RESUMEN

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

Asunto(s)

RESUMEN

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

Asunto(s)

RESUMEN

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

Asunto(s)

RESUMEN

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

RESUMEN

With the growing understanding of the relationship between oral and systemic diseases there is an increasing interest in integrating dental providers more completely into the health care delivery system. Technology advancements in traditional dental disease diagnosis and management as well as in salivary diagnostics for systemic disease provide great opportunities for the profession but numerous challenges for dental educators who are impacted by the intersection of numerous, sometimes competing interests. While economic factors play a significant role in curricular innovation and change, in the United States the accrediting process for dental education programs can be an equally vexing challenge. Movement away from primarily a competency-based assessment model to a model focusing on the establishment of Entrustable Professional Activities (EPAs) may provide a needed catalyst to enhance dental education to graduate dentists who embrace an expanded role as primary care providers.

Asunto(s)

RESUMEN

A fully integrated device for salivary detection with a sample-in-answer-out fashion is critical for noninvasive point-of-care testing (POCT), especially for the screening of contagious disease infection. Microfluidic paper-based analytical devices (µPADs) have demonstrated their huge potential in POCT due to their low cost and easy adaptation with other components. This study developed a generic POCT platform by integrating a centrifugal microfluidic disc with µPADs to realize sample-to-answer salivary diagnostics. Specifically, a custom centrifugal microfluidic disc integrated with µPADs is fabricated, which demonstrated a high efficiency in saliva treatment. To demonstrate the capability of the integrated device for salivary analysis, the SARS-CoV-2 Nucleocapsid (N) protein, a reliable biomarker for SARS-CoV-2 acute infection, is used as the model analyte. By the chemical treatment of the µPAD surface, and by optimizing the protein immobilization conditions, the on-disc µPADs were able to detect the SARS-CoV-2 N protein down to 10 pg mL-1 with a dynamic range of 10-1000 pg mL-1 and an assay time of 8 min. The integrated device was successfully used for the quantification of the N protein of pseudovirus in saliva with high specificity and demonstrated a comparable performance to the commercial paper lateral flow assay test strips.

Asunto(s)

RESUMEN

With the advantages of convenient, painless and non-invasive collection, saliva holds great promise as a valuable biomarker source for cancer detection, pathological assessment and therapeutic monitoring. Salivary glycopatterns have shown significant potential for cancer screening in recent years. However, the understanding of benign lesions at non-cancerous sites in cancer diagnosis has been overlooked. Clarifying the influence of benign lesions on salivary glycopatterns and cancer screening is crucial for advancing the development of salivary glycopattern-based diagnostics. In this study, 2885 samples were analyzed using lectin microarrays to identify variations in salivary glycopatterns according to the number, location, and type of lesions. By utilizing our previously published data of tumor-associated salivary glycopatterns, the performance of machine learning algorithm for cancer screening was investigated to evaluate the effect of adding benign disease cases to the control group. The results demonstrated that both the location and number of lesions had discernible effects on salivary glycopatterns. And it was also revealed that incorporating a broad range of benign diseases into the controls improved the classifier's performance in distinguishing cancer cases from controls. This finding holds guiding significance for enhancing salivary glycopattern-based cancer screening and facilitates their practical implementation in clinical settings.

Asunto(s)

RESUMEN

BACKGROUND: The activity of antioxidant enzymes in periodontitis is reduced, but results vary between studies and are subject to bias. In turn, the expression of genes encoding antioxidant factors has not been examined yet. OBJECTIVES: This is the first study to evaluate the expression of genes encoding superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1) and thioredoxin 1 (TXN1) in the saliva and gingival tissue of patients with periodontitis. The activity of the antioxidant enzyme protein products in the unstimulated and stimulated saliva and the gingival crevicular fluid (GCF) of patients with periodontitis was also investigated. MATERIAL AND METHODS: The prospective study involved 65 patients with periodontitis, who were divided into groups depending on the disease stage, and a control group of 31 ageand gender-matched healthy patients. RESULTS: We demonstrated that the expression of genes encoding GPX1 and TXN1 in saliva was significantly higher, and the expression of genes encoding SOD1, GPX1 and TXN1 in the gingival tissue was significantly lower in periodontitis patients as compared to the control group. We noted a lower activity of GPX1 in unstimulated saliva, of SOD1 in stimulated saliva and of both antioxidant enzymes in GCF in patients with periodontitis. CONCLUSIONS: The GPX1 transcriptome and its activity in the salivary and GCF proteome appear to be dependent on the oxidative stress related to the destructive inflammatory changes in periodontitis.

Asunto(s)

RESUMEN

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

RESUMEN

OBJECTIVES: The evaluation of salivary biomarkers has been proposed as a simple and non-invasive aid to the conventional periodontal diagnosis based on clinical-radiographic parameters. Matrix metalloproteinase-8 (MMP-8), especially in its active form, is considered one of the most reliable biomarkers of periodontitis, and point-of-care tests (POCTs) have been proposed for its clinical monitoring. In this proof-of-concept study, a novel highly sensitive POCT based on a plastic optical fiber (POF) biosensor exploiting surface plasmon resonance (SPR) to detect salivary MMP-8 is described. METHODS: A SPR-POF biosensor was functionalized with a specific antibody to develop a surface-assembled monolayer (SAM) for the detection of total MMP-8. A white light source and a spectrometer connected to the biosensor were used to quantify MMP-8 level in both buffer and real matrix (saliva) by analysing the shift of the resonance wavelength determined by the specific antigen-antibody binding upon the SAM. RESULTS: Dose-response curves by serial dilutions of human recombinant MMP-8 were realized, obtaining a limit of detection (LOD) of 40 pM (1.76 ng/ml) in buffer and 225 pM (9.9 ng/ml) in saliva and high selectivity compared to interferent analytes (MMP-2 and IL-6). CONCLUSIONS: The proposed optical fiber-based POCT was able to detect and measure total MMP-8 with high selectivity and very low LOD in both buffer and saliva. CLINICAL SIGNIFICANCE: The SPR-POF technology may be employed to create highly sensitive biosensors to monitor salivary MMP-8 levels. The possibility of specifically detecting its active, rather than total, form need to be further investigated. If confirmed and clinically validated, such a device may represent a promising tool to make an immediate, highly sensitive and reliable diagnosis of periodontitis, and to carry out a timely and targeted therapy, possibly helping to prevent the onset of local and systemic periodontitis-related complications.

Asunto(s)

RESUMEN

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

Asunto(s)

RESUMEN

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

Asunto(s)

RESUMEN

Saliva is a complex oral biofluid composed of various biological molecules secreted by major and minor salivary glands, as well as by-products of host oral cells, oral bacteria, small extracellular vesicles (sEVs), and gingival crevicular fluid. Recently, salivary small extracellular vesicles and salivary multi-omics (microbiome, transcriptome, DNA methylome and proteome) are emerging as potential diagnostic tools for oral diseases, including the highly prevalent periodontitis. Here, we describe the methodologies for how to isolate salivary sEVs using the size exclusion chromatography method, and how to perform salivaomics, which may guide future dental research.

Asunto(s)

RESUMEN

BACKGROUND: Due to their low specificity, non-enzymatic antioxidants play a significant role in the protection of organisms against free radicals. They are normally sourced from the diet, and independently react with oxidizing molecules and their products. OBJECTIVES: The study aimed to determine the concentrations of selected non-enzymatic antioxidants (uric acid (UA), reduced glutathione (GSH) and polyphenols) in the gingival fluid and saliva of patients diagnosed with periodontitis according to the current criteria. MATERIAL AND METHODS: This prospective case-control study included 50 patients with periodontitis, who were divided into 2 groups depending on disease severity, along with 25 healthy controls. Unstimulated saliva, stimulated saliva and gingival crevicular fluid (GCF) were collected from all subjects, and nonenzymatic antioxidant concentrations were determined. RESULTS: Significantly lower concentrations of all tested non-enzymatic antioxidants were observed in the gingival fluid as well as in the unstimulated and stimulated saliva of patients with periodontitis (p < 0.05). Moreover, the concentration of GSH was a parameter that differentiated the various degrees of periodontitis (p < 0.05). A significantly lower concentration of GSH was found in the stimulated saliva of patients with moderate progression as compared to those with fast progression of the disease (p < 0.05). CONCLUSIONS: The continuation of research on the GSH concentrations in the gingival fluid and saliva may be useful in the context of biomarkers for periodontitis progression.

Asunto(s)

RESUMEN

Non-coding RNAs (ncRNAs) represent a research hotspot by playing a key role in epigenetic and transcriptional regulation of diverse biological functions and due to their involvement in different diseases, including oral inflammatory diseases. Based on ncRNAs' suitability for salivary biomarkers and their involvement in neuropathic pain and tissue regeneration signaling pathways, the present narrative review aims to highlight the potential clinical applications of ncRNAs in oral inflammatory diseases, with an emphasis on salivary diagnostics, regenerative dentistry, and precision medicine for neuropathic orofacial pain.

Asunto(s)

RESUMEN

Background: The relevance of growth determination in orthodontics is driving the search for the most precise and least invasive way of tracking the pubertal growth spurt. Objectives: The aim was to explore whether minimally invasive salivary estimation of biomarkers Insulin-like growth factor (IGF-1) and Insulin-like growth factor binding protein-3 (IGFBP-3) could be used to estimate skeletal maturity with diagnostic accuracy, especially in children and adolescent age groups. Subjects and methods: The cross-sectional study was conducted on 105 participants aged 6−25 years from the out-patient Department of Preventive Dental Science at Majmaah University between the period 2 January 2021 and 12 July 2021. Each subject's lateral cephalogram radiograph was categorized based on skeletal maturity, and saliva samples were estimated for IGF-1 and IGFBP-3 using the respective ELISA kits. Two-way ANOVA with interaction was applied to examine the main effects due to cervical vertebral maturation staging (CVS), Sex and interaction effect due to CVS, and Sex on study parameters. Karl Pearson's Product Moment Correlation Coefficient was calculated for finding a significant association between IGF, IGFBP3, and the IGF-1/IGFBP3 molar ratio. Results: Highest mean salivary IGF-1 was observed in the pubertal peak stage, which coincides with cervical vertebral maturity stages 3 and 4 (CVS3 and CVS4) for both males (2.57 ng/mL) and females (1.57 ng/mL) and the lowest mean level of IGF-1 for females (0.85 ng/mL) and males (1.22 ng/mL) was observed during the prepubertal stage. There exists a significant variation in IGF-1 between males and females in the pubertal stage (p < 0.01), but the difference is very narrow in the prepubertal and post-pubertal groups (p > 0.05). There was no significant interaction effect of different skeletal stages and gender on the IGFBP3 and the IGF-1/IGFBP3 molar ratio (p > 0.05), but there exists a significant interaction effect on IGF-1 (p < 0.05). Conclusion: Estimation of the IGF-1 and the IGF-1/IGFBP3 molar ratio in saliva, being a non-invasive biological marker, could serve as an adjunctive tool along with radiographic assessment in estimating growth maturity in the adolescence age group. By initiating orthodontic treatment during the mandibular growth peak in adolescence, a positive outcome is ensured in managing skeletal deformities within the craniofacial complex.

Asunto(s)

RESUMEN

Objective: The relevance of growth determination in orthodontics is driving the search for the most precise and least invasive way of tracking the pubertal growth spurt. Our aim was to explore whether minimally invasive salivary estimation of biomarkers Insulin-like growth factor (IGF-1) and Insulin-like growth factor binding protein-3 (IGFBP-3) could be used to estimate skeletal maturity for clinical convenience, especially in children and adolescent age groups. Materials and Method: The cross-sectional study was conducted on 90 participants (56 girls and 34 males) with ages ranging from 6 to 25 years. Each subject's hand-wrist radiograph was categorized based on skeletal maturity, and saliva samples were estimated for IGF-1 and IGFBP-3 using the respective ELISA kits. Kruskal−Wallis nonparametric ANOVA was applied to compare different skeletal stages. Results: The study demonstrated low salivary IGF-1 levels at the prepubertal stage, with increase during pubertal onset and peak pubertal stage followed by a decline during pubertal deceleration to growth completion. Spearman's correlation coefficient demonstrated a strong positive association (r = 0.98 p < 0.01) between salivary IGF/IGFBP-3 ratio and different stages of skeletal maturity. Conclusion: Salivary IGF-1, IGFBP-3, and IGF/IGFBP-3 ratio could serve as a potential biochemical marker for predicting the completion of skeletal maturity.

Asunto(s)

RESUMEN

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

Asunto(s)