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Salivary proteins play an important role in repairing mechanisms of damaged tissues and the maintenance of oral health. However, there is a dearth of information in the literature regarding the concentrations of salivary proteins in caries-free (CF) and caries-active (CA) subjects. Hence, this systematic review was conducted to update our previous systematic review published in 2013 that aimed to assess the association between caries and salivary proteins by comparing CF and CA individuals. Thereby, evaluating the possibility of whether salivary proteins can be regarded as biomarkers for caries. An extensive search of studies was conducted using PubMed, EMBASE, Clarivate Analytics’ Web of Science, and Elsevier’s Scopus between July 2012 and January 2022, without any language restriction. Manual searching in Google Scholar and evaluation of bibliographies of the included studies were also undertaken. The Newcastle-Ottawa Scale was used to assess the risk of bias (RoB) within the included studies. Of 22 included studies, 1,551 human subjects (range: 30–213 participants) were recruited, of which 848 individuals (54.7%) were CA and 703 (45.3%) were CF. Regarding the utilization of DMFT as the caries index, high variability was observed across different articles. A statistically significant increase in the salivary levels of alpha-amylase, acidic proline-rich protein-1, histatin-5, lactoperoxidase, and mucin-1 was found in CA patients, while the salivary levels of carbonic anhydrase 6, proteinase-3, and statherin were observed to be significantly increased in CF subjects. Conflicting results were found regarding the salivary levels of immunoglobulin A and total proteins among CA and CF subjects. The included studies were categorized as low RoB (n = 15), medium RoB (n = 4), and high RoB (n = 3). Due to significant heterogeneity among the included studies, no meta-analysis could be performed. In conclusion, the salivary levels of protein(s) might be a useful biomarker for caries diagnosis, especially alpha-amylase, acidic proline-rich protein-1, histatin-5, lactoperoxidase, mucin-1, carbonic anhydrase 6, proteinase-3, and statherin. However, their diagnostic value must be verified by large-scale prospective studies.

Graphical abstract, Objective Viruses on environmental surfaces, in saliva and other body fluids represent risk of contamination for general population and healthcare professionals. The development of vaccines and medicines is costly and time consuming. Thus, the development of novel materials and technologies to decrease viral availability, viability, infectivity, and to improve therapeutic outcomes can positively impact the prevention and treatment of viral diseases. Methods Herein, we discuss (a) interaction mechanisms between viruses and materials, (b) novel strategies to develop materials with antiviral properties and oral antiviral delivery systems, and (c) the potential of artificial intelligence to design and optimize preventive measures and therapeutic regimen. Results The mechanisms of viral adsorption on surfaces are well characterized but no major breakthrough has become clinically available. Materials with fine-tuned physical and chemical properties have the potential to compromise viral availability and stability. Emerging strategies using oral antiviral delivery systems and artificial intelligence can decrease infectivity and improve antiviral therapies. Significance Emerging viral infections are concerning due to risk of mortality, as well as psychological and economic impacts. Materials science emerges for the development of novel materials and technologies to diminish viral availability, infectivity, and to enable enhanced preventive and therapeutic strategies, for the safety and well-being of humankind.

To assess the effectiveness of different anesthetic solutions for pain control immediately after the extraction of lower third molars. Nine databases were used to identify randomized clinical trials, without restriction of language or year of publication. The “JBI Critical Appraisal Tools for Systematic Reviews” was used to assess the risk of bias in the studies. The network meta-analysis was performed to compare the effectiveness of different anesthetics to control the pain immediately after the surgery of lower third molars, using the standardized mean difference (SMD) as the effect estimate. The GRADE approach was used to assess the certainty of evidence. The search presented 13,739 initial results, from which 45 met the eligibility criteria and presented low to moderate risk of bias. Thirteen studies were included in the meta-analysis. The 2% lidocaine + clonidine presented the lowest pain scores (SMD = − 1.44; − 2.72 to − 0.16) compared to 4% articaine + adrenaline, followed by 0.5% bupivacaine + adrenaline (SMD = − 1.36; − 2.13 to − 0.59). The certainty of evidence varied between very low to moderate. 2% lidocaine + clonidine and 0.5% bupivacaine + adrenaline were the anesthetics with the highest probability for pain control immediately after the surgical procedure of removing impacted lower third molars. The use of an adequate anesthetic with effective pain control can contribute to a more comfortable postoperative period.

To evaluate whether surface characteristics of different titanium modifications may influence the composition of the salivary pellicle on each surface by analyzing the salivary proteome through mass spectrometry–based proteomics. Titanium discs with three surfaces modifications (PT (machined titanium), SLA (sandblasted/large-grit/acid-etched), and SLActive (modified SLA)) were characterized (topography, chemistry, and energy) prior to being exposed to saliva for 2 h to form a protein pellicle. The resultant protein layer was retrieved and analyzed through mass spectrometry (nLC-ESI-MS/MS) to examine the surface specificity for protein binding, while the proteome profile of each surface was classified. The proteome analysis showed that the salivary pellicle composition was more complex on rough surfaces (SLA and SLActive). Although variability in protein composition was observed between surfaces, most proteins were detected on more than one surface, indicating a limited surface specificity for protein binding. Additionally, the salivary pellicle formed on the SLActive presented a larger number of proteins associated with immune response, biological adhesion, and biomineralization. Although topography, chemistry, and energy differed between the surfaces, they were not determinant to produce a salivary pellicle with high surface specificity. Also, we showed that several salivary proteins adsorbed on Ti surfaces are involved in biological functions important to the biointegration. This study sheds light on the necessity for the development of bioactive surfaces that favors the formation of a specific protein layer that can enhance tissue response to assist the biointegration of dental implants.

The aim of this study was to assess and compare RANK, RANKL, and OPG immunoexpression in dentigerous cyst, odontogenic keratocyst, and ameloblastoma. The protocol was registered in PROSPERO (CRD42018105543). Seven databases (Embase, Lilacs, LIVIVO, PubMed, Scopus, SciELO, and Web of Science) were the primary search sources and two databases (Open Grey and Open Thesis) partially captured the “grey literature”. Only cross sectional studies were included. The JBI Checklist assessed the risk of bias. A meta-analysis with random effects model estimated the values from the OPG and RANKL ratio reported by the individual studies and respective 95% confidence intervals. The heterogeneity among studies was assessed with I2 statistics. Only nine studies met the inclusion criteria and were considered in the analyses. The studies were published from 2008 to 2018. Two studies presented low risk of bias, while seven studies presented moderate risk. The meta-analysis showed the highest OPG>RANKL ratio for dentigerous cyst (ES=43.3%; 95% CI=14.3-74.8) and odontogenic keratocyst (ES=36.8%; 95% CI=18.8-56.7). In contrast, the highest OPG RANKL para cisto dentígero (ES=43,3%; IC95%=14,3-74,8) e ceratocisto odontogênico (ES=36,8%; IC95%=18,8-56,7). Por outro lado, a maior razão OPG

Background Dental care settings potentially carry a high risk of cross-infection between dentists and patients and among dental staff due to close contact and use of aerosol-generating procedures. This study aimed to estimate COVID-19 incidence rates among Canadian dentists over a six-month period. Methods We conducted a prospective cohort study of 644 licensed dentists across Canada, from 29th July 2020 to 12th February 2021. An online questionnaire, adopted from WHO Unity Study protocols for assessment of COVID-19 risk among healthcare workers was used to collect self-reported SARS-CoV-2 infection, every four weeks. Bayesian Poisson model was used to estimate the incidence rate and corresponding 95% credible intervals (CI). Results The median age of participants was 47 years, with the majority being female (56.4%) and general practitioners (90.8%). The median follow-up time was 188 days. Six participants reported COVID-19 infection during the study period, giving an incidence rate of 5.10 per 100,000 person-days (95% CI = 1.86 – 9.91 per 100,000 person-days). The incidence proportion was estimated to be 1084 per 100,000 dentists (95% CI = 438 to 2011 per 100,000), while it was 1864 per 100,000 persons (95% CI = 1859 to 1868 per 100,000 persons) in the Canadian population during the same period. Conclusion The low infection rate observed among Canadian dentists over the period July 2020 to February 2021 is reassuring to the dental and general community.

We explored the potential to diagnose Zika virus (ZIKV) infection by analyzing peptides in saliva during a convalescent phase of infection, long after resolution of acute disease. A 25-y-old woman clinically diagnosed with Zika fever in the first trimester was enrolled with her dizygotic twins for a 3-mo postnatal sample of saliva (9-mo after maternal infection). The female baby (A) had microcephaly while the male baby (B) was born healthy. Peptidomic analysis was completed by mass spectrometry (MS/MS), and ZIKV peptides were identified using the National Institutes of Health Zika Virus Resource database, then aligned and mapped to the ZIKV polyprotein to determine proteome coverage and phylogenetic studies. A total of 423 (mother), 607 (baby A), and 183 (baby B) unique ZIKV peptides were identified in saliva by MS/MS, providing a coverage of 67%, 84%, and 45%, respectively, of the entire ZIKV polyprotein (3,400 amino acids). All peptides were aligned to other flaviviruses that are circulating in Brazil (dengue and yellow fever) to discard false-positive matches. Nine peptides identified were highly conserved to dengue virus. Alignment of a contiguous peptide sequence for mother/babies with the 74 ZIKV sequences suggested that the virus may have entered the oral cavity through the salivary glands, leading to an infection that persists into the postnatal period (vertical transmission). Furthermore, we identified 9 sequence variations that were unique to the baby with microcephaly (not found in the mother or the twin). This sequence information could provide a template for future neuropathogenic studies. A much larger sample size is required to determine whether sequence variation in the envelope protein significantly associates with microcephaly. Finally, from a public health perspective, it will be important to determine whether viral replication is still taking place after birth and whether the virus can be transmitted through salivary contact.

Objectives Investigate the effects of a Bis-phenyl-glycidyl-dimethacrylate (BisGMA) biodegradation product, bishydroxypropoxyphenyl-propane (BisHPPP), on gene expression and protein synthesis of cariogenic bacteria. Methods Quantitative real-time polymerase chain reaction was used to investigate the effects of BisHPPP on the expression of specific virulence-associated genes, i.e. gtfB , gtfC , gbpB , comC , comD , comE and atpH in Streptococcus mutans UA159. Possible mechanisms for bacterial response to BisHPPP were explored using gene knock-out and associated complemented strains of the signal peptide encoding gene, comC . The effects of BisHPPP on global gene and protein expression was analyzed using microarray and quantitative proteomics. The role of BisHPPP in glucosyltransferase (GTF) enzyme activity of S. mutans biofilms was also measured. Results BisHPPP (0.01, 0.1 mM) up-regulated gtfB/C , gbpB , comCDE , and atpH most pronounced in biofilms at cariogenic pH (5.5). The effects of BisHPPP on the constructed knock-out and complemented strains of comC from quorum-sensing system, implicated this signaling pathway in up-regulation of the virulence-associated genes. Microarray and proteomics identified BisHPPP-regulated genes and proteins involved in biofilm formation, carbohydrate transport, acid tolerance and stress-response. GTF activity was higher in BisHPPP-exposed biofilms when compared to no-BisHPPP conditions. Significance These findings provide insight into the genetic and physiological pathways and mechanisms that help explain S. mutans adaptation to restorative conditions that are conducive to increased secondary caries around resin composite restorations and may provide guidance to clinicians’ decision on the selection of dental materials when considering the long term oral health of patients and the interactions of composite resins with oral bacteria.

To present a systematic review that provides updated information about proteins found in salivary fluid extracted strictly from ducts. The systematic review probing strategy was based on electronic databases word search (PubMed, EMBASE, LILACS, Web of Science, and Scopus). Risk of bias was assessed based on Joanna Briggs Institute Critical Appraisal Checklist for Studies Reporting Prevalence Data. After 2 rounds of scrutiny, 12 articles were included, totaling 231 individuals (125 were healthy, 41 were elder individuals with radicular caries, 56 had primary Sjogren’s syndrome, and 9 were patients who had received radiotherapy for head and neck cancer). The selected studies had no similarities among proteins found, demonstrating the need of standard reference in experimental methodology to obtain a thorough coverage of proteins. Further studies are required to better determine the relative amount of proteins described in this study. It is essential to increase the number of samples, to perform similar collection techniques, to include other analyses methods such as mass spectrometry, and to perform the validation of some proteins using immunoassay techniques such as Elisa and Western blot. Proteomic profile of saliva collected from ducts is essential to better understand the disease process, enabling the identification of biomarkers for specific clinical situations.

To assess whether salivary urea and creatinine levels accurately reflect their serum levels in blood samples of adults to detect chronic kidney disease. A systematic review was conducted in eight electronic databases. The protocol was registered in PROSPERO. Only diagnostic test studies were included. The JBI critical appraisal tools assessed the risk of bias. A meta-analysis of proportions was performed. The GRADE tool assessed the quality of evidence and strength of recommendation across the studies included. Eight studies met the eligibility criteria and were included. Six studies assessed salivary urea, and six studies assessed salivary creatinine. All studies presented moderate risk of bias. The meta-analysis depicted an overall sensitivity of 93.3% (95% CI = 88.6; 97.9) for salivary creatinine levels and 87.5% (95% CI = 83.2; 91.8) for salivary urea levels, while the overall specificity was 87.1% (95% CI = 82.8; 91.3) and 83.2% (95% CI = 65.0; 101.4) for salivary creatinine and urea levels, respectively. The overall accuracy of salivary creatinine was 5.2 percentage points higher compared with salivary urea levels (90.8% vs. 85.6%). According to the GRADE tool, the analysed outcomes were classified as having low to moderate level of certainty. Compared with blood samples, salivary urea and creatinine levels presented high diagnostic values for chronic kidney disease screening, but should not be considered equivalent to levels obtained from blood at stages three, four, or five of the disease. Chronic kidney disease patients could receive a clinically significant benefit from replacing blood with saliva for potentially monitoring renal function. Saliva collection presents greater simplicity, comfort, safety, and lower collection cost.

Objective Denture stomatitis (DS) is an oral biofilm–associated inflammation of the denture-bearing mucosa. The objective of this review was to identify and evaluate the quality of evidence on the association between the levels of salivary biomarkers and DS among adults with and without palatal DS. Materials and methods: Following the PRISMA guidelines, Medline, PubMed, EMBASE, and the Cochrane Central Register for Controlled Trials were searched for eligible studies from the beginning of the archives until December 2018. Experimental and observational studies with adult participants were included that had a control group or subgroup analysis and provided data on salivary biomarkers and DS. Articles in languages other than English or French were excluded. The level of evidence and grades of recommendation were established with the 2011 scale of the Oxford Centre for Evidence-Based Medicine. Additionally, the assessment of methodological quality was conducted with the STROBE statement (Strengthening the Reporting of Observational Studies in Epidemiology) and graded according to the Olmos scale. Results: From 1,008 citations, 9 studies were included in the systematic review (8 observational, 1 clinical trial). Seven studies suggested a statistically significant difference in the levels of salivary cytokines (IL-6, CCL3, TGF-β, CXCL8, GM-CSF, and TNF-α) between participants with DS and controls ( P < 0.05). In contrast, 2 studies concluded that the difference in the levels of several salivary cytokines (IL2, IL12, IFN-g, IL-4, IL-8, IL-10, IL-17, TNF-α, and ICAM-1) between the groups was not statistically significant. The level of evidence for the majority of studies was 3, while the grade of recommendation for all the studies was B, interpreted as “favorable.” In terms of methodological quality, most studies met 50% to 80% of STROBE criteria and were graded B. Conclusion: Palatal inflammation in DS is significantly associated with the levels of salivary cytokines. Knowledge Transfer Statement: The results of this study identified altered levels of specific salivary biomarkers associated with denture stomatitis, which may aid in the early diagnosis and treatment of this disease.

Made available in DSpace on 2021-06-25T10:55:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Canadian Institutes of Health Research Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: The aim of this in vitro study was to evaluate the effect of statherin and its naturally occurring peptides (DR9-2, DR9, GE-12, IT-32, GQ-19, IP-18) on Candida albicans metabolism and biofilm development. Design: After the killing assay, a peptide pellicle was formed on the bottom of a polystyrene plate at the IC50 of each peptide. Over the peptide pellicle, Candida albicans biofilm (48 h) was grown. The peptides antimicrobial activity after the peptides treatment was evaluated by alamarBlue, total biofilm biomass and colony forming units (CFU) counting. Results: The pellicle with statherin and the peptides (DR9-2, DR9, GE-12, IP-18, GQ-19) was able to reduce he viability of Candida albicans compared to the negative control. They also decreased cell proliferation by 20 % and total biomass. IT-32 showed the highest reduction in cell proliferation and biomass, which was similar to the positive control, histatin 5. Conclusions: These results suggest that the naturally occuring peptides from statherin are able to decrease Candida albicans colonization and biofilm proliferation. Department of Dental Materials and Prosthodontics Araraquara Dental School São Paulo State University UNESP, 1680 Rua Humaitá College of Dentistry University of Saskatchewan, 105 Wiggins Road Department of Physiology Institute of Biomedical Sciences Federal University of Uberlandia, 3Q – 2121 Av. João Naves de Ávila Department of Dental Materials and Prosthodontics Araraquara Dental School São Paulo State University UNESP, 1680 Rua Humaitá Canadian Institutes of Health Research: # 106657 FAPESP: # 2015/06016-1 Canadian Institutes of Health Research: # 400347 FAPESP: #2016/03847-2 FAPESP: #2018/13904-9

Purpose To evaluate salivary characteristics, such as flow rate, buffering capacity and amylase activity, and associate these parameters with oral health in individuals with Moebius syndrome (MS) and controls. Methods The researchers assessed caries (ICDAS), periodontal disease (PSR), and plaque (Silness Loe) indexes in both groups. Unstimulated, stimulated, and bilateral parotid saliva were also collected. The salivary flow (milliliters/minute) and α-amylase activity were measured in all samples. The α-amylase activity was determined by maltose production. The buffering capacity was measured in stimulated saliva by titration of 0.01N HCl. Results The occurrence of carious lesions, related to cut-off 2, as well as the occurrence of periodontal disease, was significantly higher (p > 0.05) in participants with MS when compared to the control group. Also, MS individuals presented decreased salivary flow, decreased buffering capacity, and amylase activity. Conclusions MS individuals present salivary alterations, which can be related to higher risk for caries.

In our recent studies, we have shown that in vivo–acquired enamel pellicle is a sophisticated biological structure containing a significant portion of naturally occurring salivary peptides. From a functional aspect, the identification of peptides in the acquired enamel pellicle is of interest because many salivary proteins exhibit functional domains that maintain the activities of the native protein. Among the in vivo–acquired enamel pellicle peptides that have been newly identified, 5 peptides are derived from statherin. Here, we assessed the ability of these statherin pellicle peptides to inhibit hydroxyapatite crystal growth. In addition, atomistic molecular dynamics (MD) simulations were performed to better understand the underlying physical mechanisms of hydroxyapatite growth inhibition. A microplate colorimetric assay was used to quantify hydroxyapatite growth. Statherin protein, 5 statherin-derived peptides, and a peptide lacking phosphate at residues 2 and 3 were analyzed. Statherin peptide phosphorylated on residues 2 and 3 indicated a significant inhibitory effect when compared with the 5 other peptides ( P < 0.05). MD simulations showed a strong affinity and fast adsorption to hydroxyapatite for phosphopeptides, whereas unphosphorylated peptides interacted weakly with the hydroxyapatite. Our data suggest that the presence of a covalently linked phosphate group (at residues 2 and 3) in statherin peptides modulates the effect of hydroxyapatite growth inhibition. This study provides a mechanism to account for the composition and function of acquired enamel pellicle statherin peptides that will contribute as a base for the development of biologically stable and functional synthetic peptides for therapeutic use against dental caries and/or periodontal disease.

Objectives For the first time, this study characterized the proteome of the acquired pellicle formed on human dentine. The changes in this proteome after exposure to lactic or citric acid were also evaluated. Methods Volunteers ( n = 9) wore a mandibular device containing 6 specimens of human root dentine. After the device remained in the volunteers’ oral cavities for 10 min or 2 h to allow the formation of the acquired pellicle in situ , the specimens were immersed in citric acid (1%, pH 2.5) or lactic acid (0.1 M, pH 4.8) or deionized water for 20 s. In sequence, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days following a crossover protocol. After harvest, proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC–ESI-MS/MS). MS/MS data were processed and submitted to Proteome Discoverer software. Searches were done using SWISS-PROT and TrEMBL databases for human proteins. Results In total, 223 distinct proteins were identified in the dentine acquired pellicle in each of the different conditions. Exposure to citric acid dramatically reduced the number of identified proteins. This did not occur for lactic acid. Acid-resistant proteins, such as mucins, were identified after pellicle was exposed to lactic or citric acid. Conclusions These proteins could be related to protective effect of tooth homeostasis. Moreover, in the future, they could be candidates to the development of a supplemental therapy for the prevention and treatment of dental caries and dental erosion. Clinical significance This study indicates some acid-resistant proteins that could be used in dental products to prevent dental caries and erosion.

Background This scoping review addressed the question ‘what do we know about stress-related changes in saliva and dental caries in general population?’ Methods The review was conducted using electronic searches via Embase, MEDLINE, PsycINFO, CINAHL and WoS. All published human studies with both observational and experimental designs were included. Two reviewers independently reviewed eligible articles and extracted the data. The studies’ quality was assessed using the Effective Public Health Practice Project Quality Assessment Tool. Results Our search identified 232 reports, of which six were included in this review. All six studies were conducted in children and used salivary cortisol as stress marker. The studies varied by design, types of stressors, children’s caries experience, methods of saliva collection. Four studies reported a positive association between saliva cortisol levels and caries (p 0.05). The quality of the included studies was weak to moderate. Conclusions There is lack of evidence about an association between stress-related changes in saliva and caries. Well-designed longitudinal studies with rigorous measurement technics for stress, saliva and dental caries are necessary. This will help to generate new insights into the multifactorial etiology of caries and provide evidence for a rational method for its control.

Proteins that have existed for millions of years frequently contain repeats of functional domains within their primary structure, thereby improving their functional capacity. In the evolutionary young statherin protein contained within the in vivo-acquired enamel pellicle (AEP), we identified a single functional domain (DR9) located within the protein N-terminal portion that exhibits a higher affinity for hydroxyapatite and more efficient protection against enamel demineralization compared to other native statherin peptides. Thus, we tested the hypothesis that multiplication of functional domains of naturally occurring pellicle peptides amplifies protection against enamel demineralization. In addition, a specific amino acid sequence from histatin 3 (RR-14) was introduced to the hybrid peptides for further testing. Enamel specimens were sectioned to 150-µm thickness and randomly grouped as follows: DR9, DR9-DR9, DR9-RR14, statherin, histatin 1, or distilled water (control). After submersion for 2 h at 37°C, the specimens were placed in 2 mL demineralization solution for 12 d at 37°C. Upon sample removal, the remaining solution was subjected to colorimetric assays to determine the amount of calcium and phosphate released from each specimen. DR9-DR9 amplified protection against enamel demineralization when compared to single DR9 or statherin. Notably, the hybrid peptide DR9-RR14 demonstrated relatively strong protection when the antimicrobial property of these peptides was tested against Candida albicans and Streptococcus mutans. DR9-RR14 was able to maintain 50% of the antifungal activity compared with RR14 for C. albicans and similar values of S. mutans killing activity. This study has pioneered the functional exploration of the natural peptide constituents of the AEP and their evolution-inspired engineered peptides. The knowledge obtained here may provide a basis for the development of stable (proteinase-resistant) synthetic peptides for therapeutic use against dental caries, dental erosion, and/or oral candidiasis.

The acquired enamel pellicle (AEP) is important for minimizing the abrasion caused by parafunctional conditions as they occur, for instance, during bruxism. It is a remarkable feature of the AEP that a protein/peptide film can provide enough protection in normofunction to prevent teeth from abrasion and wear. Despite its obvious critical role in the protection of tooth surfaces, the essential adhesion features of AEP proteins on the enamel surface are poorly characterized. The objective of this study was to measure the adhesion force between histatin 5, a primary AEP component, and hydroxyapatite (HA) surfaces. Both biotinylated histatin 5 and biotinylated human serum albumin were allowed to adsorb to streptavidin-coated silica microspheres attached to atomic force microscope (AFM) cantilevers. A multimode AFM with a Nanoscope IIIa controller was used to measure the adhesion force between protein-functionalized silica microspheres attached to cantilever tips and the HA surface. The imaging was performed in tapping mode with a Si3N4 AFM cantilever, while the adhesion forces were measured in AFM contact mode. A collection of force-distance curves (~3,000/replicate) was obtained to generate histograms from which the adhesion forces between histatin 5 or albumin and the HA surface were measured. We found that histatin 5 exhibited stronger adhesion forces (90% >1.830 nN) to the HA surface than did albumin (90% > 0.282 nN). This study presents an objective approach to adhesion force measurements between histatin 5 and HA, and provides the experimental basis for measuring the same parameters for other AEP constituents. Such knowledge will help in the design of synthetic proteins and peptides with preventive and therapeutic benefits for tooth enamel.

Aim To develop a systematic review based on the relation between salivary proteins and dental caries by comparing subjects with and without caries experience and to evaluate whether salivary proteins can be considered biomarkers for dental caries or not. Methods An electronic search was performed in the PubMed Medline, Ovid Medline, ISI Web of Science, Medline, Cochrane Library, Lilacs, Scielo, BBO, Paho and Wholis databases applying the following MeSH terms: “dental caries” OR “tooth demineralization” OR “dental caries susceptibility” OR “dental enamel solubility” AND “salivary proteins and peptides” OR “saliva” AND “proteins”. To be eligible for the systematic review, the observational controlled studies had to have groups with and without caries experience. Studies with high risk of bias were excluded. Results From a total of 188 identified studies, only seven were included in this systematic review. Four studies were classified as “low risk of bias” and three as “moderate risk of bias”. Three studies reported a relation between salivary proteins and dental caries. Conclusion There was not sufficient evidence to establish salivary proteins as a biomarker for this disease although three of the seven studies showed a relation between salivary proteins and dental caries in terms of protein phenotypes, total protein concentration and protein molecular weight.

Background: Saliva is supersaturated with respect to calcium and phosphate ions. Salivary ions may well play a role in the subsequent adsorption of proteins and consequently in the formation of the acquired enamel pellicle. Among several biological functions, the enamel pellicle forms a selectively permeable barrier that regulates demineralization processes. Aim: The aim of this study was to evaluate the importance of salivary proteins when adsorbed on enamel surface and the resultant protective effect against demineralization without the presence of salivary ions. Methods: Enamel surfaces were coated with whole saliva, parotid saliva, dialyzed whole saliva or dialyzed parotid saliva (molecular weight cutoff 1 kDa). Adsorption was allowed to proceed for a period of 2 h. Enamel specimens were then washed with deionized water and immersed into a demineralization solution of pH 4.5 for 12 days. This solution was used to measure the amount of calcium and phosphate released from enamel specimens after the demineralization period. Results: All coated specimen groups showed a significantly higher protection than those not coated with any type of saliva. In addition, undialyzed saliva (whole saliva and parotid saliva) was more effective in protecting the enamel against demineralization than dialyzed saliva. Conclusion: The present investigation indicates that the ionic composition of saliva can amplify the demineralization protection effect by reducing acid-induced enamel demineralization. Moreover, a protective effect of salivary proteins without presence of ions was demonstrated in this study.

The acquired enamel pellicle (AEP) is a thin acellular film that forms on tooth surfaces upon exposure to the oral environment. It consists predominantly of salivary proteins, but also includes non-salivary-derived proteins, carbohydrates, and lipids. Since it is the interface between teeth and the oral environment, the AEP plays a key role in the maintenance of oral health by regulating processes including lubrication, demineralization, and remineralization and shaping the composition of early microbial flora adhering to tooth surfaces. Knowledge of the 3D structure of the AEP and how that correlates with its protective functions may provide insight into several oral pathological states, including caries, erosion, and periodontal disease. This review intends to update readers about the latest discoveries related to the formation, ultrastructure, composition, and functions of the AEP, as well as the future of pellicle research, with particular emphasis on the emerging role of proteomic and microscopy techniques in oral diagnosis and therapeutics.

There is growing interest in the use of human whole saliva for diagnostics and disease monitoring as an alternative to blood samples. In contrast to blood, whole saliva is a non-sterile body fluid. Proper hand-ling and storage are required to preserve the integrity of potential biomarkers. We investigated salivary autoproteolytic degradation using a variety of approaches. We determined inhibition of protease activities by monitoring the endogenous proteome. In addition, the stability of highly protease-susceptible proteins—histatin 5, statherin, and PRP1—was assessed. Experimental variables included (a) protease inhibitors, (b) salivary pH, (c) incubation temperatures, and (d) sample heating. A cocktail containing AEBSF, aprotinin, pancreatic trypsin inhibitor, leupeptin, antipain, and EDTA could not prevent histatin 5, statherin, or PRP1 degradation in whole saliva. Among the other treatments evaluated, short-term storage of freshly collected samples on ice was effective without interfering with the chemistry of the proteome. In conclusion, whole saliva contains a unique mixture of enzymes as evidenced from their resilience to protease inhibition. Analytical evidence on protein stability is needed to ensure the validity of salivary biomarker study outcomes. Analysis of the data presented will provide help and guidance for the use of saliva samples for diagnostic purposes.

OBJECTIVE: To determine the pH over a period of 168 h and the ionic silver content in various concentrations and post-preparation times of aqueous silver nitrate solutions. Also, the possible effects of these factors on microleakage test in adhesive/resin restorations in primary and permanent teeth were evaluated. MATERIAL AND METHODS: A digital pHmeter was used for measuring the pH of the solutions prepared with three types of water (purified, deionized or distilled) and three brands of silver nitrate salt (Merck, Synth or Cennabras) at 0, 1, 2, 24, 48, 72, 96 and 168 h after preparation, and storage in transparent or dark bottles. Ionic silver was assayed according to the post-preparation times (2, 24, 48, 72, 96, 168 h) and concentrations (1, 5, 25, 50%) of solutions by atomic emission spectrometry. For each sample of each condition, three readings were obtained for calculating the mean value. Class V cavities were prepared with enamel margins on primary and permanent teeth and restored with the adhesive systems OptiBond FL or OptiBond SOLO Plus SE and the composite resin Filtek Z-250. After nail polish coverage, the permanent teeth were immersed in 25% or 50% AgNO3 solution and the primary teeth in 5% or 50% AgNO3 solutions for microleakage evaluation. ANOVA and the Tukey's test were used for data analyses (α=5%). RESULTS: The mean pH of the solutions ranged from neutral to alkaline (7.9±2.2 to 11.8±0.9). Mean ionic silver content differed depending on the concentration of the solution (4.75±0.5 to 293±15.3 ppm). In the microleakage test, significant difference was only observed for the adhesive system factor (p=0.000). CONCLUSIONS: Under the tested experimental conditions and based on the obtained results, it may be concluded that the aqueous AgNO3 solutions: have neutral/alkaline pH and service life of up to 168 h; the level of ionic silver is proportional to the concentration of the solution; even at 5% concentration, the solutions were capable of indicating loss of marginal seal in the composite restorations; the 3-step conventional adhesive system had better performance regarding microleakage in enamel on primary and permanent teeth.

This in vivo study aimed to evaluate the performance of 2 fluorescence-based methods in detecting occlusal caries lesions in primary teeth, compared with the performance of visual inspection and radiographic methods, and to propose a mathematic correction of the diagnostic parameters due to the imperfect reference standard method used in the study. Two examiners assessed the occlusal surfaces of 407 primary teeth (62 children) using visual inspection (ICDAS), radiographic, DIAGNOdent pen (pen type laser fluorescence; LFpen), and fluorescence camera (FC) methods. At the noncavitated threshold (NC) the reference standard method was the results of ICDAS, and at the dentine caries threshold (D3) teeth diagnosed with dentine caries by ICDAS or radiographic methods were subjected to operative treatment to confirm the presence of lesion. Reproducibility, sensitivity, specificity, accuracy, and the area under the ROC curve were calculated for the methods at both thresholds. At the NC threshold, LFpen had a slightly better performance compared to the FC and radiographic methods. However, at the D3 threshold, both fluorescence-based methods performed similarly. Visual inspection and radiographic methods presented higher specificities but lower sensitivities than fluorescence methods. After corrections, there was a significant decrease in some parameters. In conclusion, both fluorescence-based methods presented similar performance in detecting occlusal dentine caries lesions in primary teeth, but they usually gave more false-positive results than did the visual and radiographic methods. The correction proposed shows that the performance of the methods can be overestimated, and the correction should be validated and considered in further studies that use an imprecise reference standard method.

Objective The aim of this study was to investigate the type and the nature of peptides present in the in vivo formed human acquired enamel pellicle. Design Pellicle material was collected from 10 volunteers and subjected to sample preparations consisting of centrifugal filtration using a 10 kDa molecular weight cut-off membrane and high-resolution gel filtration chromatography. The fractions containing peptides Results 78 natural pellicle peptides with molecular weights ranging from 766.9 Da to 3981.4 Da were identified originating from 29 different proteins. Conclusions The number of peptides present in acquired enamel pellicle appears to be large and this is likely to enhance the functional spectrum of this protein film. The presence of small peptides in pellicle may be functionally important since structure/function studies of many salivary proteins have shown that specific domains within these native proteins retain or even exhibit enhanced biological activities. The data present the basis for determining the precise function of these pellicle peptides and for gaining insights into the role pellicle plays in the oral cavity.

Recent research efforts in oral biology have resulted in elucidation of the proteomes of major human salivary secretions and whole saliva. One might hypothesize that the proteome of minor gland secretions may show significantly different characteristics when compared with the proteomes of parotid or submandibular/sublingual secretions. To test this hypothesis, we conducted the first exploration into the proteome of minor salivary gland secretion. Minor gland secretion was obtained from healthy volunteers, and its components were subjected to liquid-chromatography-electrospray-ionization-tandem-mass-spectrometry. This led to the identification of 56 proteins, 12 of which had never been identified in any salivary secretion. The unique characteristics of the minor salivary gland secretion proteome are related to the types as well as the numbers of components present. The differences between salivary proteomes may be important with respect to specific oral functions.

Objectives This study characterized the proteome profile of the acquired pellicle formed in vivo on enamel. Changes in this proteome profile after exposure to lactic or citric acid were also evaluated. Methods Volunteers ( n = 8) were subjected to dental prophylaxis. After 2 h to allow the formation of the acquired pellicle, the teeth were isolated with cotton rolls and 1 mL of citric acid (1%, pH 2.5) or lactic acid (0.1 M pH 4.8) or deionized water was gently applied with a pipette on the anterior teeth (both maxillary and mandibular) for 10 s. In sequence, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days following a crossover protocol. Proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC-ESI-MS/MS). MS/MS data were processed and submitted to Proteome Discoverer software. Searches were done using SWISS-PROT and TrEMBL databases for human proteins. Results In total, seventy-two proteins were present in all groups and were submitted to quantitative analysis (SIEVE). Some of these proteins were increased more than two-fold after exposure to the acids. Among them, cystatin-B was increased 20- and 13-fold after exposure to citric and lactic acids, respectively. Additionally, some proteins were identified in only one of the groups (18, 5, and 11 proteins for deionized water, citric and lactic acids, respectively). Conclusions Our results open new insights regarding potentially acid-resistant proteins that could be added to dental products to prevent acidic dissolution of the teeth.

Summary Objective Human saliva contains a family of low molecular weight histidine-rich proteins, named histatins, characterised by bactericidal and fungicidal activities in vitro against several microbial pathogens, such as Streptococcus mutans and Candida albicans. They represent a major component of an innate host non-immune defense system. In an earlier study we described the distribution of histatins in the glandular parenchyma of human major salivary glands, confirming that all human major salivary glands are involved in the secretion of histatins into saliva. In the present study we determined the expression and localisation of histatins in human posterior deep lingual glands (von Ebner's glands) by means of immunoelectron microscopy. Design Thin sections of normal human salivary glands, embedded in Epon resin, were incubated with rabbit polyclonal antibodies specific for human histatins and successively with a gold conjugated goat anti-rabbit IgG used as secondary antibody. Sections incubated with medium devoid of primary antibody or containing non-immune serum were used as controls. Results The serous secreting cells represented the main source of histatins in the glandular parenchyma of von Ebner's glands. At the electron microscopic level, labeling was associated with rough endoplasmic reticulum, Golgi complex and secretory granules that represented the main cytoplasmic site of histatin localisation. However, variability in the intensity of labeling was observed among adjacent cells. Conclusions The present results show for the first time that human von Ebner's glands produce and represent a significant source of histatins, supporting the hypothesis of their important role in preventing microbial assaults on the tissues in the posterior region of the tongue and in the circumvallate papillae.

Study design In this study sodium, potassium, calcium, phosphorus, zinc, and magnesium ion concentration was analyzed in stimulated whole saliva in 22 children with Down syndrome aged 6 to 10 years. These children were compared with 21 healthy children of the same age. Stimulated saliva was collected by chewing a piece of parafilm for 10 minutes. The pH was measured with a portable pH meter after collection. Electrolyte concentrations were determined by inductively coupled argon plasma with atomic emission spectrometry. Results No statistically significant differences were observed in phosphorus, zinc, magnesium, and calcium concentration between the children with Down syndrome and the control children. The sodium concentration showed about 66.8% higher value in the Down syndrome than in the control group. On the other hand the flow rate and potassium concentration were lower in the Down syndrome than in the control group. Conclusion Our results suggest that there is alteration in the metabolism of the duct and/or acinar cells of salivary glands of Down syndrome children.

Dental erosion is a multifactorial condition that can result in the loss of tooth structure and function, potentially increasing tooth sensitivity. The exposure of enamel to acids from non-bacterial sources is responsible for the progression of erosion. These erosive challenges are counteracted by the anti-erosive properties of the acquired pellicle (AP), an integument formed in vivo as a result of selective adsorption of salivary proteins on the tooth surface, containing also lipids and glycoproteins. This review provides an in-depth discussion regarding how the physical structure of the AP, along with its composition, contributes to AP anti-erosive properties. The physical properties that contribute to AP protective nature include pellicle thickness, maturation time, and site of development. The pellicle contains salivary proteins embedded within its structure that demonstrate anti-erosive properties; however, rather than individual proteins, protein-protein interactions play a fundamental role in the protective nature of the AP. In addition, dietary and synthetic proteins can modify the pellicle, enhancing its protective efficiency against dental erosion. The salivary composition of the AP and its corresponding protein-profile may be employed as a diagnostic tool, since it likely contains salivary biomarkers for oral diseases that initiate at the enamel surface, including dental erosion. Finally, by modifying the composition and structure of the AP, this protein integument has the potential to be used as a target-specific treatment option for oral diseases related to tooth demineralization.

purpose of this open-tabel, crossover study was to meesure salivary pH after the oral administration of dis- tilled water, a 10% sucrose solution, and valproic acid to a population of 10 chil- dren with cerebral palsy. Salivary pH was measured just prior to giving the solutions and then every five minutes for 60 minutes. The critical pH was defined at 5.5, below which dental demineraliza- tion occurs. In this population, valproic acid decreased salivary pH to below the critical pH for an extended period of time (mean=4.32 at 20 minuted when compared to mean salivary pH after administration of distilled water (7.1 1 at 20 minutes) and a 10% sucrose solution (5.96 at 20 minutes). Recovery from this low pH to above the critical level did not occur until after 50 minutes and further- more did not recover to baseline, neutral levels, for the duration of this study (60 minutes). Our results suggest that chil- dren with cerebral palsy, a population with a tendency toward reduced salivary function, have a compromised ability to buffer the oral administration of an exogenous acid. This can result in an increased susceptibility to demineraliza- tion and caries of the teeth.

Histatins are salivary proteins that exhibit a high affinity for hydroxyapatite and contribute to the acquired enamel pellicle. Previous studies have observed that, despite the high proteolytic activity in saliva, significant numbers of histatin molecules in acquired enamel pellicle are intact. Our working hypothesis was that histatins are less susceptible to proteinases present in saliva when adsorbed on the hydroxyapatite. To test this premise, we incubated histatin 1 with hydroxyapatite and human whole saliva. Proteolytic products of this incubation were then characterized by PAGE, HPLC, and mass spectrometry. This study shows for the first time that binding to hydroxyapatite confers intact histatin 1 with resistance to proteolytic degradation.

Understanding the composition and function of the acquired enamel pellicle (AEP) has been a major goal in oral biology. The aim of this study was to test the hypothesis that intact histatins are part of the in vivo AEP and that histatins after adsorption to HA have effects on in vitro enamel demineralization. This is the first study demonstrating the presence of intact histatins in vivo in the AEP. The in vitro experiments show that all naturally occurring histatins in the AEP have the potential to provide some level of protection against acid injury.

Objective The aim was to compare the performance of different methods in detecting approximal caries lesions primary molars ex vivo. Study design One hundred thirty-one approximal surfaces were examined by 2 observers with visual inspection (VI) using the International Caries Detection and Assessment System, radiographic interpretation, and clinically using the Diagnodent pen (LFpen). To achieve a reference standard, surfaces were directly examined for the presence of white spots or cavitations, and lesion depth was determined after sectioning. The area under the receiver operating characteristic curve (A z ), sensitivity, specificity, and accuracy were calculated, as well as the interexaminer reproducibility. Results Using the cavitation threshold, all methods presented similar sensitivities. Higher A z values were achieved with VI at white spot threshold, and VI and LFpen had higher A z values at cavitation threshold. VI presented higher accuracy and A z than radiographic and LFpen at both enamel and dentin depth thresholds. Higher reliability values were achieved with VI. Conclusions VI performs better, but both radiographic and LFpen methods also show good performance in detecting more advanced approximal caries lesions.

The aim of this study was to investigate caries experience in individuals with cerebral palsy (CP) who have oromotor dysfunction and relate it to age and dietary consistency. Noninstitutionalized individuals with CP (n = 108) aged 4-19 years (mean: 10 years, 1 month +/- SD 4 years, 5 months) were recruited for this study. Subjects who were severely impaired (35.2%) had orofacial motor dysfunction most frequently, followed by those who were slightly affected (27.7%), moderately affected (20.4%), and very slightly affected (16.7%). Age was a statistically significant factor for oromotor dysfunction (p= 0.007), with the youngest individuals having the most severe oromotor problems. Dietary consistency and oromotor function were statistically significant influence on the DMF index (p= 0.0352). The highest total DMF values were measured for individuals who were severely impaired and also the youngest, as well as for those receiving liquid diets. Early rehabilitation, intervention, and prevention are important for these individuals.

Influenza A virus (IAV) is a major cause of morbidity and mortality in yearly epidemics and there is substantial concern about the development of a new pandemic because of frequent instances of transmission of avian strains to humans (33, 35). The ability of IAV strains to modify their antigenic properties through small incremental mutations and major changes resulting from the exchange of genome segments with those of animal strains (reassortment), results in continued emergence of strains for which prior adaptive immune responses are ineffective. Although essentially everyone can be infected with IAV, certain individuals and groups suffer much more severe illness after infection. It is likely that innate defense mechanisms play an important role in the first crucial days after infection, given the normal delay in development of protective antibodies or T cells. Innate mediators that are important in IAV infection include soluble components [e.g. the collectins, surfactant proteins A and D (SP-A and -D), tumor necrosis factor-α, type I interferons, natural immunoglobulin M (IgM), and complement] and cellular elements (e.g. natural killer cells, dendritic cells, macrophages, and neutrophils) (11-13, 16, 20, 32, 37)]. Although the respiratory tract is the main site of replication for IAV, the oral cavity is a potentially important route for viral transmission. There is a fairly extensive literature regarding the ability of saliva to inhibit HIV but relatively less is known about how it inhibits other viruses, including IAV. We reported that the saliva of a healthy volunteer donor inhibited IAV hemagglutination (HA) activity and infectivity (12). Innate immune factors contributed strongly to this neutralizing activity because removal of IgA only slightly reduced it. The antiviral activity of saliva differed qualitatively from that of bronchoalveolar lavage (BAL) fluid. In the case of BAL fluid, calcium-dependent lectin activity, contributed largely by SP-D, was predominant (10). We found, however, that SP-D is not present in saliva at sufficient concentrations to account for its anti-influenza activity and that calcium-dependent lectin activity makes a much smaller contribution to the antiviral activity of saliva than to BAL fluid. Salivary glycoprotein 340 (gp-340) did account for some of the antiviral activity of saliva, but activity remained after removal of gp-340 (12). The goal of this study was to characterize further the mechanisms of antiviral activity of saliva through the use of a panel of saliva donors providing samples obtained specifically from parotid and submandibular/sublingual glands, and purified salivary components.

This study evaluated the flow rate and composition of whole saliva in patients with chronic renal failure undergoing hemodialysis. In the group on dialysis (RG) (n = 15), saliva was collected just prior to hemodialysis CT1) and at completion (T2), while in the healthy subjects (HG)(n = 15) saliva was collected at the same time of day as the pre-dialysis. Saliva samples were analyzed by inductively coupled argon plasma with atomic emission spectrometry. Significant differences were found in the flow rate, potassium, magnesium and phosphorus concentrations at the RG-T1 and HG (p < 0.05). Sodium concentration at RG-T1 and RG-T2 were higher than HG (p < 0.05). Total protein concentration was higher at RG-T1 than at the other two analyses. Salivary peroxidase activity at RG-T1 and RG-T2 was lower than at HG. Our findings suggest that in patients with chronic renal failure, the saliva is altered. Hemodialysis, however, seems to help control saliva composition and flow rate.

Histatins are human salivary antifungal proteins that are prone to extensive enzymatic degradation upon their release into the oral cavity. Histatin proteolysis, leading to the disappearance of the intact protein can be expected to have functional consequences. Histatin 5, comprising 24 residues, is the smallest of the major salivary histatins and the most active in terms of its antifungal properties. The rate and mode of histatin 5 degradation were determined by incubating the protein in whole saliva supernatant for various time intervals. Fragmentation products were collected by reversed-phase high performance liquid chromatography (RP-HPLC), characterised structurally by matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry and functionally in a fungal growth inhibition assay. Of the 19 fragments identified, 16 were derived from single proteolytic cleavage events in histatin 5. A remarkable finding was the inter-subject consistency in the histatin 5 degradation pattern. Added histatin 5 disappeared from whole saliva supernatant at an average rate of 105+/-22 microg/ml/h, which in part could explain the virtual absence of histatin 5 in whole saliva. Despite the rapid proteolysis of histatin 5, the early degradation mixture was as active in antifungal assays as intact histatin 5. These data demonstrate that the oral-fluid mediated proteolysis of histatin 5 represents an intrinsic biological property of whole saliva. The data also reveal that the early proteolysis phase of histatin 5 does not abolish the antifungal properties associated with this protein.

Summary Objectives To evaluate the performance of the laser fluorescence device (DIAGNOdent) for detection and quantification of smooth-surface caries lesions in primary teeth. Methods Seventy-seven approximal surfaces with no caries or with white spot caries lesions without cavitation was selected from 51 primary molars. Laser fluorescence readings were performed and the site of higher value in each surface was indicated. After the readings, the surfaces were cut and the samples were destined for histopathological and cross-sectional microhardness evaluation. ROC analysis was conducted and the area under the ROC curve and the best cut-off points were obtained at D 1, D 2 and D 3 thresholds. With these cut-off values, the sensitivity, specificity and accuracy were calculated at these thresholds. The regression lines of laser fluorescence readings as function of lesion extension, lesion depth and cross-sectional microhardness values were plotted and the Pearson correlation coefficients were obtained. Results The best DIAGNOdent performance was obtained at D 3 threshold ( A z =0.90) and the worst performance was achieved at D 1 threshold ( A z =0.82). The correlation was better between the laser fluorescence readings and lesion depth obtained in polarized light microscope examination ( r =0.78), than that obtained with the microhardness values ( r =−0.57). Conclusion The laser fluorescence shows a good performance to predict caries lesions extension. Device presents a better correlation with the lesion depth than with mineral loss in smooth-surface caries lesions.

This study measured the flow rate, pH, and buffer capacity of saliva from children with Down syndrome aged 2–60 months. We compared 25 children with Down syndrome with 21 children without Down syndrome. The whole saliva was collected under slight suction and the pH and the buffer capacity were determined using a digital pHmeter. The buffer capacity was measured by titration with 0.01 N HCl. The Down syndrome group demonstrated higher buffer capacity than the control group in the pH ranges of 6.9–6.0, 5.9–5.0, and 4.9–4.0. The flow rate was low in the Down syndrome group. Boys from the Down syndrome group demonstrated higher buffer capacity in the pH ranges 6.9–6.0, 5.9–5.0, and 4.9–4.0 than controls, while girls from the Down syndrome group showed no difference compared with controls. Girls with Down syndrome demonstrated significant difference only in the range of pHi–pH 7.0. Conclusion. These data suggest that the Down syndrome persons present a better buffer capacity, supporting the results observed in several studies which found the low dental caries in persons with Down syndrome.

The authors report on the components of stimulated whole saliva from children with Down syndrome—including pH, flow rate, sialic acid and protein concentrations, and amylase and peroxidase activity. Saliva samples were collected from 35 children aged 6–10 years. Of the participants, 17 had Down syndrome and 18 did not. To stimulate saliva production, the children chewed a piece of parafilm for 10 minutes before the sample was collected. Soon after collecting the saliva sample, the authors measured pH using a portable pH-meter. Sialic acid levels were determined with a thiobarbituric acid assay. Protein content was determined with Folin's phenol reagent. Amylase was assayed and the authors measured the maltose produced by the breakdown of starch and peroxidase using ortho-dianisidine. No statistically significant difference was observed in levels of sialic acid (free and total) between the two groups. Protein concentration was about 36% higher in the group with Down syndrome. However, the salivary flow rate, pH, and amylase and peroxidase activities were lower among the children with Down syndrome.

Proteomic techniques have become popular in medicine and dentistry because of their widespread use in analyzing bodily fluids such as blood, saliva, urine, and gingival crevicular fluids as well as hard tissues such as enamel, dentine, and cementum. This review is a guide to proteomic techniques in general dentistry, summarizing techniques and their clinical application in understanding and diagnosing diseases and their use in identifying biomarkers of various diseases.