Search Results (1,715)

Background/Objectives: The toxic side effects and resistance-associated limitations of conventional chemotherapeutic agents necessitate the development of more effective and selective combination strategies incorporating naturally derived compounds. In this study, the cytotoxic, apoptotic, oxidative stress-associated, and immunomodulatory effects of thymoquinone (TQ), a bioactive compound derived from Nigella sativa, and docetaxel (Dos), a taxane-based chemotherapeutic agent, were investigated alone and in combination in OVCAR3 ovarian cancer cells using integrated two-dimensional (2D) and three-dimensional (3D) experimental models. Materials and Methods: Cell viability was evaluated following treatment with TQ (10–500 µM), Dos (1–500 nM), and the TQ + Dos combination, and synergistic interactions were assessed by IC₅₀⁻ and combination index-based analyses. Apoptosis and cell cycle distribution were analyzed by flow cytometry. Cytokine levels were determined using ELISA, whereas apoptosis- and cell cycle-associated gene expression profiles were evaluated by RT-qPCR. Active caspase-3 expression was assessed by immunocytochemistry. Intracellular reactive oxygen species (ROS) accumulation was examined using DCFH-DA-based fluorescence imaging and antioxidant rescue experiments using N-acetyl-L-cysteine (NAC). In addition, the antitumor activity of the combination was further evaluated in OVCAR3-derived 3D tumor spheroid models using spheroid morphology, ATP-based viability, and live/dead fluorescence imaging analyses. Results: The TQ + Dos combination demonstrated enhanced cytotoxic and apoptotic activity in OVCAR3 cells compared with single-agent treatments and induced marked G2/M cell cycle arrest. Combination treatment increased pro-apoptotic gene expression and was associated with reduced expression of anti-apoptotic markers and modulated inflammatory cytokine profiles. Fluorescence-based analyses demonstrated marked intracellular ROS accumulation following TQ + Dos treatment, whereas NAC pretreatment partially attenuated oxidative stress and restored viability, suggesting partial involvement of ROS-associated mechanisms in treatment-induced cytotoxicity. Importantly, the combination maintained stronger cytotoxic and growth-inhibitory effects than either monotherapy in 3D ovarian cancer spheroids, where combination treatment induced pronounced spheroid shrinkage, viability loss, and structural disruption. Relatively lower toxicity observed in HaCaT cells suggested partial selectivity toward cancer cells. Conclusions: Collectively, these in vitro findings suggest that the TQ + Dos combination produces greater cytotoxic, apoptotic, and growth-inhibitory effects than either agent alone in ovarian cancer models and is associated with alterations in apoptosis-, cell cycle-, and oxidative stress-related responses. The observation of these effects in 3D spheroid models supports further investigation of this combination in more advanced preclinical systems. Full article

Programmable peptide hydrogels represent advanced supramolecular biomaterials featured with customizable molecular sequences and tunable self-assembly behaviors, which can biomimetically reconstruct the structural and microenvironmental complexity of native extracellular matrix. This review systematically elaborates the molecular engineering advances of programmable peptide hydrogels following a hierarchical logic from fundamental mechanisms to translational applications. We first interpret the intrinsic self-assembly mechanisms driven by non-covalent interactions and the regulatory effects of typical external microenvironmental stimuli. On this basis, we summarize core rational design principles, covering stimuli-responsive structural optimization, biofunctional modification, and the tunable regulation of physical properties, degradability and immunogenicity. Furthermore, we correlate multi-scale structural features (nanostructures, porous architecture and mechanical properties) with their versatile biomedical functions, and comprehensively discuss their cutting-edge applications in tissue regeneration, targeted drug and gene delivery, cell-mediated therapy, immunomodulation, and anti-infective treatment. Finally, we identify critical translational barriers including batch-to-batch inconsistency, immunogenic risks, and in vivo performance instability, and highlight future directions involving multi-stimuli-responsive systems, artificial intelligence-assisted design, computational modeling, and hybrid material construction. This work systematically clarifies the structure–property–function relationship of peptide hydrogels, and underscores their great potential as next-generation platforms for precision regenerative medicine and targeted disease intervention. Full article

Ornamental fountains in the Alhambra and Generalife (Granada, Spain) constitute complex socio-ecological systems where water, stone, and biological communities interact, making them highly vulnerable to biodeterioration caused by phototrophic microorganisms such as cyanobacteria, green algae, and diatoms. Conventional chemical biocides, although widely applied, present significant drawbacks including toxicity, material degradation, ecological imbalance, and limited long-term effectiveness. In this context, this study evaluated the potential of algicidal bacteria as a sustainable alternative for controlling phototrophic growth in heritage environments. Water samples from eight ornamental fountains were analyzed using 16S ribosomal RNA (16S rRNA) gene sequencing to characterize bacterial communities and identify taxa previously reported with algicidal activity. Statistical analyses were conducted to assess relationships between microbial community structure and biofilm development. In parallel, functional screening assays using filtered fountain waters against Chlorella vulgaris were performed to evaluate intrinsic inhibitory capacity. The most active sample was selected for bacterial isolation and further validation through co-culture assays, cell density measurements, and pulse-amplitude-modulated (PAM) fluorometry. A total of 18 genera with reported algicidal capacity were detected, representing a substantial fraction of the microbiome across all samples. However, no significant association was found between these taxonomic metrics and biofilm development, highlighting a decoupling between taxonomic composition and functional activity. The most active isolate, identified as Stenotrophomonas maltophilia strain LIG25, caused a rapid decline in photosynthetic efficiency and achieved more than 98% inhibition of algal growth. These findings demonstrate that ornamental fountain microbiomes represent a reservoir of native biocontrol agents and support the development of eco-friendly strategies for cultural heritage conservation. Full article

Bone loss in the maxillofacial region arises from multiple causes, including periodontal disease, trauma, surgical procedures, infection, congenital anomalies, and cancer. Traditional treatment relies on bone grafting, either alone or in combination with biomaterials. Advances in tissue engineering have introduced synthetic or natural scaffolds to mimic the mineralized bone matrix. Natural scaffolds offer excellent biocompatibility and similarity to native tissue but often lack sufficient mechanical strength and exhibit poor degradation rates. Synthetic scaffolds provide tunable porosity and mechanical stability; however, their biological inertness makes them poor sources of osteogenic signaling. A key factor in the success of any scaffold is its interaction with the host immune system. Upon implantation, the innate immune response is initiated, with neutrophils and macrophages being the first cells to contact the scaffold. Macrophage polarization toward proinflammatory (M1) or anti-inflammatory (M2) phenotypes determines whether the microenvironment favors inflammation or remodeling. The adaptive immune response also plays a critical role: T and B lymphocytes may promote tolerance and integration through Th2/Treg pathways and antibody-mediated regulation, or they may trigger chronic inflammation and rejection through Th1/Th17 activation. This review examines the natural and synthetic materials used for bone remodeling and their biological properties. It then outlines the sequence of immune events occurring from the moment a scaffold is implanted to its potential integration or failure. Finally, this study highlights the relevance of cellular models and in vitro assays for the early evaluation of immunogenicity and biocompatibility, which are essential for optimizing scaffold design and improving outcomes in maxillofacial bone regeneration. Full article

Hydrogels constructed from peptide components often rely on β-sheet architectures for their assembly, yet the process of developing such materials in aqueous environments presents notable hurdles in the context of biological systems. To address this, a novel functional dodecapeptide has been developed, capable of self-assembling into supra-molecular hydrogels via zinc chelation interactions. Morphological observations revealed a compact meshwork structure in the hydrogel formed with 9 mM Zn²⁺, differing from the relatively sparse or excessively tangled fiber architectures seen at other zinc concentrations. Alkaline phosphatase activity, an early marker of osteoblast differentiation, was notably enhanced when MC3T3-E1 cells were cultivated for 72 h in the hydrogel extract containing 300 μg/mL of the peptide, 9 μg/mL ZnCl₂, and 18.93 μg/mL H₃BO₃. Furthermore, increased protein levels of p-p38/p38, p-ERK/ERK, and p-JNK1/2/3/JNK1/2/3 were observed in P-300-ZnB and P-300 B hydrogel-treated groups, suggesting an association with MAPK pathway activation. P-Zn-9 hydrogel also promoted MC3T3-E1 cell proliferation and demonstrated favorable biocompatibility in short-term in vitro and in vivo assays. Long-term toxicity and causal relationships via inhibitor studies remain to be investigated. These results offer a viable approach to endow zinc-chelating properties in the fabrication of assembled hydrogels, presenting an innovative and potential method for constructing injectable drug delivery systems and in situ bone repair through biomaterials in subsequent applications. Full article

Methacrylate-based materials, widely used in dentistry, must possess high biocompatibility with oral cells and tissues. Currently, to improve the integration of orthodontic devices with the biological structures, laser-assisted polymer modification is actively employed. Importantly, functionalization is required only for the surface of the material that directly interacts with the oral tissues. This study presents approaches for laser modification of polymethacrylate materials and evaluates their influence on the proliferative activity of human spleen fibroblasts. Using laser radiation, two geometric patterns were obtained on the polymer surfaces. Cell morphology and proliferation on the experimental samples were assessed using scanning electron microscopy. It was found that the polymer with a groove-textured surface (pattern 1) promoted enhanced cell adhesion and reduced material toxicity. Additionally, the antibacterial properties of the polymers were evaluated. The sample with sparsely distributed surface craters (pattern 2) demonstrated an antifouling effect against Escherichia coli. Full article

Schwann cells (SCs) are critical effectors of peripheral nerve regeneration, and their interaction with biomaterial scaffolds is a key parameter in neural tissue engineering. This pilot study described and evaluated protocols for a morphological, quantitative, and morphometric analysis of SCs seeded on electrospun polyhydroxybutyrate (PHB) scaffolds using variable-pressure scanning electron microscopy (VP-SEM) under a low vacuum, without a metal coating. Six protocols were compared, varying the number of seeded cells (50,000 or 100,000) and the method used to label the seeded face of the scaffold: no marking, graphite pencil, or permanent ink (Sharpie). Confocal microscopy confirmed SC viability and adhesion. The VP-SEM analysis revealed that seeding 100,000 cells significantly increased the number of detectable cells on the scaffold surface. Graphite labeling was associated with higher cell counts and a more stellate morphology, consistent with the biocompatibility of carbon-based materials reported in the literature. Conversely, ink labeling appeared to inhibit SC adhesion. A refined protocol for measuring SC extensions using ImageJ’s ROI Manager and segmented line tools was also established. These findings provide practical methodological insights to improve the reliability and reproducibility of SC morphological analyses on ultra-thin polymeric scaffolds, with implications for peripheral nerve regeneration research. Full article

The photothermal conversion capability of polydopamine (PDA) was exploited to load the anticancer drug doxorubicin (DOX) onto its surface via π-π stacking and hydrogen-bond interactions, yielding a PDA-DOX complex. In this study, biocompatible poly-L-lactic acid (PLLA) was employed as a shell material to fabricate multifunctional PLLA composite PDA-DOX (PLLA@PDA-DOX) nanobubbles with integrated functions of ultrasound imaging, photothermal therapy, and chemotherapy. The fabricated nanobubbles exhibited a uniform mean diameter of 489.30 ± 6.96 nm with a Polydispersity index (PDI) of 0.226 ± 0.01 and a DOX loading efficiency of 3.27%. Acute toxicity evaluation in mice revealed that the maximum tolerated dose of PLLA@PDA-DOX nanobubbles was markedly higher than the clinical equivalent dose, showing no detectable toxicity or allergic reactions. Under near-infrared (NIR) laser irradiation, the inhibition rate of HCCLM3 cells increased from 50.1% to 64.45%, indicating enhanced therapeutic efficacy through the combined effects of photothermal therapy and chemotherapy. Moreover, compared with the free DOX group, the survival rate of LX-2 cells in the composite nanobubble group significantly increased from 18.9 ± 1.56% to 68.8 ± 3.08%, suggesting that the PLLA@PDA-DOX nanobubbles effectively reduced the direct cytotoxicity of DOX by preventing its immediate contact with cells. Collectively, the results confirm that PLLA@PDA-DOX nanobubbles possess excellent biocompatibility, robust ultrasound imaging performance, and enhanced antitumor efficacy under NIR irradiation. This multifunctional nanosystem demonstrates promising potential as an integrated platform for simultaneous cancer diagnosis and therapy. Full article

Dynamic interactions among cells, including immune cells, stromal cells, endothelial cells, epithelial cells, and extracellular matrix (ECM) components, are involved in the wound healing process. In chronic wounds, particularly diabetic wounds, these interactions are hampered by prolonged inflammation and excessive reactive oxygen species generation by dysregulated immune cells, bacterial infection, and impaired angiogenesis. These pathological features have shifted the therapeutic strategies from wound coverage and antimicrobial protection toward regulation of the immune microenvironment. Polymeric and hybrid materials have emerged as promising platforms for this purpose because their versatile composition, structure, degradation behavior, mechanical properties, and drug loading capacities can be widely engineered to match the dynamic requirements of wound healing, particularly in immunomodulation strategies. In this review, we focus on the major immunological barriers and potential targets in the wound healing process using polymer-based materials. Overall, this review covers recent advances, design strategies, and challenges in immunomodulatory materials including polymer-based nanoparticles, nanofibers, hydrogels, and hybrid materials for wound repair. Full article

Wound healing is a complex, multi-stage process governed by tightly regulated molecular mechanisms. However, effective regenerative therapies remain with limitations. This study presents a novel marine-derived biocomposite, JPC-ALG-CT, designed to improve wound healing through synergistic bioactive mechanisms. The material incorporates collagen extracted from the jellyfish Rhizostoma pulmo, chitosan derived from the crab Pachygrapsus mormoratus, and polysaccharide-rich extracts from the green alga Cladophora vagabunda, all sourced from the Black Sea. The study is based on the biochemical analysis of these three marine-derived components, highlighting the collagen content of jellyfish, the polysaccharides present in algae, and the bioactive properties of chitosan. The biochemical and physico-chemical properties of each component were characterized, with particular emphasis on the structural features of jellyfish collagen and the functional bioactivity of chitosan and algal polysaccharides. The research findings are supported by the identification of the collagen type extracted from jellyfish, as well as by the characterization of chitosan and green algal extracts. The resulting composite demonstrated significant antioxidant and antimicrobial activities, indicating its potential to integrate key processes involved in wound repair, including inflammation control and microbial protection. In vitro studies using fibroblast and keratinocyte models showed that the JPC-CT-ALG biocomposite supported cell viability at lower tested concentrations and promoted scratch closure in cell monolayers, suggesting preliminary wound-relevant biological activity. These findings suggest that the combined marine-derived components interact to enchance wound healing at the cellular level. This work evidenced the potential of marine biomaterials as sources for next-generation regenerative therapies and supports further investigation into their molecular mechanisms and in vivo applications for improved wound care. Full article

Perovskite-type oxides are among the most promising cathodes for intermediate-temperature solid oxide fuel cells (IT-SOFCs) due to their mixed ionic–electronic conductivity and compositional flexibility. Many high-performance cathodes rely on Sr substitution at the A-site, often associated with surface segregation and long-term degradation. In this work, we explore an alternative strategy based on defect engineering and phase interactions in Sr-free composites. Perovskite-fluorite composites based on LaFe_0.8Co_0.2O₃ were synthesized through a one-pot route designed to promote the formation of a perovskite phase and a limited amount of fluorite-type ceria. This approach allows the introduction of small fractions of Ce into the perovskite lattice, favoring the cooperative coexistence with La-doped CeO₂. Structural, microstructural and spectroscopic characterization indicates that Ce influences the crystallization pathway and composite defect chemistry. Variations in lattice parameters and Raman features suggest modifications of perovskite structure consistent with defect formation and lattice distortion. Reduction properties and electrical conductivity measurements indicate that Ce incorporation in the perovskite and oxide interaction affect charge transport and oxygen mobility. The electrochemical results demonstrate that the optimal trade-off between activation energy (Ea) and polarization resistance (Rp) is achieved for the sample, with a nominal cerium content, Ce/(La + Ce) of 0.16. Moreover, the electrochemical properties are found to correlate with the nominal cerium content, which regulates defect chemistry and the resulting composite composition. Overall, results suggest that the one-pot synthesis promotes beneficial interactions between the perovskite and ceria phases, allowing the development of Sr-free ferrite-based materials with enhanced functional properties, minimizing the amount of ceria in the composite. Full article

Cell migration is a fundamental process in cancer progression, playing a central role in tumor invasion and metastasis. This highly coordinated behavior is regulated by dynamic interactions between cancer cells and extracellular environment. Among the different tumor types, glioblastoma (GB) represents a particularly aggressive form of cancer in which enhanced migratory capacity is a key determinant of diffuse brain infiltration, tumor recurrence, and poor prognosis. In this context, extracellular vesicles (EVs) have emerged as important mediators, regulating cell migration in several cancer types, including GB. EVs are lipid bilayer-enclosed nano- and micro-sized particles, containing various bioactive molecules that can target specific recipient cells, thereby modulating cellular properties, including the migratory behavior. Among the available methods for studying cell migration, the wound healing assay is the most widely used. Although simple, cost-effective and not requiring sophisticated equipment, its reliability and reproducibility can be affected by technical variability and the diversity of existing protocols. Here, we present an optimized protocol for executing and analyzing a cellular wound healing assay designed to assess EV-mediated migration in GB cells. The protocol incorporates the use of silicone culture inserts to enhance wound homogeneity and reproducibility, together with continuous Mitomycin C incubation to inhibit cell proliferation without inducing cytotoxicity, enabling specific assessment of cell migration. We outline a step-by-step description of the procedure, detailing all required materials and equipment and highlighting critical steps, checkpoints, and key parameters. This method provides a robust framework for reproducible wound healing assays to investigate EV effects on GB cell migration. Full article

Cu and Cd, as common heavy metals occurring in the oceans, can induce oxidative stress and toxic responses in marine organisms. Important economic cephalopods inhabit the eastern coastal areas of China, and exposure to Cu and Cd poses a threat to their normal physiological activities, resulting in serious inhibition of their growth. However, the underlying toxicological mechanisms affecting these cephalopods’ larval stages remain to be elucidated. Here, indicators of oxidative stress and transcriptomics were employed to analyze the toxicological mechanisms of S. esculenta larvae exposed to Cd and Cu. GO and KEGG analysis results indicated that material transport, cellular processes, DNA replication, and other processes were inhibited. A comprehensive analysis of a protein–protein interaction network and KEGG pathways was used to explore the mechanism underlying the toxicity of co-exposure to Cu and Cd toward S. esculenta larvae. We found that Cu and Cd induce significant damage and oxidative stress. The results showed that among 20 identified key genes, ITGA4, LAMA1, and LAMC1, which are involved in the adhesion and connection between cells and the extracellular matrix; COL6A1, COL6A3, COL6A4, and COL6A6, which maintain the integrity of the extracellular matrix; and ABCA1, ABCC5, and ABCC7, which regulate the transmembrane transport of Cu and Cd were involved in the mechanism of toxicity. We suggest that co-exposure to the metals primarily inhibits the connection and adhesion between the cells of the larvae and disrupts the structure and function of the extracellular matrix. The results provide a foundation for understanding the toxicological mechanism of S. esculenta and should be of benefit to artificial breeding programs. Full article

The increasing global frequency of harmful cyanobacterial blooms (CyanoHABs), driven by nutrient enrichment and climate change, poses a severe threat to aquatic ecosystems and public health. This study evaluates the effectiveness of novel clay–polymer nanocomposites (CPCs) that combine the charge-neutralizing capabilities of polydiallyldimethylammonium chloride (polyDADMAC) with the high clay mineral density (kaolinite and sepiolite) for rapid removal of toxic cyanobacteria from water. Laboratory experiments were performed using Microcystis aeruginosa, Aphanizomenon ovalisporum, and Chlorella sp., with treatment doses determined by particle charge detector (PCD) measurements to identify the “nominal dose” required for full charge neutralization. Results show that clay–polymer nanocomposites achieve over 95% removal of turbidity and chlorophyll in M. aeruginosa at doses significantly lower (15–20%) than the calculated nominal dose, likely due to specific physical bridging interactions with the cyanobacteria’s external exopolysaccharide fibers. In contrast, A. ovalisporum and Chlorella sp. required doses closer to full charge neutralization for optimal removal. Among the materials tested, kaolinite-based nanocomposites (DKG24) showed slightly superior, more stable performance than sepiolite-based nanocomposites. Notably, application at or above the nominal dose was associated with increased soluble microcystin levels, suggesting that excessive polymer concentrations may compromise cell integrity and lead to toxin leakage. These findings suggest that engineered nanocomposites offer highly efficient, scalable technology for CyanoHAB management, provided that operational doses are carefully optimized to maximize biomass removal while minimizing toxin release. Full article

Background and Objectives: Sjögren’s disease (SjD) is a chronic autoimmune disorder in which the immune system attacks the glands that produce tears and saliva, leading to symptoms such as dry eyes and dry mouth. If left untreated, SjD can also cause inflammation and damage to other parts of the body, including the skin, lungs, kidneys, and nervous system, and increase the risk of developing lymphoma. The human leukocyte antigen (HLA) class II molecule HLA-DR3 is strongly associated with SjD. Materials and Methods: To investigate how post-translational modifications (PTMs) influence the presentation of SjD-associated autoantigens by HLA-DR3, we employed a computational framework to determine the binding of PTM-mimic peptides to HLA-DR3. We further supported the in-silico results with in-vitro experiments. Results: Our analysis revealed that PTM-mimic substitutions at canonical anchor positions rarely improved predicted binding affinity using the Stabilized Matrix Method, with most modifications resulting in reduced affinity. However, a comprehensive analysis of full-length SjD-associated autoantigen sequences (Ro60, Ro52, La) identified discrete regions with high densities of PTM-eligible anchor sites, specifically, the Ro60 HEAT solenoid, Ro52 RING/B-box/PRY-SPRY modules, and the La motif-RRM1 region, suggesting that PTMs may alter epitope presentation in a sequence-dependent manner. Experimental validation of selected PTM-mimic peptides showed enhanced T cell responses, which were associated with increased binding affinity to HLA-DR3. Structural modeling of a representative complex revealed that PTM-mimic peptides adopt a slightly shifted backbone orientation and altered side-chain positioning, leading to a larger peptide–DR3 interaction interface. Conclusions: These findings provide new insights into the role of PTMs in shaping the immunogenicity of SjD-associated autoantigens and highlight the potential for PTM-mimic peptides to modulate T cell responses in SjD. Full article