Search Results (114)

More specific targeted drug delivery systems with low immunogenicity and toxicity are expected to increase the efficacy of therapeutic molecules. The extracellular nanovesicles have been introduced as natural delivery systems for therapeutic molecules. More recently, considerable interest has emerged in plant-derived exosomes and their natural commitment to deliver molecules of various origins. Acridine Orange (AO) is an acidophilic dye with a strong tumoricidal action following excitation with a light source at 466 nm, but its clinical use is limited by the potential systemic toxicity. In this study we investigated the ability of exosomes from Citrus Sinensis to be successfully uploaded with AO (Exo-AO). We also studied the ability of the exosomes to be uploaded into target cells, as compared to the free molecules. We found that AO was efficiently uploaded into exosomes through electroporation. In fact, Exo-AO entered into the target cells significantly better than the free molecules, leading to both a marked intracellular AO delivery into target cells and an increase in the cytotoxic effect after excitation under a fluorescence microscope. This study shows a promising new approach for a more effective and less toxic drug delivery through the use of plant-derived extracellular nanovesicles. Full article

Protease-activated receptor-1 (PAR-1) is a key regulator of mesenchymal stem cell (MSC) migration and tissue integration. Dabigatran, a direct thrombin inhibitor widely used as a non-vitamin K oral anticoagulant (NOAC), may affect PAR-1-mediated signaling pathways. This study investigated the effects of dabigatran on cell viability, apoptosis, and PAR-1 activity in adipose-derived MSCs (ADMSCs) in vitro. ADMSCs were exposed to five concentrations of dabigatran etexilate with thrombin activation. Cell viability was assessed using the MTT assay, apoptosis and morphological changes were evaluated via acridine orange/propidium iodide staining, and PAR-1 expression was analyzed by immunofluorescence. Results showed that high dabigatran concentration significantly reduced cell viability and induced apoptotic morphological changes. In contrast, lower, non-cytotoxic concentrations preserved normal fibroblastic morphology and maintained cell viability while reducing PAR-1 surface expression compared with thrombin-activated controls. These findings indicate that dabigatran at non-cytotoxic doses can modulate PAR-1 activity without compromising ADMSC survival. In conclusion, dabigatran influences MSC-related cellular functions beyond its anticoagulant properties. Full article

Pomegranate (Punica granatum L.) has attracted considerable attention for its anticancer potential; however, mechanistic studies employing bioactivity-guided fractions from geographically distinct landraces remain limited. Building on our previous report on the bioactivity and phytochemical profile of the Bidah pomegranate landrace, the present study applied bioactivity-guided fractionation to enrich biologically active constituents and investigate mitochondria-associated cellular responses in colorectal cancer cells (Caco-2 cells). A semi-polar fraction from Bidah pomegranate crude extract (B6) was evaluated for its antioxidant activity, cell viability, cell death morphology, mitochondrial membrane potential, transcriptional modulation of key regulatory genes, and phytochemical composition. High-performance liquid chromatography (HPLC) profiling of B6 revealed a chromatographic fingerprint with seven detectable peaks, including two major peaks at retention times of 7.577 and 8.602 min, together accounting for approximately 66% of the total chromatographic area, indicating the enrichment of major constituents. Consistent with this enrichment, the fraction exhibited potent DPPH radical scavenging activity at a microgram-range IC₅₀, suggesting the presence of redox-active phytochemicals. In cell-based assays, the fraction induced a dose-dependent reduction in metabolic viability, while acridine orange/propidium iodide (AO/PI) staining of Caco-2 cells revealed delayed, regulated cell death. JC-1 staining demonstrated a pronounced loss of mitochondrial membrane potential, consistent with early mitochondrial dysfunction. Gene expression analysis further revealed modulation of pro- and anti-apoptotic genes, alongside cell-cycle-associated and oxidative stress/inflammatory markers. Gas chromatography–mass spectrometry (GC–MS) profiling identified polyacetylenes, sterol derivatives, fatty acid esters, and terpenoids, providing chemical context for the observed mitochondrial perturbation. Collectively, the findings support a mitochondria-centered, regulated cell death response driven by a multi-component phytochemical matrix. This study advances mechanistic insight beyond crude extract analysis and highlights the sustainable biomedical value of the Bidah pomegranate landrace as an underutilized regional resource. Full article

A novel acridine-based fluorescent sensor (Sensor ANT) for the highly selective and sensitive detection of cyanide ions (CN⁻) was rationally designed and synthesized via the conjugation reaction of acridine-9-amine with 3-nitrophenyl isothiocyanate. The sensing mechanism is triggered by the specific interaction between exogenous CN⁻ and the hydrogen-bonding moieties within the sensor’s molecular framework, which induces a distinct fluorescence quenching response. Systematic titration experiments confirmed that Sensor ANT exhibits rapid response kinetics, excellent selectivity, and reliable qualitative/quantitative detection capabilities toward CN⁻. Complementary biocompatibility assays, including in vitro cellular imaging and in vivo zebrafish experiments, further verified the promising application potential of this sensor in practical and biological detection scenarios. The detection limit (DL) of Sensor ANT for CN⁻ was calculated to be 2.89 × 10⁻⁷ M, with a 1:1 binding stoichiometry and a binding constant of 1.95 × 10⁴ M⁻¹. These findings demonstrate that Sensor ANT represents a robust candidate for CN⁻ detection in environmental and biological systems. Full article

Background: Heterocycle compounds with acridine, quinoline, indole, and pyridine nuclei are potentially active for anticancer activity since they can promote inhibition of vital enzymes, decreasing cell survival after binding to biomolecules. However, unspecific biological interactions can result in unwanted effects, which should be defined during the synthesis and proposition of new molecules. Thus, the objective of this study was to investigate the biological and teratogenic effects of four nitrogen heterocycles proposed for anticancer therapy. Methods: Four 2-cyano-N-phenylacrylamine type derivatives containing acridine (3a), quinoline (3b), indole (3c), and pyridine (3d) nuclei were synthesized and characterized. They were evaluated for their ability to interact with DNA, physicochemical and pharmacokinetic predictions, in vitro and in silico methodologies, besides in vitro inhibition of the Topoisomerase IIα enzyme, antiproliferative activity in tumor and non-tumor cells, hemolytic activity with human erythrocytes, and in vivo toxicological studies with zebrafish embryos. Results: UV–vis absorption studies with ssDNA revealed different spectroscopic effects, with binding constants (Kb) ranging from 1.41 × 10⁵ to 6.46 × 10⁴ M⁻¹. The fluorescence quenching constant (Ksv) with ethidium bromide (EB) varied between 0.53 and 0.67 × 10³ M⁻¹. The compounds intercalated into DNA base pairs, a mechanism confirmed by molecular docking, with 3b (quinoline) showing the most substantial interaction. All derivatives exhibited antitopoisomerase IIα activity at 100 μM and were cytotoxic against MCF-7 and T47-D breast tumor cells, particularly against the more aggressive T47-D lineage. No hemolytic activity was observed in human erythrocytes. In vivo assays in zebrafish embryos showed no toxicological or cardiotoxic effects. However, all compounds altered superoxide dismutase (SOD) and catalase (CAT) enzymatic activity, requiring further studies on reactive oxygen species (ROS) generation to assess potential adverse effects. Furthermore, significant results were observed in the physicochemical and pharmacokinetic parameters of the synthesized compounds. Conclusions: The findings highlight the quinoline derivative (3b) as the most promising nitrogen heterocycle due to its antiproliferative activity and biomolecular interactions without adverse effects in zebrafish embryos, distinguishing it from clinically available agents. Full article

Background: The eco-friendly synthesis of silver nanoparticles (AgNPs) utilizing medicinal flora presents a viable strategy for the development of multifunctional agents exhibiting antimicrobial, antioxidant, anti-inflammatory, and anticancer properties. This investigation aims to elucidate the phytochemical composition of Calotropis gigantea and its contribution to the synthesis of CG-AgNPs that demonstrate efficacy against Helicobacter pylori and gastric cancer cell lines. Methods: The aqueous plant leaf extract of C. gigantea underwent comprehensive analysis via gas chromatography-mass spectrometry (GC-MS), identifying a total of 25 bioactive constituents, including oleic and oxalic acid derivatives. The fabrication and analysis of silver nanoparticles (AgNPs) were performed utilizing methodologies including ultraviolet-visible (UV–Vis) spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), high-resolution transmission electron microscopy (HR-TEM), dynamic light scattering (DLS), and assessments of zeta potential. Antibacterial efficacy was evaluated through methods including agar well diffusion, time-kill kinetics, and biofilm assays. The cytotoxic impact on AGS gastric cancer cells was investigated using MTT assays, DAPI staining, and acridine orange/ethidium bromide (AO/EtBr) staining techniques. The assessment of antioxidant potential was performed utilizing DPPH and ABTS assays. The anti-inflammatory properties were analyzed through protein denaturation and membrane stabilization tests. Results: CG-AgNPs exhibited a spherical morphology (11–17 nm) with commendable stability, denoted by using zeta potential analysis measurement of −30.2 mV. The antibacterial activity showed a significant inhibition zone of 16.00 ± 0.17 mm at a concentration of 50 µg/mL against H. pylori, in addition to notable biofilm disruption. The viability of AGS cells was reduced by 61% at a concentration of 100 micrograms per milliliter, with apoptosis being confirmed through relevant assays. The antioxidant potential varied from 18% to 83% (DPPH) and reached 74% (ABTS) at a concentration of 100 µg/mL. The anti-inflammatory assays indicated a BSA denaturation inhibition ranging from 45% to 80% and a membrane stabilization effect between 54% and 85%. Conclusions: CG-AgNPs exhibit substantial antibacterial, antioxidant, anti-inflammatory, and anticancer activities, underscoring their pharmaceutical potential, particularly for combating antibiotic-resistant pathogens and gastric malignancies. Full article

Background/Objectives: Triple-negative breast cancer (TNBC) lacks targeted therapies and has a poor prognosis. Wild ginseng (Panax ginseng) is traditionally valued for its medicinal properties, but its scarcity limits therapeutic application. Adventitious root culture technology provides a sustainable source of wild ginseng-derived bioactive compounds. This study investigated the anticancer effects of wild ginseng adventitious root extract (WGAR) on MDA-MB-231 TNBC cells and elucidated the underlying molecular mechanisms. Methods: WGAR was prepared from cultured adventitious roots of 100-year-old wild ginseng, and its chemical composition was analyzed by LC-MS/MS. Anticancer effects were evaluated using MTT assay, acridine orange/propidium iodide (AO/PI) staining, Matrigel invasion assay, Western blot analysis, and proteome profiler array. Molecular docking was performed to predict interactions between WGAR constituents and target proteins poly (ADP-ribose) polymerase (PARP)-1 and β-catenin. Results: LC-MS/MS analysis tentatively identified 17 compounds, including ginsenosides (Rg3, Rh1, Rf) and terpenoids (ursolic acid). WGAR reduced cell viability with an IC50 of 79 μg/mL at 48 h, inducing 51.2% cell death. WGAR activated the intrinsic apoptotic pathway through sequential caspase-9 and caspase-3 activation, followed by PARP cleavage, and was associated with changes in epithelial–mesenchymal transition (EMT)-related markers (reduced N-cadherin, Slug, and β-catenin) alongside decreased inhibitory Ser9 phosphorylation of GSK-3β. Proteome array analysis revealed suppression of ECM remodeling proteins (tenascin C, u-PA) and inflammatory mediators (IL-6, CXCL8). Molecular docking predicted that selected WGAR constituents, particularly terpenoid-type compounds, may potentially interact with PARP-1 and β-catenin; however, these in silico findings are hypothesis-generating and require experimental validation. Conclusions: WGAR exerts multi-target anticancer effects on TNBC cells through apoptosis induction and EMT suppression associated with modulation of GSK-3β/β-catenin signaling, suggesting its potential as a source of therapeutic agents for TNBC. Full article

Osteoarthritis (OA) is a progressive joint disorder affecting over 250 million people globally and is characterized by chronic pain and disability. Among its key pathogenic mechanisms are mitochondrial dysfunction and elevated reactive oxygen species (ROS), often triggered by inflammatory mediators such as lipopolysaccharide (LPS). This study evaluates the protective effects of curcumin on mitochondrial function, autophagy, and apoptosis in an in vitro model of OA. Human bone marrow-derived mesenchymal stem cells (BMSCs) were differentiated into chondrocytes using MesenCult™-ACF medium. Differentiation was confirmed by histological staining for Type II Collagen, Alcian Blue, and Toluidine Blue. LPS was used to induce an OA-like inflammatory response. Mitochondrial membrane potential (ΔΨm) was assessed using Rhodamine 123 staining. Autophagy and apoptosis were evaluated using Acridine orange and propidium iodide staining, respectively. Western blotting was performed to analyze the expression of pro-caspase-3, Bcl-2, Beclin-1, LC3-I/II, and GAPDH. LPS significantly impaired mitochondrial function, limited autophagy, and enhanced apoptotic signaling (reduced pro-caspase-3). Curcumin (25 µM and 100 µM) restored ΔΨm, increased Beclin-1 and LC3-II, and maintained pro-caspase-3 expression, with Bcl-2 showing a non-monotonic response (higher at 25 µM than at 100 µM). Curcumin exerted cytoprotective effects in inflamed chondrocytes by stabilizing ΔΨm, promoting autophagy, and attenuating apoptotic activation, supporting its multi-target therapeutic potential in OA. Full article

Background/Objectives: Male infertility contributes to nearly half of global infertility cases, yet conventional semen analysis is insufficient to predict assisted reproductive technology (ART) outcomes such as intracytoplasmic sperm injection (ICSI). Sperm DNA fragmentation (SDF) is a promising biomarker of genomic integrity, but clinical implementation remains hindered by methodological heterogeneity. This study compared two SDF assays—TUNEL and Acridine Orange (AO)—regarding their correlations with semen parameters and ICSI outcomes. Methods: Sixty men undergoing ICSI were prospectively enrolled. SDF was analyzed using two flow cytometric assays: TUNEL (detecting DNA strand breaks) and AO (assessing chromatin instability). Semen quality and ICSI outcomes (fertilization, cleavage, blastulation, and embryo utilization rates) were evaluated. Statistical analyses included Spearman’s rank correlation and Mann–Whitney tests. Results: Median SDF levels were significantly higher by TUNEL than AO (17.2% vs. 10.15%; p = 0.0065). Inter-assay correlation was weak (r = 0.299, p = 0.01). AO-derived SDF correlated positively with age (r = 0.311, p = 0.02), while TUNEL showed no such trend. Neither assay correlated significantly with semen parameters or ICSI outcomes, although AO tended to associate with lower motility and slightly reduced top-quality embryo rates. Conclusions: TUNEL and AO capture distinct facets of sperm DNA damage. Their limited correlation and lack of predictive value for ICSI outcomes highlight the need for assay-specific interpretation and standardization. Integration of SDF with additional biomarkers and oocyte factors may enhance its clinical utility. Full article

Charge-transfer-type fluorochromes, which exhibit shifts in fluorescence intensity and emission wavelength in response to solvent polarity changes, have been widely employed to investigate solute–solvent interactions. Humic substances (HSs) are naturally occurring macromolecular organic acids derived from plant residues, with structural properties that vary depending on their origin and environmental conditions. The polarity of HSs is closely associated with the mobility and toxicity of environmental pollutants, making their chemical characterization essential. In this study, we developed a rapid and straightforward method to characterize HS polarity using fluorescent solvatochromism. The fluorescence peak shifts of four dyes—8-anilino-1-naphthalenesulfonic acid (ANS), acridine orange (AO), methylene blue (MB), and Rhodamine B (RhB)—were evaluated in the presence of humic acids (HAs), a major component of HSs. To assess environmental variability, a total of twelve HS samples were tested, including HSs derived from soils of different origins, compost, commercial reagents, and standard reference materials. Among these, AO and MB exhibited distinct spectral shifts without overlapping with the intrinsic fluorescence of HAs. Notably, MB displayed a consistent blue shift dependent on HA concentration, with the most stable response observed at 5 mg/L. The magnitude of this shift was significantly correlated with UV–Vis parameters associated with the aromaticity, humification degree, and polarity of HSs. Overall, this study demonstrates that MB-based fluorescent solvatochromism can function as an empirical and facile indicator for assessing the structural and microenvironmental characteristics of HSs, providing a rapid and complementary screening approach for HSs extracted and purified from environmental samples. Full article

Background/Objectives: This study focused on synthesizing novel alkenyl derivatives of azinylferrocenes and evaluating their potential as Alzheimer’s disease (AD) therapeutics. Methods: 1-Azinyl-1′-acetylferrocenes were obtained by regioselective acetylation of azinylferrocenes, followed by the Wittig reaction or reduction of 1-azinyl-1′-acetylferrocenes and subsequent dehydration of the resulting alcohols. The synthesized compounds underwent the following biological activity testing relevant to AD: inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and off-target carboxylesterase (CES); antioxidant capacity (ABTS and FRAP assays); inhibition of Aβ₄₂ self-aggregation (thioflavin method); blocking AChE-induced β-amyloid aggregation (propidium displacement); and cytotoxicity in SH-SY5Y and MSC-Neu cells (MTT assay). Results: Quinoline and bipyridine derivatives demonstrated effective cholinesterase inhibition, especially quinoline 7b (AChE IC₅₀ 3.32 μM; BChE IC₅₀ 3.68 μM), while acridine derivatives were poor inhibitors. Quantum chemical (QC) calculations predicted that acridine derivatives were especially prone to form stable dimers. Molecular docking into protein targets generated by an AlphaFold3 reproduction code showed that these dimers were too bulky to access enzyme active sites, yet they could bind to protein surfaces to inhibit Aβ₄₂ self-aggregation and displace propidium from the AChE peripheral anionic site. All compounds showed high antioxidant activity in ABTS and FRAP assays, with quinoline derivatives being 2–4 times more potent than Trolox. QC calculations supported these findings. Quinoline and bipyridine derivatives also exhibited low cytotoxicity and scant CES inhibition. Conclusions: Overall, the synthesized ferrocenes, particularly the quinoline and bipyridine derivatives, appear promising for further research as multifunctional therapeutic agents targeting AD due to their anticholinesterase, antiaggregating, and antioxidant activities combined with low toxicity. Full article

Background/Objectives: Malaria is a tropical disease mainly caused by Plasmodium falciparum and represents a global public health problem, with over 200 million cases and 500 thousand deaths reported worldwide. Considering its treatment limitations, it is essential to develop new compounds against malaria. In this context, acridine derivatives are privileged structures. Methods: Thus, new spiroacridines containing N-acylhydrazone (AMTAC) and N-phenylacetamide (ACMD) were synthesized and evaluated in malaria and cytotoxicity assays, as well as in silico studies. Results: As a result, five spiroacridines showed inhibitory activity over 70% against the P. falciparum 3D7-GFP strain at 10 μM, along with an IC₅₀ range of 2–4 μM. After a brief Structure–Activity Relationship (SAR) analysis, it was observed that the spiroacridine structure must be associated with the hydrazone moiety to successfully inhibit parasite growth. In addition, these molecules presented promising resistance profile, with selectivity for the parasite. After computational studies, spiroacridines showed better affinity with dihydrofolate reductase (DHFR), overcoming the quadruple mutant resistance to pyrimethamine, with more stability in complex with the enzyme. Conclusions: Therefore, the potential of spiroacridines against malaria, with moderate resistance and selectivity profile, as well as DHFR inhibition greater than pyrimethamine, was confirmed. Full article

Alzheimer’s disease (AD) numbers among the most precipitative neurodegenerative disorders, constituting a constant subject of interest for medicinal chemistry researchers. The treatment of such disorders remains a challenge due to the complexity of their pathogenesis. Indeed, many factors are involved in the development of AD, including different enzymes such as acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), whose inhibition results in anti-AD activity, making the development of novel AChE/BuChE inhibitors a possible way to manage Alzheimer’s disease. Besides being a class of compounds with a wide spectrum of biological applications, acridine compounds also number among the most promising agents for the treatment of Alzheimer’s disease through cholinesterase inhibition. The best example to mention is the well-known anti-AD agent Tacrine. However, it was found to have a toxic effect on the liver, making the optimization of this acridine derivative a necessity. Our work was directed towards evaluating in silico a symmetrical acridine analogue as a potential cholinesterase inhibitor with controlled toxicity. A docking study was completed using Glide software (Schrodinger suites), and both AChE (pdb: 4EY6) and BuChE (4BDS) were utilized as drug targets. The molecular docking simulation resulted in satisfying docking score values alongside numerous significant interactions indicating the high stability of the investigated compound within the active sites of the enzymes studied. Additionally, ADMET prediction was carried out for the assessed acridine derivative in order to explore its drug likeness through its pharmacokinetics and toxicity profiles by employing the online servers SwissADME, MolSoft, and ProTox-II. Full article

This study explores the use of lignocellulosic Tectona grandis seeds (TGs), hydrochar (HC-230-4), and activated carbon (AC-850-5) produced via hydrothermal carbonization and followed by CO₂ activation for removing acridine yellow G (AYG) and acridine orange 14 (ABO) from water. HC-230-4 showed a rich presence of surface functional groups and irregular morphology with some sphere-like structures. In contrast, AC-850-5 exhibited a much higher surface area (729.7 m²/g), though with fewer surface functional groups than HC-230-4. The batch method was used to study the effects of contact time, pH, dye concentration, and temperature. Among the materials, AC-850-5 showed the highest adsorption capacity of 198 mg/g for AYG and 171 mg/g for ABO at 25 °C, around 12% higher than commercial activated carbon. The adsorption process was spontaneous and endothermic, fitting well to the Langmuir isotherm model, suggesting monolayer coverage. The adsorption kinetics followed the pseudo-second-order model, indicating that the rate depends on the surface site availability. Intraparticle diffusion analysis further confirmed a multi-step adsorption process. These findings show the strong potential of TG-derived activated carbon as an effective and sustainable material for removing acridine dyes from polluted water. Full article

The synthesis of heterocyclic compounds has gained significant attention in organic chemistry due to their diverse pharmacological properties. However, traditional synthetic approaches often involve hazardous chemicals, high energy consumption, and tedious workup procedures, leading to environmental concerns and low yields. In response, green chemistry strategies have emerged, emphasizing safer and more sustainable alternatives. Among these, magnetic nanoparticle (MNP)-based catalysts have shown remarkable promise in facilitating one-pot multicomponent reactions (MCRs), offering enhanced catalytic efficiency, ease of recovery, and reusability. This article provides a comprehensive overview of multicomponent reactions (MCRs) for the construction of a wide range of heterocyclic scaffolds—including chromenes, pyrazoles, phenazines, triazoles, tetrazoles, xanthenes, furans, indoles, imidazoles, pyridines, pyrimidines, oxazoles, and acridine derivatives—catalyzed by magnetic nanoparticles under sustainable and environmentally benign conditions. This review highlights recent advances (2018–2024) in the development and application of modified magnetic nanoparticles for green multicomponent synthesis. Emphasis is placed on their structural features, catalytic roles, and benefits in eco-friendly organic transformations. Full article