Search Results (272)

Annona amazonica R.E. Fries (synonyms Annona amazonica var. lancifolia R.E. Fries), popularly known in Brazil as “envireira”, is a tropical tree belonging to the Annonaceae family and is traditionally used as a food source. In this work, the in vitro and in vivo anti-liver cancer effects of essential oil (EO) from A. amazonica leaves were investigated for the first time. The chemical composition of the EO was evaluated via GC–MS and GC–FID. The alamar blue assay was used to evaluate the cytotoxicity of EOs against different cancerous and noncancerous cell lines. Cell cycle analyses, YO-PRO-1/PI staining, and rhodamine 123 staining were performed via flow cytometry in HepG2 cells treated with EO. The in vivo antitumor activity of EO was evaluated in NSG mice that were xenografted with HepG2 cells and treated with EO at a dose of 60 mg/kg. The major constituents (>5%) of the EO were (E)-caryophyllene (32.01%), 1,8-cineole (13.93%), α-copaene (7.77%), α-humulene (7.15%), and α-pinene (5.13%). EO increased apoptosis and proportionally decreased the number of viable HepG2 cells. The induction of DNA fragmentation and cell shrinkage together with a significant reduction in the ΔΨm in EO-treated HepG2 cells confirmed that EO can induce apoptosis. A significant 39.2% inhibition of tumor growth in vivo was detected in EO-treated animals. These data indicate the anti-liver cancer potential of EO from A. amazonica leaves. Full article

Background: Toxoplasma gondii is a globally widespread parasite responsible for toxoplasmosis, a zoonotic disease with significant impact on both human and animal health. The current lack of safe and effective treatments underscores the need for new drugs. Earlier, thiosemicarbazones (TSCs) and their metal complexes have shown promising activities against T. gondii. This study evaluated a gold (III) complex C3 and its TSC ligand C4 for safety in host immune cells and zebrafish embryos, followed by efficacy assessment in a murine model for chronic toxoplasmosis. Methods: The effects on viability and proliferation of murine splenocytes were determined using Alamar Blue assay and BrdU ELISA, and potential effects of the drugs on zebrafish (Danio rerio) embryos were detected through daily light microscopical inspection within the first 96 h of embryo development. The parasite burden in treated versus non-treated mice was measured by quantitative real-time PCR in the brain, eyes and the heart. Results: Neither compound showed immunosuppressive effects on the host immune cells but displayed dose-dependent toxicity on early zebrafish embryo development, suggesting that these compounds should not be applied in pregnant animals. In the murine model of chronic toxoplasmosis, C4 treatment significantly reduced the parasite load in the heart but not in the brain or eyes, while C3 did not have any impact on the parasite load. Conclusions: These results highlight the potential of C4 for further exploration but also the limitations of current approaches in effectively reducing parasite burden in vivo. Full article

Lymphoma continues to pose a serious challenge to global health, underscoring the urgent need for new therapeutic strategies. Recently, the gut microbiome has been shown to play a potential role in regulating immune responses and influencing cancer progression. However, its molecular mechanisms of action in lymphoma remain poorly understood. This study investigates the antiproliferative and apoptotic activities of gut microbiota-derived metabolites, specifically nisin (N) and urolithin B (UB), individually and in combination 7:3 (5750 μM), against the human lymphoma cell line HKB-11. Comprehensive evaluations were performed using Alamar Blue viability assays, combination index (CI) analyses, reactive oxygen species (ROS) quantification, flow cytometry for apoptosis detection, and advanced bottom-up proteomics analyses. N and UB exhibited potent antiproliferative activity, with the 7:3 combination demonstrating strong synergistic effects (CI < 1), significantly enhancing apoptosis (p < 0.01) and ROS production (p < 0.0001) compared to the untreated control. Proteomics analyses revealed substantial alterations in proteins crucial to ribosomal biogenesis, mitochondrial function, cell cycle control, and apoptosis regulation, including a marked downregulation of ribosomal proteins (RPS27; Log₂FC = −3.47) and UBE2N (Log₂FC = −0.60). These findings highlight the potential of N and UB combinations as a novel and practical therapeutic approach for lymphoma treatment, warranting further in vivo exploration and clinical validation. Full article

Gingival inflammation compromises the integrity of the gingival epithelium and the underlying tissues, highlighting the need for adjuvant therapies with immunomodulatory and healing properties. Casearia sylvestris, a medicinal plant known as guaçatonga, is traditionally used to treat inflammatory lesions. This study aimed to investigate the effects of C. sylvestris on the synthesis of pro- and anti-inflammatory, proteolytic, and antioxidant molecules and on wound healing in epithelial cells. A human telomerase-immortalized gingival keratinocyte cell line (TIGKs) was used, and cells were exposed to Escherichia coli lipopolysaccharide (LPS) in the presence and absence of C. sylvestris extract, its diterpene-concentrated fraction, and its clerodane diterpene casearin J for 24 h and 48 h. Gene expression and protein synthesis were analyzed by RT-qPCR and ELISA, respectively. Nitric oxide (NO) and NF-κB activation were analyzed by Griess reaction and immunofluorescence, respectively. Additionally, cell viability was evaluated by alamarBlue^® assay, and an automated scratch assay was used for wound healing. LPS significantly increased the expression of cytokines (TNF-α, IL-1β, IL-6, IL-8, IL-10, IL-17), proteases (MMP-1 and MMP-13), iNOS as well as NO synthesis, and triggered NF-κB nuclear translocation. It also reduced IL-4 expression, cell viability, and cellular wound repopulation. Treatment with C. sylvestris derivatives significantly abrogated all aforementioned LPS-induced effects by 80–100%. Furthermore, even at higher concentrations, C. sylvestris did not affect cell viability, thus proving the safety of its derivatives. C. sylvestris exerts anti-inflammatory, antiproteolytic, and antioxidant effects on gingival keratinocytes, highlighting its potential as a valuable adjunct in the prevention and treatment of periodontal diseases. Full article

To improve implant osseointegration while preventing infection, we developed a strontium (Sr)-doped Ti₃C₂T_x MXene coating on titanium, aiming to synergistically enhance bone integration and antibacterial performance. MXene is a family of two-dimensional transition-metal carbides/nitrides whose abundant surface terminations endow high hydrophilicity and bioactivity. The coating was fabricated via anodic electrophoretic deposition (40 V, 2 min) of Ti₃C₂T_x nanosheets, followed by SrCl₂ immersion to incorporate Sr²⁺. The coating morphology, phase composition, chemistry, hydrophilicity, mechanical stability, and Sr²⁺ release were characterized. In vitro bioactivity was assessed with rat bone marrow mesenchymal stem cells (BMSCs)—with respect to viability, proliferation, migration, alkaline phosphatase (ALP) staining, and Alizarin Red S mineralization—while the antibacterial efficacy was evaluated against Staphylococcus aureus (S. aureus) via live/dead staining, colony-forming-unit enumeration, and AlamarBlue assays. The Sr-doped MXene coating formed a uniform lamellar structure, lowered the water-contact angle to ~69°, and sustained Sr²⁺ release (0.36–1.37 ppm). Compared to undoped MXene, MXene/Sr enhanced BMSC proliferation on day 5, migration by 51%, ALP activity and mineralization by 47%, and reduced S. aureus viability by 49% within 24 h. Greater BMSCs activity accelerates early bone integration, whereas rapid bacterial suppression mitigates peri-implant infection—two critical requirements for implant success. Sr-doped Ti₃C₂T_x MXene thus offers a simple, dual-function surface-engineering strategy for dental and orthopedic implants. Full article

Lymphoma continues to pose a significant global health burden, highlighting the urgent need for novel therapeutic strategies. Recent advances in microbiome research have identified gut-microbiota-derived metabolites, or postbiotics, as promising candidates in cancer therapy. This study investigates the antiproliferative and mechanistic effects of two postbiotics, Nisin (N) and Urolithin B (UB), individually and in combination, against the human lymphoma cell line HKB-11. Moreover, this study evaluated cytotoxic efficacy and underlying molecular pathways using a comprehensive experimental approach, including the Alamar Blue assay, combination index (CI) analysis, flow cytometry, reactive oxygen species (ROS) quantification, and bottom-up proteomics. N and UB displayed notable antiproliferative effects, with IC₅₀ values of 1467 µM and 87.56 µM, respectively. Importantly, their combination at a 4:6 ratio demonstrated strong synergy (CI = 0.09 at IC₉₅), significantly enhancing apoptosis (p ≤ 0.0001) and modulating oxidative stress. Proteomic profiling revealed significant regulation of key proteins related to lipid metabolism, mitochondrial function, cell cycle control, and apoptosis, including upregulation of COX6C (Log₂FC = 2.07) and downregulation of CDK4 (Log₂FC = −1.26). These findings provide mechanistic insights and underscore the translational potential of postbiotics in lymphoma treatment. Further preclinical and clinical investigations are warranted to explore their role in therapeutic regimens. Full article

Scaffold architecture plays a significant role in regulating cellular and microbial interactions in tissue engineering applications. This study evaluates the performance of 3D-printed poly (lactic acid) (PLA) scaffolds with varying porosity levels (20%, 40%, 60%, 80%, and 100%) in mechanical strength, supporting human skin fibroblast (HSF) viability and reducing bacterial colonization of Gram-positive bacteria (Staphylococcus epidermidis, Staphylococcus aureus), and Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli). The maximum tensile strength (28 MPa) was achieved in the 100% dense scaffold. Increasing porosity drastically decreased tensile strength, where 80% PLA scaffold possessed 16 MPa strength. At greater levels of porosity (60% and 40%), tensile strengths greatly decrease (8 MPa and 4 MPa), while ductility increases, especially at high porosity levels. HSF viability, assessed using the AlamarBlue assay, showed a time-dependent increase in cell proliferation, with the highest viability observed on scaffolds with 60% and 80% porosity. SEM imaging confirmed strong cell adhesion on the 80% porous scaffold, indicating that intermediate-to-high porosity enhances cell attachment and metabolic activity. In contrast, bacterial adhesion showed species-specific responses to scaffold porosity. S. epidermidis and E. coli exhibited a progressive increase in adherence with porosity, peaking at 100%. P. aeruginosa showed maximum adhesion at 80%, suggesting a porosity “sweet spot” that favors its colonization. S. aureus adhered most strongly to scaffolds with intermediate porosities (40–60%) and significantly less at 100% porosity. The current study provides insights into scaffold design considerations, emphasizing the need for optimized scaffold architecture that balances regenerative potential with infection control in tissue engineering applications. Full article

Canine otitis externa caused by Pseudomonas aeruginosa is a relevant disease in veterinary medicine. Given P. aeruginosa’s high priority status for the development of new antimicrobials, innovative strategies like bacteriophage therapy are essential. Lytic bacteriophages are viruses with high specificity for their bacterial hosts, making them a promising therapeutic choice in both human and veterinary medicine. This study aimed to evaluate the antimicrobial potential of bacteriophages JG005 and JG024, first characterized in terms of their biofilm-forming ability and antimicrobial susceptibility profile, against P. aeruginosa isolates obtained from dogs with otitis externa,. Bacteriophages titer, host range, and activity were assessed against P. aeruginosa biofilms via microtiter assays using crystal violet and Alamar Blue. JG024 showed lytic activity against 61.2% (n = 30/49) of the isolates, while JG005 showed lytic activity against 38.8% (n = 19/49) of the isolates. Crystal violet quantification showed that JG005 can promote strong microbial suppression of 60% (n = 6/10) and 50% (n = 5/10) of the isolates at a multiplicity of infection (MOI) of 10 and 100, respectively. JG024 presented strong microbial suppression of 20% (n = 2/10) of the isolates regardless of the MOI level tested. These phages show promising potential as an innovative treatment for canine otitis externa caused by P. aeruginosa, but further studies are needed before future clinical use. Full article

Human gingival fibroblasts (HGnFs) play crucial roles in periodontal wound healing. This in vitro study examined the impact of varying concentrations of topical oxygen fluid (blue^®m) on HGnF morphology, viability, proliferation, oxidative stress and pro-inflammatory cytokine production. The attempt was to underscore the potential of blue^®m as a less cytotoxic alternative to chlorhexidine in the context of tissue-regeneration improvement. Primary HGnF cell cultures were subjected to oxygen fluid (blue^®m) at concentrations of 0.6, 1.2 and 2.4% for a duration of 1 min. The positive control was 0.12% chlorhexidine. Cell morphology as well as actin cytoskeleton were assessed using microscopy and immunofluorescence staining. Cell viability and proliferation were assessed through AlamarBlue and trypan blue assays at 1, 2, 7, 10 and 14 days. Levels of reactive oxygen species (ROS) were quantified using DCFH-DA assay. Pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, MMP-8 and TIMP-1) were assessed through ELISA. HGnF morphology and actin structure were preserved at all oxygen fluid concentrations. Cell viability and proliferation were significantly higher in the 0.6% and 1.2% groups than in the control and chlorhexidine groups (p ≤ 0.05). ROS levels were low at 0.6% and 1.2%, but increased at 2.4% and with chlorhexidine (p ≤ 0.05). Oxygen treatment reduced IL-1β, IL-6, TNF-α and TIMP-1 expression, while MMP-8 levels increased. Chlorhexidine significantly upregulated the expression of all proinflammatory cytokines (p ≤ 0.01). Oxygen fluid (blue^®m) therapy improves the viability and proliferation of gingival fibroblasts and offers anti-inflammatory and preliminary antioxidative effects at the cellular level, especially at lower concentrations (0.6% and 1.2%), indicating potential application in periodontal wound management, subject to clinical validation. Full article

This study develops and characterizes GelMA–dopamine conjugates as novel tissue adhesives, offering an alternative to sutures. GelMA was synthesized at 5%, 10%, and 15% (w/v) with medium and high dopamine (DOPA) conjugation. Adhesives were evaluated for swelling, degradation, mechanical strength, and cytocompatibility using AlamarBlue assays and F-actin staining to assess cell viability and adhesion. Our findings indicate that DOPA conjugation significantly reduced the swelling ratio while increasing the biodegradation rate, resulting in enhanced release of free methacrylate groups over time. The mechanical properties and adhesion capabilities showed a complex relationship with DOPA substitution. Notably, the formulation containing 10% GelMA with high dopamine conjugation (HD) exhibited superior adhesion and mechanical strength. All formulations demonstrated shear-thinning behavior and recovery, making them suitable for injection and bioprinting applications. Although increased DOPA levels negatively affected crosslinking, the optimal formulation achieved a balance between adhesion and gel concentration. Rapid crosslinking was achieved within five minutes, enhancing the material’s suitability for clinical applications. In vitro cell-based assays confirmed the non-cytotoxic nature of the optimal adhesives, with metabolic activity showing significant increases over a 7-day period. These advancements support the development of improved tissue adhesives, potentially reducing reliance on sutures and enhancing wound healing outcomes. Full article

This study evaluated the effects of mechanical agitation of Aloe vera Barbadensis Miller solution at different concentrations using passive ultrasonic irrigation (PUI), XP Endo Finisher (XPF), XP Clean (XPC), and Easy Clean (ECL), compared to conventional endodontic irrigation (CIE), on bond strength and adhesive failure patterns in the cervical, middle, and apical thirds of the root canal. Aloe vera solutions at 1%, 3%, and 5% were tested to reverse collagen fiber collapse induced by hypochlorous acid, a free radical released by 2.5% sodium hypochlorite, which impairs dentin hybridization and the light curing of resin cement. Fiberglass posts were cemented using an etch-and-rinse adhesive system (Ambar; FGM) and conventional dual resin cement (Allcem Core) in root dentin across all thirds. Human teeth underwent chemical–mechanical preparation, and the Aloe vera solution was agitated using the CIE, PUI, XPF, XPC, or ECL protocols. Slices from each root third were evaluated under a stereomicroscope at 10× magnification and subjected to the push-out test. Cytotoxicity was assessed by applying various Aloe vera concentrations to stem cells from the apical papilla (SCAPs) for 24 h, followed by analysis of cell metabolism (Alamar Blue), viability (Live/Dead), and proliferation (F-actin). Aloe vera demonstrated significant biological activity and enhanced bond strength, particularly at 3% and 5%, irrespective of the agitation method or root third. Thus, it can be concluded that using Aloe vera solution is an alternative for pre-treatment before the cementation of fiberglass posts with conventional dual-cure resin cement in endodontically treated dentin. Full article

UV-cured methacrylated chitosan (MCHIT) hydrogels were achieved in the presence of silanised tellurium-doped silica bioactive glass (BG-Te-Sil) to produce an antimicrobial and osteoconductive scaffold for tissue engineering applications. Methacrylation of chitosan enabled efficient crosslinking, and the curing process was evaluated by means of Fourier-transform infrared spectroscopy (FTIR) and photorheology analyses. Compressive testing on crosslinked hydrogels showed that the silanised, bioactive, doped glass increased the hydrogel’s elastic modulus by up to 200% compared to unreinforced controls. Antibacterial assays against Staphylococcus aureus ATCC 43300 revealed a significant (p < 0.05) reduction in bacterial metabolic activity for hydrogels containing 50 wt% of the Te-doped bioactive glass. In vitro cytocompatibility with human bone-marrow mesenchymal stem cells demonstrated sustained viability and uniform distribution at 72 h (live/dead staining, AlamarBlue). Under H₂O₂-induced oxidative stress, reinforced hydrogels downregulated pro-inflammatory genes (TNF-α, IFN-γ, IL-1β, and PGES-2). These results suggest that the presence of the silanised bioactive glass can significantly enhance mechanical stability, antibacterial properties, and anti-inflammatory responses without affecting cytocompatibility, making these hydrogels promising for tissue engineering applications. Full article

This study investigates the fabrication and bioactivity of monophasic octacalcium phosphate (OCP) constructs using 3D-printed calcium sulfate precursors. A single-step and a two-step process were employed, transforming calcium sulfate into OCP through a controlled phase transformation in a disodium hydrogen phosphate solution. The results revealed that a single-step process for OCP conversion in 3D printed samples was unsuccessful due to incomplete transformation and the formation of intermediate phases such as brushite and monetite. In contrast, the two-step process enabled the efficient production of monophasic OCP in a shorter timeframe. The converted OCP samples exhibited a compressive strength of 7.65 ± 0.46 MPa and a contact angle of zero, indicating adequate handling strength and high wettability. The resorbability of 3D-printed OCP in simulated body fluid (SBF) was evaluated, showing weight loss through gradual dissolution accompanied by the release of calcium and phosphorus ions, followed by the consumption of these ions for reprecipitation back into OCP without direct transformation into hydroxyapatite (HA). Biocompatibility and bioactivity testing demonstrated high cell viability (96.67 ± 0.18%) using the MTT assay, indicating that the 3D-printed OCP was not cytotoxic. Alamar blue and alkaline phosphatase (ALP) activity assay showed that 3D-printed OCP supported preosteoblast proliferation and osteogenic differentiation. Full article

Background and Objectives: Extracellular vesicles (EVs) derived from mesenchymal stem cells have gained much attention as potential therapeutic agents in many diseases, including hearing disorders such as sensorineural hearing loss (SNHL). EVs inherit similar therapeutic effects, including the stimulation of tissue regeneration from the parental cells. The aim of our study was to isolate EVs produced by MSCs and use them to treat inner ear cells in culture to evaluate their protective potential against the damaging effect of an ototoxic drug. Materials and Methods: We isolated MSC-derived EVs by precipitation and characterized them by number, size, and morphology using nanoparticle tracking analysis and TEM, evaluated the protein concentration by BCA assay and the presence of EV markers CD9, CD63, and CD81 by the Dot Blot immunoblotting method. HEI-OC1 inner ear cell line was treated with EVs either alone or followed by Cisplatin. We assessed the uptake of EVs in HEI-OC1 cells by fluorescence microscopy after PKH26 labeling, ROS production by the DCFDA (dichlorfluorescein diacetate) assay, cellular viability by Alamar Blue assay, and apoptosis with the Annexin V/Propidium Iodide method. Results: The isolated EVs had mean dimensions of 184.4 nms and the concentration of the EV suspension was 180 × 10⁶ particles/mL. TEM analysis showed intact vesicular structures with lipid-bilayer membranes having similar sizes with those measured by NTA. The PKH26-labeled EVs were observed in the HEI-OC1 cells after 24 h incubation, the amount increasing with the concentration. EVs reduced ROS production and increased the number of viable cells both alone and as pretreatment before Cisplatin, dose-dependently. Cells in early apoptosis were inhibited by EVs, while those in late apoptosis were enhanced, both with and without Cisplatin. Conclusions: EVs secreted by MSC protected HEI-OC1 cells against Cisplatin toxicity, reduced ROS production, and stimulated cell viability and the elimination of damaged cells by apoptosis, protecting the HEI-OC1 cells against Cisplatin-induced damage. Full article

Glioblastoma (GBM, isocitrate dehydrogenase wild-type) is the most common primary malignant brain tumor in adults and is associated with a severely low survival rate. Treatments offer mere palliation and are ineffective, due, in part, to a lack of understanding of the intricate mechanisms underlying the disease, including the contribution of the tumor microenvironment (TME). Current GBM models continue to face challenges as they lack the critical components and properties required. To address this limitation, we developed innovative and practical three-dimensional (3D) GBM models with structural and mechanical biomimicry and tunability. These models allowed for more accurate emulation of the extracellular matrix (ECM) and vasculature characteristics of the native GBM TME. Additionally, 3D bioprinting was utilized to integrate these complexities, employing a hydrogel composite to mimic the native environment that is known to contribute to tumor cell growth. First, we examined the changes in physical properties that resulted from adjoining hydrogels at diverse concentrations using Fourier-Transform Infrared Spectroscopy (FTIR), compression testing, scanning electron microscopy (SEM), rheological analysis, and degradation analysis. Subsequently, we refined and optimized the embedded bioprinting processes. The resulting 3D GBM models were structurally reliable and reproducible, featuring integrated inner channels and possessing tunable properties to emulate the characteristics of the GBM ECM. Biocompatibility testing was performed via live/dead and AlamarBlue analyses using GBM cells (both commercial cell lines and patient-derived cell lines) encapsulated in the constructs, along with immunohistochemistry staining to understand how ECM properties altered the functions of GBM cells. The observed behavior of GBM cells indicated greater functionality in softer matrices, while the incorporation of hyaluronic acid (HA) into the gelatin methacryloyl (gelMA) matrix enhanced its biomimicry of the native GBM TME. The findings underscore the critical role of TME components, particularly ECM properties, in influencing GBM survival, proliferation, and molecular expression, laying the groundwork for further mechanistic studies. Additionally, the outcomes validate the potential of leveraging 3D bioprinting for GBM modeling, providing a fully controllable environment to explore specific pathways and therapeutic targets that are challenging to study in conventional model systems. Full article