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This study aims to evaluate and compare the effect of fibroblastic growth factor 2 (FGF‐2) and photobiomodulation, solely or in combination, in angiogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The study comprises the following groups: control group (hPDLSCs only), FGF‐2 (50 ng/mL) group, two photobiomodulation groups with a 4 J/cm2 energy density of 808 nm diode laser (1‐Session or 2‐Session), and two groups with the combination of each 1‐Session or 2‐Session photobiomodulation with FGF‐2 (50 ng/mL). The 4′,6‐diamidino‐2‐phenylindole (DAPI) staining, and Methyl Thiazolyl Tetrazolium (MTT) assay were undertaken on days 2, 4, and 6. Quantitative Real‐time Polymerase Chain Reaction (RT‐qPCR) analysis on days 2, 4, 6, 8, and 11 was conducted to investigate VEGF‐A and ANG‐I genes. Coherently, the results of the DAPI and MTT showed the Laser (2‐Session) group had higher cell viability than others on day 6. All groups demonstrated a growth pattern in the expression of VEGF‐A and ANG‐I from day 2 to 8 and, afterward, a significant downgrowth to day 11 (p < 0.05). The most amounts of expression of VEGF‐A and ANG‐I on day 8 were seen in the Laser (2‐Session) group. Two‐time application of photobiomodulation using a diode laser with 808 nm wavelength after 2 and 4 days of cell seeding can be associated with higher cell viability and angiogenic differentiation of hPDLSCs compared to the one‐time application of photobiomodulation and administration of FGF‐2. [ABSTRACT FROM AUTHOR]

Objective: To compare the implant–abutment connection microgap between computer‐aided design and computer‐aided manufacturing (CAD/CAM) milled or laser‐sintered cobalt–chrome custom abutments with or without ceramic veneering and titanium stock abutments with or without crown cementation. Material and Methods: Six groups of six abutments each were prepared: (1) CAD/CAM cobalt–chrome custom abutments: milled, milled with ceramic veneering, laser‐sintered, and laser‐sintered with ceramic veneering (four groups: MIL, MIL‐C, SIN, and SIN‐C, respectively) and (2) titanium stock abutments with or without zirconia crown cementation (two groups: STK and STK‐Z, respectively). Abutments were screwed to the implants by applying 30 Ncm torque. All 36 samples were sectioned along their long axes. The implant–abutment connection microgap was measured using scanning electron microscopy on the right and left sides of the connection at the upper, middle, and lower levels. Data were analyzed using the Kruskal–Wallis test (p <.05). Results: Mean values (μm) of the microgap were 0.54 ± 0.44 (STK), 0.55 ± 0.48 (STK‐Z), 1.53 ± 1.30 (MIL), 2.30 ± 2.2 (MIL‐C), 1.53 ± 1.37 (SIN), and 1.87 ± 1.8 (SIN‐C). Although significant differences were observed between the STK and STK‐Z groups and the other groups (p <.05), none were observed between the milled and laser‐sintered groups before or after ceramic veneering. The largest microgap was observed at the upper level in all groups. Conclusions: Titanium stock abutments provided a closer fit than cobalt–chrome custom abutments. Neither crown cementation nor ceramic veneering resulted in significant changes in the implant–abutment connection microgap. [ABSTRACT FROM AUTHOR]

Background: Osteoarthritis (OA) is a prevalent degenerative joint disease characterized by chronic inflammation and progressive cartilage degradation, ultimately leading to joint dysfunction and disability. Oleocanthal (OC), a bioactive phenolic compound derived from extra virgin olive oil, has garnered significant attention due to its potent anti-inflammatory properties, which are comparable to those of non-steroidal anti-inflammatory drugs (NSAIDs). This study pioneers the investigation into the effects of OC on the Protease-Activated Receptor-2 (PAR-2) mediated inflammatory pathway in OA, aiming to validate its efficacy as a functional food-based therapeutic intervention. Methods: To simulate cartilage tissue in vitro, human bone marrow-derived mesenchymal stem cells (BMSCs) were differentiated into chondrocytes. An inflammatory OA-like environment was induced in these chondrocytes using lipopolysaccharide (LPS) to mimic the pathological conditions of OA. The therapeutic effects of OC were evaluated by treating these inflamed chondrocytes with various concentrations of OC. The study focused on assessing key inflammatory markers, catabolic enzymes, and mitochondrial function to elucidate the protective mechanisms of OC. Mitochondrial function, specifically mitochondrial membrane potential (ΔΨm), was assessed using Rhodamine 123 staining, a fluorescent dye that selectively accumulates in active mitochondria. The integrity of ΔΨm serves as an indicator of mitochondrial and bioenergetic function. Additionally, Western blotting was employed to analyze protein expression levels, while real-time polymerase chain reaction (RT-PCR) was used to quantify gene expression of inflammatory cytokines and catabolic enzymes. Flow cytometry was utilized to measure cell viability and apoptosis, providing a comprehensive evaluation of OC's therapeutic effects on chondrocytes. Results: The results demonstrated that OC significantly downregulated PAR-2 expression in a dose-dependent manner, leading to a substantial reduction in pro-inflammatory cytokines, including TNF-α, IL-1β, and MCP-1. Furthermore, OC attenuated the expression of catabolic markers such as SOX4 and ADAMTS5, which are critically involved in cartilage matrix degradation. Importantly, OC was found to preserve mitochondrial membrane potential (ΔΨm) in chondrocytes subjected to inflammatory stress, as evidenced by Rhodamine 123 staining, indicating a protective effect on cellular bioenergetics. Additionally, OC modulated the Receptor Activator of Nuclear Factor Kappa-Β Ligand (RANKL)/Receptor Activator of Nuclear Factor Kappa-Β (RANK) pathway, suggesting a broader therapeutic action against the multifactorial pathogenesis of OA. Conclusions: This study is the first to elucidate the modulatory effects of OC on the PAR-2 mediated inflammatory pathway in OA, revealing its potential as a multifaceted therapeutic agent that not only mitigates inflammation but also protects cartilage integrity. The preservation of mitochondrial function and modulation of the RANKL/RANK pathway further underscores OC's comprehensive therapeutic potential in counteracting the complex pathogenesis of OA. These findings position OC as a promising candidate for integration into nutritional interventions aimed at managing OA. However, further research is warranted to fully explore OC's therapeutic potential across different stages of OA and its long-term effects in musculoskeletal disorders. [ABSTRACT FROM AUTHOR]

Skin grows old due to intrinsic factors, such as age and associated hormonal changes, and external factors, like solar radiation and chemical substances to which we are exposed. With age, skin shows thinning, laxity, pallor, increased dryness, and the appearance of wrinkles, attributed to a decrease in collagen and elastin produced by fibroblasts. Several theories explain skin aging, including the free radical formation, neuroendocrine, and mitochondrial decline theories. Based on a scientific understanding of skin behavior, different in vitro methods are used to evaluate the effects of new ingredients in cosmetics. Commonly used methods include anti-collagenase, anti-elastase, and anti-hyaluronidase activity alongside techniques utilizing skin cells or 3D models. Although these methods are recognized and widely used, they lack standardization. In this review, a literature search has been conducted to examine the characteristics and variations of these methods across the laboratories. A key issue identified in this review is that many papers provide insufficient detail regarding their protocols. Moreover, the number of studies using cells is less significant than the ones determining enzyme inhibition. Our findings revealed that, in many cases, there is limited information available, underscoring the urgent need to initiate a comprehensive standardization process for the methodologies used to demonstrate anti-aging activity. [ABSTRACT FROM AUTHOR]

The present study aimed to compare the effect of photobiomodulation with different energy densities on the angiogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and stem cells from human exfoliated deciduous teeth (SHED). Photobiomodulation therapy with a 660 nm diode laser (2.4 J/cm2 and 3.9 J/cm2) on two consecutive days post-culture was applied to two types of stem cells (hPDLSCs and SHED). The Quantitative Real-time Polymerase Chain Reaction (RT-qPCR) test was undertaken to investigate Vascular Endothelial Growth Factor-A (VEGF-A) and Angiopoietin I (ANG-I) genes on days 1, 3, 5, 7, and 10 after the first session of laser application. The 4′,6-diamidino-2-phenylindole (DAPI) staining and Methyl Thiazolyl Tetrazolium (MTT) test were conducted on days 1, 3, and 5 after the first session of laser application, to assess the cell viability. The Two-way ANOVA with Tukey post hoc test was used to analyze the outcomes of the MTT and RT-qPCR tests. The results of the MTT and DAPI convergently illustrated that the groups receiving photobiomodulation with 2.4 J/cm2 had higher cell viability compared to 3.9 J/cm2. All experimental groups showed an upregulation of VEGF-A and ANG-I gene expression from day 1 to 5, followed by a downregulation from day 5 to 10. The groups with cultured hPDLSCs and SHED receiving photobiomodulation using 2.4 J/cm2 had the most amounts of VEGF-A and ANG-I gene expression on day 5, respectively. In conclusion, the 660 nm mediated photobiomodulation therapy of cultured SHED and hPDLSCs with 2.4 J/cm2 energy density may be associated with higher angiogenic differentiation (the expression of VEGF-A and ANG-I) as well as higher cell viability compared to the photobiomodulation therapy with 3.9 J/cm2. [ABSTRACT FROM AUTHOR]

Objective: To discuss the improved methods for the isolation and culture of primary chondrocytes from the neonatal rats, and to establish an efficient and economical in vitro chondrocyte culture system. Methods: The primary chondrocytes were isolated from the joints of neonatal rats and divided into overnight digestion (OD) group and rapid digestion (RD) group for separation. The chondrocytes in OD group were digested overnight by type Ⅱ collagenase, while the chondrocytes in RD group were separated by the combination of pre-digestion with physical and chemical digestion methods. The chondrocytes were cultured in modified media containing 0% (blank group 1), 1%, 2%, 4%, and 10% fetal bovine serum (FBS), 0 (blank group 2), 0. 1, 0. 2, 0. 4, 0. 8, 1. 0, and 2. 0 g·L-1 vitamin C (VC), and 0 (blank group 3), 0. 5, 1. 0, 2. 0, 4. 0, 8. 0, 10. 0 μg·L-1 poly (lactic-co-glycolic acid ) (PLGA) nanoparticles. The media containing different concentrations of FBS, VC, and PLGA were mixed with Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F12), and were divided into related groups based on the concentrations of ingredients. Cell counter was used to count the chondrocytes in various groups and the survival rates and diameters of the chondrocytes in various groups were detected; Toluidine blue staining was used to detect the morphology of the chondrocytes in various groups; CCK-8 method was used to detect the proliferative activities of the chondrocytes in various groups; cell adhesion assay was used to detect the adhesion rates of the chondrocytes in various groups; Hoechst/propidium iodide (PI) staining was used to detect the apoptosis of the chondrocytes in various groups; MTT assay was used to detect the proliferation activities of the chondrocytes in various groups after treated with modified media. The cells were divided into DMEM/F12+10%FBS group, DMEM/F12+1%FBS group, and DMEM/F12+1% FBS+0. 4 g·L-1 VC+1 μg·L-1 PLGA group. Real-time fluorescence quantitative PCR (RT-qPCR) method was used to detect the expression levels of sex-determining region Y-box 9 (SOX9), collagen type Ⅱ alpha 1 chain (Col2A1), collagen type Ⅹ alpha 1 chain (Col10A1), and matrix metallopeptidase 13 (MMP13) mRNAs in the chondrocytes in various groups after treated with modified media; immunofluorescence staining was used to detect the expressions of type Ⅱ collagen (COLⅡ) and SOX9 in the chondrocytes in various groups after treated with modified media. Results: The survival rate of primary chondrocytes in OD group was lower than that in RD group, and the average cell diameter was larger than that in RD group. The primary chondrocytes in OD group were larger and spindle-shaped, and most cells exhibited pseudopodia; in RD group, the primary chondrocytes were smaller, mostly rhomboid in shape, with only a portion of the cells showing pseudopodia. The Toluidine blue staining results showed significant coloration in both groups, but the digestion time of the chondrocytes in RD group was shorter, and compared with OD group, the actual culture time of the chondrocytes was reduced by 9-13 h, and more immature morphology of the primary chondrocytes were observed. The proliferation activity of the primary chondrocytes in OD group was slow at 24 h of culture but increased at 48 h of culture, and the proliferation activity of the primary chondrocytes was significantly higher at 48 h of culture compared with 12 h of culture (P<0. 01). Compared with 12 h of culture, the proliferation rates of the primary chondrocytes in RD group were increased at 24 and 48 h of culture (P<0. 01). At 24 and 48 h of culture, compared with OD group, the proliferation rates of the primary chondrocytes in RD group were increased (P<0. 05). The number of apoptotic chondrocytes in RD group was lower than that in OD group, and no necrotic chondrocytes were observed in either group. The proliferation activities of chondrocytes of the rats were increased with the rising of FBS concentration in the culture medium. Compared with blank group 1, the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1%, 2%, 4%, and 10% FBS were significantly increased (P<0. 05). Compared with blank group 2, the proliferative activities of chondrocytes of the rats after treated with culture mediums containing 0. 2-1. 0 g·L-1 VC were significantly increased (P<0. 05), and the highest proliferation activity was found when the concentration of VC was 0. 4 g·L-1 (P<0. 01). Compared with blank group 3, the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1-4 μg·L-1 PLGA were significantly increased (P<0. 05), and the highest proliferation activity was found after treated with culture medium containing 1 μg·L-1 PLGA (P<0. 05). Compared with DMEM/F12+10%FBS group, the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS group were significantly increased (P<0. 05 or P<0. 01). Compared with DMEM/F12+10%FBS group, the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS+0. 4 g·L-1 VC+1 μg·L-1 PLGA group were significantly increased (P<0. 01). The immunofluorescence staining results showed that the green fluorescence signal of COLⅡ and the red fluorescence signal of SOX9 were observed in some chondrocytes in DMEM/F12+10%FBS group under fluorescence microscope, and the fluorescence intensity was weak. In DMEM/F12+1%FBS group, most chondrocytes exhibited COL Ⅱ green fluorescence signal and SOX9 red fluorescence signal, and the fluorescence intensity was significantly stronger than that in DMEM/F12+10% FBS group. In DMEM/F12+1% FBS+0. 4 g·L-1 VC+1 μg·L-1 PLGA group, the COLⅡ green fluorescence signal and SOX9 red fluorescence signal were found in all the chondrocytes, and the fluorescence intensity was significantly higher than those in DMEM/F12+10%FBS and DMEM/F12+1%FBS groups. The expression levels of COLⅡ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS group were significantly higher than those in DMEM/F12+10%FBS group, and the expression levels of COLⅡ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS+ 0. 4 g·L-1 VC+ 1 μg·L-1 PLGA group were significantly higher than those in DMEM/F12+10%FBS group. Conclusion: The improved methods for the isolation and culture of primary chondrocytes of the rats can overcome the shortcomings of traditional methods, shorten the isolation time of primary chondrocytes, and improve the quality of in vitro culture of primary chondrocytes. [ABSTRACT FROM AUTHOR]

Background/Aim. The clinical success of prosthetic rehabilitation that commences after the attained implant osseointegration is dependent on the influence of several factors, of which screw loosening is a frequent one, and it is highly related to inadequate tightening (torquing) using torque wrenches. Although the wrenches are initially calibrated by the manufacturer, it is of great importance to evaluate their function after usage for a certain period of time. The aim of this study was to evaluate the accuracy and repeatability of the performance of implant torque wrenches in delivering necessary torque values before and after one year of usage. Methods. Two types of wrenches were used in the study: the beam-type and the toggle-type. Four various brands of beam-type wrench were marked as Beam 1 - Beam 4, and three various brands of toggle-type wrench were marked as Toggle 1 - Toggle 3, according to their design. Torque values delivered by wrenches were measured and analyzed using the One-Sample t-test, Independent-Samples t-test, and Mann-Whitney U test. The Bland-Altman bias test was used as an index of accuracy, whereas Forkman's comparison of datasets coefficients of variation (CV) served as an index of repeatability. Results. All wrenches except new Beam 2 and Beam 3 showed differences between the average measured torque value and target torque value. Differences were found in the average measured values between all used and new wrenches. Higher bias was observed in Toggle 1, Toggle 2, and Toggle 3 brands, whereas lower bias was recorded between used and new Beam 1 and Beam 3 wrenches. When comparing the CV for used and new wrenches, Beam 1, Beam 4, Toggle 1, and Toggle 2 revealed differences, whereas the CV for Beam 2, Beam 3, and Toggle 3 did not differ significantly. Conclusion. Compared to toggle-type, the beam-type wrenches offer greater accuracy in achieving the target torque value. The torque deteriorates in all wrenches after aging/usage and is more prominent in toggle-type devices. [ABSTRACT FROM AUTHOR]

In vitro models are invaluable tools for deconstructing the biological complexity of the periodontal ligament (PDL). Model systems that closely reproduce the 3-dimensional (3D) configuration of cell–cell and cell–matrix interactions in native tissue can deliver physiologically relevant insights. However, 3D models of the PDL that incorporate mechanical loading are currently lacking. Hence, we developed a model where periodontal tissue constructs (PTCs) are made by casting PDL cells in a collagen gel suspended between a pair of slender, silicone posts for magnetic tensile loading. Specifically, one of the posts was rigid and the other was flexible with a magnet embedded in its tip so that PTCs could be subjected to tensile loading with an external magnet. Additionally, the deflection of the flexible post could be used to measure the contractile force of PDL cells in the PTCs. Prior to tensile loading, second harmonics generation analysis of collagen fibers in PTCs revealed that incorporation of PDL cells resulted in collagen remodeling. Biomechanical testing of PTCs by tensile loading revealed an elastic response at 4 h, permanent deformation by 1 d, and creep elongation by 1 wk. Subsequently, contractile forces of PDL cells were substantially lower for PTCs under tensile loading. Immunofluorescence analysis revealed that tensile loading caused PDL cells to increase in number, express higher levels of F-actin and α–smooth muscle actin, and become aligned to the tensile axis. Second harmonics generation analysis indicated that collagen fibers in PTCs progressively remodeled over time with tensile loading. Gene expression analysis also confirmed tension-mediated upregulation of the F-actin/Rho pathway and osteogenic genes. Our model is novel in demonstrating the mechanobiological behavior that results in cell-mediated remodeling of the PDL tissue in a 3D context. Hence, it can be a valuable tool to develop therapeutics for periodontitis, periodontal regeneration, and orthodontics. [ABSTRACT FROM AUTHOR]

Lepidium meyenii Walp (LmW) or Maca, including its bioactive components such as macamides, among others, has demonstrated antioxidant effects. However, the effect size (ES) of LmW on oxidative stress has not been qualitatively described and calculated. The primary objective of this systematic review and meta-analysis was to review and qualitatively describe the studies published up to 2023 that supplemented LmW to control cellular oxidative stress; the secondary objective was to calculate the ES of the different interventions. The search was designed following the PRISMA® guidelines for systematic reviews and meta-analyses and performed in the Web of Science, Scopus, SPORTDiscus, PubMed, and MEDLINE until 2023. The selection of studies included randomized controlled trials, with tests and post-tests, both in vitro and in vivo in animals and humans. The methodological quality and risk of bias were evaluated with the CAMARADES tool. The main variables were reduced glutathione, glutathione peroxidase, superoxide dismutase, and malondialdehyde. The analysis was conducted with a pooled standardized mean difference (SMD) through Hedges' g test (95% CI). Eleven studies were included in the systematic review and eight in the meta-analysis. They revealed a small effect for reduced glutathione (SMD = 0.89), a large effect for glutathione peroxidase (SMD = 0.96), a moderate effect for superoxide dismutase (SMD = 0.68), and a moderate effect for malondialdehyde (SMD = −0.53). According to the results, the phytochemical compounds of LmW effectively controlled cellular oxidative stress, mainly macamides. It was also determined that a higher dose of LmW generated a greater antioxidant effect. However, information concerning humans is scarce. [ABSTRACT FROM AUTHOR]

Abstract Background Osteoarthritis (OA) is a prevalent degenerative joint disease characterized by chronic inflammation and progressive cartilage degradation, ultimately leading to joint dysfunction and disability. Oleocanthal (OC), a bioactive phenolic compound derived from extra virgin olive oil, has garnered significant attention due to its potent anti-inflammatory properties, which are comparable to those of non-steroidal anti-inflammatory drugs (NSAIDs). This study pioneers the investigation into the effects of OC on the Protease-Activated Receptor-2 (PAR-2) mediated inflammatory pathway in OA, aiming to validate its efficacy as a functional food-based therapeutic intervention. Methods To simulate cartilage tissue in vitro, human bone marrow-derived mesenchymal stem cells (BMSCs) were differentiated into chondrocytes. An inflammatory OA-like environment was induced in these chondrocytes using lipopolysaccharide (LPS) to mimic the pathological conditions of OA. The therapeutic effects of OC were evaluated by treating these inflamed chondrocytes with various concentrations of OC. The study focused on assessing key inflammatory markers, catabolic enzymes, and mitochondrial function to elucidate the protective mechanisms of OC. Mitochondrial function, specifically mitochondrial membrane potential (ΔΨm), was assessed using Rhodamine 123 staining, a fluorescent dye that selectively accumulates in active mitochondria. The integrity of ΔΨm serves as an indicator of mitochondrial and bioenergetic function. Additionally, Western blotting was employed to analyze protein expression levels, while real-time polymerase chain reaction (RT-PCR) was used to quantify gene expression of inflammatory cytokines and catabolic enzymes. Flow cytometry was utilized to measure cell viability and apoptosis, providing a comprehensive evaluation of OC’s therapeutic effects on chondrocytes. Results The results demonstrated that OC significantly downregulated PAR-2 expression in a dose-dependent manner, leading to a substantial reduction in pro-inflammatory cytokines, including TNF-α, IL-1β, and MCP-1. Furthermore, OC attenuated the expression of catabolic markers such as SOX4 and ADAMTS5, which are critically involved in cartilage matrix degradation. Importantly, OC was found to preserve mitochondrial membrane potential (ΔΨm) in chondrocytes subjected to inflammatory stress, as evidenced by Rhodamine 123 staining, indicating a protective effect on cellular bioenergetics. Additionally, OC modulated the Receptor Activator of Nuclear Factor Kappa-Β Ligand (RANKL)/Receptor Activator of Nuclear Factor Kappa-Β (RANK) pathway, suggesting a broader therapeutic action against the multifactorial pathogenesis of OA. Conclusions This study is the first to elucidate the modulatory effects of OC on the PAR-2 mediated inflammatory pathway in OA, revealing its potential as a multifaceted therapeutic agent that not only mitigates inflammation but also protects cartilage integrity. The preservation of mitochondrial function and modulation of the RANKL/RANK pathway further underscores OC’s comprehensive therapeutic potential in counteracting the complex pathogenesis of OA. These findings position OC as a promising candidate for integration into nutritional interventions aimed at managing OA. However, further research is warranted to fully explore OC’s therapeutic potential across different stages of OA and its long-term effects in musculoskeletal disorders.

Abstract The present study aimed to compare the effect of photobiomodulation with different energy densities on the angiogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and stem cells from human exfoliated deciduous teeth (SHED). Photobiomodulation therapy with a 660 nm diode laser (2.4 J/cm2 and 3.9 J/cm2) on two consecutive days post-culture was applied to two types of stem cells (hPDLSCs and SHED). The Quantitative Real-time Polymerase Chain Reaction (RT-qPCR) test was undertaken to investigate Vascular Endothelial Growth Factor-A (VEGF-A) and Angiopoietin I (ANG-I) genes on days 1, 3, 5, 7, and 10 after the first session of laser application. The 4′,6-diamidino-2-phenylindole (DAPI) staining and Methyl Thiazolyl Tetrazolium (MTT) test were conducted on days 1, 3, and 5 after the first session of laser application, to assess the cell viability. The Two-way ANOVA with Tukey post hoc test was used to analyze the outcomes of the MTT and RT-qPCR tests. The results of the MTT and DAPI convergently illustrated that the groups receiving photobiomodulation with 2.4 J/cm2 had higher cell viability compared to 3.9 J/cm2. All experimental groups showed an upregulation of VEGF-A and ANG-I gene expression from day 1 to 5, followed by a downregulation from day 5 to 10. The groups with cultured hPDLSCs and SHED receiving photobiomodulation using 2.4 J/cm2 had the most amounts of VEGF-A and ANG-I gene expression on day 5, respectively. In conclusion, the 660 nm mediated photobiomodulation therapy of cultured SHED and hPDLSCs with 2.4 J/cm2 energy density may be associated with higher angiogenic differentiation (the expression of VEGF-A and ANG-I) as well as higher cell viability compared to the photobiomodulation therapy with 3.9 J/cm2.

Background: Plumbago zeylanicum, commonly known as `Chitramula’ and `Chitrak’ and belonging to the family Plumbaginaceae, is a rambling perennial under shrub grown in most parts of India. The present work deals with the pharmacognostical and preliminary phytochemical studies on the roots of Plumbago zeylanica Linn. Objectives: This study aimed to look at the preliminary phytochemical screening and pharmacognostic characteristics of P. zeylanicum roots. Material and Methods: Macroscopy, microscopy, phytochemical screening, physicochemical parameters, and the production of thin-layer chromatograms were used to evaluate the pharmacognostic properties of the Plumbago zeylanicum root. Results: Macroscopic examination revealed brown roots with an unpleasant flavor. Transverse cuts displayed visible features like medullary rays, arteries, secondary xylem, and starch grains. Powder microscopy further revealed secondary xylem elements, trichomes, fibers, and starch grains. Physicochemical properties of Plumbago zeylanicum were determined, including thin-layer chromatography fingerprinting, ash levels, extractive values, and fluorescence analysis, indicating diverse phytoconstituents in the root extract. Conclusion: The extracts contain steroids, alkaloids, saponins, glycosides, terpenoids, flavonoids, and carbohydrates, suggesting potential medicinal applications. The method is a superior alternative to time-consuming and complex steam distillation-based isolation techniques.

Objective To investigate the effect of miR-181c-5p on vasculogenic mimicry (VM) in ovarian cancer stem-like cells (OCS-LCs) and its mechanism. Methods OVCAR3 cells were induced into OCS-LCs by serum-free suspension culture. OVCAR3 cells were divided into groups A, B, and C and were transfected with NC-miR-181c-5p, siRNA-miR-181c-5p, and pRNA-miR-181c-5p, respectively. The sphere-forming experiment was used to assess the sphere formation ability of groups A, B, and C. RT-qPCR was used to measure the relative expression level of miR-181c-5p in cells of groups A, B, and C, and Western blot was used to measure the relative protein expression levels of Oct-4, Nanog, HIF-1α, and VEGF in cells of groups A, B, and C. CCK-8 assay was used to measure the viability of cells in groups A, B, and C, and the three-dimensional culture experiment was used to measure the angiogenesis rate of groups A, B, and C. Results OVCAR3 cells were successfully induced into OCS-LCs. RT-qPCR showed that group B had a significantly lower relative expression level of miR-181c-5p than group A, and group C had a significantly higher relative expression level than group A (t=2.25,8.68,P

Bioactive peptides derived from native proteins modulate physiological processes in the metabolic pathways. Given that multiple protocols in the literature mimic the digestion of dietary components, gathering studies that use such models directed at protein digestion processes is critical. This systematic review aimed to gather evidence that adopted adequate experimental models to simulate human protein digestion. The databases searched were PubMed, Web of Science, ScienceDirect, Embase, Virtual Health Library, and Scopus. A total of 1985 articles were found, resulting in 20 eligible in vitro studies. The Office of Health Assessment and Translation was used to evaluate methodological quality. Seven studies used plant-based protein sources, twelve used animal protein sources, and one used both. The duration of the oral phase varied, although 60% of the studies employed a protein digestion period of 120 min. Amylase, pepsin, and pancreatin enzymes were utilized in 40% of the studies, with pH levels of 7, 3, and 7, respectively, during the oral, gastric, and intestinal phases. The INFOGEST harmonized static model was adopted by 65% of the studies; INFOGEST is the most effective model for simulating gastrointestinal protein processes in humans and can be used to answer several research questions because it describes experimental conditions close to the human physiological situation. [ABSTRACT FROM AUTHOR]

Wounds represent a growing global issue demanding increased attention. To expedite wound healing, technologies are under development, and light emitting diode (LED) devices of varying wavelengths are being explored for their stimulating influence on the healing process. This article presents a systematic literature review aiming to compile, organize, and analyze the impacts of LED devices on wound healing. This review is registered on the PROSPERO platform [CRD42023403870]. Two blinded authors conducted searches in the Pubmed, Web of Science, Scopus, Embase, and ScienceDirect databases. In vitro and in vivo experimental studies assessing LED utilization in the wound healing process were included. The search yielded 1010 studies, of which 27 were included in the review. It was identified that LED stimulates different healing pathways, promoting enhanced cell proliferation and migration, angiogenesis stimulation, increased collagen deposition, and modulation of the inflammatory response. Thus, it can be concluded that the LED stimulates cellular and molecular processes contingent on the utilized parameters. The effects depend on the standards used. Cell migration and proliferation were better influenced by green and red LED. The extracellular matrix components and angiogenesis were regulated by all wavelengths and the modulation of inflammation was mediated by green, red, and infrared LEDs. Significance statement: Several authors have studied the different mechanisms of action that justify the effect of light emitting diode (LED) on healing. However, no analysis was carried out combining these results. This systematic review analyzed and discussed the effects of LED based on the results of different studies, with an analysis of the action of wavelengths. LED acts in all phases of wound healing through different mechanisms. Cell migration, cell proliferation, and inflammation appear to be most influenced by green and red wavelengths. The extracellular matrix and angiogenesis can be influenced by all wavelengths. The results of this study may encourage the construction of new devices with wavelengths and enhance healing effects. [ABSTRACT FROM AUTHOR]

Copyright of Cumhuriyet Dental Journal is the property of Cumhuriyet University, Faculty of Dentistry and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Odontologica Latinoamericana: AOL is the property of Acta Odontologica Latinoamericana and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background and Objectives: Limited diabetes drug availability and frequent adverse reactions to oral medications highlight the need for improved drug delivery. This study suggests optimizing drug efficiency through process modifications, focusing on anti-diabetic medications like metformin hydrochloride. Matrix-type formulations are utilized to overcome current drug targeting and dosage management limitations. Methods: This study aims to improve anti-diabetic drugs, focusing on metformin hydrochloride, via process modifications in drug delivery. We developed a sustainable drug release process with matrix-type formulations, comparing hydroxyl propyl methylcellulose (HPMC) and guar gum as rate-controlling polymers. The optimized formulation was then compared with Glyciphage SR 500 to identify similarities and differences in drug release profiles. Results: We utilized matrix-type formulations to administer metformin hydrochloride. It develops a sustainable drug release process with HPMC and guar gum as rate-controlling polymers. Next, we conducted comparative drug release studies, systematically optimized the formulation, and compared it to Glyciphage SR 500 using appropriate analytical methods. Results: Comparative dissolution data, including a high similarity factor (f2) of 75.33 with Glyciphage SR 500, emphasizes the promising resemblance between metformin hydrochloride tablet (MT5) and the market sample, warranting further stability studies for potential upscaling. Conclusion: The study's novelty lies in successfully formulating sustained-release MT5 using HPMC K100M and guar gum via wet granulation, demonstrating a potential reduction in dosing frequency and adverse effects. MT5's optimal physical and chemical parameters and sustained drug release exceeded 99% at 12 hours.

Abstract Vascular injury such as central venous stenosis (CVS) is a common complication in hemodialysis patients with central venous catheters (CVCs), yet the impact of the microstructure and partial physic characteristics of catheter surface on the chronic injury of central vein has not been elucidated. In this study, the microscopic morphology of tips and bodies of six different brands of polyurethane CVCs was observed and their roughness was assessed. Subsequently, an in vitro model was established to measure the coefficients of friction (COF) between CVCs (tips and bodies) and the vena cava intima of Japanese rabbits under the same condition in a linear reciprocating mode, and changes in the intima of vessels after friction were observed. The study found that there was a significant variation in surface roughness among different brands of CVCs (tips P

Objectives: To simulate the abrasive potential of an interdental brush when applied with toothpastes and prophylactic gels/solutions in a novel laboratory brushing simulation set‐up. Methods: A brushing device was customized to treat dentin samples mimicking a simplified interdental space with an interdental brush (ISO 2). The brushing, that is, 7200 strokes for 1 h, was performed with artificial saliva (control), a povidone‐iodine solution, and slurries of chlorhexidine and fluoride gels as well as three toothpastes with different RDA values ranging from 29 to 100, respectively. The loss of dentin was profilometrically assessed and compared with ANOVA and Fishers LSD. Results: While artificial saliva as control, the solution and the gel slurries showed no measurable dentin loss, toothpastes resulted in a measurable linear surface damage with respect to the actual intrinsic RDA values and ranged from 12.6 to 26.5 μm (p < 0.001). Conclusions: Any interdental cleaning product should be tailored and carefully instructed. Any over‐ and misuse should be avoided, which applies especially to the use of interdental brushes in combination with abrasive toothpastes. [ABSTRACT FROM AUTHOR]

Zinc oxide nanoparticles (ZnO NPs) are metal oxide nanomaterials, which are important for several applications: antibacterial, anthelmintic, antiprotozoal and antitumoral, among others. These applications are mainly related to the ability to spontaneously produce and induce the production of reactive oxygen species that are important components for the destruction of pathogens and tumor cells. While trying to potentiate ZnO NPs, studies have associated these NPs with silver oxide (AgO) or silver (Ag) NPs. It has already been reported that this combination (Ag-ZnO/AgO NPs) is able to enhance the microbicidal potential. Although possessing much potential for several purposes, it is important to evaluate whether this association also poses the risk of toxicity to cells and experimental models. Therefore, this work aimed to evaluate the toxicity of various Ag-ZnO/AgO NP nanocomposites, in vitro and in vivo. Accordingly, ZnO nanocrystals and nanocomposites with various concentrations of AgO (ZnO:5Ag, ZnO:9Ag or ZnO:11Ag) were used in different cytotoxicity models: Galleria mellonella (G. mellonella), cell lines (VERO and RAW 264.7) and C57BL/6 mice. In the G. mellonella model, four concentrations were used in a single dose, with subsequent evaluation of mortality. In the case of cells, serial concentrations starting at 125 µg/mL were used, with subsequent cytotoxicity assessment. Based on the safe doses obtained in G. mellonella and cell models, the best doses were used in mice, with subsequent evaluations of weight, biochemistry as also renal and liver histopathology. It was observed that the toxicity, although low, of the nanocomposites was dependent upon the concentration of AgO used in association with ZnO NPs, both in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Introduction: Microorganism infiltration through the im-plant-abutment interface causes oral health problems such as periimplantitis, leading to implant loss. Materials and Methods: A feasible new method to quantify the Streptococcus mutans (S. mutans) infiltration through the implant-abutment interface gap is introduced in the present work. Internal hexagon (IH; n = 10), external hexagon (EH; n = 10), Morse taper (MT; n = 10), and a control for each group (n = 1) were tested. Bacteria suspension was prepared at 1.5x108 CFU/mL (CFU: colony forming units), and the implants were individually submerged up to the connection level, allowing the bacteria to contact it. The abutment was removed, and bacteria count was performed. Results: The implant sets were tested under normal bacterial growth and early and late biofilm growth conditions. Colony-forming units per mL were obtained, and the results were compared among groups. Differences in bacterial count between the MT and EH (p

Objective: To evaluate the influence of opacity and the layering technique on the fluorescence of different composite resins. Materials and Methods: Two opacities (enamel and dentin) and the layering technique (enamel + dentin) of the composite resins: Filtek® Z350 and Palfique LX5 were evaluated in vitro. Composite resin discs were fabricated using a preformed matrix of 10 mm diameter and 0.5 mm thick for the single opacity groups and 10 mm thick for the layering technique groups, using 2 layers of 0.5 mm thickness of each opacity (n = 5). Specimens were analyzed using the Raman spectroscopy method. Data were analyzed using the Kruskall-wallis and Mann-Whitney U tests. Results: When evaluating the intensity of fluorescence, no statistically significant difference was found when comparing the layering technique and enamel opacity (p2> 0.05) and an increase in the dentin opacity value for both brands of composite resin. Regarding wavelength, no statistically significant difference was found when comparing the layering technique with enamel opacity and dentin opacity for both Filtek® Z350 and Palfique LX5® composite resins (p2 > 0.05). Conclusions: The fluorescence intensity of the layering technique is similar to enamel opacity for both composite resins. Likewise, the wavelength of the layering technique is similar to the enamel opacity and dentin opacity for both brands.

Abstract Objectives This in vitro study assessed the potential of tooth discoloration by aerosols generated from three heated tobacco products (HTPs) with different specifications: in‐direct heating tobacco system platform 1.0a (IT1.0a), in‐direct heating tobacco system platform 2.0a (IT2.0a), and direct heating tobacco system platform 3.0a (DT3.0a). In addition, three flavor types (regular, menthol, and berry menthol) were selected for each HTP to characterize the effect of flavor types on tooth discoloration. Material and Methods Six bovine tooth samples were exposed directly to aerosols generated from one pack of each HTP: 350 puffs for IT1.0a, 325 puffs for IT2.0a, and 220 puffs for DT3.0a. Six bovine tooth samples were also exposed to air (350 puffs) and smoke generated from one pack of cigarettes (160 puffs) as negative and positive controls, respectively. The color of each tooth sample was measured before and after exposure. The overall color changes were assessed using overall color differences (ΔE) calculated according to the Commission International de I'Eclairage color system. A one‐way analysis of variance followed by Tukey's post hoc test was used to compare ΔE among bovine tooth samples exposed to air, cigarette smoke, and aerosols generated from each HTP. Results ΔE values for tooth samples exposed to air and aerosols generated from the three HTPs (IT1.0a, IT2.0a, and DT3.0a) were significantly lower than ΔE value for tooth samples exposed to cigarette smoke. ΔE values obtained with DT3.0a were significantly higher than those obtained with air‐exposed control samples. However, ΔE values obtained with IT1.0a and IT2.0a were not significantly different from that obtained with air‐exposed control samples. No HTPs showed significant differences in ΔE values among the three flavor types. Conclusions This study showed that HTP aerosols reduce tooth discoloration potential compared with cigarette smoke, regardless of flavor types, and the tooth discoloration potential of the product may depend on product specifications.

Skin grows old due to intrinsic factors, such as age and associated hormonal changes, and external factors, like solar radiation and chemical substances to which we are exposed. With age, skin shows thinning, laxity, pallor, increased dryness, and the appearance of wrinkles, attributed to a decrease in collagen and elastin produced by fibroblasts. Several theories explain skin aging, including the free radical formation, neuroendocrine, and mitochondrial decline theories. Based on a scientific understanding of skin behavior, different in vitro methods are used to evaluate the effects of new ingredients in cosmetics. Commonly used methods include anti-collagenase, anti-elastase, and anti-hyaluronidase activity alongside techniques utilizing skin cells or 3D models. Although these methods are recognized and widely used, they lack standardization. In this review, a literature search has been conducted to examine the characteristics and variations of these methods across the laboratories. A key issue identified in this review is that many papers provide insufficient detail regarding their protocols. Moreover, the number of studies using cells is less significant than the ones determining enzyme inhibition. Our findings revealed that, in many cases, there is limited information available, underscoring the urgent need to initiate a comprehensive standardization process for the methodologies used to demonstrate anti-aging activity.

The endosomal sorting complex required for transport (ESCRT) machinery is centrally involved in the repair of damage to both the plasma and lysosome membranes. ESCRT recruitment to sites of damage occurs on a fast time scale, and Ca2+ has been proposed to play a key signaling role in the process. Here, we show that the Ca2+-binding regulatory protein ALG-2 binds directly to negatively charged membranes in a Ca2+-dependent manner. Next, by monitoring the colocalization of ALIX with ALG-2 on negatively charged membranes, we show that ALG-2 recruits ALIX to the membrane. Furthermore, we show that ALIX recruitment to the membrane orchestrates the downstream assembly of late-acting CHMP4B, CHMP3, and CHMP2A subunits along with the AAA+ ATPase VPS4B. Finally, we show that ALG-2 can also recruit the ESCRT-III machinery to the membrane via the canonical ESCRT-I/II pathway. Our reconstitution experiments delineate the minimal sets of components needed to assemble the entire membrane repair machinery and open an avenue for the mechanistic understanding of endolysosomal membrane repair.

Objective: To evaluate the penetration of hydrogen peroxide (HP) into the pulp chamber and the color change of different bleaching varnishes in low concentrations used for at‐home bleaching. Materials and Methods: Ninety healthy premolars were used, randomly distributed into nine groups (n = 10) according to bleaching varnish (PL, PolaLuminate; VS, VivaStyle Paint On Plus; CA, Cavex Bite&White whitening pen and; AW AlignerWhite) and time (10 and 30 min), and a control group (no bleaching). The penetration of HP was evaluated by UV–Vis spectroscopy. To evaluate the color change (ΔEab, ΔE00, ΔWID) a digital spectrophotometer was used (α = 0.05). Results: The AW group in 10 min and the control group showed similar and lower HP penetration in the pulp chamber when compared to the other groups (p = 0.003). Increasing the application time to 30 minutes elevated the amount of HP inside the pulp chamber for all groups (p = 0.003), except for PL (p > 0.05). When applied for 30 min all bleaching varnishes showed higher color change (ΔWID) when compared to 10 min (p = 0.04). Conclusions: For all bleaching varnishes evaluated, PolaLuminate applied for 30 min showed lower penetration into the pulp chamber and higher bleaching effects. Clinical Significance: The use of bleaching varnishes seems promising for teeth bleaching, but it varies according to user product and protocol. [ABSTRACT FROM AUTHOR]

Bone density at the implant site is correlated to the success of osseointegration. The objective of this in vitro study was to evaluate the efficacy of osseodensification burs in increasing bone density using a solid polyurethane foam block model. The osseodensification burs kit was used to perform 48 osteotomies on a rigid polyurethane foam test ground. Burs were utilized on a TMM2 implant motor for data collection. The osteotomies were divided into two study groups (A and C) in which implant sites, extended 12 and 14 mm deep, respectively, were prepared using the drills to a compaction rotation; two control groups, B and D, represented the osteotomies for which the drills were used in cutting direction. A 3.8 × 12 mm conical implant was inserted into each site; for each implant, data were collected on the peak torque (Cp), mean torque (Cm), and integral depth curve (I). The implants underwent resonance frequency analysis (RFA) to assess the implant stability quotient (ISQ). Correlation analysis was performed between I, Cm, Cp and ISQ. One-way analysis of variance (ANOVA) was used to identify statistically significant differences between groups. Group C, representing osteotomies prepared at 14 mm with osseodensification burs, showed a significantly higher value for each parameter. Implants at sites obtained with osteocondensation drills and prepared at greater depth for autologous particle grafting showed significant increases in each implant stability parameter. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) is a representative malignant brain tumor characterized by a dismal prognosis, with survival rates of less than 2 years and high recurrence rates. Despite surgical resection and several alternative treatments, GBM remains a refractory disease due to its aggressive invasiveness and resistance to anticancer therapy. In this report, we explore the role of fibronectin type III domain containing 3B (FNDC3B) and its potential as a prognostic and therapeutic biomarker in GBM. GBM exhibited a significantly higher cancer-to-normal ratio compared to other organs, and patients with high FNDC3B expression had a poor prognosis (p < 0.01). In vitro studies revealed that silencing FNDC3B significantly reduced the expression of Survivin, an apoptosis inhibitor, and also reduced cell migration, invasion, extracellular matrix adhesion ability, and stem cell properties in GBM cells. Furthermore, we identified that FNDC3B regulates PTEN/PI3K/Akt signaling in GBM cells using MetaCore integrated pathway bioinformatics analysis and a proteome profiler phospho-kinase array with sequential western blot analysis. Collectively, our findings suggest FNDC3B as a potential biomarker for predicting GBM patient survival and for the development of treatment strategies for GBM. [ABSTRACT FROM AUTHOR]

Objectives: This in vitro study assessed the potential of tooth discoloration by aerosols generated from three heated tobacco products (HTPs) with different specifications: in‐direct heating tobacco system platform 1.0a (IT1.0a), in‐direct heating tobacco system platform 2.0a (IT2.0a), and direct heating tobacco system platform 3.0a (DT3.0a). In addition, three flavor types (regular, menthol, and berry menthol) were selected for each HTP to characterize the effect of flavor types on tooth discoloration. Material and Methods: Six bovine tooth samples were exposed directly to aerosols generated from one pack of each HTP: 350 puffs for IT1.0a, 325 puffs for IT2.0a, and 220 puffs for DT3.0a. Six bovine tooth samples were also exposed to air (350 puffs) and smoke generated from one pack of cigarettes (160 puffs) as negative and positive controls, respectively. The color of each tooth sample was measured before and after exposure. The overall color changes were assessed using overall color differences (ΔE) calculated according to the Commission International de I'Eclairage color system. A one‐way analysis of variance followed by Tukey's post hoc test was used to compare ΔE among bovine tooth samples exposed to air, cigarette smoke, and aerosols generated from each HTP. Results: ΔE values for tooth samples exposed to air and aerosols generated from the three HTPs (IT1.0a, IT2.0a, and DT3.0a) were significantly lower than ΔE value for tooth samples exposed to cigarette smoke. ΔE values obtained with DT3.0a were significantly higher than those obtained with air‐exposed control samples. However, ΔE values obtained with IT1.0a and IT2.0a were not significantly different from that obtained with air‐exposed control samples. No HTPs showed significant differences in ΔE values among the three flavor types. Conclusions: This study showed that HTP aerosols reduce tooth discoloration potential compared with cigarette smoke, regardless of flavor types, and the tooth discoloration potential of the product may depend on product specifications. [ABSTRACT FROM AUTHOR]

ABSTRACT Objective To evaluate the in vitro and in vivo toxicities of polyethylene glycol-coated gold nanoparticles synthesized using a one-step process. Methods Gold nanoparticles were prepared via a co-precipitation method using polyethylene glycol, and the synthesis product was characterized. For the in vitro evaluation, a flow cytometry analysis with Annexin V and iodide propidium staining was used to assess cytotoxicity in MG-63 cells labeled with 10, 50, and 100µg/mL of nanoparticle concentration. For the in vivo evaluation, nanoparticles were administered intraperitoneally at a dose of 10mg/kg dose in 10-week-old mice. Toxicity was assessed 24 hours and 7 days after administration via histopathological analysis of various tissues, as well as through renal, hepatic, and hematopoietic evaluations. Results Synthesized nanoparticles exhibited different hydrodynamic sizes depending on the medium: 51.27±1.62nm in water and 268.12±28.45nm (0 hour) in culture medium. They demonstrated a maximum absorbance at 520nm and a zeta potential of -8.419mV. Cellular viability exceeded 90%, with less than 3% early apoptosis, 6% late apoptosis, and 1% necrosis across all labeling conditions, indicating minimal cytotoxicity differences. Histopathological analysis highlighted the accumulation of nanoparticles in the mesentery; however, no lesions or visible agglomeration was observed in the remaining tissues. Renal, hepatic, and hematopoietic analyses showed no significant differences at any time point. Conclusion Polyethylene glycol-coated gold nanoparticles exhibit extremely low toxicity and high biocompatibility, showing promise for future studies.

Lepidium meyenii Walp (LmW) or Maca, including its bioactive components such as macamides, among others, has demonstrated antioxidant effects. However, the effect size (ES) of LmW on oxidative stress has not been qualitatively described and calculated. The primary objective of this systematic review and meta-analysis was to review and qualitatively describe the studies published up to 2023 that supplemented LmW to control cellular oxidative stress; the secondary objective was to calculate the ES of the different interventions. The search was designed following the PRISMA® guidelines for systematic reviews and meta-analyses and performed in the Web of Science, Scopus, SPORTDiscus, PubMed, and MEDLINE until 2023. The selection of studies included randomized controlled trials, with tests and post-tests, both in vitro and in vivo in animals and humans. The methodological quality and risk of bias were evaluated with the CAMARADES tool. The main variables were reduced glutathione, glutathione peroxidase, superoxide dismutase, and malondialdehyde. The analysis was conducted with a pooled standardized mean difference (SMD) through Hedges’ g test (95% CI). Eleven studies were included in the systematic review and eight in the meta-analysis. They revealed a small effect for reduced glutathione (SMD = 0.89), a large effect for glutathione peroxidase (SMD = 0.96), a moderate effect for superoxide dismutase (SMD = 0.68), and a moderate effect for malondialdehyde (SMD = −0.53). According to the results, the phytochemical compounds of LmW effectively controlled cellular oxidative stress, mainly macamides. It was also determined that a higher dose of LmW generated a greater antioxidant effect. However, information concerning humans is scarce.

Bioactive peptides derived from native proteins modulate physiological processes in the metabolic pathways. Given that multiple protocols in the literature mimic the digestion of dietary components, gathering studies that use such models directed at protein digestion processes is critical. This systematic review aimed to gather evidence that adopted adequate experimental models to simulate human protein digestion. The databases searched were PubMed, Web of Science, ScienceDirect, Embase, Virtual Health Library, and Scopus. A total of 1985 articles were found, resulting in 20 eligible in vitro studies. The Office of Health Assessment and Translation was used to evaluate methodological quality. Seven studies used plant-based protein sources, twelve used animal protein sources, and one used both. The duration of the oral phase varied, although 60% of the studies employed a protein digestion period of 120 min. Amylase, pepsin, and pancreatin enzymes were utilized in 40% of the studies, with pH levels of 7, 3, and 7, respectively, during the oral, gastric, and intestinal phases. The INFOGEST harmonized static model was adopted by 65% of the studies; INFOGEST is the most effective model for simulating gastrointestinal protein processes in humans and can be used to answer several research questions because it describes experimental conditions close to the human physiological situation.

Immune checkpoint inhibitors targeting programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have revolutionized the treatment of melanoma patients. Based on early studies addressing the mechanism of action, it was assumed that PD-1 blockade mostly influences T cell responses at the tumor site. However, recent work has demonstrated that PD-1 blockade can influence the T cell compartment in peripheral blood. If the activation of circulating, tumor-reactive T cells would form an important mechanism of action of PD-1 blockade, it may be predicted that such blockade would alter either the frequency and/or the breadth of the tumor-reactive CD8 T cell response. To address this question, we analyzed CD8 T cell responses toward 71 melanoma-associated epitopes in peripheral blood of 24 melanoma patients. We show that both the frequency and the breadth of the circulating melanoma-reactive CD8 T cell response was unaltered upon PD-1 blockade. In contrast, a broadening of the circulating melanoma-reactive CD8 T cell response was observed upon CTLA-4 blockade, in concordance with our prior data. Based on these results, we conclude that PD-1 and CTLA-4 blockade have distinct mechanisms of action. In addition, the data provide an argument in favor of the hypothesis that anti-PD-1 therapy may primarily act at the tumor site.

Genome-wide association studies revealed that loss-of-function mutations in protein tyrosine phosphatase non-receptor type 2 (PTPN2) increase the risk of developing chronic immune diseases, such as inflammatory bowel disease (IBD) and celiac disease. These conditions are associated with increased intestinal permeability as an early etiological event. The aim of this study was to examine the consequences of deficient activity of the PTPN2 gene product, T cell protein tyrosine phosphatase (TCPTP), on intestinal barrier function and tight junction organization in vivo and in vitro. Here, we demonstrate that TCPTP protected against intestinal barrier dysfunction induced by the inflammatory cytokine IFN-γ by 2 mechanisms: it maintained localization of zonula occludens 1 and occludin at apical tight junctions and restricted both expression and insertion of the cation pore-forming transmembrane protein, claudin-2, at tight junctions through upregulation of the inhibitory cysteine protease, matriptase. We also confirmed that the loss-of-function PTPN2 rs1893217 SNP was associated with increased intestinal claudin-2 expression in patients with IBD. Moreover, elevated claudin-2 levels and paracellular electrolyte flux in TCPTP-deficient intestinal epithelial cells were normalized by recombinant matriptase. Our findings uncover distinct and critical roles for epithelial TCPTP in preserving intestinal barrier integrity, thereby proposing a mechanism by which PTPN2 mutations contribute to IBD.

CGG expansions between 55 and 200 in the 5'-untranslated region of the fragile-X mental retardation gene (FMR1) increase the risk of developing the late-onset debilitating neuromuscular disease Fragile X-Associated Tremor/Ataxia Syndrome (FXTAS). While the science behind this mutation, as a paradigm for RNA-mediated nucleotide triplet repeat expansion diseases, has progressed rapidly, no treatment has proven effective at delaying the onset or decreasing morbidity, especially at later stages of the disease. Here, we demonstrated the beneficial effect of the phytochemical sulforaphane (SFN), exerted through NRF2-dependent and independent manner, on pathways relevant to brain function, bioenergetics, unfolded protein response, proteosome, antioxidant defenses, and iron metabolism in fibroblasts from FXTAS-affected subjects at all disease stages. This study paves the way for future clinical studies with SFN in the treatment of FXTAS, substantiated by the established use of this agent in clinical trials of diseases with NRF2 dysregulation and in which age is the leading risk factor.

Objective To investigate the effect of lenvatinib on the autophagy and migration of cancer-associated fibroblasts (CAFs) in hepatocellular carcinoma (HCC) tissue. Methods Primary CAFs from HCC patients were isolated and cultured and were then treated with an equal volume of DMSO (DMSO group) or 46 nmol/L lenvatinib (lenvatinib group) for 72 h. Transcriptomic sequencing and the omics analysis of drug targets were performed for the two groups, and GO functional enrichment analysis and KEGG pathway enrichment analysis were performed for differentially expressed genes and peptides. Transmission electron microscopy was used to observe the effect of lenvatinib on autophagy of CAFs, and Transwell assay and wound hea-ling assay were used to observe the effect of lenvatinib on the migration ability of CAFs. Results The differentially expressed genes and peptides between the DMSO group and the lenvatinib group were mainly enriched in the functions and pathways such as cell migration and autophagy. Transmission electron microscopy showed that the lenvatinib group had a significantly higher number of autophagosomes than the DMSO group (t=5.376,P

The elimination of biofilms during root canal therapy continues to pose a challenge due to complex anatomies and uninstrumented portions of the root canal system. The incorporation of nanoparticles in endodontic sealers is an area of interest for potentially enhancing antimicrobial activity and improving treatment outcomes. This systematic review evaluated the antimicrobial effects of various nanoparticles in endodontic sealers. Comprehensive literature review was conducted using the electronic Embase, Web of Science, and PubMed databases followed by citation searching for articles eligible per the inclusion criteria. A total of 1845 citations were screened, of which 13 articles met the inclusion criteria and were included in this review. All included articles were in vitro studies with low-to-moderate quality assessment scores. The incorporation of select nanoparticles was associated with significant enhancement of antibacterial effects in planktonic and/or biofilm forms, whereas other nanoparticles were not. The incorporation of certain types and concentrations of nanoparticles into endodontic sealers displayed antimicrobial effects in vitro. The need for well-designed clinical studies translating in vitro findings into clinical practice is warranted. The incorporation of nanoparticles may enhance the antimicrobial properties of endodontic sealers and may improve treatment outcomes. [ABSTRACT FROM AUTHOR]

Background/Aim. One of the disadvantages of the cement-retained fixed implant-supported restorations is the residual cement, which is found on the superstructure after the cementation procedure and has been identified as a risk factor for the occurrence of peri-implantitis. The aim of the study was to examine the influence of cementation techniques on the amount of residual cement at different levels of demarcation of the abutment in relation to the gingiva in the process of cementing restorations on implants. Methods. The research was conducted in in vitro conditions on casts obtained after implant placement. The abutments are milled at the level of the gingiva, 1.5 mm subgingivally and 3 mm subgingivally. Zirconium dioxide ceramic restorations were cemented using a standard cementation technique, a cementation technique using Teflon tape, and a precementation method using a silicone replica of the abutment and a 3D printed replica. The amount of residual cement was measured by photograph analysis. Adobe Photoshop was used for software analysis of photographs and determination of cement surface. Statistical data processing was performed in the SPSS program, and the Kruskal-Wallis test was used for data analysis. Results. A comparative analysis of the effectiveness of cementation techniques at the gingival margin level revealed a statistically significant difference in the amount of residual cement in relation to the cementation technique (p < 0.001). Analyzing the cementation technique effectiveness at the level of the finish line, 1.5 mm subgingivally, it was established that there was a statistically significant difference in the amount of residual cement compared to the cementation technique (p = 0.001). Comparing the effectiveness of cementation techniques at the 3 mm subgingival finish line level, it was established that there was a statistically significant difference in the amount of residual cement compared to the cementation technique (p < 0.001). Conclusion. Subgingival localization significantly affects the amount of residual cement in fixed prosthetic restorations on implants. Applying precementation techniques significantly reduces the amount of residual cement. [ABSTRACT FROM AUTHOR]

CD8+ T cell responses restricted by MHC-E, a nonclassical MHC molecule, have been associated with protection in an SIV/rhesus macaque model. The biological relevance of HLA-E-restricted CD8+ T cell responses in HIV infection, however, remains unknown. In this study, CD8+ T cells responding to HIV-1 Gag peptides presented by HLA-E were analyzed. Using in vitro assays, we observed HLA-E-restricted T cell responses to what we believe to be a newly identified subdominant Gag-KL9 as well as a well-described immunodominant Gag-KF11 epitope in T cell lines derived from chronically HIV-infected patients and also primed from healthy donors. Blocking of the HLA-E/KF11 binding by the B7 signal peptide resulted in decreased CD8+ T cell responses. KF11 presented via HLA-E in HIV-infected cells was recognized by antigen-specific CD8+ T cells. Importantly, bulk CD8+ T cells obtained from HIV-infected individuals recognized infected cells via HLA-E presentation. Ex vivo analyses at the epitope level showed a higher responder frequency of HLA-E-restricted responses to KF11 compared with KL9. Taken together, our findings of HLA-E-restricted HIV-specific immune responses offer intriguing and possibly paradigm-shifting insights into factors that contribute to the immunodominance of CD8+ T cell responses in HIV infection.

In the postnatal brain, neurogenesis occurs only within a few regions, such as the hippocampal sub-granular zone (SGZ). Postnatal neurogenesis is tightly regulated by factors that balance stem cell renewal with differentiation, and it gives rise to neurons that participate in learning and memory formation. The Kv1.1 channel, a voltage-gated potassium channel, was previously shown to suppress postnatal neurogenesis in the SGZ in a cell-autonomous manner. In this study, we have clarified the physiological and molecular mechanisms underlying Kv1.1-dependent postnatal neurogenesis. First, we discovered that the membrane potential of neural progenitor cells is highly dynamic during development. We further established a multinomial logistic regression model for cell-type classification based on the biophysical characteristics and corresponding cell markers. We found that the loss of Kv1.1 channel activity causes significant depolarization of type 2b neural progenitor cells. This depolarization is associated with increased tropomyosin receptor kinase B (TrkB) signaling and proliferation of neural progenitor cells; suppressing TrkB signaling reduces the extent of postnatal neurogenesis. Thus, our study defines the role of the Kv1.1 potassium channel in regulating the proliferation of postnatal neural progenitor cells in mouse hippocampus.

Human stem-cell-derived models provide the promise of accelerating our understanding of brain disorders, but not knowing whether they possess the ability to mature beyond mid- to late-fetal stages potentially limits their utility. We leveraged a directed differentiation protocol to comprehensively assess maturation in vitro. Based on genome-wide analysis of the epigenetic clock and transcriptomics, as well as RNA editing, we observe that three-dimensional human cortical organoids reach postnatal stages between 250 and 300 days, a timeline paralleling in vivo development. We demonstrate the presence of several known developmental milestones, including switches in the histone deacetylase complex and NMDA receptor subunits, which we confirm at the protein and physiological levels. These results suggest that important components of an intrinsic in vivo developmental program persist in vitro. We further map neurodevelopmental and neurodegenerative disease risk genes onto in vitro gene expression trajectories to provide a resource and webtool (Gene Expression in Cortical Organoids, GECO) to guide disease modeling.

Objective To establish and validate a drug in vitro absorption model of MDCK-MDR1 cells for research on the mechanism of oral drug absorption and transport. Methods MDCK-MDR1 cells with different concentrations (group L with 1.0×108/L, group M with 2.5×108/L, and group H with 5.0×108/L) were inoculated onto a 24-well Transwell plate and cultured for 1-7 d. Optical density (OD) was measured to plot the growth curve of MDCK-MDR1 cells, and cell morphology was observed at different time points of culture. Transepithelial electrical resistance (TEER) was measured at different time points to determine the optimal cell inoculation concentration and culture time for the formation of a monolayer structure. Lucifer yellow transfer assay was used to validate the monolayer structure formed at different concentrations and time points. Results The groups L, M, and H showed the formation of the monolayer structure on days 5, 3, and 1, respectively, after inoculation, and OD reached the peak value on days 5, 4, and 3, respectively, For group L, TEER reached 300 Ω·cm2 on day 5 of inoculation and tended to be stable on days 5-7. Therefore, 1.0×108/L was selected as the optimal cell inoculation concentration, with an optimal culture time of 5 d. Lucifer yellow transfer assay showed that the monolayer structure had a Lucifer yellow apparent permeability coefficient of 4.27×10-7 cm/s, which was lower than 5.0×10-7 cm/s determined by the permeability test. Conclusion The MDCK-MDR1 cell model established in this study has validated monolayer structural integrity and permeability and can be used as an in vitro model to simulate the absorption and transport mechanisms of oral drugs.

Abstract Objectives To examine the dimensional changes of endodontic sealers during 18 months using three‐dimensional (3D) surface scanning and subtraction radiography in a novel in vitro sealer‐extrusion model. Material and Methods Fifty endodontically instrumented acrylic teeth were randomly allocated to five groups (n = 10) filled with Apexit Plus, AH Plus, BioRoot RCS, TubliSeal EWT, and gutta‐percha (control). Freshly mixed sealers were intentionally extruded during obturation. All teeth were immersed in a physiologic solution for up to 18 months. Blinded 3D surface scans (resolution: ~10 μm) and digital radiographs of the teeth were obtained at baseline (immediately after obturation); and then after 1 week, and at 1, 3, and 18 months. For blinded assessment of sealer dimensional change, 3D models and radiographs were superimposed using specific software. Volumetric differences, root mean square (RMS), and area change from subtraction radiography measured at each period within each sealer group were thereafter calculated. Repeated measures analyses were done with Bonferroni adjustment for multiple comparisons; standard errors, p values, and 95% confidence intervals (CI) were reported. Results Analyses of the volumetric data confirmed significant, progressive material loss for Apexit Plus when compared to the other investigated sealers or the control group (p ≤ 0.02). Immersion period significantly influenced the volumetric dimensional changes of Apexit Plus already after 1 month (p

Glucagon-like peptide-1 (GLP-1) is an incretin hormone that potentiates glucose-stimulated insulin secretion. GLP-1 is classically produced by gut L cells; however, under certain circumstances α cells can express the prohormone convertase required for proglucagon processing to GLP-1, prohormone convertase 1/3 (PC1/3), and can produce GLP-1. However, the mechanisms through which this occurs are poorly defined. Understanding the mechanisms by which α cell PC1/3 expression can be activated may reveal new targets for diabetes treatment. Here, we demonstrate that the GLP-1 receptor (GLP-1R) agonist, liraglutide, increased α cell GLP-1 expression in a β cell GLP-1R-dependent manner. We demonstrate that this effect of liraglutide was translationally relevant in human islets through application of a new scRNA-seq technology, DART-Seq. We found that the effect of liraglutide to increase α cell PC1/3 mRNA expression occurred in a subcluster of α cells and was associated with increased expression of other β cell-like genes, which we confirmed by IHC. Finally, we found that the effect of liraglutide to increase bihormonal insulin+ glucagon+ cells was mediated by the β cell GLP-1R in mice. Together, our data validate a high-sensitivity method for scRNA-seq in human islets and identify a potentially novel GLP-1-mediated pathway regulating human α cell function.