Search Results (3,163)

Glioblastoma (GBM) is one of the most aggressive human cancers, with therapeutic failure driven by pronounced intratumoral heterogeneity, microenvironmental plasticity, immune suppression, blood–brain barrier (BBB)-related pharmacological constraints, and adaptive resistance mechanisms. A major limitation in GBM research is the lack of a human-relevant experimental system able to reproduce these dynamic features while generating interpretable, multimodal datasets. In this context, we propose a testable organ-on-chip (OoC)-extracellular vesicle (EV)-deep learning (DL) framework in which patient-derived GBM cells, endothelial cells, astrocytes, pericytes, stromal cells, and immune components are organized within perfused microphysiological systems. EVs are selectively and temporally harvested from defined compartments, and imaging, barrier-function, sensor, and EV-cargo data are integrated through modality-specific and multimodal DL architectures. This framework is intended not as an immediately validated clinical tool but as an experimental roadmap for linking EV-mediated communication to measurable phenotypes such as BBB disruption, invasion, immune reprogramming, and drug response. We critically discuss the technical requirements of BBB-on-chip systems, EV source attribution, immune-component integration, DL model selection, data scarcity, overfitting, batch effects, domain shift, regulatory barriers, cost, throughput, and reproducibility. By repositioning OoC-EV-DL integration as a staged translational strategy rather than a clinically established solution, this work aims to define a realistic and biologically grounded route for advancing precision oncology in GBM. Full article

Background: Precise implant placement is crucial during inferior alveolar nerve (IAN) bypass in the posterior mandible where bone height above the IAN is limited. This in vitro study compared the accuracy of static computer-assisted implant surgery (sCAIS) and dynamic computer-assisted implant surgery (dCAIS) for implant placement during IAN bypass. Methods: Two cone-beam computed tomography (CBCT) mandibular models with deficient bone height (<7 mm) above the IAN canal, classified as clinical scenario I and clinical scenario II, were used as an in vitro setting. Thirty models per clinical scenario were prepared, after which 60 dental implants were placed in the edentulous area of tooth no. 47. Software-based analysis compared planned and actual implant placements by postoperative CBCT. The two models were compared for deviation in distance to the inferior alveolar nerve (DIAN), entry-3D deviation, entry-2D deviation, apex-3D deviation, apex-vertical deviation, and angular deviation by comparative statistical analysis. Results: Both sCAIS and dCAIS showed less deviation from planned implant position in both scenarios. No statistically significant differences were detected except for angular deviation (sCAIS: 1.73° vs. dCAIS: 1.19°, p = 0.004), including clinical scenario I (sCAIS: 1.65° vs. dCAIS: 1.19°, p = 0.033) and II (sCAIS: 1.98° vs. dCAIS: 1.2°, p = 0.033). Conclusions: Both approaches showed minor deviation in both IAN bypass models, while dCAIS showed better angular control, requiring future in vitro and in vivo research in complex clinical environments. Full article

Objectives: A customizable 3D-bioprinted core-in-shell platform was developed for time-dependent oral colon delivery of live microorganisms. The system conveyed Lacticaseibacillus paracasei as a model bacterial species within a monolithic core, which was surrounded by a swellable hydroxypropyl cellulose barrier, imparting a lag phase of programmable duration, and by an enteric outer layer, protecting the dosage form during unpredictable gastric residence. Methods: Pastes of different compositions were investigated to shape the core. Core and core-in-shell units were fabricated from digital models using a bioprinter equipped with a high-precision plunger dispenser and pressure-based thermoplastic printhead. The printed units were characterized in terms of mass, dimensions, mechanical properties and release performance using paracetamol as a reference tracer. Bacterial viability was evaluated during screening of the formulation components and after each processing step by manual counting of colony-forming units. Results: A mannitol-based formulation was selected for fabrication of the core, offering a favorable balance of printability, physico-technological properties, release behavior and ability to preserve bacterial viability. Two-layer core-in-shell systems were manufactured via a dual-printing operating mode. The desired in vitro performance was attained, with no release under acidic conditions, a lag phase in pH 6.8 fluid and a subsequent release profile comparable with that generated by the core as such. Viability studies demonstrated that compounding, core printing, shell deposition and drying did not adversely affect L. paracasei survival. Conclusions: 3D bioprinting was proved to be a versatile technique for the manufacturing of oral colon delivery systems containing probiotics or live biotherapeutics. Full article

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

In this study, we evaluated the impact of different in vitro 3D culture modelling methods on the activity of doxorubicin (DOX) and 5-fluorouracil (5-FU) in human melanoma spheroids. Human melanoma A375 and IGR39 spheroids were generated using the hanging drop and non-adhesive surface methods. Spheroid growth dynamics were assessed by measuring changes in spheroid diameter. To compare the effects of anticancer drugs in spheroids of different sizes, spheroids of approximately 200 and 400 µm were formed. Drug activity was evaluated based on spheroid growth and cell viability using the MTT assay. A375 spheroids formed using the non-adhesive surface method were more sensitive to DOX than spheroids formed using the hanging drop method. In smaller A375 spheroids, 10 µM 5-FU reduced cell viability more effectively in spheroids formed using the hanging drop method. In contrast, IGR39 spheroids formed by the hanging drop method were more resistant than those formed on a non-adhesive surface. However, in IGR39 spheroids, the effects of DOX and 5-FU on growth and viability did not significantly differ between formation methods. In conclusion, A375 spheroid growth was not significantly influenced by the formation method, whereas IGR39 spheroid growth depended on the method used. A375 spheroids formed on non-adhesive surfaces were more sensitive to DOX, whereas 5-FU activity depended on drug concentration and spheroid size. In IGR39 spheroids, the effects of DOX and 5-FU on growth and viability were largely independent of the spheroid formation method. Full article

The accuracy of full-arch digital impressions remains a topic of debate despite advancements in intraoral scanning technologies. This in vitro study aimed to evaluate the effect of different intraoral scanner generations (TRIOS 3, TRIOS 5, and TRIOS 6) on full-arch digital impression accuracy by comparing dentate and fully prepared models. A dentate and a fully prepared maxillary model were digitized using a high-accuracy desktop scanner to create reference datasets. The models were fabricated with a 3D printer, and a total of 60 digital impressions were obtained, with 10 repeated scans for each scanner–model combination. Accuracy was assessed by comparing STL datasets with the reference models using root mean square (RMS) deviation values. The results showed that TRIOS 3 and TRIOS 5 demonstrated similar accuracy performance across both model types, whereas TRIOS 6 exhibited higher deviation values, particularly in the fully prepared model. Furthermore, model type had a significant effect on accuracy, and a statistically significant interaction was observed between scanner type and model type. These findings indicate that digital impression accuracy is influenced not only by scanner generation but also by surface morphology and reference characteristics. Full article

The number of patients who require endoprosthetic treatment related to osteoporosis has increased in recent years. Vitamin D3 supplementation has long been standard practice in osteoporosis treatment, while vitamin K2 has gained importance. Using our in vitro model, we compared the osteogenic behaviour of primary healthy human osteoblasts (hOBs) and primary osteoporotic human osteoblasts (hopOBs) under unchanged conditions, with vitamin K2, vitamin D3 and the combined addition. Fluorescence microscopy examinations on a plastic surface and a rough titanium surface structure revealed morphological differences. A quantitative analysis of mineralisation and differentiation was performed using an alizarin red S assay and analysis of alkaline phosphatase activity. It was shown that the hopOBs behave differently morphologically on the titanium surface, while hopOBs are particularly noticeable due to the higher number of cell–cell interactions with vitamin K2. The rough surface led to more pronounced mineralisation of the hopOBs. This effect was pronounced under vitamin K2. Vitamin D3 had an effect in the initial phase of hopOB differentiation. Overall, vitamin K2 had a greater influence on the mineralisation of hopOBs than expected. It must be assumed that vitamin K2 plays a significantly greater role in the metabolism of hopOBs than previously assumed. Full article

Amphotericin B (AmB) is a broad-spectrum antifungal agent and a long-standing standard of care; however, its clinical use is compromised by poor solubility, off-target tissue distribution and severe dose-limiting toxicity. AmB-loaded poly (D, L-lactide-co-glycolide) nanoparticles (PLGA-AmB-NPs) were developed and characterized with respect to their physicochemical properties, antifungal activity against A. fumigatus biofilms, in vivo efficacy in the Galleria mellonella infection model, and hemolytic toxicity in vitro. Blank PLGA nanoparticles (PLGA-NPs) and PLGA-AmB-NPs exhibited mean diameters of 165 nm (PDI 0.075) and 120 nm (PDI 0.210), respectively, with negative zeta potential values consistent with colloidal stability in aqueous media. PLGA-AmB-NPs showed significantly enhanced activity against 24 h A. fumigatus biofilms compared with free AmB at concentrations of 5 and 10 µg/mL, while unloaded PLGA-NPs were inactive. Infected G. mellonella larvae treated with PLGA-AmB-NPs displayed markedly improved survival over a 5-day period relative to those receiving equivalent doses of free AmB. Furthermore, PLGA-AmB-NPs induced substantially lower hemolysis of human red blood cells than free AmB across all tested concentrations (5–20 µg/mL). PLGA-AmB-NPs represent a promising polymeric platform for the treatment of A. fumigatus infections. Full article

Irradiation (IR) can cause intestinal epithelial cell death, damage to crypt stem cells, and mucosal barrier dysfunction, which are the features of radiation-induced intestinal injury (RIII). Our study first discovered a natural small-molecule alkaloid Orychophragine D (OV16) with an obvious radiation protection effect. This study aims to investigate the radiation protection effect of OV16 on RIII and its potential molecular mechanism. The results showed that in vitro OV16 exhibited a significant protective effect on an irradiated human small intestinal epithelial cell-6 (HIEC-6) model. Then, transglutaminase 2 (TGM2), which is the key protein for OV16 to exert its anti-RIII protective effect, was identified as a crucial cellular target of OV16 using drug affinity responsive target stability (DARTS), molecular docking, molecular dynamics simulation, cell thermal shift assay (CETSA), and microscale thermophoresis (MST). Moreover, OV16 can upregulate the expression level of TGM2 in the nucleus of HIEC-6. TGM2 can reduce radiation-induced damage by enhancing the proliferation ability and migration ability of HIEC-6 and reducing the generation of γ-H2AX. Collectively, our study first identified TGM2 as a previously unreported therapeutic target for RIII, and provided a future drug design direction for TGM2 allosteric activators using OV16 as a novel molecular template. Full article

Cyathula officinalis Kuan, a medicinal plant used in traditional medicine, remains underexplored as a source of structurally defined wound-repair polysaccharides. In this study, a water-soluble polysaccharide fraction, CoPS, was isolated from C. officinalis roots and structurally characterized using methylation analysis and 1D/2D NMR spectroscopy. Purified CoPS had a total carbohydrate content of 94.8%, a weight-average molecular weight (M_w) of 7.491 kDa, and a narrow dispersity (M_w/M_n = 1.04). Monosaccharide composition analysis showed that CoPS was mainly composed of fructose and glucose at a molar ratio of 95.60:4.40. Structural analyses identified CoPS as a branched levan-type fructan with a β-(2→6)-linked fructofuranosyl backbone and β-(2→1)-linked branching motifs. CoPS was incorporated into a Carbomer/alginate topical formulation, termed CoPS-CPG, and evaluated in vitro and in vivo. CoPS-CPG showed good cytocompatibility and promoted HaCaT keratinocyte migration, reducing the residual scratch area to 48.10% at 12 h compared with 70.13% in the control group and 65.18% in the vehicle (Blank-CPG) group. In a murine full-thickness excisional wound model, CoPS-CPG reduced the residual wound area to 8.70 ± 1.20% on day 14, compared with 24.83 ± 1.51% in the control group and 14.20 ± 0.72% in the vehicle group. Histological and immunological analyses further indicated improved tissue reconstruction, a reduced inflammatory burden, enhanced CD206-associated macrophage signals, increased CD31-associated vascular structures, improved α-SMA-associated perivascular coverage, and lower late-stage HIF-1α expression. These findings identify CoPS as a structurally defined plant-derived levan-type fructan that supports cutaneous wound repair. Full article

Aloysia triphylla (lemon verbena, LV) herbal tea is a rich source of phenolic compounds with recognized antioxidant and antiglycoxidant properties, although their stability during digestion remains insufficiently understood. This study evaluated the impact of simulated gastrointestinal digestion on the phenolic composition and bioactivity of LV infusion using a standardized in vitro model. Total phenolic, flavonoid, and phenolic acid contents were determined spectrophotometrically, while individual compounds were analyzed by HPLC. Antioxidant activity was assessed using complementary assays (DPPH•, ABTS•, FRAP, ORAC, and nitric oxide scavenging), and antiglycation activity was evaluated using a BSA/D-ribose model. Digestion did not significantly affect total phenolic and phenolic acid contents, whereas flavonoids moderately decreased during the intestinal phase. Verbascoside underwent partial degradation, leading to increased levels of isoverbascoside and caffeic acid. Despite these transformations, antioxidant capacity was maintained or enhanced, particularly in ABTS and ORAC assays, suggesting a contribution of digestion-derived metabolites. Antiglycation activity remained stable after digestion. Pre-column HPLC analyses identified verbascoside and its derivatives as the main contributors to radical scavenging and methylglyoxal trapping activities. These findings indicate that LV infusion retains its bioactive potential after digestion and supports its relevance as a functional beverage. Full article

The intracellular accumulation of amyloid beta 42 (iAβ42) has been proposed as an early pathological indicator of familial Alzheimer’s disease (FAD). DJ-1 is a multifunctional protein sensitive to oxidative stress (OS) that has been associated with neurodegeneration; however, its role in iAβ42 pathology is unclear. In this study, we examined whether oxidized (sulfonic) DJ-1 (Cys¹⁰⁶-SO₃H) drives iAβ42 accumulation using postmortem brain samples and in vitro 3D iPSC-derived cerebral organoids (COs) or 2D induced pluripotent stem cells (iPSC)-derived ChLNs (cholinergic-like neurons) models from a PSEN1 E280A patient and a healthy volunteer (as a control sample). Post-mortem analyses of the temporal and frontal cortices and hippocampus from FAD PSEN1 E280A patients revealed strong intracellular co-localization of sulfonic DJ-1 and iAβ42, which was absent in control samples. To validate these findings, we generated COs from an iPSC PSEN1 E280A FAD patient and a healthy donor. In these organoids, we observed the co-localization of oxidized DJ-1 and Aβ42 in the absence of extracellular fibrils or plaques, as confirmed by BTA-1 staining. To further support these observations, 2D iPSC PSEN1 E280A-derived ChLNs cultures showed that intracellular Aβ42 accumulates progressively in direct correlation with increasing DJ-1 oxidation, as demonstrated by immunofluorescence microscopy and Western blotting analysis. These results indicate that DJ-1 oxidation accompanies the earliest intracellular stages of Aβ42 pathology. Furthermore, complementary in silico molecular docking analysis revealed a higher affinity between Aβ42 and oxidized sulfonic DJ-1 (DJ-1 Cys¹⁰⁶-SO₃H) compared to sulfenic (DJ-1 Cys¹⁰⁶-SOH) or sulfinic acid (DJ-1 Cys¹⁰⁶-SO₂H) forms. Likewise, ELISA tests and seeding assays confirmed that oxidized DJ-1 binds to and decelerates Aβ42 aggregation kinetics. Together, our results identify DJ-1 oxidation as a critical molecular event in the accumulation of iAβ42 in FAD. These findings suggest that oxidized DJ-1 represents not only a potential early biomarker of intracellular pathology but also a pharmacological target. Preventing the oxidation of DJ-1 or its pathological aggregation could provide new biomarkers and therapeutic strategies for reducing the intracellular accumulation of Aβ42 and neurodegeneration in FAD. Full article

The seeds of Impatiens balsamina L. have been traditionally used in East Asian medicine and are known to contain bioactive compounds with antioxidant properties. However, studies focusing on seed-derived callus remain limited. This study aimed to comparatively evaluate the antioxidant activities and lipid accumulation-inhibitory effects of 70% ethanol extracts from seeds (IB) and seed-derived callus (IBC) of I. balsamina. Callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Antioxidant activities were evaluated using DPPH radical scavenging, superoxide anion scavenging, deoxyribose-based hydroxyl radical scavenging, DNA nicking, lipid peroxidation, and relative electrophoretic mobility (REM) assays, along with the determination of total phenolic, flavonoid, and tannin contents. Cell viability and lipid accumulation were assessed in FFA-treated HepG2 cells. In silico network and transcription factor (TF) enrichment analyses were performed to explore underlying mechanisms. Callus induction was most effective at 1 mg/L 2,4-D. Both IB and IBC exhibited antioxidant activities across all assays, with IB showing higher activity and greater phytochemical content than IBC. Both extracts reduced lipid accumulation in FFA-treated HepG2 cells at non-cytotoxic concentrations. Network analysis identified enrichment in pathways related to oxidative stress, inflammation, and lipid metabolism, and TF enrichment analysis identified NFKB1 and ATF3 as major upstream regulators. Both IB and IBC exhibited antioxidant activities across multiple in vitro assays, with IB showing higher activity attributable to its more complex phytochemical content. The lipid accumulation-inhibitory effects observed in FFA-treated HepG2 cells suggest a potential association between antioxidant capacity and lipid regulation, although the underlying mechanisms remain to be experimentally validated. Seed-derived callus may serve as a useful in vitro model for studying plant-derived bioactive compounds, pending further optimization. Full article

The lack of accessible in vitro 3D bone tissue models puts developing countries at a disadvantage in terms of research capacity and healthcare access. In this study, a decellularized corn cob scaffold was functionalized with GTMAC by adding quaternary ammonium groups and coating it with an alginate–gelatin hydrogel to promote mesenchymal stem cell adhesion, offering a low-cost platform for bone disease modeling. Systematic characterization of the scaffold demonstrated that decellularization reduced DNA content by over 80%. Chemical treatment affected the mechanical properties of the matrix, while the hydrogel coating and the functionalized surface of the scaffold promoted cell adhesion and morphology comparable to that observed in 3D cell cultures. MTT analysis showed a subtle reduction in metabolic signal in functionalized scaffolds, which may reflect changes in cell distribution and adhesion within the 3D matrix rather than cytotoxic effects. Full article

Trimethyl chitosan (TMC)-coated alginate/dextran sulfate (ADS) nanoparticles were developed as mucoadhesive nanocarriers for oral insulin delivery using a Quality-by-Design strategy. In a first screening step, a two-level factorial design was applied to evaluate the influence of ADS concentration, TMC concentration, insulin concentration, and poloxamer^® concentration on particle size and encapsulation efficiency. The screening design identified the ADS-TMC pair as the main formulation parameter for particle size, while TMC and poloxamer^® were the most influential factors for encapsulation efficiency. In a second step, formulation optimization was performed using a three-factor, three-level Box–Behnken design in which ADS concentration, TMC concentration, and the degree of quaternization (DQ) of TMC were investigated as critical material attributes. Particle size, zeta potential, and in vitro mucoadhesion were selected as critical quality attributes. Across the Box–Behnken design, the experimental formulations showed particle sizes ranging from 316 to 1340 nm, zeta potentials between +17 and +39 mV, and mucin-binding values from 7 to 87%. Numerical optimization by Design-Expert^® desirability analysis identified an optimal formulation composed of 0.096% (w/v) ADS and 0.700% (w/v) TMC with 60% DQ. The model predicted a particle size of 316.24 nm, a zeta potential of +38.43 mV, and an in vitro mucoadhesion of 87.14%. Experimental confirmation yielded values of 330.79 nm, +37.09 mV, and 84.61%, respectively, with prediction errors below 5% for all responses. In simulated gastric medium, partial insulin leakage was observed during the first 120 min, whereas cumulative insulin release reached 54% after 5 h in simulated intestinal medium. These results demonstrate the usefulness of a QbD framework combined with desirability-based optimization for defining robust formulation conditions for mucoadhesive TMC-coated ADS nanoparticles intended for oral insulin delivery. Full article