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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.
Subject(s)
Biofilms , Dental Caries , Dental Enamel , Lacticaseibacillus casei , Streptococcus mutans , Humans , Streptococcus mutans/physiology , Dental Caries/microbiology , Dental Caries/therapy , Dental Enamel/microbiology , Dental Enamel/chemistry , Lacticaseibacillus casei/physiology , Time Factors , Reproducibility of Results , Streptococcus sobrinus/physiology , Spectrum Analysis, Raman , Analysis of Variance , Microscopy, Polarization , Statistics, Nonparametric , Tooth Remineralization/methods , Reference Values , Saliva/microbiology , Saliva/chemistry , Tooth Demineralization/microbiology , FluorescenceABSTRACT
Abstract Creating artificial caries-like lesions that mimic the complex changes observed in natural caries is essential for properly evaluating new strategies, dental materials, and devices designed to arrest their progression and avoid more costly and invasive treatments. 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.
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Objectives: This study verified the possibility of cementing fiberglass-reinforced posts using a flowable bulk-fill composite (BF), comparing its push-out bond strength and microhardness with these properties of 3 luting materials. Materials and Methods: Sixty endodontically treated bovine roots were used. Posts were cemented using conventional dual-cured cement (CC); self-adhesive cement (SA); dual-cured composite (RC); and BF. Push-out bond strength (n = 10) and microhardness (n = 5) tests were performed after 1 week and 4 months of storage. Two-way repeated measures analysis of variance (ANOVA), 1-way ANOVA, t-test, and Tukey post-hoc tests were applied for the push-out bond strength and microhardness results; and Pearson correlation test was applied to verify the correlation between push-out bond strength and microhardness results (α = 0.05). Results: BF presented higher push-out bond strength than CC and SA in the cervical third before aging (p < 0.01). No differences were found between push-out bond strength before and after aging for all the luting materials (p = 0.84). Regarding hardness, only SA presented higher values measured before than after aging (p < 0.01). RC and BF did not present 80% of the maximum hardness at the apical regions. A strong positive correlation was found between the luting materials' push-out bond strength and microhardness (p < 0.01, R2 = 0.7912). Conclusions: The BF presented comparable or higher push-out bond strength and microhardness than the luting materials, which indicates that it could be used for cementing resin posts in situations where adequate light curing is possible.
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Leukotrienes (LTs) are derived from arachidonic acid metabolism by the 5-lipoxygenase (5-LO) enzyme. The production of LTs is stimulated in the pathogenesis of rheumatoid arthritis (RA), osteoarthritis, and periodontitis, with a relevant contribution to bone resorption. However, its role in bone turnover, particularly the suppression of bone formation by modulating the function of osteoclasts and osteoblasts, remains unclear. We investigated the effects of LTs on bone metabolism and their impact on osteogenic differentiation and osteoclastogenesis using a 5-LO knockout (KO) mouse model. Results from micro-computed tomography (µCT) analysis of femur from 8-week-old 5-LO-deficient mice showed increased cortical bone and medullary region in females and males and decreased trabecular bone in females. In the vertebra, we observed increased marrow area in both females and males 5-LO KO and decreased trabecular bone only in females 5-LO KO. Immunohistochemistry (IHC) analysis showed higher levels of osteogenic markers tissue-nonspecific alkaline phosphatase (TNAP) and osteopontin (OPN) and lower expression of osteoclastogenic marker tartrate-resistant acid phosphatase (TRAP) in the femurs of 5-LO KO mice versus wild-type (WT). Alkaline phosphatase activity and mineralization assay results showed that the 5-LO absence enhances osteoblasts differentiation and mineralization but decreases the proliferation. Alkaline phosphatase (ALP), Bglap, and Sp7 gene expression were higher in 5-LO KO osteoblasts compared to WT cells. Eicosanoids production was higher in 5-LO KO osteoblasts except for thromboxane 2, which was lower in 5-LO-deficient mice. Proteomic analysis identified the downregulation of proteins related to adenosine triphosphate (ATP) metabolism in 5-LO KO osteoblasts, and the upregulation of transcription factors such as the adaptor-related protein complex 1 (AP-1 complex) in long bones from 5-LO KO mice leading to an increased bone formation pattern in 5-LO-deficient mice. We observed enormous differences in the morphology and function of osteoclasts with reduced bone resorption markers and impaired osteoclasts in 5-LO KO compared to WT osteoclasts. Altogether, these results demonstrate that the absence of 5-LO is related to the greater osteogenic profile. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).
Subject(s)
Bone Resorption , Osteogenesis , Male , Female , Mice , Animals , Alkaline Phosphatase/metabolism , X-Ray Microtomography , Proteomics , Osteoclasts/metabolism , Osteoblasts/metabolism , Bone Resorption/pathology , Cell Differentiation , Mice, Knockout , Leukotrienes/metabolism , Leukotrienes/pharmacologyABSTRACT
OBJECTIVES: To compare the biodegradability, mechanical behavior, and physicochemical features of the collagen-rich extracellular matrix (ECM) of artificial caries-affected dentin (ACAD), natural caries-affected dentin (NCAD) and sound dentin (SD). METHODS: Dentin specimens from human molars were prepared and assigned into groups according to the type of dentin: ACAD, NCAD, or SD. ACAD was produced by incubation of demineralized SD with Streptococcus mutans in a chemically defined medium (CDM) with 1% sucrose for 7 days at 37 °C under anaerobic conditions. Specimens were assessed to determine collagen birefringence, biodegradability, mechanical behavior, and chemical composition. Data were individually processed and analyzed by ANOVA and post-hoc tests (α = 0.05). RESULTS: CDM-based biofilm challenge reduced loss, storage, and complex moduli in ACAD (p < 0.001), while the damping capacity remained unaffected (p = 0.066). Higher red and lower green birefringence were found in ACAD and NCAD when compared with SD (p < 0.001). Differently to ACAD, SD and NCAD presented higher biodegradability to exogenous proteases (p = 0.02). Chemical analysis of the integrated areas of characteristic bands that assess mineral quality (carbonate/phosphate and crystallinity index), mineral to matrix (phosphate/amide I) and post-translational modifications (amide III/CH2, pentosidine/CH2, and pentosidine/amide III) (p<0.05) showed that NCAD was significantly different from SD while ACAD exhibited intermediate values. CONCLUSIONS: CDM-based biofilm challenge produced a dentin ECM with decreased mechanical properties and increased collagen maturity. The compositional and structural conformation of the ACAD suggested that CDM-based biofilm challenge showed potential to produce artificial lesions by revealing a transitional condition towards mimicking critical features of NCAD. CLINICAL SIGNIFICANCE: This study highlights the importance of developing a tissue that mimics the features of natural caries-affected dentin ECM for in vitro studies. Our findings suggested the potential of a modified biofilm challenge protocol to produce and simulate a relevant substrate, such as caries-affected dentin.
Subject(s)
Dental Caries , Dentin-Bonding Agents , Humans , Dentin-Bonding Agents/chemistry , Dentin/chemistry , Dental Caries Susceptibility , Dental Caries/pathology , Collagen/analysis , Phosphates , Amides/analysisABSTRACT
Pseudomonas aeruginosa is an opportunist pathogen usually associated with life threatening infections and exhibits a set of intrinsic and acquired antimicrobial mechanisms. Although resistance to penicillins-like compounds is commonly associated with the chromosomal Pseudomonas-derived cephalosporinases ß-lactamase, the real contribution of OXA-50, a second chromosomally encoded ß-lactamase, remains unclear. In this study, we characterized the biochemical properties of OXA-50, OXA-488, and OXA-494. Both oxacilinases differ from OXA-50 in two amino acids each. The blaOXA-50, blaOXA-488, and blaOXA-494 were cloned into pET26b+ that was transformed into Escherichia coli DH5α strain, expressed in E. coli BL21 strain, and then purified for obtaining the hydrolytic parameters. Benzylpenicillin was the preferential substrate instead of oxacillin. Besides, OXA-488 showed a threefold increase in catalytic efficiency for benzylpenicillin, and it was twofold more efficient in hydrolyzing imipenem, compared with OXA-50, although such carbapenemase activity was considered weak. In addition, OXA-488 and OXA-494 showed an increased affinity for penicillins, which contributed to the increased catalytic efficiency against ampicillin, especially OXA-488. Chromosomally encoded resistance mechanisms are usually overshadowed by acquired mechanisms. However, understanding their real contribution is essential to comprehend the versatile profiles verified in P. aeruginosa isolates. Such information can help to choose the best therapy in a scenario of limited options.
Subject(s)
Pseudomonas aeruginosa , beta-Lactams , Anti-Bacterial Agents/pharmacology , Cloning, Molecular , Escherichia coli/genetics , Escherichia coli/metabolism , Hydrolysis , Kinetics , Microbial Sensitivity Tests , Oxacillin , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , beta-Lactamases/metabolism , beta-Lactams/metabolism , beta-Lactams/pharmacologyABSTRACT
OBJECTIVE: The study evaluated the effect of solutions containing fluoride (F) and/or sodium trimetaphosphate (TMP) and F/TMP on the inhibition of MMP-2 and MMP-9, and on dentin remineralization in vitro. DESIGN: Bovine root dentin blocks were prepared, and caries-like lesions were induced in two thirds of the surface. Blocks were then randomly divided into 13 groups/solutions (n = 10): Placebo; 0.3 %, 1 % and 3 % NaOH-hydrolyzed TMP; 0.3 %, 1 % and 3 % TMP; 250, 500 and 1100 ppm F; 250 ppm F + 0.3 % TMP; 500 ppm F + 1 % TMP and 1100 ppm F + 3 % TMP. One third of each specimen was treated with the respective solutions in pH-cycling. The mineral concentration (gHAp × cm-3 × µm) was determined by computed X-ray microtomography, and data submitted to ANOVA and Student-Newman-Keuls' test (p < 0.05). The ability of the solutions to inhibit MMP-2 and MMP-9 activity was assessed by zymography. RESULTS: F/TMP association led to less mineral loss in the deeper region of the lesion and reduced the depth of lesions when compared to its counterpart without TMP (p < 0.001). 3 % TMP (hydrolyzed or not), 500 ppm F and 1100 ppm F completely inhibited MMP-2 activity, while for MMP-9 such effects were only achieved by treatment with 1100 ppm F + 3 % TMP. CONCLUSION: Treatment with 1100 ppm F + 3 % TMP fully inhibits the gelatinolytic activity of MMPs-2 and - 9 and shows greater remineralizing capacity in artificial caries lesions in dentin. However, hydrolyzing TMP does not improve its anti-proteolytic activity and its remineralizing capacity.
Subject(s)
Dental Caries , Fluorides , Animals , Cariostatic Agents/pharmacology , Cattle , Dental Caries/drug therapy , Dental Caries/prevention & control , Dentin , Fluorides/pharmacology , Humans , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Minerals , Sodium Fluoride/pharmacology , Tooth Remineralization/methodsABSTRACT
Objective: To compare optical, morphological, chemical, and physical aspects of the sound enamel and white spot lesions (WSL) classified as ICDAS 2. Material and Methods: Seventeen human molars with one surface presenting WSL and a sound surface (2 x 2 mm window) were characterized by Quantitative light-induced fluorescence (QLF ®), Optical coherence tomography (OCT), microhardness, and Raman spectroscopy. The ANOVA and Tukey's test were used at 5% significance level. Results: The QLF comparison between distinct substrates yielded decreased ∆Q (integrated fluorescence loss) of -15,37%mm2 and -11,68% ∆F (fluorescence loss) for WSL. The OCT detected mean lesion depth of 174,43 µm. ANOVA could not detect differences in the optical attenuation coefficient between the substrates (p>0.05). Lower microhardness measures were observed in WSL than on sound enamel (p<0.05). The Raman spectra showed four vibrational phosphate bands (v1, v2, v3, v4), where the highest peak was at 960.3 cm-1(v1) for both substrates. However, a 40% decrease in phosphate (v1) was detected in WSL. The peak at 1071 cm-1 was higher for sound enamel, indicating the presence of a phosphate band instead of the B-type carbonate. The spectra showed higher intensity of the organic composition at 1295 cm-1 and 1450 cm -1 for WSL. Conclusion: Non-invasive QLF, OCT and Raman spectroscopy were able to distinguish differences in fluorescence, optical properties, and organic/inorganic components, respectively, between sound enamel and WSL, validated by the destructive microhardness analysis. (AU)
Objetivo: Comparar os aspectos ópticos, morfológicos, químicos e físicos do esmalte sadio e das lesões de mancha branca naturais, classificada como ICDAS 2. Material e métodos: Dezessete molares humanos com uma face apresentando uma lesão de mancha branca natural e outra face o esmalte hígido (2 x 2 mm) foram caracterizados utilizando a Fluorescência quantitativa induzida pela luz (QLF ®), Tomografia de coerência óptica (OCT), Microdureza e Espectroscopia Raman. A ANOVA e o teste de Tukey foram utilizados ao nível de significância de 5%. Resultados: A comparação entre os substratos distintos, utilizando o QLF ® demonstrou uma diminuição no ∆Q (perda de fluorescência integrada) de -15,37%mm2 e -11,68% de ∆F (Perda de fluorescência) para a lesão de mancha branca. O OCT detectou uma profundidade média de lesão de 174,43µm. A ANOVA não detectou diferenças no coeficiente de atenuação óptica entre os substratos (>0,05). Microdureza significantemente menor foi detectada nas lesões de mancha branca do que no esmalte sadio (p<0,05). Os espectros Raman mostraram quatro bandas vibracionais do fosfato (v1,v2,v3,v4), onde o maior pico foi em 960,3cm-1para ambos os substratos. No entanto, uma diminuição de 40% no fosfato (v1) foi detectada na lesão. O pico em 1071cm-1foi maior para o esmalte hígido, demonstrando tratar-se da banda do fosfato, ao invés do carbonato tipo B. Os espectros apresentaram maior intensidade da composição orgânica em 1295cm-1e 1450 cm-1para a lesão de mancha branca. Conclusão:Os métodos não invasivos QLF, OCT e espectroscopia Raman foram capazes dediferenciar a fluorescência, propriedades ópticas e conteúdo orgânico/inorgânico do esmalte sadio comparado com esmalte com lesões de mancha branca, sendo validado pela análise de microdureza. (AU)
Subject(s)
Spectrum Analysis, Raman , Dental Caries , Dental Enamel , DiagnosisABSTRACT
Cannabis products that contain the tetrahydrocannabinol (THC) cannabinoid are emerging as promising therapeutic agents for the treatment of medical conditions such as chronic pain. THC elicits psychoactive effects through modulation of dopaminergic neurons, thereby altering levels of dopamine in the brain. This case report highlights the complexity associated with medicinal cannabis and the health risks associated with its use. A 57-year-old male with Parkinson's disease was experiencing worsening tremors and vivid hallucinations despite therapy optimization attempts. It was discovered that the patient took cannabis for chronic back pain, and a pharmacogenomics (PGx) test indicated the presence of variants for the COMT and HTR2A genes. These variants could increase dopamine levels and predispose patients to visual hallucinations. Once the cannabis was discontinued, the patient's hallucinations began to slowly dissipate. Cannabis use continues to expand as it gains more acceptance legally and medicinally, but cannabis can affect the response to drugs. This patient case suggests that cannabis use in combination with dopamine-promoting drugs, especially in a patient with genetic variants, can increase the risk for vivid hallucinations. These conditions support the importance of considering herb-drug interactions and PGx data when performing a medication safety review.
Subject(s)
Cannabis , Parkinson Disease , Cannabis/adverse effects , Dopamine Agents , Dronabinol/adverse effects , Hallucinations/chemically induced , Humans , Middle Aged , Parkinson Disease/complications , Parkinson Disease/drug therapyABSTRACT
Pharmacogenomic (PGx) information can guide drug and dose selection, optimize therapy outcomes, and/or decrease the risk of adverse drug events (ADEs). This report demonstrates the impact of a pharmacist-led medication evaluation, with PGx assisted by a clinical decision support system (CDSS), of a patient with multiple comorbidities. Following several sub-optimal pharmacotherapy attempts, PGx testing was recommended. The results were integrated into the CDSS, which supported the identification of clinically significant drug-drug, drug-gene, and drug-drug-gene interactions that led to the phenoconversion of cytochrome P450. The pharmacist evaluated PGx results, concomitant medications, and patient-specific factors to address medication-related problems. The results identified the patient as a CYP2D6 intermediate metabolizer (IM). Duloxetine-mediated competitive inhibition of CYP2D6 resulted in phenoconversion, whereby the patient's CYP2D6 phenotype was converted from IM to poor metabolizer for CYP2D6 co-medication. The medication risk score suggested a high risk of ADEs. Recommendations that accounted for PGx and drug-induced phenoconversion were accepted. After 1.5 months, therapy changes led to improved pain control, depression status, and quality of life, as well as increased heart rate, evidenced by patient-reported improved sleep patterns, movement, and cognition. This case highlights the pharmacist's role in using PGx testing and a CDSS to identify and mitigate medication-related problems to optimize medication regimen and medication safety.
Subject(s)
Pharmaceutical Preparations , Pharmacogenetics , Cytochrome P-450 CYP2D6 , Humans , Pharmacists , Quality of LifeABSTRACT
OBJECTIVE: To describe the neurological and neurodevelopmental outcomes of children with Congenital Zika Syndrome (CZS) associated microcephaly beyond 2 years of age. METHOD: We followed children with CZS-associated microcephaly in an outpatient clinic in Salvador, Brazil. Neurological and neurodevelopmental assessments were performed using the Hammersmith Infant Neurological Examination (HINE) and Bayley Scales of Infant and Toddler Neurodevelopment (Bayley-III) respectively. RESULTS: Of the 42 children included, 19 were male (45.2%); median (interquartile range) age at neurological evaluation was 28 (25-32) months, and 36 (85.7%) had severe microcephaly. HINE and Bayley-III results were completed for 35/42 (83.3%) and 33/42 (78.5%) children respectively. Bayley-III identified a severe developmental delay in 32/33 (97.0%) children while 1/33 (3.0%) had only a mild delay. In the multivariable analysis, we found that Bayley-III and HINE scores were correlated. Better HINE scores were associated with higher Bayley-III cognitive raw scores (ß = 0.29; CI 95% = 0.02-0.57) and motor raw scores (ß = 0.43; CI 95% = 0.04-0.82) after adjusting for head circumference, prematurity, and age at neurodevelopmental evaluation. Furthermore, we found that greater head circumference at follow up was associated with higher cognitive (ß = 1.27; CI 95% = 0.01-2.53) and motor raw scores (ß = 2.03; CI 95% = 0.25-3.81). CONCLUSION: Children with CZS-associated microcephaly demonstrate severe neurodevelopmental delays and slower growth rates than their peers over time. Still, they have remarkably heterogeneous neurodevelopmental profiles according to neurological exam scores which correlate with their long-term outcomes. We found that HINE scores effectively captured the heterogeneity of neurological capabilities among these children and could be predictive of cognitive and motor development progress.
Subject(s)
Developmental Disabilities/diagnosis , Microcephaly/diagnosis , Microcephaly/epidemiology , Zika Virus Infection/diagnosis , Brazil/epidemiology , Cephalometry , Child, Preschool , Developmental Disabilities/physiopathology , Developmental Disabilities/virology , Female , Humans , Infant , Infant, Newborn , Male , Microcephaly/etiology , Microcephaly/virology , Neurologic Examination , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/physiopathology , Pregnancy Complications, Infectious/virology , Zika Virus/pathogenicity , Zika Virus Infection/complications , Zika Virus Infection/virologySubject(s)
Biomedical Research/trends , Clinical Pharmacy Information Systems/trends , Health Care Sector/trends , Pharmacogenetics/trends , Precision Medicine/trends , Biomedical Research/standards , Clinical Pharmacy Information Systems/standards , Health Care Sector/standards , Humans , Pharmacogenetics/standards , Precision Medicine/standardsABSTRACT
Bone marrow-derived mesenchymal stem/stromal cells (BMSCs) are fundamental to bone regenerative therapies, tissue engineering, and postmenopausal osteoporosis. Donor variation among patients, cell heterogeneity, and unpredictable capacity for differentiation reduce effectiveness of BMSCs for regenerative cell therapies. The cell surface glycoprotein CD24 exhibits the most prominent differential expression during osteogenic versus adipogenic differentiation of human BMSCs. Therefore, CD24 may represent a selective biomarker for subpopulations of BMSCs with increased osteoblastic potential. In undifferentiated human BMSCs, CD24 cell surface expression is variable among donors (range: 2%-10%) and increased by two to fourfold upon osteogenic differentiation. Strikingly, FACS sorted CD24pos cells exhibit delayed mineralization and reduced capacity for adipocyte differentiation. RNAseq analysis of CD24pos and CD24neg BMSCs identified a limited number of genes with increased expression in CD24pos cells that are associated with cell adhesion, motility, and extracellular matrix. Downregulated genes are associated with cell cycle regulation, and biological assays revealed that CD24pos cells have reduced proliferation. Hence, expression of the cell surface glycoprotein CD24 identifies a subpopulation of human BMSCs with reduced capacity for proliferation and extracellular matrix mineralization. Functional specialization among BMSCs populations may support their regenerative potential and therapeutic success by accommodating cell activities that promote skeletal tissue formation, homeostasis, and repair.
Subject(s)
Biomarkers/metabolism , CD24 Antigen/genetics , Cell Differentiation/genetics , Cell Proliferation/genetics , Membrane Glycoproteins/genetics , Mesenchymal Stem Cells/metabolism , Adipogenesis/genetics , CD24 Antigen/metabolism , Cells, Cultured , Flow Cytometry/methods , Gene Expression Profiling/methods , Humans , Membrane Glycoproteins/metabolism , Mesenchymal Stem Cells/cytology , Osteoblasts/cytology , Osteoblasts/metabolism , Osteogenesis/genetics , RNA-Seq/methods , Time FactorsABSTRACT
This study investigated the effect of irradiation with an erbium-doped yttrium aluminium garnet (Er:YAG) laser and coating with silica on the surface characteristics, bond strength, and flexural strength of dental zirconia. Three hundred and forty-three standard zirconia specimens were created, and 49 were assigned to each of seven surface treatment groups: (i) no treatment; Er:YAG laser (80 mJ/2 Hz) with pulse widths of 50 µs (ii), 100 µs (iii), 300 µs (iv), or 600 µs (v); or tribochemical silica coating at the partially sintered stage (vi) or after sintering was complete (vii). All specimens were sintered after the surface treatments, except for the group in which specimens were sintered before treatment. The study outcomes were roughness, surface loss, microshear bond strength (µSBS), and biaxial flexural strength (BFS). Mean roughness and surface loss values were significantly higher in specimens from irradiated groups than in those from silica-coated groups. Regarding µSBS, after aging, specimens from all experimental groups presented very low and similar µSBS values, irrespective of the surface treatment. Silica coating after sintering yielded the highest BFS (1149.5 ± 167.6 MPa), while coating partially sintered specimens with silica resulted a BFS (826.9 ± 60.9 MPa) similar to that of the untreated control group (794.9 ± 101.7 MPa). Laser treatments, irrespective of pulse width used, significantly decreased the BFS. In the group treated with laser at 300 µs pulse width, specimens exhibited the lowest BFS value (514.1 ± 71.5 MPa). Adhesion to zirconia was not stable after aging, regardless of the surface treatment implemented.
Subject(s)
Dental Bonding , Flexural Strength , Ceramics , Materials Testing , Resin Cements , Shear Strength , Silicon Dioxide , Surface Properties , ZirconiumABSTRACT
Although it was demonstrated that curcumin-mediated antimicrobial photodynamic therapy (aPDT) is effective for reducing the viability of microbial cells and the vitality of oral biofilms, the cytotoxicity of this therapeutic approach for host cells has not been yet elucidated. Hence, the aim of this study was to evaluate the cytotoxicity and apoptotic effects of curcumin-mediated aPDT on mouse fibroblasts. Cells were treated with 0.6 or 6 µmol.L-1 curcumin combined with 0.075 or 7.5 J.cm-2 LED at 455 nm. Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while quantitative reverse transcriptase-PCR (qRT-PCR) was used to assess the expression of Bax, Bad, Bcl-2, VDAC-1, cytochrome C, and Fas-L genes for apoptosis. The differences between groups were detected by Kruskal-Wallis and post hoc Dunn's tests for MTT and CV assays and by ANOVA and post hoc Tukey test for qRT-PCR (P < 0.05). The effect of 0.6 µmol.L-1 curcumin plus 0.075 J.cm-2 LED (minimum parameter) did not differ statistically from control group; however, the combination of 0.6 µmol.L-1 curcumin plus 7.5 J.cm-2 LED reduced viable cells in 34%, while the combinations of 6 µmol.L-1 curcumin plus 0.075 and 7.5 J.cm-2 LED reduced viable cells in 47% and 99%, respectively. aPDT increased significantly the relative expression of Bax/Bcl-2, cytochrome C, VDAC-1, and Fas-L genes, without influence on the ratio Bad/Bcl-2. Therefore, curcumin-mediated aPDT activated Bcl-2 apoptosis signaling pathways in mouse fibroblasts regarding present conditions, reducing the viability of cells with the increase of curcumin concentrations and light energies.
Subject(s)
Anti-Infective Agents/pharmacology , Apoptosis/drug effects , Curcumin/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Photochemotherapy , Signal Transduction/drug effects , Animals , Apoptosis/radiation effects , Fibroblasts/radiation effects , Mice , Signal Transduction/radiation effectsABSTRACT
PURPOSE: To investigate the influence of implant surface decontaminated and uncontaminated on osteoblast-like cell adhesion and proliferation MATERIALS AND METHODS: Commercially available implants of different brands and surface characteristics were selected: Biomet 3i® Nanotite (NT) and Osseotite (OT), Straumann® SLActive (SLA), and Neodent® Acqua Drive (ACQ) and Neoporos Drive CM (CM). Physical and chemical properties of the implants were investigated by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and wettability analysis (WETT). Implants were previously contaminated with Aggregatibacter actinomycetemcomitans strains; after that, samples were decontaminated by different chemical methods. Decontaminated (test group; n = 15/type of implant) and uncontaminated (control group; n = 5/type of implant) samples were analyzed according to the number of human osteoblastic osteosarcoma cells (Saos-2) adhered on the implant surface after 24 h and 72 h in SEM images. RESULTS: ACQ was found to be highly hydrophilic, and NT was the most hydrophobic implant. Increased variation of Saos-2 cell adhesion and proliferation were observed on all test and control groups. Controversially, at the proliferation analysis in 72 h, CM implant was the only implant that showed no significant difference between test and group (p = 0.2833; Tukey's multiple comparisons test). NT implants showed the greater value of cell proliferation when compared with all types of implant surface (p = 0.0002; Tukey's multiple comparisons test). CONCLUSIONS: These findings suggest that decontaminated surfaces were able to impair the counting of osteoblast-like cell adhesion and proliferation.
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BACKGROUND: To evaluate the physicochemical properties and cytotoxicity of AH Plus, MTA Fillapex and TotalFill BC Sealer. Volumetric changes were also evaluating using micro-computed tomography (micro-CT). MATERIAL AND METHODS: Radiopacity and flow were evaluated in accordance with the ISO 6876, while setting time was evaluated in accordance with the ASTM- C266-08 specifications. The release of Ca2+ ions and pH were measured with spectrophotometer and pH meter, respectively, after different time intervals (1h, 3h, 24h, 72h, 168h, and 360h). Cytotoxicity was evaluated by MTT reduction assay to check 3T3 cells viability at 24, 48 and 72 hours. Volumetric change was evaluated by micro-CT, by using 30 acrylic teeth, filled with gutta-percha cones and the tested root canal sealer. The samples were evaluated after 168h, 360h and 720h of immersion in distilled water. Data were statistically analyzed by one-way ANOVA and Tukey test or by Kruskal-Wallis and Dunn tests (P<0.05). RESULTS: MTA Fillapex and TotalFill BC Sealer showed lower radiopacity than AH Plus (P<0.05). The MTA Fillapex showed the highest flow, while AH Plus showed the lowest flow (P<0.05). The initial and final setting time of AH Plus were lower than MTA Fillapex and TotalFill BC Sealer (P<0.05). In general, TotalFill BC Sealer presented higher Ca2+ ion release and pH than the other tested sealers. TotalFill BC Sealer also showed overall lower cytotoxicity when compared to the other sealers. Volumetric change of AH Plus and TotalFill BC Sealer was lower than MTA Fillapex (P<0.05). CONCLUSIONS: AH Plus, MTA Fillapex and TotalFill BC Sealer showed slight differences in the physicochemical properties and cytotoxicity, but all suitable for an endodontic sealer. However, AH Plus and TotalFill BC Sealer showed low volumetric changes when compared to MTA Fillapex. Key words:Calcium silicate, cytotoxicity, physicochemical properties, micro computed tomography.
ABSTRACT
Polypharmacy is a common phenomenon among adults using opioids, which may influence the frequency, severity, and complexity of drug-drug interactions (DDIs) experienced. Clinicians must be able to easily identify and resolve DDIs since opioid-related DDIs are common and can be life-threatening. Given that clinicians often rely on technological aids-such as clinical decision support systems (CDSS) and drug interaction software-to identify and resolve DDIs in patients with complex drug regimens, this narrative review provides an appraisal of the performance of existing technologies. Opioid-specific CDSS have several system- and content-related limitations that need to be overcome. Specifically, we found that these CDSS often analyze DDIs in a pairwise manner, do not account for relevant pharmacogenomic results, and do not integrate well with electronic health records. In the context of polypharmacy, existing systems may encourage inadvertent serious alert dismissal due to the generation of multiple incoherent alerts. Future technological systems should minimize alert fatigue, limit manual input, allow for simultaneous multidrug interaction assessments, incorporate pharmacogenomic data, conduct iterative risk simulations, and integrate seamlessly with normal workflow.
ABSTRACT
PURPOSE: This study aims to evaluate the effect of erosive, abrasive, and erosive/abrasive challenges on the glaze layer of ceramic materials. METHODS: Ninety-five samples of monolithic zirconia (MZ) (LuxaCam Zircon HT-Plus) and lithium disilicate (LD) (IPS e.max CAD) were divided according to the response variables: Surface roughness and surface loss (n = 10), evaluated with optical profilometry; surface topography, with scanning electron microscopy SEM (n = 3); and biofilm deposition, with microbiological assay (n = 5). The evaluations were performed in three different time evaluations: (a) Sintered, (b) Glaze, and (c) Challenge (Erosion, Abrasion, and Erosion/Abrasion). Erosion consisted in immersing specimens in HCl solution, abrasion was performed with brushing machine, and erosion/abrasion consisted of a combination of the two previous protocols. Data were analyzed with parametric tests (P < 0.05). RESULTS: MZ glaze layer presented significantly higher surface roughness (P = 0.00), surface loss (P = 0.03), and biofilm deposition (P = 0.00) than LD. Abrasion and erosion/abrasion showed similar outcomes, generating significantly higher surface roughness (P = 0.00), surface loss (P = 0.00), and biofilm deposition (P = 0.01) than erosion. CONCLUSIONS: Glaze layer properties were altered by the challenges, with abrasion and erosion/abrasion generating higher surface roughness, surface loss, and biofilm deposition than erosion. A significant correlation was found between the surface roughness and biofilm deposition. CLINICAL SIGNIFICANCE: The glaze layer is susceptible to challenges, especially to abrasion and erosion/abrasion, which generated greater surface roughness and surface loss than erosion. The greater surface roughness lead to a greater biofilm deposition on the glaze layer.
Subject(s)
Ceramics , Dental Porcelain , Materials Testing , Microscopy, Electron, Scanning , Surface Properties , ToothbrushingABSTRACT
PURPOSE: To investigate whether interfacial enzymatic activity and adhesion receptiveness of artificial caries-affected dentin (ACAD) simulate those of natural caries-affected dentin (NCAD). MATERIALS AND METHODS: Thirty dentin specimens were prepared from human molars to determine interfacial gelatinolytic activity using in situ zymography and adhesion experiments (micropermeability and bond strength [µTBS]). Groups were formed according to the type of dentin: artificial caries-affected dentin (ACAD), natural caries-affected dentin (NCAD), or sound dentin. ACAD was produced by incubating dentin with Streptococcus mutans in a chemically defined medium (CDM) with 1% sucrose for 7 days at 37°C under anaerobic conditions. CDM was replaced daily, and the sterility as well as the pH of the culture was monitored. Adhesion experiments employed Single Bond Universal (3M Oral Care) in self-etch mode. Data were individually processed and analyzed using ANOVA and post-hoc tests (α = 0.05). RESULTS: The enzymatic activity of ACAD was similar to that of sound dentin, but was lower than that of NCAD, which elicited the highest activity (p < 0.05). Interfacial micropermeability intensity at the hybrid layer or in underlying dentin (5 µm below the interface) was similar in all types of dentin (p > 0.05). On the other hand, substrate permeability was higher for NCAD than for ACAD. The highest sealing ability was detected in sound dentin. Bond strengths to ACAD were higher than to NCAD. However, the highest µTBS was observed in sound dentin (p < 0.05). CONCLUSION: Artificial caries-affected dentin simulated neither the gelatinolytic activity nor bonding receptiveness of natural caries-affected dentin.