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Objectives To study which salivary proteins form the protective bound mucosal pellicle and to determine the role of transglutaminase in pellicle development. Materials and Methods Oral epithelial cells were collected and underwent washes of different strengths, followed by homogenisation. SDS- PAGE, western blotting, IgA ELISAs and amylase activity assays were completed on cell homogenates and compared to saliva samples to confirm which salivary proteins were bound to cell surfaces. Results Salivary mucins, MUC5B and MUC7, were strongly retained on the oral epithelial cell surface. Other bound proteins including cystatin S, carbonic anhydrase VI, secretory component and IgA could be washed off. IgA was present in concentrated levels in the bound mucosal pellicle compared to amounts in saliva. Amylase, one of the most abundant proteins present in saliva, showed minimal levels of binding. Transglutaminase 3 presence was confirmed, but proteins that it catalyses cross-links between, statherin and proline-rich proteins, showed minimal presence. Conclusion Some protective salivary proteins including mucins and IgA become concentrated on oral surfaces in the bound mucosal pellicle, through specific interactions. Concentration of mucins would contribute to lubrication to prevent abrasion damage to soft tissues, whilst increased IgA could create an 'immune reservoir' against mucosal infection. [ABSTRACT FROM AUTHOR]

Introduction: This longitudinal study assessed the association between salivary protein composition and the clinical onset/severity of oral mucositis (OM) in patients with head and neck tumours treated with intensity-modulated-radiotherapy (IMRT)., Methods: Saliva samples/clinical data were obtained from 40 head and neck cancer patients treated at Guy's Hospital before -IMRT(T0) and after-IMRT (T1 = 6 m, T2 = 12 m) (ethics approval/consent). Salivary flow rate, total protein concentration, and secretion rate were determined from saliva samples and compared with pre-treatment values. OM was assessed, total/specific salivary proteins, including mucin 5B and 7, IgA, cystatin-S, albumin, and α-amylase, were quantified., Results: 95% patients experienced OM during IMRT, with 33 subjects reaching grade 2&3. At T1, there was a significant reduction in salivary flow rate, total protein secretion rate, α-amylase and cystatin-S compared to baseline. Remarkably IMRT did not significantly alter mucin 5B and 7, or the IgA secretion rate at any time point. At T1, all the analyzed proteins were associated with the OM outcomes. In addition, there was a significant inverse correlation between IgA concentration at T0 and the severity of OM during IMRT., Conclusion: This study revealed significant associations between several salivary proteins and OM in patients with head and neck cancer undergoing IMRT. Further longitudinal studies are needed to confirm these results., Clinical Significance: The study contributes to the understanding of certain salivary proteins association with OM. This could be the first step towards identifying potential salivary markers that could offer perspectives for personalized medicine approaches to improve their quality of life (QoL)., Research Question: What is the association between salivary proteins and the occurrence and severity of OM in head and neck cancer patients?, Aim: To assess the association between salivary protein composition with the clinical onset/severity of oral mucositis (OM) in head and neck cancer patients treated with intensity modulated radiotherapy., Null Hypothesis: There is no association between salivary proteins and onset/severity of OM in HNC patients., (© 2024. The Author(s).)

This study aimed to compare the metabolic profile of unstimulated (US) and stimulated (SS) saliva samples from pregnant women with/without obesity and periodontitis. Ninety-six pregnant women were divided into: obesity + periodontitis (OP = 20); obesity/no periodontitis (OWP = 27); normal BMI + periodontitis (NP = 20); and normal BMI/no periodontitis (NWP = 29). US and SS samples were collected by expectoration and chewing of sterilized parafilm gum, respectively, and samples were individually analyzed by Proton Nuclear Magnetic Resonance ( 1 H-NMR). Univariate ( t test and correlations) and multivariate (Principal Component Analysis-PCA, and Partial Least Square-Discriminant Analysis-PLS-DA with Variance Importance Projection-VIP scores) and Metabolite Set Enrichment Analysis were done ( p < 0.05). Metabolites commonly found in all groups in elevated concentration in US samples were 5-Aminopentoate, Acetic acid, Butyric acid, Propionic acid, Pyruvic acid, and Succinic acid. They were mainly related to the butyrate metabolism, citric acid cycle, amino sugar metabolism, fatty acids biosynthesis, pyruvate metabolism, glutamate metabolism, and Warburg effect. Metabolites commonly found in all groups that were in elevated concentration in SS samples were Citrulline, Fumaric acid, Histidine, N -acetyl glutamine, N -acetylneuraminic acid, para-hydroxyphenylacetic acid, Proline, Tyrosine. Although some differences were found between unstimulated and stimulated saliva samples from pregnant women with/without obesity and periodontitis, stimulated saliva collection seems adequate, demonstrating similar metabolic pathways to unstimulated saliva samples when groups are compared.

Mucus reduces friction between epithelial surfaces by providing lubrication in the boundary and mixed regime. Mucins, the main macromolecule, are heavily glycosylated proteins that polymerise and retain water molecules, resulting in a hydrated biogel. It is assumed that positively charged ions can influence mucin film structure by screening the electrostatic repulsions between the negatively charged glycans on mucin moieties and draw in water molecules via hydration shells. The ionic concentration can vary significantly in different mucus systems and here we show that increasing the ionic concentration in mucin films leads to an increase in lubrication between two polydimethylsiloxane surfaces at sliding contact in a compliant oral mimic. Mucins were found to bind sodium ions in a concentration-dependent manner and increased ionic concentration appears to cause mucin films to swell when assessed by Quartz Crystal hiMicrobalance with Dissipation (QCM-D) analysis. Furthermore, we determined that the removal of negatively charged sialic acid moieties by sialidase digestion resulted in reduced adsorption to hydrophilic surfaces but did not affect the swelling of mucin films with increasing ionic concentrations. Moreover, the coefficient of friction was increased with sialic acid removal, but lubrication was still increased with increasing ionic concentrations. Taken together this suggests that sialic acids are important for lubrication and may exert this through the sacrificial layer mechanism. Ionic concentration appears to influence mucin films and their lubrication, and sialic acids, at least partly, may be important for ion binding., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Abby Weston reports financial support was provided by Medical Research Council. Abby Weston reports financial support was provided by Mars Wrigley Confectionery Ltd. Abby Weston reports a relationship with Medical Research Council that includes: funding grants and travel reimbursement. Abby Weston reports a relationship with Mars Wrigley Confectionery Ltd that includes: funding grants., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Dental erosion is a disease of the oral cavity where acids cause a loss of tooth enamel and is defined as having no bacterial involvement. The tooth surface is protected from acid attack by salivary proteins that make up the acquired enamel pellicle (AEP). Bacteria have been shown to readily degrade salivary proteins, and some of which are present in the AEP. This study aimed to explore the role of bacteria in dental erosion using a multi-omics approach by comparing saliva collected from participants with dental erosion and healthy controls., Results: Salivary proteomics was assessed by liquid-chromatography mass spectrometry (LC-MS) and demonstrated two altered AEP proteins in erosion, prolactin inducible protein (PIP), and zinc-alpha-2 glycoprotein (ZAG). Immunoblotting further suggested that degradation of PIP and ZAG is associated with erosion. Salivary microbiome analysis was performed by sequencing the bacterial 16S rRNA gene (V1-V2 region, Illumina) and showed that participants with dental erosion had a significantly (p < 0.05) less diverse microbiome than healthy controls (observed and Shannon diversity). Sequencing of bacterial mRNA for gene expression (Illumina sequencing) demonstrated that genes over-expressed in saliva from erosion participants included H + proton transporter genes, and three protease genes (msrAB, vanY, and ppdC). Salivary metabolomics was assessed using nuclear magnetic resonance spectrometry (NMR). Metabolite concentrations correlated with gene expression, demonstrating that the dental erosion group had strong correlations between metabolites associated with protein degradation and amino acid fermentation., Conclusions: We conclude that microbial proteolysis of salivary proteins found in the protective acquired enamel pellicle strongly correlates with dental erosion, and we propose four novel microbial genes implicated in this process. Video Abstract., (© 2023. The Author(s).)

This study analyzed the salivary proteomics, adiponectin and albumin, related to weight loss and periodontitis in patients undergoing bariatric surgery. This study included fourteen patients with morbid obesity (body mass index, BMI > 40 kg/m 2 ) who underwent bariatric surgery Roux-en-Y gastric bypass (RYGB) in System Health Public in Brazil. Data on demographic and anthropometric measures were extracted from medical records preoperatively and 6 and 12 months post-surgery. The variables assessed were: probing pocket depth (PPD), clinical attachment loss (CAL), bleeding on probing (BOP), and stimulated whole-mouth saliva. In this study, saliva samples were analyzed by electrophoresis and immunoblotting. The ELISA kit was used to measure the MMP8 levels to determine potential markers for obesity. Adiponectin and albumin levels were also evaluated. Weight loss was associated with significant changes in patients' periodontal clinical data. Although 7 out of 10 periodontal patients showed an increase in salivary adiponectin levels after root planning treatment, when analyzed by Western blotting, the increase was not statistically significant (21.1 ± 4.8 to 26.3 ± 9.4 arbitrary units, p > 0.99). There was no correlation between albumin levels and salivary adiponectin pre-surgery, nor 6 months or 12 months after surgery. Weight loss was not improved by low-grade inflammation in bariatric patients, since albumin levels were similar between periods. Periodontitis is an inflammatory disease that is modulated by several factors, among which adiponectin plays an important role for the treatment of periodontal disease.

Introduction: Space closure is a challenging and time-consuming phase of orthodontic treatment with fixed appliances. This systematic review evaluated canine retraction duration using fixed appliances after maxillary first premolar extraction., Methods: Unrestricted systematic literature searches were conducted in 8 databases for randomized clinical trials, assessing the duration and rate of maxillary canine retraction using fixed appliances with or without treatment adjuncts published up to July 2021. Study selection, data extraction, and risk of bias evaluation were conducted independently and in duplicate. Random-effects meta-analyses of average rates or mean differences (MD) and 95% confidence intervals (CI) were conducted at α = 5%, followed by sensitivity and Grading of Recommendations Assessment, Development, and Evaluation analysis., Results: Fifty randomized clinical trials (6 parallel and 44 split-mouth designs) covering 811 participants (mean age 19.9 years; 34% male) were included. The estimated average pooled duration to achieve complete canine retraction was 4.98 months (2 trials; 95% CI, -2.9 to 12.88 months). Pooled average canine retraction was 0.97 mm at months 0-1 (23 trials; 95% CI, 0.79-1.16), 1.83 mm at months 0-2 (20 trials; 95% CI, 1.52-2.14), 2.44 mm at months 0-3 (23 trials; 95% CI, 2.10-2.79), 3.49 mm at months 0-4 (6 trials; 95% CI, 1.81-5.17) and 4.25 mm at months 0-5 (2 trials; 95% CI, 0.36-8.14). Surgically-assisted orthodontics was associated with greater canine retraction at all time points: months 0-1 (10 trials; MD, 0.52 mm; P = 0.004), months 0-2 (8 trials; MD, 0.53 mm; P = 0.04), months 0-3 (8 trials; MD, 0.67 mm; P = 0.01), and months 0-4 (3 trials; MD, 1.13 mm; P = 0.01), whereas subgroup analyses indicated significant effects of anchorage reinforcement method and bracket slot size on canine retraction., Conclusions: The average time to achieve complete retraction of the maxillary canine using fixed appliances was around 5.0 months. Most studies used split-mouth randomization to investigate canine retraction for around 1-3 months, with substantial heterogeneity across studies. At 3 months of treatment, high-quality evidence supported greater canine retraction with surgically-assisted orthodontics., (Copyright © 2022. Published by Elsevier Inc.)

Orthodontic tooth movement (OTM) occurs through proteolytic remodelling within the periodontium following the application of external force to the tooth. This study describes the first characterization of the salivary peptidome and protease profile during the alignment stage of fixed appliance orthodontic treatment. Unstimulated whole mouth saliva from 16 orthodontic patients (10 males, 6 females, mean (SD) age 15.2 (1.6) years) was collected prior to fixed appliance placement (T1), 1-h (T2), 1-week (T3) following fixed appliance placement and on completion of mandibular arch alignment (T4). Salivary peptides were extracted using filtration followed by mass spectrometry to identify amino acid sequences. Protease prediction was carried out in silico using Proteasix and validated with gelatin zymography and enzyme-linked immunosorbent assay. A total of 2852 naturally-occurring peptides were detected, originating from 436 different proteins. Both collagen and statherin-derived peptide levels were increased at T2. Proteasix predicted 73 proteases potentially involved in generating these peptides, including metalloproteinases, calpains and cathepsins. Changes in predicted activity of proteases over time were also observed, with most metalloproteinases showing increased predicted activity at T2-T3. Increased gelatinolytic activity and MMP8/MMP9 levels were detected at T3. Collectively, multiple protein targets and changes in protease-predicted activity during OTM have been identified., (© 2023. The Author(s).)

Obesity and pregnancy may have synergistic effects on periodontal condition, and proteomics could be an ideal approach to highlight the pathophysiological mechanisms associated with these outcomes. This study analyzed the salivary proteomics related to obesity and periodontitis in women during pregnancy (T1) and after delivery (T2). Initially, 126 women were recruited and forty were allocated into groups: with obesity and periodontitis (OP); with obesity, but without periodontitis (OWP); with normal BMI, but with periodontitis (NP); with normal BMI and without periodontitis (NWP). Whole-mouth saliva was collected in T1 and T2, and proteins were extracted and individually processed by label-free proteomics ( nLC-ESI-MS/MS ). The up-regulations of Heat shock 70 kDa protein 1A , 1B , and 1-like were related to both obesity and periodontitis, separately. Albumin and Thioredoxin were up-regulated in periodontitis cases, while Cystatins (mainly S , SA , SN ) and Lactotransferrin were down-regulated. The high abundances of Submaxillary gland androgen-regulated protein 3B , Protein S100-A8 , Matrix metalloproteinase-9 , Heat shock 70 kDa protein 2 and 6 , Putative Heat shock 70 kDa protein 7 , Heat shock 71 kDa protein , Haptoglobin and Plastin-1 were significant in the combination of obesity and periodontitis. Obesity and periodontitis remarkably altered the proteome of the saliva during pregnancy with substantial alterations after delivery.

Objectives: The aim of this study was to compare the intra-oral bacterial profile of normal-weight and obese adolescents prior to orthodontic treatment with fixed appliances., Materials and Methods: Nineteen adolescent patients were recruited into two groups based upon body mass index (BMI) and classified as normal-weight or obese. Unstimulated whole mouth saliva was obtained for 5 minutes. Bacterial DNA extraction was performed from saliva, and 16S rRNA gene sequencing of the V1-2 variable regions was undertaken followed by analysis using the mothur pipeline., Results: Saliva from a total of 19 adolescent patients with mean (SD) age 15.6 (1.8) years were divided into 10 normal-weight with mean BMI of 19.4 (2.2) kg/m 2 and 9 obese with mean BMI of 30.2 (3.5) kg/m 2 . A total of 156 783 sequences were obtained from the 19 samples with no significant differences in richness or diversity between sample groups by obesity status or gender (AMOVA). The bacterial community in both groups was dominated by bacterial genera characteristic of the human mouth, which included Streptococcus, Porphyromonas, Veillonella, Gemella, Prevotella, Fusobacterium and Rothia., Conclusion: There were no differences in alpha or beta diversity of oral bacterial communities between normal-weight and obese orthodontic patients. Obese adolescents attending for orthodontic treatment had a similar microflora to their normal-weight counterparts., (© 2022 The Authors. Orthodontics & Craniofacial Research published by John Wiley & Sons Ltd.)

This study aimed to elucidate the metabolomic signature associated with obesity and periodontitis during pregnancy in plasma and saliva biofluids. Ninety-eight pregnant women were divided into: with obesity and periodontitis (OP = 20), with obesity but without periodontitis (OWP = 27), with normal BMI but with periodontitis (NP = 21), with normal BMI and without periodontitis (NWP = 30). Saliva and plasma were analyzed by 1H-NMR for metabolites identification. Partial Least Squares-Discriminant Analysis (PLS-DA), Sparse PLS-DA (sPLS-DA), and Variable Importance of Projection (VIP) were performed. ANOVA and Pearson’s correlation were applied (p < 0.05). Plasmatic analysis indicated the levels of glucose (p = 0.041) and phenylalanine (p = 0.015) were positively correlated with periodontal parameters and BMI, respectively. In saliva, periodontitis was mainly associated with high levels of acetic acid (p = 0.024), isovaleric acid, butyric acid, leucine, valine, isoleucine, and propionic acid (p < 0.001). High salivary concentrations of glycine (p = 0.015), succinic acid (p = 0.015), and lactate (p = 0.026) were associated with obesity. Saliva demonstrated a more elucidative difference than plasma, indicating the glucose-alanine cycle, alanine metabolism, valine, leucine and isoleucine degradation, glutamate metabolism, and Warburg effect as the main metabolic pathways.

Escherichia coli is a Gram-negative bacterium that colonises the human intestine and virulent strains can cause severe diarrhoeal and extraintestinal diseases. The protein SslE is secreted by a range of pathogenic and commensal E. coli strains. It can degrade mucins in the intestine, promotes biofilm maturation and it is a major determinant of infection in virulent strains, although how it carries out these functions is not well understood. Here, we examine SslE from the commensal E. coli Waksman and BL21 (DE3) strains and the enterotoxigenic H10407 and enteropathogenic E2348/69 strains. We reveal that SslE has a unique and dynamic structure in solution and in response to acidification within mature biofilms it can form a unique aggregate with amyloid-like properties. Furthermore, we show that both SslE monomers and aggregates bind DNA in vitro and co-localise with extracellular DNA (eDNA) in mature biofilms, and SslE aggregates may also associate with cellulose under certain conditions. Our results suggest that interactions between SslE and eDNA are important for biofilm maturation in many E. coli strains and SslE may also be a factor that drives biofilm formation in other SslE-secreting bacteria., (© 2022. The Author(s).)

Introduction: A key goal of orthodontic treatment with fixed appliances is alignment of the dentition, and this remains a commonly selected outcome in clinical studies investigating orthodontic tooth movement. This systematic review has evaluated treatment duration to achieve alignment of the mandibular dentition using fixed appliances., Methods: Systematic literature searches without restrictions were undertaken in 9 databases for randomized clinical trials (RCTs) assessing duration and rate of tooth alignment using fixed appliances with or without treatment adjuncts published up to January 2021. After duplicate study selection, data extraction, and risk of bias assessment according to Cochrane, random-effects meta-analyses of aggregate data, and individual patient data were conducted., Results: Thirty-five trials were included with 2258 participants (39% male; mean age 17.8 years), giving a pooled duration to achieve whole-arch alignment of the mandibular dentition of 263.0 days (4 trials; 95% confidence interval [CI], 186.7-339.4 days) and incisor alignment in the mandibular arch of 100.7 days (9 trials; 95% CI, 84.1-117.4 days). Surgical-assisted orthodontics was associated with reduced duration of incisor alignment: mean difference of 44.3 days less (4 trials; 95% CI, 20.0-68.9 days; P <0.001; high quality of evidence), whereas subgroup and meta-regression analyses indicated significant effects of baseline crowding and premolar extractions. Individual patient data analysis from 3 RCTs indicated that for each additional participant age year, whole-arch alignment of the mandibular dentition took 13.7 days longer (3 trials; 95% CI, 7.7-17.7 days; P <0.001) and for each additional mm of irregularity, 17.5 days more were needed (2 trials; 95% CI, 9.8-25.2 days; P <0.001)., Conclusions: Patient and treatment-related characteristics can significantly affect the duration of tooth alignment and should be taken into account both clinically and when designing trial outcomes. Future research studies investigating rates of orthodontic tooth alignment would benefit from adequate sample sizes and a more consistent methodology in outcome assessment. Data in this systematic review provides a basis for appropriate trial design for future RCTs investigating the rate of orthodontic tooth alignment with fixed appliances., (Copyright © 2021 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This prospective clinical cohort study investigated the potential influence of obesity on orthodontic treatment outcome., Methods: A prospective cohort of adolescent patients undergoing routine fixed appliance treatment were recruited into normal-weight or obese groups based upon body mass index (BMI) centile and followed up until the completion of treatment. Primary outcome was treatment duration, and secondary outcomes included treatment outcome (occlusal change measured using peer assessment rating [PAR]), appointment characteristics, and compliance measures., Results: A total of 45 patients mean age 14.8 (1.6) years were included in the final analysis. The normal-weight group included 23 patients with mean BMI 19.4 (2.4) kg/m2 and the obese group 22 patients with mean BMI 30.5 (3.8) kg/m2. There were no significant differences in baseline demographics between groups, except for BMI and pre-treatment PAR. The normal-weight group had a mean pre-treatment PAR of 25.6 (8.3) and the obese 33.3 (11.8) giving the obese group a more severe pre-treatment malocclusion (P = 0.02). There were no significant differences in treatment duration between groups (P = 0.36), but obese patients needed less time per each additional baseline PAR point compared to normal weight (P = 0.02). Obese patients also needed less appointments compared to normal-weight patients (P = 0.02). There were no significant differences between groups for appointment characteristics or compliance. Finally, obese patients were more likely to experience a great PAR reduction than normal-weight patients (relative risk = 2.6; 95% confidence interval = 1.2-4.2; P = 0.02)., Conclusions: There were no significant differences in treatment duration between obese and normal-weight patients. Obesity does not appear to be a risk factor for negative orthodontic treatment outcome with fixed appliances., (© The Author(s) 2020. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Within the mouth bacteria are starved of saccharides as their main nutrient source between meals and it is unclear what drives their metabolism. Previously oral in vitro biofilms grown in saliva have shown proteolytic degradation of salivary proteins and increased extracellular proline. Although arginine and glucose have been shown before to have an effect on oral biofilm growth and activity, there is limited evidence for proline. Nuclear magnetic resonance (NMR) spectroscopy was used to identify extracellular metabolites produced by bacteria in oral biofilms grown on hydroxyapatite discs. Biofilms were inoculated with stimulated whole mouth saliva and then grown for 7 days using sterilized stimulated whole mouth saliva supplemented with proline, arginine or glucose as a growth-medium. Overall proline had a beneficial effect on biofilm growth-with significantly fewer dead bacteria present by biomass and surface area of the biofilms ( p < 0.05). Where arginine and glucose significantly increased and decreased pH, respectively, the pH of proline supplemented biofilms remained neutral at pH 7.3-7.5. SDS-polyacrylamide gel electrophoresis of the spent saliva from proline and arginine supplemented biofilms showed inhibition of salivary protein degradation of immature biofilms. NMR analysis of the spent saliva revealed that proline supplemented biofilms were metabolically similar to unsupplemented biofilms, but these biofilms actively metabolized proline to 5-aminopentanoate, butyrate and propionate, and actively utilized glycine. This study shows that in a nutrient limited environment, proline has a beneficial effect on in vitro oral biofilms grown from a saliva inoculum., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Cleaver, Moazzez and Carpenter.)

The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse which can be ascribed to the variety of niches available in the mouth. Previous studies have focused on the microflora in disease-either caries or periodontitis-and only recently have they considered factors that maintain the normal microflora. This has led to the perception that the microflora proliferate in nutrient-rich periods during oral processing of foods and drinks and starves in between times. In this review, evidence is presented which shows that the normal flora are maintained on a diet of salivary factors including urea, lactate, and salivary protein degradation. These factors are actively secreted by salivary glands which suggests these factors are important in maintaining normal commensals in the mouth. In addition, the immobilization of SIgA in the mucosal pellicle indicates a mechanism to retain certain bacteria that does not rely on the bacterial-centric mechanisms such as adhesins. By examining the salivary metabolome, it is clear that protein degradation is a key nutrient and the availability of free amino acids increases resistance to environmental stresses.

Metabolomics has been identified as a means of functionally assessing the net biological activity of a particular microbial community. Considering the oral microbiome, such an approach remains largely underused. While the current knowledge of the oral microbiome is constantly expanding, there are several deficits in knowledge particularly relating to their interactions with their host. This work uses nuclear magnetic resonance spectroscopy to investigate metabolic differences between oral microbial metabolism of endogenous (i.e., salivary protein) and exogenous (i.e., dietary carbohydrates) substrates. It also investigated whether microbial generation of different metabolites may be associated with host taste perception. This work found that in the absence of exogenous substrate, oral bacteria readily catabolize salivary protein and generate metabolic profiles similar to those seen in vivo. Important metabolites such as acetate, butyrate, and propionate are generated at relatively high concentrations. Higher concentrations of metabolites were generated by tongue biofilm compared to planktonic salivary bacteria. Thus, as has been postulated, metabolite production in proximity to taste receptors could reach relatively high concentrations. In the presence of 0.25 M exogenous sucrose, increased catabolism was observed with increased concentrations of a range of metabolites relating to glycolysis (lactate, pyruvate, succinate). Additional pyruvate-derived molecules such as acetoin and alanine were also increased. Furthermore, there was evidence that individual taste sensitivity to sucrose was related to differences in the metabolic fate of sucrose in the mouth. High-sensitivity perceivers appeared more inclined toward continual citric acid cycle activity postsucrose, whereas low-sensitivity perceivers had a more efficient conversion of pyruvate to lactate. This work collectively indicates that the oral microbiome exists in a complex balance with the host, with fluctuating metabolic activity depending on nutrient availability. There is preliminary evidence of an association between host behavior (sweet taste perception) and oral catabolism of sugar.

One of the most accepted mechanisms of astringency consists of the interaction between polyphenols and some specific salivary proteins. This work aims to obtain further insights into the mechanisms leading to a modulation of astringency elicited by polyphenols. The effect of the presence of different chemical species (present in food and beverages as food additives) on the polyphenol-protein interaction has been evaluated by means of techniques such as sodium dodecyl sulfate polyacrylamide gel electrophoresis and cell cultures using a cell-based model of the oral epithelium. Results obtained showed that several chemicals, particularly sodium carbonate, seem to inhibit polyphenol binding to salivary proteins and to oral epithelium. These results point out that polyphenol-saliva protein interactions can be affected by some food additives, which can help to better understand changes in astringency perception.

Objectives: To investigate the effects of obesity on biomarker levels within lower incisor gingival crevicular fluid (GCF) in subjects undergoing routine fixed appliance orthodontic treatment., Materials and Methods: This was a cross-sectional clinical cohort study. GCF was collected from normal-weight and obese subjects at completion of alignment at least 1 month after placement of 0.019 × 0.025-inch stainless-steel archwires. The primary outcome was the difference in GCF biomarker levels between groups. Secondary outcomes included differences in clinical parameters of plaque and gingival indices, unstimulated whole-mouth saliva, and GCF flow rates., Results: Thirty-eight subjects (18 male, 20 female) with a mean age of 25.6 (SD, 6.3) years and mean body mass index (BMI) of 22.6 (1.6) in normal-weight and 32.4 (2.2) kg/m 2 in obese groups were investigated. Apart from BMI ( P < .0001), there were no statistically significant differences in essential demographics between groups. Significantly increased levels of the adipokine leptin ( P < .009) and the tissue-remodeling biomarker matrix metalloproteinase-9 (MMP9; P < .020) were identified in the obese cohort. For the remainder of the biomarkers, including the RANKL bone-remodeling marker and several inflammatory markers, there were no significant differences between groups. No correlation was observed between plaque index or gingival index for any GCF biomarker for either group ( P = .07-1.00)., Conclusions: This study investigated the GCF biochemical profile of obese and normal-weight subjects undergoing fixed-appliance orthodontic treatment. Significantly increased levels of the adipokine leptin and the tissue-remodeling biomarker MMP9 were identified in the obese group. These data provide evidence of differences in GCF biochemistry between obese and normal-weight subjects undergoing fixed appliance orthodontic treatment.

The phosphorylation of (+) alpha tocopherol produces adhesive nanostructures that interact with oral biofilms to restrict their growth. The aim of this work was to understand if these adhesive (+) alpha tocopheryl phosphate (α-TP) nanostructures could also control macrophage responses to the presence of oral bacteria. The (+) α-TP planar bilayer fragments (175 nm ± 21 nm) formed in a Trizma®/ethanol vehicle swelled when exposed to the cell lines (maximum stabilized size = 29 μm). The swelled (+) α-TP aggregates showed selective toxicity towards THP-1 macrophages (LD 50  = 304 μM) compared to human gingival fibroblasts (HGF-1 cells; LD 50  > 5 mM), and they inhibited heat killed bacteria stimulated MCP-1 production in both macrophages (control 57.3 ± 18.1 pg/mL vs (+) α-TP 6.5 ± 3.2 pg/mL) and HGF-1 cells (control 673.5 ± 133 pg/mL vs (+) α-TP - 463.9 ± 68.9 pg/mL)., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Taste loss is a significant problem in older adults, affecting quality of life and nutrition. Altered salivary rheology and loss of mucin function may contribute to taste loss by reducing mucosal defences in the oral cavity, impairing sensitivity to oral stimulants. This study aimed to investigate the effects of salivary rheology on taste loss in ageing. Salivary mucin glycosylation and binding to the oral epithelium was investigated in older and younger adults. A cell-based model was utilised to consider the role of saliva in taste loss., Methods: Human subjects aged >60 years ( n = 25) and 18-30 ( n = 30) provided saliva samples which were analysed for viscosity, mucin composition and mucin binding to oral epithelial cells (TR146/MUC1). Oral epithelial cells (TR146/MUC1 and SCC090) provided models for taste receptor activation., Results: Reduced levels and sialylation of MUC7 were evident in saliva of older adults which may lead to reduced viscoelasticity, while viscosity is unaffected. Impaired muco-adhesion of saliva from older adults was also observed. Saliva from older adults facilitated the bitter taste receptor activation less well than saliva from younger adults. The causes of taste dysfunction in older adults are unknown, but this study supports a role of saliva in facilitating the activation of taste receptors.

Oral biofilms have not been studied using both metabolome and protein profiling concurrently. Bacteria produce proteases that lead to degradation of functional salivary proteins. The novel protocol described here allows for complete characterisation of  in vitro  oral biofilms, including proteolytic, metabolic, and microbiome analysis. Biofilms were grown on hydroxyapatite discs from whole mouth saliva, using sterilised saliva as a growth-medium, in different growth environments. Salivary protein degradation was assessed from spent saliva growth-medium using SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and metabolic activity by nuclear magnetic resonance (NMR). Discs were assessed for depth and coverage of biofilms by confocal laser scanning microscopy (CLSM), and biofilms were collected at the end of the experiment for  16S rRNA  gene sequence analysis. There was a significant difference in biofilm viability, salivary protein degradation, and metabolites identified between biofilms grown aerobically and biofilms exposed to an anaerobic environment. Bacterial  16S rRNA  gene sequencing showed the predominant genus in the 7-day aerobic biofilms was  Streptococcus , in aerobic-anaerobic and anaerobic 7-day biofilms  Porphyromonas , and in aerobic-anaerobic and anaerobic 13-day biofilms  Fusobacterium . This data suggests new growth requirements and capabilities for analysing salivary biofilms  in vitro , which can be used to benefit future research into oral bacterial biofilms.

Fungiform papillae house taste buds on the anterior dorsal tongue. Literature is inconclusive as to whether taste perception correlates with fungiform papillae density (FPD). Gustatory reflexes modulate the amount and composition of saliva subsequently produced, and thus may be a more physiologically objective measure of tastant-receptor interactions. Taste perception fluctuates with time but the stability of individual fungiform papillae is unclear. This study followed ten healthy volunteers longitudinally at baseline, one and six months. FPD, diameter and position were measured and participants rated intensity perception of sucrose, caffeine, menthol and capsaicin solutions. Salivary flow rate, protein concentration and relative changes in protein composition were measured following each tastant. FPD, diameter and position were unchanged at six months. FPD did not correlate with intensity rating for any taste. FPD did correlate with changes in salivary protein output following sucrose (ρ = 0.72, p = 0.02) and changes in levels of proline-rich protein and mucin 7 following capsaicin (ρ = 0.71, p = 0.02, ρ = 0.68, p = 0.04, respectively). These results suggest that over six months fungiform papillae are anatomically stable, playing a greater role in mediating the physiological salivary response to stimuli rather than determining the perceived intensity of taste.

Background: Salivary metabolomics is rapidly advancing., Aim and Methods: To determine the extent to which salivary metabolites reflects host or microbial metabolic activity whole-mouth saliva (WMS), parotid saliva (PS) and plasma collected contemporaneously from healthy volunteers were analysed by 1 H-NMR spectroscopy. Spectra underwent principal component analysis and k-means cluster analysis and metabolite quantification. WMS samples were cultured on both sucrose and peptide-enriched media. Correlation between metabolite concentration and bacterial load was assessed., Results: WMS contained abundant short-chain fatty acids (SCFAs), which were minimal in PS and plasma. WMS spectral exhibited greater inter-individual variation than those of PS or plasma (6.7 and 3.6 fold, respectively), likely reflecting diversity of microbial metabolomes. WMS bacterial load correlated strongly with SCFA levels. Additional WMS metabolites including amines, amino acids and organic acids were positively correlated with bacterial load. Lactate, urea and citrate appeared to enter WMS via PS and the circulation. Urea correlated inversely with WMS bacterial load., Conclusions: Oral microbiota contribute significantly to the WMS metabolome. Several WMS metabolites (lactate, urea and citrate) are derived from the host circulation. WMS may be particularly useful to aid diagnosis of conditions reflective of dysbiosis. WMS could also complement other gastrointestinal fluids in future metabolomic studies., Competing Interests: Disclosure statement No potential conflict of interest was reported by the authors.

Taste and smell perceptions diminish in older age, impacting upon quality of life and nutrition, yet the causes of taste loss are largely unknown. Transient receptor potential channels (TRP) found on the oral mucosa are also involved in oral sensations including cooling and burning and may contribute to the eating experience of older people. Older adults often have reduced salivary flow and the physical properties of saliva may change, but the role of saliva in oral sensations of older adults is yet to be elucidated. Here, the effect of older age on subjective (perception) and objective (stimulated salivary response) measures of TRP stimulants, odors, and basic tastants was investigated. Whole mouth saliva was collected from younger (mean age 24 years) and older adults (mean age 72 years) following stimulation of taste [mono sodium glutamate (MSG) and caffeine], olfaction (menthol), and TRP receptors (capsaicin). Participants rated perceived intensity of each stimulus, and salivary properties were assessed. Older age was associated with 15% lower umami taste and 26% lower menthol odor perception, coupled with 17% lower salivary response to MSG. Interestingly, there were no differences for perception of TRP stimulants, so chemo-sensation was not affected by age. Younger adults had four times greater elasticity (Spinnbarkeit) with MUC7 levels almost double and 66% greater resting salivary flow rate. Stimulated salivary responses in the younger group were also higher compared to the older group, with changes in protein and viscoelasticity in response to taste and TRP stimulation. These results show the impact of older age upon taste and smell sensation which may lead to changes in the physical and compositional properties of saliva in response to taste/odor stimulation. Measurement of stimulated salivary flow and rheology provides an objective measure of taste in addition to subjective perceptions which can be influenced by participant bias. Chemo-sensation may be retained with age and trigeminal stimuli such as chili could be employed in future studies to enhance meals for an age group at risk of malnutrition. Alteration in salivary properties due to advanced age could impact on ability to taste due to poor diffusion of tastants and reduced oral surface protection.

Background: Salivary gland dysfunction is one of the main clinical features of Sjögren's syndrome (SS), manifested by xerostomia with subsequent complications and well-established effects on the person's quality of life., Objectives: To determine firstly whether selected tests of salivary gland function and structure, unstimulated whole salivary flow rate (UWSFR), parotid flow rate (PFR), clinical oral dryness score (CODS) and ultrasound score (USS), can discriminate SS from non-SS sicca patients and secondly whether these tests can differentiate between patients in different subgroups of SS., Method: Unstimulated whole salivary flow rate, PFR, CODS and USS were determined in 244 patients comprised of SS patients (n = 118), SS patients at higher risk of lymphoma (n = 30) or with lymphoma (n = 26), and non-SS sicca disease controls (n = 70)., Results: All assessments showed a significant difference between the overall SS group and the disease control group, attributed mainly to the lymphoma subgroups of SS (p < 0.0001 for all parameters). There was a significant correlation (Spearman r = 0.7, p value <0.0001) and 87.3% agreement between USS and the histology focus scores of 119 patients., Conclusion: The results suggest that salivary gland tests including USS can aid in differentiating between SS and non-SS dry mouth, especially the subgroups of SS with lymphoma or at higher risk of developing lymphoma., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

The extracellular polymer substances (EPS) generated by biofilms confers resistance to antimicrobial agents through electrostatic and steric interactions that hinder molecular diffusion. This resistance mechanism is particularly evident for antibacterial nanomaterials, which inherently diffuse more slowly compared to small organic antibacterial agents. The aim of this study was to determine if a biofilm's resistance to antibacterial nanomaterial diffusion could be diminished using electrolytes to screen the EPS's electrostatic interactions. Anionic (+) alpha-tocopherol phosphate (α-TP) liposomes were used as the antimicrobial nanomaterials in the study. They self-assembled into 700 nm sized structures with a zeta potential of -20 mV that were capable of killing oral bacteria (S. oralis growth inhibition time of 3.34 ± 0.52 h). In a phosphate (-ve) buffer the -ve α-TP liposomes did not penetrate multispecies oral biofilms, but in a Tris (hydroxymethyl)aminomethane (+ve) buffer they did (depth - 12.4 ± 3.6 μm). The Tris did not modify the surface charge of the α-TP nanomaterials, rather it facilitated the α-TP-biofilm interactions through electrolyte screening (Langmuir modelled surface pressure increase of 2.7 ± 1.8 mN/ m). This data indicated that EPS resistance was mediated through charge repulsion and that this effect could be diminished through the co-administration of cationic electrolytes., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Sjögren's syndrome (SS) is an autoimmune inflammatory disease that affects the exocrine glands. The absence of early diagnostic markers contributes to delays in its diagnosis. Identification of changes in the protein profile of saliva is considered one of the promising strategies for the discovery of new biomarkers for SS., Objective: To identify salivary protein biomarkers with potential for use in discriminating between different lymphoma risk subgroups of SS., Method: Parotid and whole mouth saliva samples were collected from patients with SS, including those in subgroups at higher risk of developing or with confirmed lymphoma, non-SS sicca disease controls and healthy subjects. An initial proteomics analysis by mass spectrometry (LCMSMS) identified S100A8/A9 as a biomarker and was followed by validation with an enzyme-linked immunosorbent assay (ELISA)., Results: Significant differences were found in levels of S100A8/A9 in parotid saliva but not whole mouth saliva between patients with SS compared with healthy and disease control subjects (P = 0.001 and 0.031, respectively). Subgroups of patients with SS based on lymphoma risk showed significant differences in salivary levels of S100A8/A9., Conclusion: The results suggest that salivary levels of S100A8/A9 can aid in differentiating between SS, disease control and healthy control subjects, especially the subgroups of SS with lymphoma or at higher risk of lymphoma., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Saliva is a complex fluid produced by 3 pairs of major salivary glands and by hundreds of minor salivary glands. It comprises a large variety of constituents and physicochemical properties, which are important for the maintenance of oral health. Saliva not only protects the teeth and the oropharyngeal mucosa, it also facilitates articulation of speech, and is imperative for mastication and swallowing. Furthermore, saliva plays an important role in maintaining a balanced microbiota. Thus, the multiple functions provided by saliva are essential for proper protection and functioning of the body as a whole and for the general health. A large number of diseases and medications can affect salivary secretion through different mechanisms, leading to salivary gland dysfunction and associated oral problems, including xerostomia, dental caries and fungal infections. The first part of this review article provides an updated insight into our understanding of salivary gland structure, the neural regulation of salivary gland secretion, the mechanisms underlying the formation of saliva, the various functions of saliva and factors that influence salivary secretion under normal physiological conditions. The second part focuses on how various diseases and medical treatment including commonly prescribed medications and cancer therapies can affect salivary gland structure and function. We also provide a brief insight into how to diagnose salivary gland dysfunction., (© 2018 John Wiley & Sons Ltd.)

Introduction: We have investigated orofacial pain in a prospective cohort of obese and normal-weight subjects undergoing fixed-appliance orthodontic treatment., Methods: Fifty-five subjects (27 males, 28 females) mean age 15.1 (1.6) years and mean body mass index 30.2 (3.5) in obese and 19.4 (2.2) kg/m2 in normal-weight groups were followed for 1 week after appliance placement. Primary outcome was maximum-pain measured using a 100-mm visual analogue scale. Secondary outcomes included mean pain and oral analgesic consumption., Results: Mean maximum pain for the total sample was 73.7 (standard deviation 14.8; 95% confidence interval 69.8-77.7) mm with no significant differences among groups (P = 0.247). However, mean maximum pain was higher at all time-points for the obese group and significant at 72 hours (P = 0.034). Total analgesia consumed by the obese group was also significantly higher than normal weight (P = 0.041). Multivariable regression found the only significant predictor for mean pain was time. After adjusting for confounding, obesity was associated with higher (+4.47 mm) mean pain at each time-point (P = 0.018). A significant association existed between obesity and total analgesic consumption (univariable-analysis, P = 0.035; multivariable analysis, P = 0.023). After accounting for confounders, obese patients were associated with taking a higher quantity of oral analgesics., Conclusions: We found a trend towards increased mean pain and an association with increased analgesic consumption in obese subjects during the first week following fixed-appliance placement.

Metabolic profiling by 1 H NMR spectroscopy is an underutilized technology in salivary research, although preliminary studies have identified promising results in multiple fields (diagnostics, nutrition, sports physiology). Translation of preliminary findings into validated, clinically approved knowledge is hindered by variability in protocol for the collection, storage, preparation, and analysis of saliva. This study aims to evaluate the effects of differing sample pretreatments on the 1 H NMR metabolic profile of saliva. Protocol considerations are highly varied in the current literature base, including centrifugation, freeze-thaw cycles, and different NMR quantification methods. Our findings suggest that the 1 H NMR metabolite profile of saliva is resilient to any change resulting from freezing, including freezing of saliva prior to centrifuging. However, centrifugation was necessary to remove an unidentified broad peak between 1.24 and 1.3 ppm, the intensity of which correlated strongly with saliva cellular content. This peak obscured the methyl peak from lactate and significantly affected quantification. Metabolite quantification was similar for saliva centrifuged between 750 g to 15 000 g. Quantification of salivary metabolites was similar whether quantified using internal phosphate-buffered sodium trimethylsilyl-[2,2,3,3- 2 H 4 ]-propionate (TSP) or external TSP in a coaxial NMR tube placed inside the NMR tube containing the saliva sample. Our results suggest that the existing literature on salivary 1 H NMR will not have been adversely affected by variations of the common protocol; however, use of TSP as an internal standard without a buffered medium appears to affect metabolite quantification, notably for acetate and methanol. We include protocol recommendations to facilitate future NMR-based studies of saliva.

The extensional rheology (ER) of saliva is a property associated with its ability to coat surfaces and is important for the maintenance of a normal mouth feeling. Transient receptor potential (TRP) channels are expressed in the oral cavity and this study investigated how the sensory effects of TRP channel agonists modify the ER of saliva. Healthy volunteers rinsed with solutions containing a TRP agonist. Unstimulated whole mouth saliva (WMS) was collected prior to rinsing and WMS was collected during the first and second minutes after the mouth rinse. The Spinnbarkeit of the collected saliva was measured using a Neva Meter. The nonivamide (TRPV1) mouth rinse increased WMS ER from 37.0 (± 6.3) mm to 49.3 (± 5.1) mm when compared with the vehicle control, which itself had no effect on WMS ER. However, this effect was short-lived and ER of WMS was not increased in the second minute after the nonivamide mouth rinse. The menthol (TRPM8) mouth rinse resulted in an increase up to 57.8 (± 7.8) mm in WMS ER from the vehicle control and returned to control levels in the second minute. The cinnamaldehyde (TRPA1) mouth rinse resulted in no change in WMS ER. It can be concluded that nonivamide and menthol mouth rinsing has a short-term effect of increasing WMS ER, an effect not observed after cinnamaldehyde rinsing. We hypothesize that the activation of some TRP channels in the oral cavity results in changes in the salivary protein composition that in turn alters WMS ER., Practical Applications: Identifying compounds that modify the physical properties of saliva in a desirable way is important in developing treatments for conditions associated with changes in the physical properties of saliva such as xerostomia (also known as dry mouth). Furthermore, understanding the rheology of saliva contributes to the elucidation of food oral processing which is of importance to food manufacturers., (© 2017 Wiley Periodicals, Inc.)

Obesity is a widespread chronic inflammatory disorder characterized by an increased overall disease burden and significant association with periodontitis. The aim of this prospective clinical cohort study was to investigate the effect of obesity on orthodontic tooth movement. Fifty-five adolescent patients (27 males, 28 females) with a mean (SD) age of 15.1 (1.7) years and mean (SD) body mass index (BMI) of 30.2 (3.5) kg/m 2 in obese and 19.4 (2.2) kg/m 2 in normal-weight groups were followed from start of treatment to completion of tooth alignment with fixed orthodontic appliances. Primary outcome was time taken to complete tooth alignment, while secondary outcomes included rate of tooth movement and change in clinical parameters (plaque/gingival indices, unstimulated whole-mouth salivary flow rate, gingival crevicular fluid biomarkers). Data collection took place at baseline (start of treatment: appliance placement), 1 h and 1 wk following appliance placement, and completion of alignment. Results were analyzed by descriptive statistics followed by generalized estimating equation regression modeling. There were no significant differences between groups in time taken to achieve tooth alignment (mean [SD] 158.7 [75.3] d; P = 0.486). However, at 1 wk, initial tooth displacement was significantly increased in the obese group ( P < 0.001), and after adjusting for confounders, obese patients had a significantly higher rate of tooth movement compared with normal-weight patients (+0.017 mm/d; 95% confidence interval, 0.008-0.025; P < 0.001) over the period of alignment. Explorative analyses indicated that levels of the adipokines leptin and resistin, the inflammatory marker myeloperoxidase (MPO), and the cytokine receptor for nuclear factor kappa-B ligand (RANKL) were significantly different between obese- and normal-weight patients and associated with observed rates of tooth movement. This represents the first prospective data demonstrating a different response in obese patients compared with normal-weight patients during early orthodontic treatment. These differences in the response of periodontal tissues to orthodontic force in the presence of obesity have potential short- and long-term clinical implications.

Objective: to investigate the effects of a high fat diet (HFD) on salivary glands in vivo, in a mouse model. In particular, whether it will induce the appearance of fat cells in salivary glands, alterations related to autophagy, mTOR pathway and sympathetic innervation., Design: 27 adult female ICR mice were separated in six groups. Three groups fed with (HFD) containing 55% fat, for one, two and three month and another three groups fed with normal diet (2.7% of fat), for the same time periods. The submandibular glands and liver were dissected and part homogenized for protein analyses and part fixed in formalin for histological analyses., Results: After three months the HFD fed mice total body weight fold change increased compared to controls. The Oil Red O staining showed no fat cells deposit in salivary gland however a large increase was observed in liver after three months of HFD. Adiponectin levels were significantly decreased in the HFD group after three months. The group fed with HFD for three months showed increased conversion of the LC3 autophagy marker in salivary gland. mTOR showed no activation regarding the time point studied. Tyrosine hydroxylase significantly decreased after two and three month of HFD., Conclusion: HFD caused several changes after three months however the earliest change was noticed after two months regarding sympathetic innervation. This suggests neural alteration may drive other diet induced changes in salivary glands. These early changes may be the starting point for longer term alterations of salivary glands with alterations in diet., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. Despite the implications of this prevalent disease, diagnosis currently takes years, partly due to the diversity in patient presentation. Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)

This study aimed to investigate masticatory function and taste and their possible relationship with salivary flow in young adults with good oral health. The study also examined whether anthropometric measurements and gender could influence the variables studied. A total of 171 subjects were selected (125 females, 46 males). Masticatory performance was evaluated with the sieve method, and perceived masticatory ability was measured using the visual analogue scale. Taste was evaluated using the drop test with four different flavors in three different concentrations, and unstimulated and stimulated saliva flows were measured. The anthropometric variables measured included body mass index (BMI) and waist circumference (WC). The independent variables studied could not predict masticatory performance. The independent variables, BMI, WC, and gender, predicted 14% of perceived masticatory ability, and BMI predicted 5% of taste. Masticatory performance was not related to salivary flow or anthropometric parameters in young healthy adults. Perceived masticatory ability was related to BMI, WC, and gender, whereas taste was only weakly related to BMI. The flow rate did not exhibit a statistically significant difference between males and females for the anthropometric groups. (J Oral Sci 58, 391-399, 2016).

Saliva is vital for the maintenance of normal oral physiology and mucosal health. The loss of salivary function can have far-reaching consequences, as observed with dry mouth, which is associated with increased orodental disease, speech impairment, dysphagia, and a significant negative effect on quality of life. The timely diagnosis of oral dryness is vital for the management of orodental disease and any associated often-undiagnosed systemic disease (e.g., Sjögren syndrome). Our aim was to investigate differences in mucin glycoproteins and saliva rheological properties between sufferers and nonsufferers of dry mouth in order to understand the relationship between saliva composition, rheological properties, and dryness perception and provide additional potential diagnostic markers. All patients exhibited objective and subjective oral dryness, irrespective of etiology. Over half of the patients (n = 20, 58.8%) had a saliva secretion rate above the gland dysfunction cutoff of 0.1 mL/min. Mucin (MUC5B and MUC7) concentrations were generally similar or higher in patients. Despite the abundance of these moisture-retaining proteins, patients exhibited reduced mucosal hydration (wetness) and significantly lower saliva spinnbarkeit (stringiness), suggesting a loss of the lubricating and retention/adhesion properties of saliva, which, at least partially, are associated with mucin glycoproteins. Over 90% of patients with dry mouth (DMPs) consistently had unstimulated whole mouth saliva (UWMS) spinnbarkeit below the proposed normal cutoff (10 mm). Further analysis of mucins revealed the reduced glycosylation of mucins in DMPs compared to healthy controls. Our data indicate that UWMS mucin concentrations are not reduced in dry mouth but that the mucin structure (glycosylation) is altered. UWMS from DMPs had reduced spinnbarkeit, the assessment of which, in conjunction with sialometry, could improve sensitivity for the diagnosis of dry mouth. Additionally, it may be useful to take into consideration the altered mucin glycosylation and saliva rheological properties when designing synthetic or purified mucins for saliva substitutes and dry mouth therapy., (© International & American Associations for Dental Research 2015.)

The oral mucosal pellicle is a layer of absorbed salivary proteins, including secretory IgA (SIgA), bound onto the surface of oral epithelial cells and is a useful model for all mucosal surfaces. The mechanism by which SIgA concentrates on mucosal surfaces is examined here using a tissue culture model with real saliva. Salivary mucins may initiate the formation of the mucosal pellicle through interactions with membrane-bound mucins on cells. Further protein interactions with mucins may then trigger binding of other pellicle proteins. HT29 colon cell lines, which when treated with methotrexate (HT29-MTX) produce a gel-forming mucin, were used to determine the importance of these mucin-mucin interactions. Binding of SIgA to cells was then compared using whole mouth saliva, parotid (mucin-free) saliva and a source of purified SIgA. Greatest SIgA binding occurred when WMS was incubated with HT29-MTX expressing mucus. Since salivary MUC5B was only able to bind to cells which produced mucus and purified SIgA showed little binding to the same cells we conclude that most SIgA binding to mucosal cells occurs because SIgA forms complexes with salivary mucins which then bind to cells expressing membrane-bound mucins. This work highlights the importance of mucin interactions in the development of the mucosal pellicle.

The bound salivary pellicle is essential for protection of both the enamel and mucosa in the oral cavity. The enamel pellicle formation is well characterised, however the mucosal pellicle proteins have only recently been clarified and what drives their formation is still unclear. The aim of this study was to examine the salivary pellicle on particles with different surface properties (hydrophobic or hydrophilic with a positive or negative charge), to determine a suitable model to mimic the mucosal pellicle. A secondary aim was to use the model to test how transglutaminase may alter pellicle formation. Particles were incubated with resting whole mouth saliva, parotid saliva and submandibular/sublingual saliva. Following incubation and two PBS and water washes bound salivary proteins were eluted with two concentrations of SDS, which were later analysed using SDS-PAGE and Western blotting. Experiments were repeated with purified transglutaminase to determine how this epithelial-derived enzyme may alter the bound pellicle. Protein pellicles varied according to the starting salivary composition and the particle chemistry. Amylase, the single most abundant protein in saliva, did not bind to any particle indicating specific protein binding. Most proteins bound through hydrophobic interactions and a few according to their charges. The hydrophobic surface most closely matched the known salivary mucosal pellicle by containing mucins, cystatin and statherin but an absence of amylase and proline-rich proteins. This surface was further used to examine the effect of added transglutaminase. At the concentrations used only statherin showed any evidence of crosslinking with itself or another saliva protein. In conclusion, the formation of the salivary mucosal pellicle is probably mediated, at least in part, by hydrophobic interactions to the epithelial cell surface., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

Objectives: To evaluate the effects of acidic functional monomers with different hydrophilicity and spacer carbon chain length on the degree of conversion (DC), wettability (contact angle), water sorption (WS) and ultimate tensile strength (UTS) of experimental one-step self-etch adhesives (1-SEAs)., Methods: A series of standard resin blends was prepared with each formulation containing 15mol% of each acidic monomer. The structural variations of the acidic monomers were MEP (spacer chain with 2 carbons), MDP (10-carbons), MDDP (12-carbons), MTEP (more hydrophilic polyether spacer) and CAP-P (intermediate hydrophilicity ester spacer). Dumbbell-shaped and disc specimens were prepared and tested for UTS and WS, respectively. DC was assessed by FTIR, while the wettability of each 1-SEA was evaluated on glass slides and flat dentine surfaces. Results were analysed with one-way ANOVA and Tukey's test (p<0.05)., Results: The outcomes showed lower UTS for CAP-P, control blend and MEP than MTEP, MDDP and MDP (p<0.05). The degree of conversion was statistically similar for all resins (p=0.122). On dentine, the wettability was higher (lower contact angle) with the most hydrophilic monomer MTEP. Higher WS was attained using MTEP. Different lengths of the spacer chains did not result in different wettability and WS (p>0.05)., Conclusion: At similar molar percentage, different acidic functional monomers induced similar degree of conversion and different UTS when included in a 1-SEA. However, the inclusion of highly hydrophilic monomer may increase the wettability on dentine and the WS., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Salivary gland atrophy is a frequent consequence of head and neck cancer irradiation therapy but can potentially be regulated through the mammalian target of rapamycin (mTOR). Excretory duct ligation of the mouse submandibular gland provokes severe glandular atrophy causing activation of mTOR. This study aims to discover the effects of blocking mTOR signaling in ligation-induced atrophic salivary glands. Following 1 week of unilateral submandibular excretory duct ligation: gland weights were significantly reduced, 4E-BP1 and S6rp were activated, and tissue morphology revealed typical signs of atrophy. However, 3 days following ligation with rapamycin treatment, a selective mTOR inhibitor, gland weights were maintained, 4E-BP1 and S6rp phosphorylation was inhibited, and there were morphological signs of recovery from atrophy. However, following 5 and 7 days of ligation and rapamycin treatment, glands expressed active mTOR and showed signs of considerable atrophy. This evidence suggests that inhibition of mTOR by rapamycin delays ligation-induced atrophy of salivary glands.

Maintenance of a film of saliva on oral surfaces is dependent upon nerve-mediated, reflex salivary gland secretion. Afferent signalling arises from taste, olfaction and mastication and is modified by signalling from other centres in the central nervous system before efferent signals are delivered to salivary glands in autonomic nerves. Salivary fluid secretion is largely dependent upon cholinergic signalling from parasympathetic nerves whilst the protein content of saliva is additionally dependent upon signalling by neuropeptides and, in the major (parotid, submandibular and sublingual) salivary glands, by sympathetic nerves and the release of noradrenaline. There have been significant recent advances in our understanding of the membrane transport proteins involved in intracellular calcium signalling in salivary acinar cells in response to nerve stimulation and of the ion transport proteins responsible for acinar cell secretion of saliva. Salivary glands retain an ability to regenerate following extreme atrophy, and autonomic nerves have an important role in both gland development and maintenance of long-term normal function. Continued advances in the understanding of the nerve-mediated regulation of salivary glands should help in the development of strategies for preventing chronic oral dryness resulting from drugs or atrophic disease associated with inflammation and irradiation.

Saliva has one of the most difficult roles to perform in the body. It must facilitate the taste and detection of foods nutritious to the body but also defend the mucosa from infection by the ever-present microbiota present in the mouth. It achieves these roles by having a complex composition and versatile physical properties. The protein and ion components make a solution that is 99% water into a viscoelastic solution capable of many roles, such as acting as a lubricant and an antimicrobial, preventing the dissolution of teeth, aiding digestion, and facilitating taste. This review describes the neural regulation of salivary secretion in terms of fluid, protein, and ion secretion. It then describes some of the components and physical properties of saliva and attempts to relate them to the functions that saliva must perform.

Human salivary statherin was purified from parotid saliva and adsorbed to bare hydrophilic (HP) mica and STAI-coated hydrophobic (HB) mica in a series of Surface Force Balance experiments that measured the normal (F(n)) and friction forces (F(s)*) between statherin-coated mica substrata. Readings were taken both in the presence of statherin solution (HP and HB mica) and after rinsing (HP mica). F(n) measurements showed, for both substrata, monotonic steric repulsion that set on at a surface separation D ~20 nm, indicating an adsorbed layer whose unperturbed thickness was ca 10 nm. An additional longer-ranged repulsion, probably of electrostatic double-layer origin, was observed for rinsed surfaces under pure water. Under applied pressures of ~1 MPa, each surface layer was compressed to a thickness of ca 2 nm on both types of substratum, comparable with earlier estimates of the size of the statherin molecule. Friction measurements, in contrast with F(n) observations, were markedly different on the two different substrata: friction coefficients, μ ≡ ∂F(s)*/∂F(n), on the HB substratum (μ ≈ 0.88) were almost an order of magnitude higher than on the HP substratum (μ ≈ 0.09 and 0.12 for unrinsed and rinsed, respectively), and on the HB mica there was a lower dependence of friction on sliding speed than on the HP mica. The observations were attributed to statherin adsorbing to the mica in multimer aggregates, with internal re-arrangement of the protein molecules within the aggregate dependent on the substratum to which the aggregate adsorbed. This internal re-arrangement permitted aggregates to be of similar size on HP and HB mica but to have different internal molecular orientations, thus exposing different moieties to the solution in each case and accounting for the very different friction behaviour.

Rat submandibular gland can regenerate following ligation-induced atrophy, eventually recovering its normal morphology and function. Previous studies have suggested that the regeneration process implies both self-proliferation of existing acini and formation of new acinar cells. One hypothesis is that new acinar cells may differentiate from the ductal cells in a similar fashion to the process of cytodifferentiation occurring during submandibular glandular development. In this study atrophy was induced, under recovery anaesthesia, by applying a metal clip on the main duct of the submandibular gland without including the chorda lingual nerve. After 2 weeks the duct was deligated for 3, 5 or 7 days or 8 weeks and the glands collected. Tissue was prepared for immunohistochemistry, biochemical analysis and RNA extraction. The histology of the regenerated glands shows several normal-looking acini, which have regained their glycoprotein content (AB/PAS positive), data also confirmed by biochemical analysis (SDS-PAGE/PAS). Regenerating tissue was characterized by the presence of embryonic-like branched structures ending with AB/PAS positive acinar cells. The proteins SMG-B and PSP are normally expressed in acinar cell precursors during development but only by intercalated ductal cells in the adult stage. In the adult regenerating gland mRNA levels of both SMG-B and PSP were found to be up-regulated compared to ligated glands and SMG-B expression localized to acinar cells whilst the ductal cells were negative. This study of rat submandibular gland regeneration suggests new acinar cells have differentiated from ducts and express markers of acinar cell precursors in a similar manner to the cytodifferentiation process occurring during glandular development., (2009 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)

Salivary gland atrophy is a common consequence of pathology, including Sjögren's syndrome, irradiation therapy and obstructive sialadenitis. During severe atrophy of the rat submandibular gland caused by excretory duct ligation, the majority of acinar cells disappear through apoptosis, whereas ductal cells proliferate and dedifferentiate; yet, the gland can survive in the atrophic state almost indefinitely, with an ability to fully recover if deligated. The control mechanisms governing these observations are not well understood. We report that ~10% of acinar cells survive in ligation-induced atrophy. Microarray and quantitative real-time PCR analysis of ligated glands indicated sustained transcription of acinar cell-specific genes, whereas ductal-specific genes were reduced to background levels. After 3 days of ligation, activation of the mammalian target of rapamycin (mTOR) pathway and autophagy occurred as shown by phosphorylation of 4E-BP1 and expression of autophagy-related proteins. These results suggest that activation of mTOR and the autophagosomal pathway are important mechanisms that may help to preserve acinar cells during atrophy of salivary glands after injury.