Search Results (3,870)

The fate of heavy metals in agricultural soils is governed by organo-mineral–microbial interactions, which predictive models often fail to capture. The competitive sorption mechanisms of Cd(II) and Cu(II) on montmorillonite/kaolinite composites (Mont/Kao) functionalized with humic acid (HA) and Pseudomonas putida (P. p), a model system representative of contaminated agricultural soils, were investigated. Batch experiments, XRD, FTIR, and thermodynamic analysis reveal that metal retention is a non-additive function of competing interfacial processes. Bacterial biomass dominated sorption, accounting for >50% of total metal uptake, with capacity ranked as: P. p > Mont/Kao-P. p > Mont/Kao-HA-P. p > Mont/Kao-HA > Mont/Kao. Humic acid exerts a dual, concentration-dependent role: Low levels enhanced adsorption via mineral dispersion, while high levels induced surface masking, suppressing bacterial binding sites. Competition was highly asymmetric: Cd(II) reduced Cu(II) maximum adsorption capacity by 75.5% in the Mont/Kao-HA system by preferentially occupying montmorillonite interlayer sites, whereas Cu(II) inhibited Cd(II) below pH 6. Single-metal sorption was characterized by positive ΔS° (32.96–58.89 J·mol⁻¹·K⁻¹), indicative of inner-sphere complexation, while negative ΔS° under competitive conditions signals a transition to outer-sphere complexation. This work provides mechanistic insights into site masking, competitive displacement, and ternary cation bridging controlling metal immobilization in organo-mineral assemblages. Full article

The development of nanoparticle (NP)-based drug delivery systems that combine passive tumor targeting, physiological stability, and therapeutic efficacy remains a key challenge in cancer nanomedicine. Here, we report a pH-responsive peptide-functionalized poly(lactic-co-glycolic acid) (PLGA) NP system designed for cancer targeting. The PLGA core is coated with a short glutamic acid–lysine–histidine–phenylalanine x3 (EKHFFF) peptide shell, enabling tunable surface charge modulation around its isoelectric point and promoting environmental responsiveness. Physicochemical characterization confirms spherical NPs (~70–75 nm) with good colloidal stability, serum compatibility, and ion-dependent stability in physiological conditions. The peptide coating also provides pH-dependent modulation of the zeta potential. Evaluation of the NPs in ovarian cancer (OvCA) models, including immortalized and patient-derived cell lines (PDCLs), demonstrates efficient uptake across OvCA cell lines, with significantly enhanced internalization in PDCLs compared to immortalized cells. The EKHFFF nanoparticle (EKHFFF NP) induced minimal reactive oxygen species and nitric oxide production in macrophages, indicating low immunogenicity and favorable biocompatibility. Upon platinum loading (EKHFFF-Pt NP), the system exhibits potent cytotoxicity in both platinum-sensitive and platinum-resistant OvCA cell lines, outperforming carboplatin and showing comparable or improved efficacy relative to cisplatin in several cell lines. In vivo studies further demonstrate preferential tumor accumulation, sustained intratumoral retention, and measurable systemic circulation with a half-life of approximately 35 min. Full article

Cadmium (Cd) is a highly toxic non-essential heavy metal that severely impairs plant growth, compromises crop yield and quality, and threatens food safety and human health. Jasmonic acid (JA) and salicylic acid (SA) are well-characterized endogenous phytohormones that serve as central regulators in modulating plant adaptive responses to Cd stress. This review comprehensively synthesizes current knowledge on the involvement of JA and SA in regulating Cd accumulation and tolerance in plants, including their Cd-induced biosynthetic dynamics and signaling transduction pathways, as well as their functions in restricting Cd uptake and translocation, modulating chelation and sequestration, reinforcing antioxidant defense systems and protecting photosynthetic apparatus. Moreover, we analyze the antagonistic and synergistic crosstalk between JA and SA, and discuss how this interplay shapes Cd resilience in plants. Finally, the application potential of JA and SA in developing Cd-safe crops and phytoremediation in Cd-contaminated farmland is explored. This review provides a systematic analysis of the regulatory roles of JA and SA in plant responses to Cd stress, along with an in-depth discussion of their crosstalk. These insights contribute to the rational development of hormone-based breeding strategies for Cd-safe crops and the optimization of agronomic management practices. Full article

Background: Pathogenic variants in SLC6A1, which encodes the γ-aminobutyric acid (GABA) transporter GAT-1, cause developmental and epileptic encephalopathies (DEEs) through reduced GABA uptake, impaired transporter trafficking, and functional haploinsufficiency. 4-phenylbutyrate (PBA) is a clinically available small molecule with chemical-chaperone and histone-deacetylase-inhibitor activities that can rescue misfolded GABAergic proteins, but variant-level rescue data are needed to guide precision treatment. Methods: We report a novel de novo missense mutation p.Ala305Val in GAT-1 encoding SLC6A1, in a patient with myoclonic-atonic epilepsy and a developmental and epileptic encephalopathy phenotype. Ala305Val was compared with the residue-matched comparator p.Ala305Thr (Ala305Thr). Variant effects were evaluated by (i) protein-structure prediction across nine stability-prediction algorithms using the cryo-EM-derived human GAT-1 template (PDB 7Y7W); (ii) 3H-GABA uptake assays in HEK293T cells and in human iPSC-derived astrocytes and cortical neurons; (iii) live-cell confocal microscopy of ER colocalization; (iv) pharmacologic rescue with PBA, TUDCA and salubrinal (v) and GAT-1 cDNA gene-augmentation, alone and in combination with PBA. Results: AI-based stability predictors uniformly indicated destabilization of GAT-1 p.Ala305Val and GAT-1 p.Ala305Thr. GAT-1 p.Ala305Val reduced 3H GABA uptake across HEK293Ts, astrocytes, and neurons. The mutant transporter accumulated within the endoplasmic reticulum (ER), with ER colocalization rising from approximately 30% in wildtype to ~80% in GAT-1 p.Ala305Val; PBA reduced ER retention to approximately ~40% and restored total GAT-1 fluorescence toward wildtype levels. Pharmacochaperones (PBA, TUDCA) restored GABA uptake for the mutant transporters. Wildtype GAT-1 gene augmentation improved GABA uptake in the heterozygous condition but combined PBA plus wildtype allele augmentation produced rescue greater than either intervention alone in the available dose-response ranges. Conclusions: GAT-1 p.Ala305Val is a trafficking-impaired, loss-of-function variant whose dysfunction is amenable to two convergent therapeutic axes: pharmacologic correction of folding and trafficking, and augmentation of functional transporter expression. These findings support a two-pronged precision-medicine framework for SLC6A1-related DEEs in which PBA increased the transporter function augmented by genetic approaches. Full article

Breast cancer cells exhibit a reversed pH gradient and metabolic plasticity that promote proliferation, invasion, and resistance to therapy. Natural products such as chlorogenic acid (CGA) and cinnamaldehyde (CA) have shown emerging anticancer potential. However, their effects on intracellular pH and metabolic transport systems remain undefined. Therefore, the aim of this study was to characterize these parameters in breast cancer and non-tumorigenic breast cells. This study evaluated the physiochemical properties of CGA and CA using LC–MS, under pH conditions (pH 1.2, 7.4, and 9.0) mimicking the gastrointestinal track (GIT). Additionally, LC–MS-based human liver microsome (HLM) assays with NADPH were used to evaluate susceptibility to CYP-mediated metabolism to evaluate first-pass metabolic stability. Intracellular uptake kinetics were quantified at multiple time points using LC–MS. Following CGA:CA treatment, intracellular pH (pH_i) was measured in cancerous MDA-MB-231 and non-tumorigenic MCF-10A breast cell lines using SNARF-1 targeted ratio-metric fluorescence approach. Expression of OATP1B1, GLUT1, and MCT1 were analyzed by Western and immunofluorescence respectively, to assess potential cellular uptake of CGA:CA through OATP1B1 and their effects on glucose uptake and lactate and proton transport. Physiochemical results demonstrated that the compounds ranged from fully stable (pH 1.2 and 7.4) to completely unstable (pH 9.0). HLM incubation indicated no CYP-mediated hepatic metabolism. Treatment results showed that there was rapid intracellular uptake of CGA and CA in cancer cells and that CGA:CA lowered pH_i in both MDA-MB-231 and MCF-7 cells, while pH_i remained mostly unchanged in MCF-10A cells. Protein analysis revealed that CGA:CA treatment downregulated GLUT1 and MCT1 expression in cancer cells, suggesting impaired glycolytic activity and lactate shuttling. OATP1B1 expression was significantly suppressed in cancer cells, suggesting feedback inhibition of the solute carrier protein. Collectively, these findings indicate that CGA and CA exhibit favorable biochemical stability and disrupt intracellular pH regulation and metabolic transporter expression in breast cancer cells. Importantly, normal cells are not significantly affected. Thus, CGA:CA demonstrates therapeutic potential for breast cancer through pH_i and metabolic modulation. Full article

Immunotherapy can reduce treatment-related side effects but shows limited efficacy in “cold tumors,” whose immunosuppressive tumor immune microenvironment is characterized by abundant M2 macrophages and poor T cell infiltration. Because biopsy-based qualitative assessment of the tumor microenvironment is invasive and conventional imaging lacks functional information, this study aimed to develop an M2 macrophage-targeted theranostic agent enabling non-invasive photoacoustic (PA) imaging and pH-triggered cytotoxicity. A pullulan-based nanogel conjugated with mannose and near-infrared dye (IR-820) was further functionalized with the pH-responsive doxorubicin (DOX) prodrug, Aldoxorubicin, to develop Pullulan-mannose-IR820-Aldoxorubicin (PMID) nanogel. PMID was successfully synthesized, and the resulting self-assembled nanogels (<100 nm) exhibited a highly negative ζ-potential, near-infrared absorption peaks at 780 and 850 nm, and PA contrast comparable to IR-820 at 850 nm excitation. Dialysis studies demonstrated suppressed drug release at neutral pH (~20%) but accelerated release under acidic conditions, reaching ~80% within 48 h at pH 5.5, consistent with hydrazone hydrolysis and supporting tumor/lysosome-activated delivery. In RAW264.7 macrophages, PMID nanogel showed preferential uptake by M2-poralized versus M1-polarized macrophages, outperforming non-mannosylated PID nanogel and IR-820, and produced the strongest PA signal in M2 macrophage pellets. PMID nanogel also induced the highest concentration-dependent cytotoxicity in M2 macrophages, and microscopy indicated lysosomal accumulation of the nanogel with partial nuclear localization of released DOX. These findings support the use of PMID nanogel as M2 macrophage-targeted PA contrast agents and pH-responsive drug carriers with the potential to deplete immunosuppressive macrophages, modulate cold tumor microenvironments, and improve precision cancer theranostics. Full article

Metabolic dysfunction-associated fatty liver disease (MAFLD) is the most prevalent chronic liver disease worldwide, affecting 37–52% of the population and potentially progressing to cirrhosis and hepatocellular carcinoma. Effective treatments remain limited, making nutraceuticals such as flavonoids promising therapeutic alternatives. Opuntia ficus-indica (L.) (OFI) has demonstrated beneficial effects against MAFLD, although its active compounds remain unclear. In this study, the complexity of the OFI sample was addressed through fractionation by sequential exhaustive extraction (SEE), resulting in six fractions enriched with different compound families, and metabolic markers related to steatosis were evaluated. Butanol (BT), ethanol (ET), and water (WA) extracts reduced lipid accumulation by 18–21%. BT and WA fractions also improved glucose uptake and decreased ketone body and reactive oxygen species production. UPLC-MS analysis showed that the WA extract was rich in piscidic acid and exhibited the highest antioxidant activity, whereas the BT displayed the strongest antisteatotic effect. Glycosylated flavonoids, representing 78% of the BT, were the predominant compounds. Isorhamnetin-glucosyl-rhamnosyl-rhamnoside and isorhamnetin-glucosyl-pentoside were inversely associated with triglyceride release, oxidative stress, and ketone body production. These findings identify isorhamnetin derivatives as key bioactive compounds underlying OFI’s beneficial effects against hepatic steatosis. Full article

Metabolic syndrome (MetS) is a clinical condition defined by the coexistence of interconnected cardiometabolic risk factors, including central obesity, dyslipidemia, elevated blood pressure, and impaired glucose regulation, which collectively increase the risk of type 2 diabetes mellitus and cardiovascular disease. Beyond these clinical diagnostic features, MetS is characterized by complex pathophysiological alterations involving systemic dysregulation of metabolic signaling across adipose tissue, skeletal muscle, liver, vascular endothelium, and the immune system. Key molecular alterations include impaired insulin receptor substrate (IRS)–Akt signaling, chronic nuclear factor kappa B (NF-κB) activation, mitochondrial dysfunction, and oxidative stress. Physical exercise is recognized as a pleiotropic biomedical intervention capable of restoring metabolic homeostasis through coordinated modulation of intracellular signaling pathways and inter-organ communication. Exercise activates AMP-activated protein kinase (AMPK), enhances peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α)-mediated mitochondrial biogenesis, and stimulates nuclear factor erythroid 2-related factor 2 (Nrf2)-dependent antioxidant responses. These adaptations improve glucose uptake, enhance fatty acid oxidation, and reduce ectopic lipid accumulation across metabolically active tissues. At the systemic level, skeletal muscle functions as an endocrine organ by releasing myokines such as irisin, interleukin-6 (IL-6), and fibroblast growth factor 21 (FGF21), which contribute to metabolic regulation across the liver, adipose tissue, and vasculature. These exercise-induced signals promote immune modulation, reduce pro-inflammatory cytokine production, and improve endothelial function. Different exercise modalities including aerobic, resistance, and high-intensity interval training (HIIT) activate both common and modality-specific molecular pathways, supporting individualized exercise strategies. Collectively, exercise targets the multi-organ pathophysiology of MetS and provides a mechanistic foundation for precision exercise medicine in cardiometabolic disease management. Full article

Mesenchymal stem/stromal cells (MSCs) are promising candidates in regenerative medicine, but their effectiveness is significantly influenced by the surrounding metabolic and nutritional conditions. Increasing evidence suggests that lipids act not only as energy sources but also as regulators of MSC fate. This review explores how lipid metabolism influences the balance among stemness, immunomodulation, and differentiation into adipogenic or osteogenic lineages. It does so through mechanisms such as fatty acid uptake, β-oxidation, de novo lipogenesis, and membrane remodeling, all orchestrated by CD36, carnitine palmitoyltransferase 1A, PPARγ, AMP-activated protein kinase, and the PI3K/AKT/mTOR pathway. We then examine how diet reshapes the MSC lipidome: obesity and high-fat diets promote adipogenesis and senescence, while omega-3 fatty acids, caloric restriction, micronutrients, and a balanced microbiota help preserve regenerative capacity. Lastly, we discuss how combining lipidomics with multi-omics could uncover lipid-metabolic signatures and regulatory nodes that connect diet to MSC function. Overall, the diet–lipid–MSC axis emerges as a modifiable determinant of MSC function. Full article

Selective recovery of gold from cyanide leach liquors is hindered by the co-dissolution of iron minerals that generate ferrocyanide complexes which strongly compete with [Au(CN)₂]⁻ at ion-exchange sorbents. Here, we investigate mixed-bed interpolymer systems (IPS) composed of a strong-acid sulfonated polystyrene–divinylbenzene cation exchanger (TC007, Na⁺ form) and a strong-base quaternary ammonium anion exchanger (AV-17-8, Cl⁻ form) as a charge-selective platform for gold cyanide recovery. IPS compositions spanning cation-to-anion molar ratios from 6:0 to 0:6 were evaluated in batch contact with binary model solutions containing 30 mg L⁻¹ each of [Au(CN)₂]⁻ and [Fe(CN)₆]⁴⁻ at pH 10 and 25 °C. The optimal 1:5 IPS achieved an [Au(CN)₂]⁻ extraction degree of 79.88% and a selectivity coefficient β = D_Au/D_Fe = 4.95 at 48 h, whereas the pure AV-17-8 anion exchanger (0:6) reached only 37.55% Au extraction at 48 h, following an atypical delayed-uptake kinetic profile rather than the rapid, near-quantitative capture expected of an unmodified strong-base resin. Sorption kinetics were best described by a pseudo-second-order model (R² = 0.9992), confirming ion exchange at quaternary ammonium sites as the dominant rate-controlling step, with a ~30-fold increase in k₂ for [Au(CN)₂]⁻ in the 1:5 IPS relative to AV-17-8 alone. FTIR spectroscopy and TGA-DSC revealed the incorporation of metal cyanide complexes into the IPS matrix, with diagnostic C≡N stretching bands at 2108.7 and 2034.1 cm⁻¹ and an additional thermal event at 200–280 °C. These findings establish compositionally tunable IPS based on commercially available resins as a charge-selective sorbent platform demonstrating a capacity to regenerate under single-cycle elution conditions for gold cyanide recovery from ferrocyanide-containing process streams while highlighting the need for further evaluation under industrial Fe:Au ratios. Full article

Aronia melanocarpa contains abundant phenolic compounds, including procyanidins, flavonols, flavan-3-ols, and phenolic acids, but its pronounced astringency limits its broader use in food applications. Catechol and other oxygen-donor motifs in plant phenolics are known to associate with Fe³⁺ and can support metal–phenolic assembly. In this study, ultrasound-assisted cellulase–pectinase extraction was used to recover an A. melanocarpa polyphenol extract, which was subsequently combined with Fe³⁺ to construct a plant-derived hair dye coating on keratin fibers. Under optimized extraction conditions, the total phenolic yield reached 82.315 mg gallic acid equivalents (GAE)/g dry fruit. A limited targeted LC–MS/MS panel was used to evaluate 12 selected non-anthocyanin phenolic analytes, including procyanidin dimers, a flavan-3-ol, flavonols, and phenolic acids; the contribution of the pH-sensitive anthocyanin fraction was not resolved. Among the tested metal ions, Fe³⁺ produced the strongest chromogenic response. The optimized L-cysteine pretreatment–Fe³⁺ mordanting–polyphenol dyeing sequence produced a ΔE* of 55.42. Adsorption experiments empirically described the uptake behavior of the polyphenol extract, whereas complementary surface, spectroscopic, elemental, diffraction, thermal, and wettability analyses were consistent with the presence of a relatively uniform Fe-containing polyphenol coating at the hair fiber interface. Neither the adsorption-model fits nor the individual characterization techniques uniquely resolved the underlying molecular mechanism. Preliminary HET-CAM, single-exposure dermal, and acute eye irritation assessments showed no obvious acute irritation under the tested conditions. An exploratory image-based analysis examined whether standardized hair tress photographs could approximate a predefined colorimetric score. Because only 30 independent original samples were available and no external validation was performed, this analysis was treated solely as a proof of concept. This laboratory proof-of-concept extends established Fe³⁺–polyphenol assembly chemistry to a compositionally complex A. melanocarpa extract for hair fiber coloration. Further work is required to clarify anthocyanin behavior, simplify the sequential protocol, and benchmark its performance against representative commercial hair dyes. Full article

Objectives: This study aimed to explore the cellular injury phenotypes induced by outer membrane vesicles (OMVs) from uropathogenic Escherichia coli (E. coli, UPEC) and Proteus mirabilis (P. mirabilis, P. m) in human renal tubular epithelial HK-2 cells in vitro. Introduction: In urological practice, UPEC and P. mirabilis are representative pathogens of uncomplicated and complicated UTIs, respectively. Both are Gram-negative bacteria, and a notable feature of these bacteria is their ability to secrete OMVs. OMVs are nanoscale vesicles containing lipopolysaccharides (LPS), phospholipids, peptidoglycan, nucleic acids, and various virulence factors such as proteases and toxins. OMVs serve as effective carriers, delivering these toxic substances to host cells, triggering inflammatory responses, cellular damage, and ultimately cell death. Methods: In this study, we cultured standard pathogenic strains of E. coli and P. mirabilis in lysogeny broth (LB) medium until they reached the logarithmic growth phase. OMVs were isolated and identified. The uptake of OMVs by HK-2 cells was observed in vitro, and we evaluated the cytotoxic effects of both types of OMVs by measuring cell viability, oxidative stress, membrane permeability, mitochondrial function, and organelle morphology. Results: The results demonstrated that OMVs from both bacterial strains were efficiently internalized by HK-2 cells. Both OMVs induced oxidative stress, decreased antioxidant capacity, disrupted membrane permeability, and damaged mitochondrial function, leading to cell injury. Interestingly, while the two types of OMVs caused comparable cytotoxicity in terms of cell viability, oxidative stress and mitochondrial membrane potential, they produced distinct patterns of mitochondrial ultrastructural injury: P. mirabilis OMVs primarily caused mitochondrial structural deformation and blurred cristae, while E. coli OMVs led to mitochondrial swelling, cristae breakage, and vacuolization. These findings provide phenotypic evidence and experimental clues for understanding OMV-associated renal tubular epithelial injury caused by different uropathogens. Full article

Cellulose acetate (CA) is a promising bio-derived membrane material for water treatment; however, its limited availability of active functional sites can restrict its affinity toward dissolved pollutants. In this study, a cellulose acetate membrane was fabricated by phase inversion and subsequently functionalized through novel direct graft copolymerization with bio-based itaconic acid (IA) using potassium persulfate (KPS) as an initiator in an aqueous medium. The grafting approach introduced carboxylic functional groups into the CA matrix, providing additional active sites for pollutant removal. The successful grafting was confirmed by UV–Vis and 1H NMR spectroscopy, while XRD indicated changes in the structural organization of the polymer matrix. SEM/EDS characterization further revealed morphological changes associated with grafting, and cross-sectional SEM showed the development of finger-like, continuous pore channels within the modified membrane. The effects of reaction time, IA concentration, and KPS concentration on the grafting percentage were systematically evaluated, with grafting increasing up to an optimum range before declining at excessive monomer or initiator concentrations. Thermal analysis demonstrated improved stability after grafting, with the 6.6% grafted CA-g-IA membrane exhibiting an initial decomposition temperature of 351 °C and a reduced weight loss of 85% at 500 °C, compared with 344 °C and 91%, respectively, for pristine CA. The 6.6% CA-g-IA membrane was subsequently evaluated for the removal of Cu(II) and methylene blue (MB) from aqueous solutions. Cu(II) uptake was strongly influenced by contact time, solution pH, initial concentration, and grafting percentage, with the highest performance observed around pH 6 and 240 min contact time. The membrane also maintained its Cu(II)-binding performance over four regeneration cycles following HNO₃ treatment. Overall, direct IA grafting provides a simple bio-based functionalization strategy for enhancing the pollutant-binding functionality of cellulose acetate membranes, demonstrating potential for the removal of metal ions and cationic dyes from contaminated water. Full article

Several studies have suggested potential mechanisms through which Sutherlandia frutescens exerts antidiabetic effects, yet much of its therapeutic potential remains unexplored. This study aimed to provide greater insights into the antidiabetic capabilities and limitations of S. frutescens through the use of a comprehensive in vitro screening platform incorporating target-directed assays with automated image cytometry and analysis, where relevant. The antidiabetic effects of a crude hot aqueous extract of S. frutescens were assessed across well-characterised targets representing prominent hallmarks of diabetes: postprandial hyperglycaemia, insulin sensitivity, β-cell dysfunction, chronic inflammation and adipose tissue dysfunction. The findings revealed S. frutescens as a multicomponent therapeutic, with its individual target activities being relatively subtle and a few appearing novel. Notably, however, S. frutescens inhibited multiple targets relevant to postprandial hyperglycaemia—including carbohydrate digestion (29.1% and 26.4% inhibition for sucrose and maltose, respectively, at 500 µg/mL), intestinal glucose uptake in Caco-2 cells (between 9.51% and 20.92% reduction from 100 to 500 µg/mL) and protein glycation (23.2% inhibition at 200 µg/mL)—activities not previously documented in vivo. Glucose consumption in C3A hepatocytes showed a dose-dependent increase in glucose consumption from 12.5 to 100 µg/mL. Glucose consumption was reduced in palmitic acid-induced insulin-resistant L6 skeletal muscle cells from 100% to 69.57%, while 50 µg/mL S. frutescens restored it to 97.84%. S. frutescens also enhanced INS-1 β-cell survival under oxidative stress at concentrations as low as 12.5 µg/mL. The integration of automated image cytometry and analysis provides a novel approach with which to characterise its antidiabetic properties and elucidate its potential molecular mechanisms. Overall, S. frutescens is shown to impact several interrelated mechanisms simultaneously, suggesting that the coordinated modulation of multiple pathways may contribute to its overall biological activity. Whether these effects are additive or synergistic remains to be determined. Full article

Background/Objectives: The buccal mucosa offers a promising non-invasive route for systemic drug delivery, particularly for hydrophilic compounds like captopril (CAP), which exhibit low permeability and are subject to gastrointestinal instability and first-pass metabolism. This study aimed to develop and characterize captopril-loaded, chitosan-based mucoadhesive buccal films with different permeation enhancers and to evaluate their physicochemical properties, drug release, cytocompatibility, and in vitro transport across a TR146 buccal epithelial cell model. Methods: Films were prepared by the solvent-casting method using chitosan as the film-forming polymer. Different enhancers were investigated, including organic acid salts of chitosan (ascorbate, citrate, and lactate) and chemical permeation enhancers (sodium lauryl sulfate, polyethylene glycol 400, Span 20, and EDTA). Results: The resulting films exhibited acceptable thickness, moisture content, appropriate mechanical properties, and good mucoadhesive strength. In vitro dissolution studies demonstrated rapid CAP release, with >50% released within 15 min and near-complete release by 180 min across all formulations. Cytotoxicity assessment via a Neutral Red uptake assay in TR146 cells confirmed high cell viability (>81%) after 4 h of exposure, indicating good biocompatibility. In vitro permeation experiments revealed that films prepared with chitosan ascorbate and chitosan lactate enhanced CAP transport compared to other formulations, achieving the highest flux and apparent permeability coefficients. Conclusions: These findings demonstrate that chitosan ascorbate and lactate salts effectively improve the buccal permeability of captopril while maintaining good film properties and biocompatibility. This work highlights the potential of chitosan ascorbate- and lactate-based mucoadhesive films as an efficient platform for the buccal delivery of CAP. Full article