Search Results (9,650)

Background/Objectives: This study aimed to improve the biological profile of oleanolic acid (OA) through structural modification at the C-17 carboxyl group and the C-3 hydroxyl group, with a focus on the design of short-chain alkyl esters and 3-O-furoyl hybrids. Methods: Two series of OA derivatives were synthesized and characterized using spectroscopic methods, including ¹H NMR, ¹³C NMR and MS. In silico structure–activity relationship (SAR) analysis, ADMETox profiling, and molecular docking to the epidermal growth factor receptor (EGFR) tyrosine kinase domain were performed as predictive and hypothesis-generating tools. Anticancer activity was evaluated in vitro using the MTT assay against human cancer cell lines, including HeLa, MCF-7, A-549, SKBR-3, PC-3 and SKOV-3, as well as non-malignant human dermal fibroblasts (HDFs). Antioxidant properties were assessed using cell-free CUPRAC and DPPH assays. Results: The C-17 esterification markedly enhanced cytotoxic potency compared to the parent OA, while the introduction of the 3-O-furoyl moiety further improved antiproliferative activity in several derivatives. Selected compounds showed low-micromolar IC₅₀ values and moderate selectivity toward cancer cells. Molecular docking suggested favorable accommodation of selected derivatives within the EGFR ATP-binding pocket, mainly through hydrophobic and π-related interactions; however, these results do not confirm direct EGFR binding and require experimental validation. The CUPRAC and DPPH assays provided preliminary insight into chemical redox behavior but should not be directly extrapolated to intracellular antioxidant or pro-oxidant activity. Predicted ADMETox profiles indicated moderate permeability and relatively low predicted risk for selected toxicity endpoints, while also highlighting high lipophilicity, poor aqueous solubility and potential metabolic liabilities. Conclusions: Overall, the results identify several OA derivatives as promising anticancer lead compounds for further optimization and mechanistic investigation. Full article

Background: Bougainvillea species have been used in traditional Mexican medicine, but their bioactive compounds and mechanisms of action are insufficiently studied. This is the first comprehensive evaluation of fractions from the acetone extract of Bougainvillea × buttiana Holtum & Standl (var. Rose), combining phytochemical profiling with in vitro multitarget bioactivity assessment. Methods: Eleven fractions were analyzed for total phenolic (TPC) and flavonoid content (TFC) and antioxidant capacity using the DPPH assay. The most active fractions were further tested for nitric oxide (NO) scavenging, protection of erythrocytes and bovine serum albumin (BSA) from oxidative damage, inhibition of enzymes involved in inflammation (PLA₂, COX, LOX) and carbohydrate metabolism (α-glucosidase, α-amylase, tyrosinase), cytoprotective effects in L929 fibroblasts exposed to hydrogen peroxide, and their main metabolites were qualitatively identified by HPLC-UV-Vis. Results: All fractions showed significant TPC and TFC and concentration-dependent antioxidant activity. The fractions with the highest antioxidant indices were F5, F7, and F9. These effectively scavenged NO, protected erythrocytes and L929 cells (maintaining viability at 82.0%, 75.6%, and 72.0%, respectively), and inhibited key enzymes. Seven major compounds, mainly flavonoids, were identified. Conclusions: These findings showed that flavonoid-enriched fractions from B. × buttiana exhibit coordinated antioxidant, anti-inflammatory, antidiabetic, and cytoprotective effects, suggesting potential to treat oxidative stress-related disorders. Full article

Background/Objectives: Oxidative stress is a major cause of impaired oocyte quality and early embryo development, a challenge that still needs to be addressed in assisted reproduction. Mesenchymal stem cell secretomes have been investigated as cell-free therapeutics with antioxidant activity and relevant anti-apoptotic effects. This study aimed to evaluate the effects of human amniotic membrane-derived mesenchymal stem cell-conditioned saline (AMSC-CS) as an injectable formulation on oxidative stress–related markers in ovarian tissue and preimplantation developmental outcomes. Methods: AMSC-CS was administered intravenously to female mice in a dose-dependent manner. Safety assessments were conducted to evaluate systemic and target organ toxicity within the dosage range. In vitro fertilization (IVF) outcomes and oxidative status in ovaries, oocytes, and embryos were evaluated following treatment with low, medium, and high doses of AMSC-CS (1, 3, and 5 μL/g). Results: As an injectable formulation, the safety assessments did not reveal systemic or target organ toxicity of AMSC-CS within the dosage range. Medium-to-high doses of AMSC-CS improved the expression of folliculogenesis-related genes and decreased oxidative stress and apoptosis signaling in ovarian tissue. At the high dose, AMSC-CS promoted preimplantation embryo development to the blastocyst and hatched blastocyst stages, along with improved blastocyst quality and reduced oxidative stress in oocytes and blastocysts. Conclusions: These findings suggest that AMSC-CS at medium-to-high doses, as an injectable formulation with antioxidant activity, may be a promising adjunct for assisted reproductive technologies. Full article

Neutrophils are essential for antimicrobial defense through reactive oxygen species (ROS) production, tumor necrosis factor-alpha (TNF-α) release, and neutrophil extracellular trap formation. Lipoic acid, a redox-active antioxidant, modulates activation in human neutrophils. Neutrophils isolated from healthy donors were pretreated with lipoic acid and then exposed to lipopolysaccharide (LPS) or whole Escherichia coli, according to the specific assay. ROS production, NET formation, TNF-α release, bacterial killing, metabolic activity, and cell death were assessed using fluorometric assays, enzyme-linked immunosorbent assay, colony-forming unit assays, MTT reduction, and Annexin V-FITC/propidium iodide flow cytometry. Lipoic acid significantly reduced ROS production and NET formation induced by LPS and Escherichia coli. at 0.5 mM, lipoic acid also reduced E. coli-induced NET formation by approximately 50% and attenuated TNF-α release at early stimulation times. In colony-forming unit assays, lipoic acid did not significantly reduce neutrophil-mediated killing of Escherichia coli or Staphylococcus aureus. Although only neutrophil preparations with high baseline viability were used, Escherichia coli challenge markedly reduced cell viability during the assay; under this condition, lipoic acid pretreatment limited bacteria-induced necrosis and preserved a higher proportion of viable neutrophils. These findings indicate that lipoic acid dampens excessive oxidative and inflammatory neutrophil responses while maintaining measurable bactericidal capacity in vitro. Full article

Efficient micropropagation systems are essential for the large-scale production of uniform and high-quality plant material in aromatic species. In this study, the effects of gibberellic acid (GA₃; 1.4, 2.8, and 5.6 µM) and methyl jasmonate (MeJA; 2.2, 4.4, and 8.8 µM) were evaluated on the in vitro performance of Artemisia pontica across two successive subcultures. Morphological, physiological, and biochemical parameters were assessed, and the most effective treatment was further investigated through scanning electron microscopy (SEM) to evaluate leaf trichome characteristics. GA₃ treatments significantly enhanced shoot growth, shoot number, and relative growth rate, with the strongest response observed at 2.8 µM. This concentration also promoted higher chlorophyll content and antioxidant activity, indicating improved physiological status and metabolic performance of plantlets. In contrast, MeJA treatments, particularly at 8.8 µM, reduced growth performance and pigment accumulation, suggesting a less favorable physiological status for micropropagation. Multivariate analysis (PCA and hierarchical clustering) revealed a clear separation among treatments, with GA₃ at 2.8 µM associated with a coordinated increase in growth-related and antioxidant traits. SEM analysis showed that GA₃ influenced leaf epidermal structure, increasing the density of T-shaped, non-glandular trichomes and the diameter of glandular secreting trichomes, suggesting structural adjustments linked to metabolic activity. Overall, the results indicate that GA₃ at 2.8 µM represents the most effective supplementation under the tested conditions, promoting a balanced improvement in shoot proliferation, physiological performance, antioxidant activity, and selected structural traits. Full article

Lipopolysaccharide (LPS) impairs the function of ovine follicular granulosa cells (GCs), representing a primary cause of follicular atresia. Selenium (Se), an essential trace element, possesses anti-inflammatory and cytoprotective properties; however, its effects on GC ultrastructure remain largely unknown. In this study, primary ovine GCs were exposed to LPS (10 µg/mL) and treated with sodium selenite (25 nM). Transmission electron microscopy (TEM), JC-1 staining, enzyme-linked immunosorbent assay (ELISA), reactive oxygen species (ROS) detection, flow cytometry, and quantitative real-time PCR (qRT-PCR) were employed to evaluate cellular ultrastructure, mitochondrial membrane potential (ΔΨm), and downstream physiological processes. LPS induced severe mitochondrial pyknosis, cristae loss, and reduced ΔΨm, accompanied by inflammation, oxidative stress, apoptosis, and impaired steroidogenesis. Se intervention markedly ameliorated these ultrastructural injuries, preserving mitochondrial morphology and ΔΨm. Functionally, Se suppressed the release of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β); enhanced the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) while attenuating ROS accumulation; inhibited apoptosis by upregulating BCL-2 and downregulating BAX and CASPASE-3; and restored E₂ and P₄ secretion via upregulation of STAR and NR5A1. This study provides direct morphological evidence that Se protects ovine GCs from LPS-induced damage by repairing mitochondrial ultrastructure. This structural restoration is central to its integrated anti-inflammatory, antioxidant, anti-apoptotic, and steroidogenic effects. These in vitro findings suggest that Se may serve as a promising nutritional strategy for mitigating inflammation-driven follicular atresia, pending further in vivo validation. Full article

Traditional methods for screening high-melanin-yielding Auricularia heimuer strains are time-consuming and environmentally unfriendly. To address this issue, fifteen A. heimuer strains were used to determine the mycelial biomass, absorbance at 500 nm, CIE L*a*b* colorimetric values, and melanin yield of the fermentation broth. Pearson correlation analysis was performed to clarify the correlations among these indicators, and a regression equation was fitted to establish a rapid screening method. A total of 84 A. heimuer strains were used to verify this method, of which one high-melanin-yielding strain was obtained. The structural characterization and in vitro antioxidant activities of A. heimuer melanin (AHM) were determined. The results showed that the melanin yields of fifteen A. heimuer strains were extremely significantly positively correlated with absorbance at 500 nm (r = 0.880, p < 0.01). The fitted linear regression equation was Y = 0.0246X + 0.00094 (R² = 0.8756, p < 0.01). When 84 tested strains were investigated with this method, 8 strains (53.33%) exhibited relative differences below 10%, which is consistent with the satisfactory accuracy of the absorbance-based screening method. Finally, a high-melanin-yielding strain HMCC50028 was obtained, with a melanin yield of 0.0540 g/100 mL. The results of UV-Vis spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM) of AHM indicated that the melanin exhibited structural characteristics consistent with fungal melanins, belonging to the natural melanin family. In vitro assays demonstrated that AHM possessed excellent superoxide anion radical scavenging activity and ferric reducing power. Full article

Congenital toxoplasmosis, caused by the obligatory intracellular apicomplexan protozoan Toxoplasma gondii, can lead to severe complications during pregnancy, including fetal malformations and spontaneous abortion. In the present study, the anti-T. gondii effects of the natural flavonoid quercetin were evaluated using in vitro, ex vivo, and in silico models. Cell viability and intracellular proliferation of the parasite were determined via colorimetric assays. The lipid droplet assay was analyzed using Nile Red staining, the antioxidant and oxidative stress parameters were determined by biochemical assays, and the cytokine levels were quantified by immunoassays. Our results demonstrated that non-cytotoxic concentrations of quercetin (CC₅₀ > 100 μM) significantly inhibited parasite proliferation (IC₅₀ = 14.10 ± 2.83 μM; SI > 7.09) in an irreversible manner. Quercetin impairs parasite adhesion, invasion, and reinfection capacity. In parallel, quercetin reduced lipid droplet accumulation, restored antioxidant balance by modulating redox biomarkers, and regulated cytokine production, notably increasing IL-4, IL-6, and IL-8 levels. Corroborating the in vitro findings, quercetin significantly reduced T. gondii proliferation in human placental villous explants while preserving tissue architecture and viability. In silico analyses revealed that quercetin binds to the active site of T. gondii hypoxanthine-guanine phosphoribosyltransferase (TgHGPRT) and exhibits favorable pharmacokinetic and drug-likeness properties. Full article

In traditional Chinese medicine, Symplocos chinensis (Lour.) Druce is a well-known herbal remedy prescribed to treat malaria, snakebites, and thermal injuries (burns and scalds). The objective of this study was to characterize the volatile profile of S. chinensis essential oil (EO) and evaluate its therapeutic potential through antioxidant, acetylcholinesterase (AChE), and β-lactamase inhibitory assays. The major components of S. chinensis EO included squalene (12.08%), octanol (7.01%), edulan III (6.81%), n-Hexadecanoic acid (6.81%), geranylacetone (4.80%), β-longipinene (4.60%), 2-methyl-1-octene (2.55%), (E, E)-2,4-decadienal (2.15%), and linalool (2.00%). Antioxidant evaluation revealed that the S. chinensis EO possessed relatively moderate radical scavenging properties, achieving 39.6% inhibition of DPPH radicals at 10 mg/mL, while its activity against ABTS radicals yielded an IC₅₀ of 6.85 ± 1.97 mg/mL. Its reducing power, as determined by the FRAP assay, was further quantified at 175.50 ± 23.25 µmol/g. Furthermore, the EO exhibited inhibitory activities against AChE (IC₅₀ = 149.40 ± 16.92 μg/mL) and β-lactamase (IC₅₀ = 30.20 ± 0.84 μg/mL). These results demonstrate that S. chinensis EO exhibits antioxidant, AChE-inhibitory, and β-lactamase-inhibitory activities in vitro, warranting further comprehensive, systematic studies to evaluate its biological properties and potential applications. Full article

The influence of natural and synthetic blue food colorants on the human body, including the cardiovascular system, is a complex and not fully understood topic. Considering that various papers have demonstrated that oxidative stress is a crucial step in the development of cardiovascular diseases (CVDs), our experiments on the pro- or antioxidant action of three blue food colorants (one natural colorant—genipin—and two synthetic colorants—brilliant blue FCF and patent blue V) focused on two aspects that are important for the development of CVDs: the level of biomarkers of oxidative stress induced by H₂O₂/Fe²⁺ (the donor of hydroxyl radicals—one of the most aggressive reactive oxygen species produced in humans) in human blood platelets and human plasma, as well as the arachidonic acid cascade in blood platelets stimulated by thrombin (in vitro). Our results demonstrated that two tested blue colorants—genipin and brilliant blue FCF (at four used concentrations: 2, 10, 20, and 200 µM)—reduced plasma lipid peroxidation induced by H₂O₂/Fe²⁺. Moreover, all tested blue colorants (genipin, brilliant blue FCF, and patent blue V; at the concentrations 2, 10, 20, and 200 µM) inhibited lipid peroxidation in blood platelets treated with H₂O₂/Fe²⁺. In contrast, only genipin (at the highest used concentration—200 µM) statistically significantly reduced plasma protein carbonylation induced by H₂O₂/Fe²⁺ (inhibition of this process: about 25%). However, all tested food colorants decreased blood platelet protein carbonylation stimulated by H₂O₂/Fe²⁺, but their action was not always statistically significant. In addition, we noted that all used blue food colorants (1–200 µM) have protector effects on the change in the level of thiol groups in plasma proteins stimulated by H₂O₂/Fe²⁺, but these tested colorants change the level of thiol groups in blood platelets treated with H₂O₂/Fe²⁺ only at the highest used concentration—200 µM. In conclusion, the present study provides the first data on the antioxidant potential of genipin, brilliant blue FCF, and patent blue V in selected elements of blood treated with H₂O₂/Fe²⁺. Earlier and current studies have indicated the promising potential of these blue food colorants, especially genipin (without cytotoxicity toward human blood platelets), which can modify the oxidative stress of platelets and plasma in vitro at concentrations (1–200 µM) which can be obtained in blood during its administration. However, the presented results have limitations, especially concerning the mechanistic clarity surrounding the antioxidant properties of the tested blue food colorants. Therefore, further in vivo experiments are needed to provide a better understanding of their antioxidant potential. Full article

Background: Artemisia absinthium (A. absinthium) is a perennial plant valued for its antibacterial, antioxidant, and anti-inflammatory properties, exhibiting broader therapeutic potential. Given the need to deliver low doses of A. absinthium extract, mesoporous silica nanoparticles have attracted considerable attention as promising nanocarriers due to their distinctive physical and chemical properties. Methods: Physicochemical characterization of the materials was performed and biological assays were conducted to investigate the ROS, antibacterial and antioxidant activity of A. absinthium extract encapsulated within cerium- and calcium-doped mesoporous silica nanoparticles (MNSiCaCe) against both aerobic and anaerobic bacteria. Results: FTIR, SEM, and BET analysis confirmed successful synthesis of the MNSiCaCe. Phytochemical profiling of Artemisia absinthium extract using HPLC revealed the presence of artemisinin and a rich composition of phenolic and flavonoid constituents, with a total phenolic content of 182 ± 3.6 mg GAE/100 g dry plant material and a total flavonoid content of 42.5 ± 0.6 mg QE/100 g. Quantitative drug loading profiling demonstrated that while plain MNSi nanocarriers achieved a loading capacity of 16.96%, the MNSiCaCe enhanced this threshold to 43.11%. The in vitro controlled-release kinetics exhibited a highly prolonged and slow-release profile of the MNSiCaCe. The materials demonstrated excellent hemocompatibility and high mitochondrial activity with human periodontal ligament cells (hPDLCs). Elevated ROS generation was observed under conditions where antibacterial activity was most pronounced. While the artemisinin-doped nanoparticles showed notable antibacterial effects, the complete Artemisia absinthium-loaded nanoparticles achieved a significantly greater reduction in bacterial viability probably due to the synergistic interaction between artemisinin and the extract’s rich polyphenol profile. Conclusions: These findings highlight MNSiCaCe as a promising and safe nanocarrier system for drug delivery, with strong antibacterial potential, offering valuable applications in antibacterial therapies. Full article

The aim of this study was to evaluate the effect of adding white mulberry (Morus alba L.) fruit to extruded corn snacks on their polyphenol profile, antioxidant properties, acetylcholinesterase (AChE) inhibitory activity and the preservation of phenolic compounds in an in vitro digestion model. Mixtures of corn grits with 0, 10, 15 and 20% dried mulberry fruit were extruded at temperatures of 100, 120 and 140 °C, and then the total polyphenol content (TPC) and antioxidant activity (IC₅₀ for DPPH) were determined. For selected samples (0%, 140—3E; 15% mulberry, 140—9E; mulberry—13E), further antioxidant tests (FRAP, CUPRAC, Fe²⁺ chelation) were performed, the phenolic compound profile (UHPLC) and AChE inhibition were assessed, and a two-step in vitro digestion was conducted. The addition of mulberry significantly increased TPC- and free-radical-scavenging capacity compared to the control sample, with snacks containing 15% mulberry extruded at 140 °C showing approximately a 3.5-fold higher TPC than the control, while dried mulberry fruit itself exhibited about a five-fold higher TPC than this enriched snack. Among the snacks, the most favorable DPPH-radical-scavenging effect was obtained for the variant with 20% mulberry at 120 °C (IC₅₀ = 0.176 mg/mL), whereas the mulberry fruit extract reached an IC₅₀ of 0.0926 mg/mL. In a two-step in vitro digestion model, the mulberry-enriched snack with 15% fruit retained 69.3% of its initial TPC after the gastric phase and 33.3% after the intestinal phase, compared with 55.0% and 20.0%, respectively, for the control snack, confirming a partial but meaningful preservation of phenolic compounds under simulated gastrointestinal conditions. UHPLC analysis confirmed that mulberry and the enriched snacks are a rich source of chlorogenic acids and their isomers, as well as quercetin and kaempferol glycosides, which largely survived the two-step in vitro digestion, despite an observed decrease in TPC after the gastric stage and a further reduction after the intestinal stage. At the same time, mulberry extract and mulberry-enriched snacks exhibited high antioxidant activity in all tests conducted and in vitro AChE inhibitory activity, suggesting that Morus alba L. fruit has the potential to be used as a natural functional ingredient in the production of gluten-free snacks with antioxidant and potentially neuroprotective properties. Full article

Plant extracts in vegetable oils are foundational and eco-responsible for skin care, combining their emollient properties with other additional benefits, derived from their antioxidant, antimicrobial and UV-absorbing activity. The present research conducted a complex investigation of Usnea barbata extract in Karanja oil (KO), aiming for its further incorporation into various cosmetic formulations. The lichen extract (UBKO) was obtained through cold maceration. Phytochemical screening was performed using the Folin–Ciocalteu method and Graphite Furnace Atomic Absorption Spectrophotometry (GFAAS). Physicochemical properties were evaluated via Fourier Transform Infrared Spectroscopy (FTIR) and Atomic Force Microscopy (AFM). The rheological behavior and oxidative stability of the oil samples, UBKO and KO, were also investigated. UBKO had a slightly lower density (0.827 vs. 0.955) and pH (4.22 vs. 4.86) than KO, and a slightly higher oxidative resistance, quantified as the induction period (IP) value (6.45 vs. 6.00). The total phenolic-equivalent content (TPC, µg GAE/mL oil sample) was significantly greater in UBKO than in KO (567.16 ± 14.96 vs. 433.26 ± 22.96, p = 0.001). The values of minimum inhibitory concentration (MIC, mg/mL) indicated significantly higher antibacterial effect against S. aureus and antifungal effect against C. albicans for UBKO than KO (9.62 ± 2.87 vs. 31.25 ± 18.75, p = 0.049, and, respectively, 5.06 ± 1.68 vs. 37.50 ± 12.50, p = 0.01). Finally, our results showed that UBKO had an estimated sun-protective factor (SPF) of 30.9, slightly higher than 29.8 for the base oil formulation, KO; these findings represent baseline in vitro UV-absorbing trends. All of these results suggest that U. barbata extract in Karanja oil may exhibit complementary bioactive properties with potential applications in skincare. Full article

Chronic infected wounds are often characterized by persistent bacterial colonization, biofilm formation, excessive oxidative stress, and prolonged inflammation, which severely impair tissue regeneration. To address these challenges, a multifunctional wound dressing capable of antibacterial activity and microenvironment modulation was developed. In this study, amide-modified hyaluronic acid (HA-ADH) was used as the matrix, and a dynamic coordination network was constructed via Cu²⁺-hydrazide interactions to form an in situ HA-Cu hydrogel. Curcumin-loaded DSPE-PEG2000 micelles were further incorporated to obtain a pH-responsive composite hydrogel (HA-Cu/Cur). The prepared hydrogel exhibited a porous interconnected structure, along with favorable injectability, self-healing capability, tissue adhesiveness, moderate swelling, controllable degradability, and pH-responsive behavior under acidic conditions. In vitro antibacterial assays demonstrated that both HA-Cu and HA-Cu/Cur effectively inhibited the growth and biofilm formation of Escherichia coli and Staphylococcus aureus. The antibacterial activity was associated with disruption of bacterial morphology, depletion of intracellular ATP, and induction of reactive oxygen species, while HA-Cu/Cur showed enhanced performance in antibiofilm activity and oxidative stress-related effects compared with HA-Cu. Cytocompatibility studies revealed that the hydrogel extracts exhibited negligible cytotoxicity toward L929 fibroblasts and RAW 264.7 macrophages, while promoting fibroblast migration and significantly reducing the expression of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) in lipopolysaccharide-stimulated RAW 264.7 cells, with HA-Cu/Cur showing a more pronounced anti-inflammatory effect. In summary, the HA-Cu/Cur hydrogel integrates the antibacterial and pro-healing properties of Cu²⁺ with the antioxidant and anti-inflammatory activities of curcumin. The hydrogel effectively inhibited the growth and biofilm formation of both E. coli and S. aureus, reduced the expression of TNF-α, IL-6, and IL-1β in LPS-stimulated macrophages, and promoted fibroblast migration, demonstrating its potential as a multifunctional wound dressing for the management of infected wounds. Full article

Background/Objectives: Drug repurposing has emerged as a promising strategy for identifying novel anticancer agents among clinically established drugs. Fexofenadine, a second-generation H1 antihistamine, has been proposed as a candidate for repurposing in oncology; however, the molecular mechanisms underlying its biological activity remain insufficiently characterized. This study investigated the effects of fexofenadine on oxidative stress, mitochondrial function, apoptosis, and pro-survival signaling pathways in cervical and lung cancer cells. Methods: HeLa and A549 cancer cells, as well as non-tumorigenic Beas-2B epithelial cells, were exposed to fexofenadine under in vitro conditions. Cell viability, apoptosis, reactive oxygen species generation, mitochondrial membrane potential, DNA damage, autophagy-associated responses, and PI3K/AKT and MAPK/ERK pathway activation were assessed using flow cytometry, fluorescence microscopy, electron microscopy, and biochemical assays. Three-dimensional spheroid cultures and N-acetyl-L-cysteine rescue experiments were additionally employed to evaluate biological relevance and the contribution of oxidative stress. Results: Fexofenadine induced concentration-dependent accumulation of reactive oxygen species, mitochondrial membrane depolarization, Bcl-2 inactivation, caspase-3/7 activation, DNA damage, and apoptotic cell death in HeLa and A549 cells. Antioxidant pretreatment with N-acetyl-L-cysteine significantly reduced oxidative stress, attenuated mitochondrial dysfunction, and partially suppressed apoptosis. Fexofenadine was associated with reduced PI3K/AKT and MAPK/ERK pathway activation and promoted autophagy-associated responses. In three-dimensional spheroid cultures, treatment disrupted spheroid integrity and increased apoptotic cell death. Non-tumorigenic Beas-2B cells exhibited lower sensitivity to treatment than malignant cells. Conclusions: Fexofenadine disrupts redox homeostasis and is associated with reduced activation of pro-survival signaling pathways, resulting in oxidative stress-associated mitochondrial dysfunction and apoptosis in cancer cells. These findings provide mechanistic support for further evaluation of fexofenadine as a candidate for anticancer drug repurposing, while additional pharmacokinetic and in vivo studies are required to determine its translational relevance. Full article