Comparison of oral metabolome profiles of stimulated saliva, unstimulated saliva, and mouth-rinsed water

Saliva includes a substantial amount of biological information, which has enabled us to understand the relationship between oral metabolites and various oral and systemic disorders. However, collecting saliva using a controlled protocol is time-consuming, making saliva an unsuitable analyte in large cohort studies. Mouth-rinsed water (MW), the water used to rinse the mouth, can be collected easily in less time with less difference between subjects than saliva and could be used as an alternative in oral metabolome analyses. In this study, we investigated the potential of MW collection as an efficient alternative to saliva sample collection for oral metabolome profiling. MW, stimulated saliva, and unstimulated saliva were collected from 10 systemically healthy participants. The samples were subjected to metabolome analysis using capillary electrophoresis time-of-flight mass spectrometry, and the types and amounts of metabolites in the samples were compared. Qualitatively, MW contained the same metabolites as unstimulated and stimulated saliva. While the quantity of the metabolites did not drastically change between the sampling methods, all three reflected individual differences, and the features of MW were the same as those of the unstimulated saliva. Overall, these results suggest that MW may be an appropriate alternative to saliva in oral metabolome profile analysis.

Effect of Temperature on Tooth Staining by 0.12% Chlorhexidine Gluconate

Introduction: Natural color of dentin is affected by many variables, including anatomical variations, age, how much dentin is exposed, or how much enamel is covering the dentin. Chlorhexidine gluconate (CHX) has been observed to cause tooth staining, especially of exposed dentin. Risk factors for CHX staining include the amount of time for CHX utilization amongst others. Interestingly, the temperature of the rinse when used has been identified as a risk factor. However, no evidence of the effect of temperature is available in the literature. The purpose of this study was to determine the effect of temperature on dentin staining due to CHX exposure.Methods: Two studies were done. The first a pilot study at room temperature to determine the time needed to establish staining solutions, a method to evaluate stain intensity, and establish the time needed to stain dentin samples in vitro. The second study exposed dentin samples on a twice daily basis to a 1 min soak in CHX at different temperatures, followed by a period in an unstimulated saliva mixed with black tea mixture. Temperatures tested were 4, 23, 37 and 50°C. Control samples were exposed to only black tea and saliva (no CHX) and tested at 23°C.Results: The pilot study found that the combination of CHX and black tea causes dentin staining. From this data the sample size needed for the second experiment was calculated, requiring 12 samples per group. Sixty dentin samples were divided amongst 5 groups. The data from this study showed significant darkening of the dentin samples over 18 days. The 4 and 23°C CHX rinses resulted in significant staining compared to the control samples. The 37 and 50°C CHX rinses did not stain significantly more than the control samples.Conclusions: Chlorhexidine has the ability to cause tooth staining in the presence of chromogens such as those in black tea. Significant darkening was observed at lower temperatures (4 and 23°C) over 18 days, therefore dental professionals may wish to advise gently warming the CHX rinse toward 37°C prior to use to reduce the risk of staining.

Unstimulated Parotid Saliva Is a Better Method for Blood Glucose Prediction

Objective: Saliva glucose has been widely used in diagnosing and monitoring diabetes, but the saliva collection method will affect saliva glucose concentration. So, this study aims to identify the ideal saliva collection method. Method: A total amount of six saliva collection methods were employed in 80 healthy participants in the morning. Besides, three unstimulated saliva methods were employed in another 30 healthy participants in the morning; in the meantime the blood glucose of these 30 participants was detected with a Roche blood glucose meter. The glucose oxidase method with 2, 4, 6-tribromo-3-hydroxybenzoic acid (TBHBA) as the chromogen has been improved to be suitable for healthy people, through the selection of the optimal pH value and ionic strength of the reaction system. This method was used for the detection of saliva glucose. Results: The improved method obtained absorbance at the wavelength of 520 nm, and the optimized parameter combination was pH 6.5 and 5 mg/dL NaCl. The lower limit of glucose detection was 0.1 mg/dL. Unstimulated saliva glucose concentration was higher than stimulated saliva glucose concentration. Unstimulated parotid saliva glucose concentration was the highest. Besides, unstimulated saliva glucose has a better normal distribution effect. Meantime, it was found that unstimulated parotid saliva was the most highly correlated with blood glucose (R2 = 0.707). Conclusions: the saliva collection method was an important factor that affected saliva glucose concentration. Unstimulated parotid saliva was the most highly correlated with blood glucose, which provided a reference for prediction of diabetes mellitus.

Descriptive Statistical Analysis Report on the Results of Questionnaire and Observations on Patients With Physiological Halitosis Who Visited the KUMC Halitosis Clinic- A Retrospective Study

Background: We initiated to prepare the basic data that could help us to understand the characteristics of patients with physiological halitosis symptoms in Korea and aid in explaining the causes of these symptoms. Methods: We collected questionnaire and dietary analysis results from the data of 246 adult patients diagnosed with physiological halitosis symptoms, collected the data from the patients’ salivary measurements and the analysis data of patients’ tongue and throat management status. Using the SPSS (IBM SPSS™, Ver. 22.0) program, we performed a frequency analysis, calculating the correlation coefficient Phi with an independence test.Results: The mean age and one standard deviation of 246 patients with physiological halitosis was 41.85±11.63 years. The proportion of women, the proportion of patients who replaced breakfast with fast food or skipped breakfast, with beverage drinking habits such as coffee, tea, or carbonated drink, with irregular or insufficient consumption of water, who recognized their own oral malodor, suffering anxiety or stress over oral malodor, with an introverted personality, with insufficient unstimulated salivation volume, with incorrect tongue management, and with PND (Postnasal drip) or PTC (Posterior Tongue Coating) were all statistically analyzed to be significant (p < 0.05). As a result of the correlation analysis, the Phi values of the seven pairs of the bivariate variables were from 0.137 to 0.196, and the Phi values of the five pairs of the bivariate variables - the Phi value between the breakfast pattern variable and the regular diet variable, the Phi value between the stimulating menu variable and the drinking variable, the Phi value between the unstimulated saliva variable and the stimulated saliva variable, the Phi value between the unstimulated saliva variable and the sedimentation ratio of saliva variable, and the Phi value between the stimulated saliva variable and the sedimentation ratio of saliva variable - were from 0.203 to 0.394 (p < 0.05). Conclusion: We could suggest the 10 potential common risk factors related to physiological halitosis symptom, and the five pairs of variables with Phi values ranging from 0.20 to 0.40 could be judged to have a moderate correlation.

Salivary Biomarkers (Opiorphin, Cortisol, Amylase, and IgA) Related to Age, Sex, and Stress Perception in a Prospective Cohort of Healthy Schoolchildren

Background. The use of easily accessible biomarkers for assessing young patients’ health is weighty. This cohort study is aimed at measuring stress/immune biomarkers in the saliva of healthy school-age children and comparing subgroups according to age, sex, and stress perception. Material and Methods. 503 children under 12 years old ( 8.7 ± 1.3 ) were included with anthropometric evaluation (height, waist, hip circumference, body weight, and body mass index (BMI)). Levels of opiorphin (OPI), free cortisol, alpha-amylase (sAA), and secreted immunoglobulin (sIgA) were determined by quantitative assays (ELISA) in unstimulated saliva. Unpaired t -test, Welch test, and Mann–Whitney U test were applied for appropriate group comparisons, and the correlation between variables was analyzed with Spearman’s rank coefficient. Results were considered significant at p < 0.05 . Results. sIgA and sAA exhibited significant differences depending on age and sex: IgA (ng/mL): 86 ± 68.6 vs. 104.9 ± 72.1 for (6-7 y.o.) and (8-11 y.o.), respectively, and 108.1 ± 80.1 vs. 94.6 ± 62.2 for male and females, respectively; sAA (U/mL): 78.9 ± 54.4 vs. 100.5 ± 81.2 for (6-7 y.o.) and (8-11 y.o.). No difference related to age or sex between groups was observed for cortisol and OPI. However, OPI levels were higher and correlated to prior stress exposure in children ( 0.31 ± 0.4 vs. 0.26 ± 0.5 ng / mL , p = 0.031 ). sAA was negatively correlated to low mood self-declaration in children in the last two weeks ( r = − 0.10 , p = 0.045 ). Conclusions. sIgA and sAA can be used as sex- and age-related biomarkers in children 6-12 y.o., which is not the case for free cortisol and opiorphin. However, OPI reflected previous exposure to stress, suggesting its use for evaluating stress-related changes in children

Salivary Atopy Biomarkers in Patients with Geographic Tongue

Objectives Geographic tongue is the mucositis of the dorsal surface of the tongue with no definite etiology, although it may be associated with atopic conditions. In this study, the salivary immunoglobulin E (IgE) and eosinophilic cationic protein (ECP) levels were assessed in patients with geographic tongue to examine the relationship between this condition and allergy-related biomarkers. Materials and Methods In this case-control study, unstimulated saliva was collected from 45 geographic tongue patients and 45 controls, and the salivary IgE and ECP levels were assessed. The data were analyzed with SPSS 20.0 using the Mann–Whitney test with a significance level of p < 0.05. Results The salivary IgE level in the case and control groups was 123.76 ± 8.1 and 74.34 ± 6.2 IU/mL, respectively, and the salivary ECP level was 9.4 ± 6.0 and 7.6 ± 3.2 ng/mL, respectively. There was a statistically significant difference between the two groups in terms of salivary IgE (p = 0.007) and ECP (p = 0.001) levels. Conclusion Salivary IgE and ECP levels increase in patients with geographic tongue. They can, therefore, be used for the initial diagnostic workup and to investigate the possible association of geographic tongue with allergic reactions.

The Use of Saliva as a Biosample in the Light of COVID-19

Saliva is easy to collect and a biofluid that is readily available without the need for special equipment for its collection. The collection process, which is non-invasive and inexpensive, leads to obtaining a biomaterial that can serve as a source of information for molecular diagnostics of diseases in general medicine, genetics and dentistry. Unfortunately, many of the salivary methodologies are lacking important parameters to provide for not only the safety of the operator, but also the quality and reproducibility of the research. Since the COVID-19 pandemic, salivary diagnostics demonstrate a great potential for research of SARS-CoV 2. In this review, good practice for unstimulated saliva collection and patient preparation was provided, based on the latest literature and available guidelines. Schemes for saliva collection procedures were presented following an extended literature search. Descriptions of salivary probes/cups, techniques of saliva collection, and the use of specific buffering solutions for the stability of collected samples for SARS-CoV-2 detection were also evaluated.

The estimation of the viable count of mutans streptococcus in waterpipe smokers and cigarette smokers

Background: Waterpipe and cigarette are two products of tobacco consumption, tobacco use has detrimental effects on the oral cavity, numerous studies around the world have reported a significant relationship between smoking and increase dental caries and viable count of cariogenic bacteria, Materials and Methods: unstimulated saliva was collected from 84 subjects and divided equally into three groups waterpipe smokers, cigarette smokers, and non-smokers all of the participants are adult male aged between 25-60 years, dental caries was measured by use DMFT index, while S.mutans and S.sobrinus were isolated by using a selective medium SB 20M (Sugar bacitracin-20 modified) agar Results: this present study showed a significant (p≤0.01). higher DMFT, DT, MT, and FT among cigarette smokers group than both waterpipe smokers and non-smokers groups. The viable count of S mutans was significantly (p≤0.01).higher in the cigarette smokers group followed by the waterpipe smokers group and then the non-smoker's group, while the viable count of S sobrinus showed no statistical differences (P>0.05) between groups. The correlation of DMFT with S.mutans, and S sobrinus count, were significantly positive (p ≤0.05), in the cigarette smokers group only Conclusion: Dental caries increase in cigarette smokers, where the DMFT and S mutans viable count are less affected by waterpipe than cigarette smoking, a positive correlation is found between DMFT and .mutans streptococcus count only in cigarette smokers

Evaluation of Salivary Leptin Levels and Its Correlation with Class I, Class II, and Class III Facial Skeletal Pattern: A Prefatory Study

Materials and Methods A sample of 62 patients were selected prior to the orthodontic treatment from a population that attended the International Islamic University Malaysia Specialist Orthodontic Clinic. Based on the lateral cephalometric analysis, the subjects were grouped into Class I, Class II, and Class III facial skeletal patterns, according to Eastman and Wits appraisal. Subsequently, unstimulated saliva samples were taken and purified to undergo leptin enzyme-linked immunosorbent assay analysis to determine the levels of leptin hormone. Statistical analysis using the Kruskal–Wallis test was used to analyze the data obtained. Results The results showed that there was a significant difference between the levels of leptin hormone between Class I and Class II skeletal patterns and between Class I and Class III facial skeletal patterns. No statistical difference was noted between the levels of leptin of Class II and Class III facial skeletal patterns. Conclusion Salivary leptin hormone levels are higher in patients with Class II and Class III facial skeletal patterns compared with Class I.