary biosensors in the general population, followed by the application of saliva as a diagnostic tool in the pediatric population. We searched the literature for pediatric applications of salivary biomarkers, more specifically, in children from 0 to 18 years old. The use of those biomarkers spans autoimmune, developmental disorders, oncology, neuropsychiatry, respiratory illnesses, gastrointestinal disorders, and oral diseases. Four major applications of salivary proteins as biomarkers are: (1) dental health (caries, stress from orthodontic appliances, and gingivitis); (2) gastrointestinal conditions (eosinophilic esophagitis, acid reflux, appendicitis); (3) metabolic conditions (obesity, diabetes); and (4) respiratory conditions (asthma, allergic rhinitis, small airway inflammation, pneumonia). Genomics, metabolomics, microbiomics, proteomics, and transcriptomics, are various other classifications for biosensing based on the type of biomarkers used and reviewed here. Lastly, we describe the recent advances in pediatric biosensing applications using saliva. This work guides scientists in fabricating saliva-based biosensors by comprehensively overviewing the potential markers and techniques that can be employed. [ABSTRACT FROM AUTHOR]

Background: The oral cavity is the most important link between the human body and the external environment and is well-equipped to protect the body against invading pathogens. Emerging data on the bidirectional relationship between oral health and the severity of COVID-19 symptoms has prompted us to reexamine our role as the guardians of the oral cavity - the true gateway to the body. Recent studies have shown that the oral microbiome is a complex ecological system and the interactions between its residents and the host cells influence human health and disease. This narrative review elaborates on the oral-systemic connection from an oral microbiome standpoint and discusses the future role of oral health professionals. Methods: A PubMed literature search using the keywords "Oral Microbiome and Diseases" and "Oral microbiome and Health" for a range of ten years between 2012-2022 was completed. Meta-Analyses, Reviews, Randomized Control Trials, Clinical Trials, and books were included in the search. Results: "Oral microbiome and Diseases" yielded 6392 results and "Oral microbiome and Health" yielded 4287 results. Conclusion: Oral health professionals need to be trained on the oral-systemic connection periodically to appreciate the recent advances in this field. Practical Implications: A deeper understanding of the impact of existing treatment modalities on the oral microbiome could expand the role of oral health professionals in preventing and maintaining patients' overall health. Implementing point-of-care salivary diagnostics to detect salivary biomarkers for oral and non-oral diseases and conditions may be a significant leap toward this goal. [ABSTRACT FROM AUTHOR]

Saliva is a multifaceted biological fluid that plays a pivotal role in oral health and overall well-being. It is primarily produced by major salivary glands, with additional contributions from minor glands. Saliva is essential for various physiological functions, including oral lubrication, digestion, and defense against pathogens. Its intricate composition comprises proteins, electrolytes, enzymes, hormones, and microbial DNA, enabling it to act as a dynamic indicator of both local and systemic health. A literature search was conducted using PubMed, Web of Science, and Google Scholar to identify relevant studies published up to June 2024. The included studies involved human participants and provided original data or comprehensive reviews on salivary biomarkers. The findings indicate that salivary diagnostics show promise in diagnosing and monitoring systemic conditions such as diabetes and cardiovascular diseases, with salivary glucose levels correlating well with blood glucose levels. Biomarkers such as C-reactive protein (CRP) have been linked to cardiovascular risk, while saliva has been explored for cancer detection, including pancreatic and prostate cancers. Advances in techniques such as enzyme-linked immunosorbent assay (ELISA), saliva omics, and single-cell sequencing have furthered salivary diagnostics, providing insights into disease mechanisms. Additionally, quantitative mass spectrometry (qMS) and Raman spectroscopy (RS) contribute to non-invasive diagnostics for various conditions, including cancer. Collecting saliva samples from healthy individuals is crucial for early disease detection and evaluating treatment efficacy. This review underscores the growing importance of salivary tests in dental practice and their potential for diagnosing various health conditions. Further research is essential to address challenges related to variability and standardization. Dentists and healthcare professionals should consider incorporating salivary tests into clinical decision-making to enhance patient outcomes., Competing Interests: Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2025, Albagieh et al.)

Salivary diagnostics has great potential to be used in the early detection and prevention of many cancerous diseases. If implemented with rigour and efficiency, it can result in improving patient survival times and achieving earlier diagnosis of disease. Recently, extraordinary efforts have been taken to develop non-invasive technologies that can be applied without complicated and expensive procedures. Saliva is a biofluid that has demonstrated excellent properties and can be used as a diagnostic fluid, since many of the biomarkers suggested for cancers can also be found in whole saliva, apart from blood or other body fluids. The currently accepted gold standard methods for biomarker development include chromatography, mass spectometry, gel electrophoresis, microarrays and polymerase chain reaction-based quantification. However, salivary diagnostics is a flourishing field with the rapid development of novel technologies associated with point-of-care diagnostics, RNA sequencing, electrochemical detection and liquid biopsy. Those technologies will help introduce population-based screening programs, thus enabling early detection, prognosis assessment and disease monitoring. The purpose of this review is to give a comprehensive update on the emerging diagnostic technologies and tools for the early detection of cancerous diseases based on saliva.

OBJECTIVE: Zinc insufficiency has been proposed to play a role in taste and smell impairment in Coronavirus disease 2019 (COVID-19) patients. Therefore, this study aimed at comparing salivary zinc levels in COVID-19 patients with and without dysgeusia. PATIENTS AND METHODS: A total of 127 participants were recruited for this study. The patients were divided into three groups based on their COVID-19 test results and taste impairment. Groups I and II were COVID-19 positive with and without taste loss, respectively. Group III included the negative control participants. Salivary zinc levels were measured at baseline in all groups and three months after baseline in groups I and II. Wilcoxon signed-rank test was used to compare the zinc levels between baseline and three months after baseline within each group. Mann-Whitney U test was used to compare zinc levels between groups with different degrees of taste loss. RESULTS: Salivary zinc levels were significantly lower in the COVID-19 positive group with taste loss compared to levels in the group without taste loss or the negative controls (p<0.005). Three months after baseline, salivary zinc levels were significantly elevated in both COVID-19 positive groups (p<0.001). CONCLUSIONS: COVID-19 patients with dysgeusia had significantly lower levels of salivary zinc than positive and negative controls. Zinc levels were elevated after recovery, which may indicate that salivary zinc is directly associated with taste abnormalities and COVID-19 outcomes. This study showed that taste impairment is associated with lower salivary zinc levels in COVID-19 patients. [ABSTRACT FROM AUTHOR]

Exosomes are small membrane vesicles of endocytic origin that are secreted by most cells and detected in saliva. Pathophysiological roles for salivary exosomes are beginning to be recognized in diseases including cancer, highlighting potential biomarkers and biological functions. Since early detection of cancer is vital for successful treatment, salivary exosomes would be advantageous in achieving a better survival rate due to their ready availability and noninvasiveness. The use of salivary exosomes may therefore be promising in the accurate detection of premalignant lesions and early-stage cancers, also for better our understanding of the molecular basis of tumorigenesis. In this chapter, we review our current knowledge of salivaomics, focusing on nucleic acids and proteins in saliva as potential cancer biomarkers. Since salivaomics is a rapidly evolving field, we hope to expand frameworks toward salivary exosomes, integrate new and existing information, and bridge salivaomics with other biomedical researches. Furthermore, we would like to coin the term "saliva-exosomics" as the next-generation salivaomics. Our goal in this chapter is to provide the most updated information on cancer-derived exosomes in the saliva as natural carriers of biomarkers and signaling molecules. Major advances include definitive structure analysis and molecular characterization of salivary exosomes. We also highlight the exosome biogenesis and cargo trafficking mechanisms in which recent animal studies have expanded our understanding of exosome-mediated transfer of cancer-derived products from distal tumor to salivary gland. The potential roles of the salivary exosomes in cancer progression and immune surveillance are also addressed.

Salivary diagnostics is a rapidly advancing field that offers clinicians and patients the potential of rapid, noninvasive diagnostics with excellent accuracy. In order for the complete realization of the potential of saliva, however, extensive profiling of constituents must be conducted and diagnostic biomarkers must be thoroughly validated. This article briefly overviews the process of conducting a study of salivary biomarkers in a patient cohort and highlights the studies that have been conducted on different classes of molecules in the saliva. Emerging frontiers in salivary diagnostics research that may significantly advance the field will also be highlighted.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also currently known as CoV disease 2019 (COVID-19), has created a global health upheaval. The saliva of an infected person serves as a source for the transmission of CoV. The angiotensin-converting enzyme-2 receptors serve as the host receptor cells for CoV which is expressed in high numbers in salivary glands, oral mucosa, and gingiva. The salivary gland is a potential reservoir for COVID-19 even in asymptomatic but infected carriers. This knowledge could be used to employ salivary sampling as a noninvasive diagnostic method. IgA, IgM antibodies detected in self-collected saliva show more stability than RNA and may aid in the identification of asymptomatic patients. Saliva shows high sensitivity and specificity in the diagnosis of COVID-19 with >90% concordance reported between the saliva and nasopharyngeal swabs. Standardization with respect to sample collection, storage, and transport media can help in validation of diagnostic application of salivary-based testing.

Introduction: Laboratory testing for the SARS‐CoV‐2 virus and the consequent respiratory coronavirus disease 2019 (COVID‐19) is categorized into methods that detect the viral presence and methods that detect antibodies produced in the host as a response to infection. Methods that detect viral presence into the host excretions measure current infection by SARS‐CoV‐2, whereas the detection of human antibodies exploited against SARS‐CoV‐2 evaluates the past exposure to the virus. Objective: This review provides a comprehensive overview for the use of saliva as a specimen for the detection of SARS‐CoV‐2, the methods for the salivary diagnostics utilized till very recently, and the arisen considerations for the diagnosis of COVID‐19 disease. Conclusion: The major advantage of using saliva as a specimen for the detection of SARS‐CoV‐2 is that saliva collection is a non‐invasive method which produces no discomfort to the patient and permits the patients to utilize home self‐sampling techniques in order to protect health providers from the exposure to the pathogen. There is an urgent need to increase the active research for the detection of SARS‐CoV‐2 in the saliva because the non‐invasive salivary diagnostics may provide a reliable and cost‐effective method suitable for the fast and early detection of COVID‐19 infection. [ABSTRACT FROM AUTHOR]

Background/Aim: Human saliva offers many advantages over blood-based biochemical assays, therefore, becomes the biological fluid of interest. Once antiseptic solutions react with saliva, both fluids undergo significant changes of their biophysical properties, consequently, those changes have an impact on their principal function.

Saliva contains a diverse array of biomarkers indicative of various diseases. Saliva testing has been a major advancement towards non-invasive point-of-care diagnosis with clinical significance. However, there are challenges associated with salivary diagnosis from sample treatment and standardization. This review highlights the biomarkers in saliva and their role in identifying relevant diseases. It provides an overview and discussion about the current practice of saliva collection and processing, and advancements in saliva detection systems from in vitro methods to wearable oral devices. The review also addresses challenges in saliva diagnostics and proposes solutions, aiming to offer a comprehensive understanding and practical guidance for improving saliva-based detection in clinical diagnosis. Saliva diagnosis provides a rapid, effective, and safe alternative to traditional blood and urine tests for screening large populations and enhancing infectious disease diagnosis and surveillance. It meets the needs of various fields such as disease management, drug screening, and personalized healthcare with advances in saliva detection systems offering high sensitivity, fast response times, portability, and automation. Standardization of saliva collection, treatment, biomarker discovery, and detection between different laboratories needs to be implemented to obtain reliable salivary diagnosis in clinical practice., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Salivary biomarkers are increasingly being used as an alternative to diagnose and monitor the progression of various diseases due to their ease of use, on site application, non-invasiveness, and most likely improved patient compliance. Here, we highlight the role of salivary biosensors in the general population, followed by the application of saliva as a diagnostic tool in the pediatric population. We searched the literature for pediatric applications of salivary biomarkers, more specifically, in children from 0 to 18 years old. The use of those biomarkers spans autoimmune, developmental disorders, oncology, neuropsychiatry, respiratory illnesses, gastrointestinal disorders, and oral diseases. Four major applications of salivary proteins as biomarkers are: (1) dental health (caries, stress from orthodontic appliances, and gingivitis); (2) gastrointestinal conditions (eosinophilic esophagitis, acid reflux, appendicitis); (3) metabolic conditions (obesity, diabetes); and (4) respiratory conditions (asthma, allergic rhinitis, small airway inflammation, pneumonia). Genomics, metabolomics, microbiomics, proteomics, and transcriptomics, are various other classifications for biosensing based on the type of biomarkers used and reviewed here. Lastly, we describe the recent advances in pediatric biosensing applications using saliva. This work guides scientists in fabricating saliva-based biosensors by comprehensively overviewing the potential markers and techniques that can be employed.

Salivary glands are considered important targets of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Recent evidence suggests that along with angiotensin converting enzyme 2, certain cell surface sialic acids (Sia) may function as receptors for binding SARS-CoV-2 spike protein. Over 50 forms of Sia have been identified in nature, with N-acetylneuraminic acid (Neu5Ac) being the most abundant. We explored the Human Protein Atlas repository to analyze important enzymes in Neu5Ac biosynthesis and propose a hypothesis that further highlights the significance of salivary glands in coronavirus disease 19 (COVID-19). This work may facilitate research into targeted drug therapies for COVID-19. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also currently known as CoV disease 2019 (COVID-19), has created a global health upheaval. The saliva of an infected person serves as a source for the transmission of CoV. The angiotensin-converting enzyme-2 receptors serve as the host receptor cells for CoV which is expressed in high numbers in salivary glands, oral mucosa, and gingiva. The salivary gland is a potential reservoir for COVID-19 even in asymptomatic but infected carriers. This knowledge could be used to employ salivary sampling as a noninvasive diagnostic method. IgA, IgM antibodies detected in self-collected saliva show more stability than RNA and may aid in the identification of asymptomatic patients. Saliva shows high sensitivity and specificity in the diagnosis of COVID-19 with >90% concordance reported between the saliva and nasopharyngeal swabs. Standardization with respect to sample collection, storage, and transport media can help in validation of diagnostic application of salivary-based testing. [ABSTRACT FROM AUTHOR]

Exosomes are microvesicular structures that play a mediating role in intercellular communication. It is of interest to study the internal cargo of exosomes to determine if they carry disease discriminatory biomarkers. For performing exosomal analysis, it is necessary to develop a method for extracting and analyzing exosomes from target biofluids without damaging the internal content. Electric field-induced release and measurement (EFIRM) is a method for specifically extracting exosomes from biofluids, unloading their cargo, and testing their internal RNA/protein content. Using an anti-human CD63 specific antibody magnetic microparticle, exosomes are first precipitated from biofluids. Following extraction, low-voltage electric cyclic square waves (CSW) are applied to disrupt the vesicular membrane and cause cargo unloading. The content of the exosome is hybridized to DNA primers or antibodies immobilized on an electrode surface for quantification of molecular content. The EFIRM method is advantageous for extraction of exosomes and unloading cargo for analysis without lysis buffer. This method is capable of performing specific detection of both RNA and protein biomarker targets in the exosome. EFIRM extracts exosomes specifically based on their surface markers as opposed to size-based techniques. Transmission electron microscopy (TEM) and assay demonstrate the functionality of the method for exosome capture and analysis. The EFIRM method was applied to exosomal analysis of 9 mice injected with human lung cancer H640 cells (a cell line transfected to express the exosome marker human CD63-GFP) in order to test their exosome profile against 11 mice receiving saline controls. Elevated levels of exosomal biomarkers (reference gene GAPDH and protein surface marker human CD63-GFP) were found for the H640 injected mice in both serum and saliva samples. Furthermore, saliva and serum samples were demonstrated to have linearity (R = 0.79). These results are suggestive for the viability of salivary exosome biomarkers for detection of distal diseases.

Oral cancer is a major contributor to death worldwide. This life-threatening disease still lacks inexpensive screening. Novel strategies for early detection and therapy response are required. Saliva, as “mirror of the body,” reflects body's physiological and pathological state. Saliva is the epitome of a noninvasive, readily available, and easy to collect, transport, and store “biofluid.” Since the emphasis is switching more toward high impact personalized medicine, pioneering “Point-of-care salivary technologies” are being developed. These facilitate biomarker identification, as an alternative to invasive tissue biopsy. The concept that a patient can take a single drop of their own saliva and test it using portable wireless devices like biosensors, actuators, microelectronics, etc., like Oral Fluid NanoSensor Test (OFNASET) for oral cancer detection is being introduced. An upcoming contemporary tool that integrates both therapeutics and diagnostics is “Liquid biopsy.” It is a PCR-based sequencing technique that detects circulating tumor cells (CTCs) and fragments of tumor DNA shed into the bloodstream by cells undergoing apoptosis or necrosis. This review focuses on promising futuristic salivary technologies, which shall bring breakthroughs in terms of precision and efficiency. Oral physicians and oncologists will have ease in detecting and shaping diagnosis of oral cancer in near decade and beyond.

Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm of the oral cavity. Detection of OSCC is currently based on thorough clinical oral examination combined with biopsy for histological analysis. Most cases of OSCC are not detected until the cancer has developed into advanced stages; thus, a reliable early stage diagnostic marker is needed. This literature review presents an overview of the status of current advances in salivary diagnostics for OSCC. Though many protein and mRNA salivary biomarkers have been identified that can detect OSCC with high sensitivity and specificity, the most discernable findings occur with the use of multiple markers. Studies that incorporate proteomic, transcriptomic, and potentially additional "omics", including methylomics, need to be initiated to bring technology to clinical applications and allow the best use of saliva in diagnosing OSCC.

Abstract Background Differences in the expression of oxidative stress (OS) markers between female patients with temporomandibular disorders (TMD) and healthy individuals indicate that OS plays a role in the pathogenesis of TMD. Because chronic exposure to stress generates oxidative damage during continuous stimulation of the hypothalamic-pituitary-adrenal axis, we expected that higher levels of cortisol might be associated with higher oxidative damage. Our aim was to test the association between OS markers, stress perception, and salivary cortisol (SC) in chronic, female TMD patients. We tracked changes in OS markers and SC during occlusal splint therapy in order to evaluate the influence of treatment on oxidative status. We hypothesized that the effects of TMD therapy would differ among individuals depending on the source and intensity of pain. Methods Sixteen female patients were recruited, and 12 finished the study. Clinical assessment and saliva sampling were performed at the baseline and follow-up appointments. Repeated measures analysis of variance and Pearson’s correlation were used for analyzing the data. Results After 3 months, a significant reduction in afternoon total antioxidant capacity (TAC) was observed (p

Samuel Marshall,1 Alec J McCann,1 Tina L Samuels,1 Amy Blair,2 Valerie Bonne,2 Nikki Johnston,1,3 Jamie Koufman41Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Milwaukee, WI, USA; 2Department of Medicine, Medical College of Wisconsin, Milwaukee, WI, USA; 3Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, USA; 4Voice Institute of New York, New York, NY, USAObjective: Asthma and gastric reflux disease are widespread and often coexisting diseases with complex interactions, leading some to suspect that asthma symptoms of patients with reflux may improve with anti-reflux therapy. The objective of this study was to determine whether pepsin in saliva, indicative of airway reflux, could be detected in patients with asthma of varying severity and test the requirement of citric acid as a pepsin preservative.Methods: Saliva samples were collected in the clinic (with/without citric acid) and upon waking the following morning from 25 asthmatic patients. Enzyme-linked immunosorbent assay was performed for pepsin and interleukin-8 (IL-8), an inflammatory cytokine induced by pepsin in other airway epithelia. Pepsin induction of IL-8 was tested in a lung epithelial cell culture model.Results: Pepsin was detected in saliva from 14/25 patients (56%; mean concentration of pepsin in specimens where observed ±SD =80.3±87.5 ng/mL); significant agreement was found between samples collected in the presence/absence of citric acid. No significant associations were found with pepsin and clinical measures of asthma severity. IL-8 was detected in saliva from 22/25 patients (88%; mean IL-8 in all specimens where observed =3.27±3.91 ng/mL). IL-8 was significantly upregulated in human lung epithelial cells exposed to pepsin at pH7 in vitro (P=0.041).Conclusion: In summary, more than half of the asthma patients in this study were found to have pepsin in their saliva, indicative of airway reflux. These data support the use of salivary pepsin as a noninvasive tool for future investigation of airway reflux in a larger cohort. The data further suggest that collection in citric acid as a sample preservative is not warranted and that pooling of multiple saliva samples collected at various timepoints may improve sensitivity of pepsin detection and reduce costs incurred by multiple sample analysis in future studies.Keywords: extraesophageal reflux, laryngopharyngeal reflux, asthma, salivary diagnostics, IL-8