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Frontiers in Molecular Biosciences 2024Human saliva was used to develop non-invasive liquid biopsy biomarkers to establish saliva as an alternate to blood and plasma in translational research. The present...
Human saliva was used to develop non-invasive liquid biopsy biomarkers to establish saliva as an alternate to blood and plasma in translational research. The present study focused on understanding the impact of sample storage conditions on the extraction of RNA from saliva and the RNA yield, to be applied in clinical diagnosis. In this study, genes related to asthma were used to test the method developed. Salivary RNA was extracted from three subjects using the Qiazol based method and quantified by both spectrophotometric (NanoDrop) and fluorometric (Qubit) methods. RNA integrity was measured using a bioanalyzer. Quantitative PCR was used to monitor the impact of storage conditions on the expression of housekeeping genes: and β-actin, and the asthma related genes: and . In addition, an independent cohort of 38 asthmatics and 10 healthy controls were used to validate the expression of and as mRNA salivary biomarkers. Approximately 2 µg of total RNA was obtained from the saliva stored at 40°C without any preservative for 2 weeks showing consistent gene expression with RNA stored at room temperature (RT) for 48 h with RNA. Although saliva stored with RNA showed a substantial increase in the yield (110 to 234 ng/μL), a similar Cq (15.6 ± 1.4) for the 18s rRNA gene from saliva without preservative showed that the RNA was stable enough. Gene expression analysis from the degraded RNA can be performed by designing the assay using a smaller fragment size spanning a single exon as described below in the case of the and genes in the asthma cohort. This study showed that samples stored at room temperature up to a temperature of 40°C without any preservative for 2 weeks yielded relatively stable RNA. The methodology developed can be employed to transport samples from the point of collection to the laboratory, under non-stringent storage conditions enabling the execution of gene expression studies in a cost effective and efficient manner.
PubMed: 38948078
DOI: 10.3389/fmolb.2024.1363897 -
Journal of Conservative Dentistry and... May 2024The purpose of this study is to comparatively evaluate the effect of discoloration of nanohybrid composite by four different phytopigments.
OBJECTIVE
The purpose of this study is to comparatively evaluate the effect of discoloration of nanohybrid composite by four different phytopigments.
MATERIALS AND METHODS
Fifty disk-shaped samples of nanohybrid (3M Filtek Z350) resin composites were prepared using an acrylic template of dimension 5 mm × 3 mm. They were randomly divided into five groups and immersed in solutions of tomato powder, beetroot powder, java plum powder, and turmeric powder. Distilled water was used as the control group. The samples were placed in respective solutions for 3 h daily and stored in artificial saliva for the rest of the day for 28 days. Color values (L*, a*, b*) were measured by colorimeter using the CIE L*a*b* system at the end of the 7 and 28 days of immersion. Color differences ΔE*ab were statistically analyzed.
RESULTS
All the samples showed a change in color of nanohybrid composite resin to varying degrees. The mean ΔE*ab value obtained with beetroot solution was the highest among all the groups at the end of the 7 and 28 days, depicting that beetroot solution showed maximum mean color variation, followed by java plum solution, turmeric solution, and tomato solution.
CONCLUSION
All the phytopigments used in this study have the potential to discolor the nanohybrid composite resin, with beetroot causing the most severe discoloration.
PubMed: 38939539
DOI: 10.4103/JCDE.JCDE_169_24 -
Journal of Pharmaceutical and... Jun 2024An instrumental-free, high-throughput assay has been developed for the quantification of thiocyanate in human saliva. The proposed green method is based on the rapid...
An instrumental-free, high-throughput assay has been developed for the quantification of thiocyanate in human saliva. The proposed green method is based on the rapid reaction of the analyte with Fe(III) under acidic pH in a microplates format to form a colored complex that is captured as an image by an overhead book scanner. Optimization included the effects of the amount concentration of Fe(III), acidity and reaction time / complex stability using a total volume of 300 μL per well. Validation towards the matrix effect was focused on the specific application and was performed using both artificial and human saliva. The linearity of the developed assay was up to 500 μM thiocyanate offering a lower limit of quantification (LLOQ) of 30 μM. The green potentials were evaluated by both the Green Analytical Procedure (GAPI) and Blue Applicability Grade (BAGI) indexes. The thiocyanate content in the saliva of non-smoking volunteers ranged between 750 and 1350 μΜ, while elevated concentrations were verified in smoking individuals (1860-3080 μΜ). Statistical agreement with a corroborative method was assessed using the Bland-Altman plot.
PubMed: 38936170
DOI: 10.1016/j.jpba.2024.116317 -
Indian Journal of Dental Research :... Jan 2024Wet oral environment may have deleterious effects on performance of the composites due to influences of water sorption and solubility. The study evaluated the hydrolytic...
INTRODUCTION
Wet oral environment may have deleterious effects on performance of the composites due to influences of water sorption and solubility. The study evaluated the hydrolytic degradation caused because of water sorption and solubility of silorane and methacrylate-based dental composites.
METHODS
Ten disc samples (2 mm × 10 mm) were prepared. Samples were analyzed for water solubility and sorption according to ISO 4049:2000 regulations and tested for mass gain or loss following immersion in water or in artificial saliva at 1 day, 15 days, and 30 days period. Student's 't' test, repeated measures analysis of variance (ANOVA), and Tukey's post-hoc tests determined statistical significance of the experimental results with global significance set at P = 0.05.
RESULTS
Considerable sorption and solubility was observed with time in both materials on immersion. Silorane composites showed lower water sorption and solubility than methacrylate-based composite (MBC). Artificial saliva demonstrated higher sorption and solubility compared to distilled water.
CONCLUSION
Silorane composites display enhanced hydrolytic stability even after a month of immersion in contrast to conventional methacrylate-based composites (MBCs), making it a better alternative to MBC resins clinically.
Topics: Solubility; Methacrylates; Composite Resins; Water; Silorane Resins; Saliva, Artificial; Materials Testing; Hydrolysis; Dental Materials
PubMed: 38934754
DOI: 10.4103/ijdr.ijdr_195_23 -
Journal of Biomedical Materials... Jul 2024The objective of this in vitro study was to assess the efficiency of incorporating Biosilicate particles (30 and 50 mg) into an experimental orodispersible film and...
OBJECTIVES
The objective of this in vitro study was to assess the efficiency of incorporating Biosilicate particles (30 and 50 mg) into an experimental orodispersible film and its efficacy in the remineralization process of bovine dental enamel under cariogenic and erosive challenges.
METHODS
Ninety-nine intact incisors, devoid of cracks or fractures, yielding 198 samples (6 × 6 × 2 mm) via vestibular sectioning using a low-speed diamond disc under water cooling. After flattening the enamel surface with 600, 1200, and 2000 grit sandpaper, the samples were divided into two groups based on the challenges they underwent: cariogenic (0.1 M lactic acid at pH 5.0) or erosive (0.05 M citric acid solution at pH 2.3). Samples from each challenge were further categorized into 11 groups (n = 9) according to the duration of cariogenic (3, 7, and 14 days) or erosive (3, 7, and 10 days) challenge, along with positive control groups (fragments untreated with challenges and treated with different Biosilicate concentrations) and negative controls (fragments treated with artificial saliva for the same periods established for cariogenic and erosive challenges). Treatments with orodispersible films containing Biosilicate (30 and 50 mg) were administered for 2 min per day for 15 days.
RESULTS
The highest remineralizing potential was observed in samples treated with Biosilicate after 14 days of cariogenic challenge, irrespective of the concentration tested. For samples subjected to erosive challenge, erosion time did not affect Biosilicate's remineralizing potential.
CONCLUSION
Biosilicate shows promise in terms of remineralizing potential in enamel subjected to cariogenic challenge due to its ability to form hydroxycarbonapatite in mineralized tissues.
Topics: Dental Enamel; Animals; Cattle; Tooth Remineralization; Dental Caries; Tooth Erosion
PubMed: 38932619
DOI: 10.1002/jbm.b.35446 -
Sensors (Basel, Switzerland) Jun 2024Salivary 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...
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.
Topics: Colorimetry; Printing, Three-Dimensional; Hydrogen-Ion Concentration; Saliva; Hydrogels; Humans; Biosensing Techniques
PubMed: 38931525
DOI: 10.3390/s24123740 -
Archives of Oral Biology Jun 2024To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro.
OBJECTIVES
To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro.
DESIGN
Biofilms were grown on bovine enamel specimens in artificial saliva (AS) for seven days. After 24 h of formation, the AS was supplemented or not with fluoride (F) using sodium fluoride (0.005 or 0.008 ppm F), and the biofilms were exposed or not to a 20 % sucrose solution (reproducing a cariogenic challenge) once/day. On the seventh day, the biofilms were harvested and had their extracellular polysaccharides (EPS) and inorganic components analyzed. The specimens were subjected to computed X-ray microtomography analysis to determine their mineral concentration. Data were compared using two-way analyses of variance, followed by Fisher's LSD or Student-Newman-Keuls tests (p < 0.05).
RESULTS
Biofilms exposed to the cariogenic challenge had significantly higher EPS concentrations than those not exposed, regardless of the presence of F. For biofilms grown with 0.008 ppm F, those exposed to the cariogenic challenge had lower F levels than those not exposed. For biofilms exposed to the cariogenic challenge, those grown with 0.008 ppm F had lower lesion depths and integrated mineral loss, and higher outer layers than those grown without F.
CONCLUSIONS
The dual biofilm model assessed was able to create subsurface caries lesions in bovine enamel in vitro, which was influenced by the presence of F in the culture medium and exposure to sucrose.
PubMed: 38924874
DOI: 10.1016/j.archoralbio.2024.106029 -
Journal of Applied Oral Science :... 2024This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models. (Comparative Study)
Comparative Study
OBJECTIVE
This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models.
METHODOLOGY
In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey's post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001).
RESULTS
In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols.
CONCLUSION
The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth.
Topics: Biofilms; Humans; Streptococcus mutans; Dental Caries; Dental Enamel; Lacticaseibacillus casei; Time Factors; Reproducibility of Results; Streptococcus sobrinus; Spectrum Analysis, Raman; Analysis of Variance; Microscopy, Polarization; Statistics, Nonparametric; Tooth Remineralization; Reference Values; Saliva; Tooth Demineralization; Fluorescence
PubMed: 38922241
DOI: 10.1590/1678-7757-2023-0458 -
Acta Odontologica Latinoamericana : AOL Apr 2024Glass ceramics are widely used to manufacture esthetic veneers, inlays, onlays, and crowns. Although the clinical survival rates ofglass-ceramic restorations...
UNLABELLED
Glass ceramics are widely used to manufacture esthetic veneers, inlays, onlays, and crowns. Although the clinical survival rates ofglass-ceramic restorations arefavorable,fractures or chips are common. Certain cases can be repaired with direct composite.
AIM
The aim of this study was to investigate the interaction effect of different designs and surface treatments on the load-to-failure of lithium disilicate glass-ceramic repaired with nanofilled composite.
MATERIALS AND METHOD
Lithium-disilicate glass-ceramic slabs (IPS e.max Press, Ivoclar Vivadent) with three different designs of the top surface (flat, single plateau, or doubleplateau) (n=U) received 'no treatment', '5% HF etching', or "AI2O3 sandblasting". HF-etched and sandblasted slabs also received silane and universal one-step adhesive application. All slabs were incrementally repaired with nanofilled composite (Filtek Z350, 3M ESPE) up to6 mm above the highest ceramic top plateau. Specimens were stored in artificial saliva at 37 °C for 21 days and then subjected to 1,000 thermocycles between 5 and 55 °C. The interface composite-ceramic of each specimen was tensile tested until failure in a universal testing machine and the mode of failure was determined under a stereomicroscope. The ceramic surface morphology of one representative tested specimen from each subgroup (design/surface treatment) was observed through scanning electron microscopy (SEM).
RESULTS
Regardless of ceramic design, the absence of surface treatment resulted in significantly lower load-to-failure values. No significant differences in load-to-failure values were observed between HF-etched and sandblasted specimens for the flat design; however, HF etching resulted in significantly higher load-to-failure values than sandblasting for both single plateau and double plateau designs. The majority (60%) of HF-etched specimens with single plateau or double plateau presented mixed failures. SEM photomicrographs showed that HF-etched specimens had smoother surfaces than sandblasted specimens.
CONCLUSION
The surface treatment of a defective lithium disilicate glass-ceramic restoration has more influence than its macroscopic design on the retention of the composite repair. HF etching seems to provide higher bond strength to the composite repair.
Topics: Ceramics; Surface Properties; Dental Porcelain; Materials Testing; Dental Restoration Failure; Composite Resins; Dental Stress Analysis; Dental Prosthesis Design
PubMed: 38920130
DOI: 10.54589/aol.37/1/88 -
Acta Odontologica Latinoamericana : AOL Apr 2024Coloured compounds (anthocyanins) in açaí can stain resin-modified glass-ionomer cement (RMGIC) due to its low staining resistance.
UNLABELLED
Coloured compounds (anthocyanins) in açaí can stain resin-modified glass-ionomer cement (RMGIC) due to its low staining resistance.
AIM
The aim of this study was to assess whether açaí compromises the surface colour and roughness of RMGIC in vitro.
MATERIALS AND METHOD
Disc-shaped specimens (2 mm thick, 8 mm in diameter) of Vitremer™ (3M ESPE, St Paul, MN, USA) were prepared according to the manufacturer 's instructions. The mixture was inserted into a silicone mouldplaced between two mylar strips, and light cured. Specimens were randomly divided into three groups (n=25) according to the solutions to be used for chemical degradation: artificial saliva (control), açaí sorbet and açaí juice. A spectrophotometer CM-2600d/2500d (Konica Minolta, Tokyo, Japan) was used to analyse the colour (CIELa*b* scale). Surface roughness (Ra, mm) was measuredusing theprofilometer Surfcorder SE 1700 (Kosaka Corp, Tokyo, Japan). The specimens were subjected to three daily soaks (6 ml, 15 minutes) for 14 days at 37°C. They were washed in distilled water and placed in fresh saliva (30 minutes in the interval). After the third soak in a day, they were stored in fresh saliva overnight. Outcomes were analysed at baseline (L*, a*, b*, Ra) and after degradation (L'*, a'*, b'*, Ra').
RESULTS
The pH values of saliva, sorbet, and juice were 7.0, 3.8, and 4.9, respectively. ΔE* values were 6.6 for saliva, 6.9 for sorbet and 7.8 for juice. There was a significant ΔE* difference between saliva (p=0.005) and juice (p=0.002), and between juice and sorbet (p=0.019), but none between saliva and sorbet (p=0.401). There was no significant Δb* difference between the solutions. No difference between juice and sorbet was observed for Δa*, but they were significantly different from saliva (p<0.001). Brightness (L*) changed significantly. Juice showed the highest ΔE* (7.8) and ΔL* (7.7). No significant change was observed for roughness and there was no difference between the solutions for ARa.
CONCLUSIONS
Açaí and saliva led to unacceptable staining, but no significant roughness changes in the resin-modified glass-ionomer cement.
Topics: Glass Ionomer Cements; Color; Materials Testing; Surface Properties; Carbonated Beverages
PubMed: 38920125
DOI: 10.54589/aol.37/1/40