Search Results (131)

Exosomes are nano-sized particles with a structure similar to cells, and they are attracting attention as a premium cosmetic raw material because they can be effectively absorbed through skin pores and delivered without decomposing the active ingredients. In this study, the effects of exosome extracts derived from Zanthoxylum piperitum, Lagerstroemia indica, and Pinus densiflora on skin barrier enhancement and moisturizing were evaluated using HaCaT cells. Cell viability was confirmed through MTS assay, and the skin barrier improvement effect was evaluated by analyzing interleukin (IL)-6 expression in an inflammatory response induced by TNFα/IFN-γ. In addition, procollagen, matrix metalloproteinase (MMP)-1, hyaluronic acid, collagenase inhibitory activity, and elastase inhibitory activity were measured to verify the moisturizing effect. The results of the study show that exosome treatment did not affect the viability of HaCaT cells, and the skin barrier improvement effect was confirmed by decreasing IL-6 expression, which increased due to TNF-α/IFN-γ treatment. In addition, after exosome treatment, the expression of procollagen and hyaluronic acid increased, the expression of MMP-1 decreased, and significant improvements in collagenase and elastase inhibitory activities were observed, suggesting a skin moisturizing effect. The results of this study indicate that exosome extracts derived from Pinus densiflora, Zanthoxylum piperitum, and Lagerstroemia indica can contribute to enhancing the skin barrier and moisturizing, providing basic data for the development of exosome-based cosmetic raw materials. Full article

Background: Pneumococcal diseases remain a global threat due to the serotype-specific limitations of polysaccharide vaccines. This study evaluated a recombinant protein-based pneumococcal vaccine (PBPV) combining three PspA variants (PRX1/Family1Clade2, P3296/Family2/Clade3, P5668/Family2/Clade4) and detoxified pneumolysin (PlyLD). PspA targets conserved surface epitopes to block immune evasion and achieve broad coverage, while PlyLD neutralizes pore-forming toxins and enhances adaptive immunity. Methods: We evaluated the safety and immunogenicity of the PBPV in animal models. Acute toxicity studies were conducted by administering a single intramuscular injection to ICR mice, whereas chronic toxicity and immunogenicity studies were performed in cynomolgus monkeys via repeated intramuscular injections, with an equal number of male and female animals in both groups. Immune responses were assessed using ELISA, multiplexed opsonophagocytic killing assays (MOPAs), and neutralizing antibody assays. Results: Acute toxicity studies in ICR mice showed no signs of abnormal toxicity or irritation at one-dose levels. In the chronic toxicity study, cynomolgus monkeys received repeated intramuscular injections once every 3 weeks for a total of four administrations, at doses of one dose/monkey and five doses/monkey, followed by a 4-week recovery period. No significant systemic toxic reactions were observed, and the safe dose was determined to be five doses/monkey. In the immunogenicity study of monkey serum, both low-dose and high-dose groups demonstrated significant increases in antigen-specific IgG titers against each component; opsonophagocytic killing activity against pneumococcal strains from Clades 2, 3, and 4 from PspA Families 1 and 2; and neutralization antibody titers against pneumolysin post-vaccination. Conclusions: The recombinant protein-based pneumococcal vaccine exhibited a favorable safety profile and potent immunogenicity in animal models, indicating promise for broad protection against pneumococcal disease. These findings support the further development of PBPVs as a viable alternative to conventional polysaccharide-based vaccines. Full article

Folate receptor alpha is expressed at low levels in normal tissues, but is elevated in aggressive breast cancer types and can be utilized for targeted nanoparticle delivery. Hence, we prepared a hybrid nanocarrier based on in-house synthesized mesoporous silica nanoparticles (MSNs) which were further lipid-coated and reinforced with folic acid (FA). Thorough physicochemical evaluation was performed including dynamic light scattering (DLS), powder x-ray diffraction (PXRD), thermogravimetric analysis (TGA), and nitrogen physisorption. In vitro dissolution of the model drug doxorubicin was carried out in release media with pH 7.4 and pH 5.5. The cytotoxic potential and cellular uptake were investigated in MCF-7 breast cancer cells via the MTT assay, doxorubicin fluorescence measurement, and microscopy. The potential amelioration of doxorubicin’s cardiotoxicity was evaluated in vitro on the H9c2 cell line. The results showed MSNs with significant pore volume (1.38 cm³/g) and relatively small sizes (98.05 ± 1.34 nm). The lipid coat and FA attachment improved the physicochemical stability and sustained release pattern over 24 h. MSNs were non-toxic, while when doxorubicin-loaded, they caused moderate cytotoxicity. The highest cytotoxic activity was observed with folate-functionalized, doxorubicin-loaded nanoparticles (NPs). Even though non-loaded folate-functionalized NPs exhibited significant cytotoxicity, their physical mixture with doxorubicin was inferior in MCF-7 cytotoxicity as opposed to the corresponding loaded nanocarrier. Fluorescence-based quantification showed a higher intracellular accumulation of doxorubicin when delivered via NPs. These results demonstrate the potential to use folate-functionalized NPs as carriers for doxorubicin delivery in breast cancer cells. Its cardiotoxicity was significantly reduced in the case of loading onto the folic acid-functionalized lipid-coated MSNs. All these findings provide a promising proof-of-concept, although further experimental validation, particularly regarding targeting selectivity and safety, is required. Full article

Bone regeneration relies on the coordinated interplay between mechanical and biological cues. Piezoelectric composites, capable of converting mechanical strain into electrical signals, offer a promising approach to stimulate osteogenesis. This study aimed to develop and characterize polycaprolactone (PCL) and barium titanate (BaTiO₃) composite scaffolds fabricated through thermally induced phase separation (TIPS), and to systematically evaluate the effects of polymer concentration and ceramic incorporation on scaffold morphology, porosity, mechanical properties, and cytocompatibility were systematically evaluated. The resulting scaffolds exhibited a highly porous, interconnected architecture, with 9% PCL formulation showing the most uniform morphology and consistent mechanical and biological behavior. Incorporation of BaTiO₃ did not alter pore structure or compromise cytocompatibility but slightly enhanced stiffness and surface uniformity. SEM-based image analysis confirmed homogeneous BaTiO₃ dispersion across all formulations. MTT assays and confocal microscopy demonstrated robust pre-osteoblast adhesion and spreading, particularly on denser composite scaffolds, confirming that the inclusion of BaTiO₃ supports a favorable environment for cell proliferation. Overall, optimizing polymer concentration and ceramic dispersion enables fabrication of structurally coherent, cytocompatible scaffolds. The findings establish structure–property–biology relationships that serve as a baseline for future investigations into the electromechanical behavior of PCL/BaTiO₃ scaffolds and their potential to promote osteogenic differentiation under physiological loading. Full article

Background/Objectives: Chagas disease, caused by Trypanosoma cruzi, remains a major neglected tropical disease with limited therapeutic options restricted to benznidazole and nifurtimox, both associated with significant toxicity and reduced efficacy during chronic infection. Seeking novel, safe, and sustainable chemotherapeutic candidates, two new saturated cardanol-derived phospholipid analogs—LDT10 and LDT119—were rationally designed based on the molecular scaffold of miltefosine and biosourced from cashew nut shell liquid (CNSL). This study aimed to evaluate the pharmacokinetic properties of these compounds in silico and assess their antiparasitic activity, cytotoxicity, and morphological and ultrastructural effects on all developmental forms of T. cruzi in vitro. Materials and Methods: In silico ADMET predictions (SwissADME, pkCSM) were performed to determine bioavailability, pharmacokinetic behavior, CYP inhibition, mutagenicity, and hepatotoxicity. Antiproliferative activity was evaluated in epimastigotes, trypomastigotes, and intracellular amastigotes using dose–response assays and flow cytometry. Cytotoxicity was assessed in HEPG2 and HFF-1 cells using resazurin-based viability assays. Morphological and ultrastructural alterations were investigated through scanning (SEM) and transmission (TEM) electron microscopy. Reactive oxygen species (ROS) generation was quantified with H₂DCFDA after 4 h and 24 h of exposure. Results: In silico analyses indicated favorable drug-like profiles, high intestinal absorption (>89%), absence of mutagenicity or hepatotoxicity, and non-penetration of the blood–brain barrier. LDT10 was not a P-gp substrate, and LDT119 acted as a P-gp inhibitor, suggesting reduced efflux and higher intracellular retention. Both compounds inhibited epimastigote proliferation with low IC₅₀ values (LDT10: 0.81 µM; LDT119: 1.2 µM at 48 h) and reduced trypomastigote viability (LD₅₀ LDT10: 2.1 ± 2 µM; LDT119: 1.8 ± 0.8 µM). Intracellular amastigotes were highly susceptible (IC₅₀ LDT10: 0.48 µM; LDT119: 0.3 µM at 72 h), with >90% inhibition at higher concentrations. No cytotoxicity was observed in mammalian cells up to 20 µM. SEM revealed membrane wrinkling, pore-like depressions, rounded cell bodies, and multiple flagella, indicating cell division defects. TEM showed Golgi disorganization, autophagic vacuoles, mitochondrial vesiculation, and abnormal kinetoplast replication, while host cells remained structurally preserved. Both compounds induced significant ROS production in trypomastigotes after 24 h in a dose-dependent manner. Conclusions: LDT10 and LDT119 exhibited potent and selective in vitro activity against all developmental stages of T. cruzi, with low micromolar to submicromolar IC₅₀/LD₅₀ values, minimal mammalian cytotoxicity, and extensive morphological and ultrastructural damage consistent with disruption of phospholipid biosynthesis pathways. Combined with favorable in silico pharmacokinetic predictions, these CNSL-derived phospholipid analogs represent promising candidates for future Chagas disease chemotherapy and warrant further in vivo evaluation. Full article

Functionalized carbon-based materials have recently attracted attention for the efficient removal of complex pollutants, including dyes. In this study, agave bagasse, an abundant by-product of the Mexican tequila industry, was used as biomass to produce H₂SO₄-modified hydrochar (HC) for the removal of Reactive Orange 84 (RO84). FTIR-ATR analysis revealed characteristic signals of –SO₃H groups in the functionalized HC. BET characterization showed a broad range of surface areas and pore volumes, with pore radio indicating nano-, micro-, and mesoporous structures, depending on the acid concentration used during synthesis. TGA and XRD analyses indicated that higher acid concentration promoted the depolymerization of biomass components. Adsorption assays demonstrated that increasing H₂SO₄ concentrations enhanced dye removal. Post-adsorption FTIR-ATR analysis revealed signal shifts consistent with interactions between sulfonated groups and dye RO84. Statistical and mathematical analyses showed that optimal results were achieved by combining high acid concentrations during HC synthesis with high HC dosages during adsorption. Full article

Activated carbons were synthesized from petroleum-based pitch and evaluated for the removal of trace siloxanes and hydrogen sulfide (H₂S) from gas streams. Oxidative stabilization followed by steam activation produced high specific surface area with enlarged mesoporosity (BET up to 1620.9 m² g⁻¹), as confirmed by N₂ sorption (BET/PSD), SEM, and elemental analysis. A GC/MS-based fixed-bed assay using 5 g of adsorbent, a 100 mL min⁻¹ challenge flow, and a 30 min readout was employed to quantify performance under consistent conditions. Under these tests, siloxanes were not detected at 30 min, and H₂S decreased to 0.38 ppm. Samples with greater mesopore volume while retaining high surface area showed higher 30 min removal. Surface-chemistry analysis indicated that oxygen functionalities introduced during stabilization facilitated pore development during subsequent steam activation without substantial loss of area. Taken together, the textural and adsorption results present a coherent picture in which a micro/mesopore architecture supports siloxane and H₂S control under the stated test conditions. The study records the key testing parameters and performance values to enable practical comparison of petroleum-pitch-derived activated carbons for gas purification. Full article

Wharton’s Jelly-derived mesenchymal stem cell (WJ-MSC) secretome contains diverse bioactive factors with potential for skin regeneration, but its clinical efficacy is limited by poor transdermal delivery. In this study, we developed a dual-delivery system by nanoencapsulating WJ-MSC secretome and coating it onto marine sponge-derived spicules. Physicochemical characterization, in vitro assays (fibroblast and keratinocyte proliferation, keratinocyte migration, type I procollagen secretion, and antioxidant activity), and in vivo penetration studies were conducted. A single-arm clinical trial evaluated dermal absorption, pore characteristics, skin texture, wrinkles, and pigmentation following topical application. Transdermal penetration efficiency was significantly higher in the nano-coated spicule group than in the uncoated secretome control. In vitro, secretome treatment promoted fibroblast and keratinocyte activity, accelerated wound closure, and increased collagen synthesis. Clinically, a single application enhanced dermal absorption and significantly reduced pore number, while two weeks of treatment decreased wrinkles and pigmentation. Spicule-based nanoencapsulation effectively overcomes the skin barrier, enhances the regenerative activity of WJ-MSC secretome, and induces measurable clinical improvements in skin rejuvenation. This platform represents a promising cosmetic and therapeutic strategy in dermatology. Full article

Exosomes are nanoscale extracellular vesicles that carry valuable biomolecular information. However, their characterization still depends on complex and costly techniques such as flow cytometry. In this study, a paper-based Vertical Flow Assay (VFA) specifically designed for the detection and profiling of exosomes derived from metastatic breast cancer cell lines is presented. The assay operates in an ELISA-like format, targeting exosomal surface proteins (CD9, CD63, CD81, and EGFR1) with specific antibodies and a secondary antibody conjugated to alkaline phosphatase. Upon reaction with the NBT/BCIP substrate, an insoluble indigo precipitate forms on the nitrocellulose membrane, generating a visual signal that can be further quantified by smartphone imaging. The VFA was optimized for membrane type, pore size, and blocking agents, reaching a detection limit of ~6 × 10⁷ exosomes µL⁻¹ in less than 20 min. Comparative studies with bead-based flow cytometry confirmed consistent biomarker expression profiles, demonstrating the reliability of the method. By enabling exosome biomarker profiling in a simplified and low-cost format, this approach provides a promising alternative to flow cytometry and other applications required for exosome characterization. Full article

Rice is one of the world’s most consumed foods, and the cereal that most efficiently uptakes and accumulates As, contributing to human health risk. Flooded rice fields alter Eh-pH conditions and, consequently, the proportion of As(III)/As(V), favoring their accumulation in the crop. The use of algae in paddy soils can improve fertility and C-stock and affect chemical conditions and As availability. This study aimed to evaluate the effect of algae application on: As adsorption capacity in paddy soils from Sado, Portugal, changes in pH-Eh conditions in the soil–water environment, and consequent As speciation. Batch-based As adsorption assays were performed with different solid–solution ratios and Chlorella minutissima algae application, and fitted to the Freundlich and Langmuir linear models. In semi-continuous column assays, simulating rice field conditions, the effect of algae on the pH-Eh of soil pore water was evaluated. The soil quality assessment showed pseudo-total contents of As and other elements higher than Portuguese agriculture limits (11 mg As kg⁻¹), but their availability was low, posing no environmental risk. The studied soils had great As adsorption, which increased with algae application (1.07 mg g⁻¹). Algae application favored oxygenation, increasing Eh values, and maintaining As(V) species. This indicated a potential approach to reducing As(III) mobility. Full article

Chronic wounds remain a persistent clinical challenge due to delayed healing, recurrent infections, and limited effectiveness of conventional dressings. To address these unmet needs, we designed a multifunctional hydrogel system based on poly(vinyl alcohol) (PVA) and alginate (ALG), incorporating hyaluronic acid (HA)-loaded dipalmitoylphosphatidylcholine (DPPC) liposomes for regenerative stimulation and Aronia-mediated silver nanoparticles (Ag_Aro) for antimicrobial protection. Physicochemical analyses (DLS, SEM, FTIR) confirmed the successful assembly of the system and demonstrated distinct particle sizes, pore morphologies, and structural interactions. Swelling and degradation studies revealed favorable hydration capacity and stability under physiologically relevant conditions. In vitro assays with HaCaT keratinocytes indicated excellent biocompatibility, with HA-liposomes enhancing cell viability to ~190% and Ag_Aro showing minimal cytotoxicity, likely due to polyphenolic surface capping. The combined formulation achieved a balanced swelling profile, controlled degradation, and the highest biocompatibility (~195% viability), underscoring the synergistic benefits of the dual-agent design. This study introduces, to our knowledge, the first PVA–ALG bilayer hydrogel integrating HA-liposomes and phytosynthesized AgNPs, offering a promising platform for advanced wound management. Further in vivo studies are warranted to validate its therapeutic performance. Full article

Background/Objectives: Temporizin-1, a hybrid antimicrobial peptide derived from the combination of Temporin A, Gramicidin peptide, and a poly-leu sequence, has strong trypanocide activity against Trypanosoma cruzi and moderate cytotoxicity towards mammalian cells. In this study, we investigated the mode of action of the peptide upon interaction with protozoan and eukaryotic membranes. Methods: To this end, we conducted a series of biophysical assays using liposomes as biomimetic models, along with fluorescence-based experiments such as lipid mixing, membrane leakage, and assays involving Thioflavin and Laurdan. Results: Temporizin-1 displayed potent membranolytic activity on protozoan and eukaryotic membranes, causing significant membrane fusion and leakage with consequent pore formation. In addition, we also performed structural studies on liposome interaction, where we observed a helical structure that is conserved during membrane interaction. The NMR study confirms all the data obtained, providing both the structure of free Temporizin-1 in solution and the way it interacts with micelles. Moreover, Temporizin-1 demonstrated high selectivity against intracellular forms of T. cruzi and exhibited an additive effect when combined with benznidazole, highlighting its promising therapeutic activity. Conclusions: In conclusion, elucidating the mechanism of action of Temporizin-1 is essential for optimizing its structure and improving target selectivity, and driving the rational design of next-generation antimicrobial peptides by applying chemical strategies and delivery system’s conjugation. Full article

Scaffold architecture is a key determinant of cell behavior and tissue regeneration in bone tissue engineering, yet the influence of pore size under dynamic culture conditions remains incompletely understood. This study aimed to evaluate the effects of scaffold pore size on osteogenic differentiation of porcine bone marrow-derived mesenchymal stem cells (pBMSCs) cultured in a rotational oxygen-permeable bioreactor system (ROBS). Three-dimensionally (3D) printed beta-tricalcium phosphate (β-TCP) scaffolds with pore sizes of 500 µm and 1000 µm were seeded with pBMSC and cultured for 7 and 14 days under dynamic perfusion conditions. Gene expression analysis revealed significantly higher levels of osteogenic markers (Runx2, BMP-2, ALP, Osx, Col1A1) in the 1000 µm group, particularly at the early time point, with the later-stage marker Osteocalcin (Ocl) rising faster and higher in the 1000 µm group, after a lower expression at 7 days. ALP activity assays corroborated these findings. Despite having lower mechanical strength, the 1000 µm scaffolds supported a homogeneous cell distribution and high viability across all regions. These results suggest that larger pore sizes enhance early osteogenic commitment by improving nutrient transport and fluid flow in dynamic culture. These findings also support the use of larger-pore scaffolds in bioreactor-based preconditioning strategies and underscore the clinical importance of promoting early osteogenic differentiation to reduce in vitro culture time, an essential consideration for the timely preparation of implantable grafts in bone tissue engineering. Full article

Bacterial-mediated infections represent a major risk factor for chronic wounds. Numerous polymeric dressings have been proposed to reduce this incidence and promote wound healing. In the present investigation, chitosan/collagen/honey-based sponges were prepared by freeze-drying. The effect of honey incorporation at different concentrations on the physicochemical and antibacterial properties of the sponges was evaluated. The SEM images showed that the surface and cross-sections of all samples had a porous structure. The pore size gradually increased in the range of 78.14 to 126.9 μm due to the increase in honey content in the sponges. This property resulted in considerably higher porosity degrees (79.90–90.13%) and absorption rates (ranges of 1357–1665% in deionized water and 865–1938% in PBS solution) in honey-loaded systems. Conversely, the honey composite formulations exhibited a reduction in permeability, with WVTR values ranging from 131.01 to 99.39 gh⁻¹m⁻² and values of WVP from 0.3255 to 0.2118 gm⁻¹d⁻¹mm Hg⁻¹. The mechanical properties showed that adding honey made the sponges more flexible (12.49–7.95% MPa) but decreased elongation rates in the sponges (16.36–7.56%) due to higher pore heterogeneity. The antibacterial tests indicated that all treatments had inhibitory effects against S. aureus, P. aeruginosa, E. coli, and L. monocytogenes. The results from cells viability assays and in vitro healing models using human keratinocytes demonstrate that chitosan/collagen/honey sponges represent a potential alternative for applications such as wound dressings to help treat skin ulcers. The physicochemical, antibacterial, and biocompatibility properties of chitosan/collagen/honey sponges indicated their potential as a promising alternative for clinical use. Full article

The aim of this study was to evaluate the haemostatic potential and biocompatibility of a newly developed composite material for its use in blood-contacting applications. Based on promising reports on polylactide (PLA), sodium alginate (ALG), and bioactive additives such as hardystonite (HT) and zinc sulphide (ZnS), a melt-blown PLA nonwoven was modified via dip-coating using an ALG solution as a matrix for incorporating HT and ZnS particles, resulting in the PLA-ALG-ZnS-HT composite. The material was characterised in terms of surface morphology, specific surface area, pore volume, average pore size, and zeta potential (pH~7.4). Haemostatic activity was assessed by measuring blood coagulation parameters, while biocompatibility was evaluated through the viability of human peripheral blood mononuclear (PBM) cells and human foreskin fibroblasts (Hs68). Genotoxicity was analysed using the comet assay and plasmid relaxation test. Results confirmed a uniform alginate coating with dispersed HT and ZnS particles on PLA fibres. The modification increased the surface area and pore volume and caused a shift toward less negative zeta potential. Haemostatic testing showed prolonged activated partial thromboplastin time (aPTT), likely due to Zn²⁺ interactions with clotting factors. Biocompatibility tests showed high cell viability and no genotoxic effects. Our findings suggest that the PLA-ALG-ZnS-HT composite is safe for blood and skin cells and may serve as an anticoagulant material. Full article