Search Results (769)

The rapid expansion of molecular point-of-care (POC) diagnostics into decentralized settings, including emergency departments, retail pharmacies, and home environments, has shifted the burden of diagnostic performance from laboratory professionals to heterogeneous, often non-expert users. While traditional evaluation frameworks focus on analytical and clinical validity, they often overlook the impact of human-system interactions on real-world reliability. This review introduces the concept of Operational Validity: the ability of a diagnostic system to preserve its intended performance when operated by intended users within the constraints of real-world workflows and environments. To establish a rigorous foundation for this concept, this study provides a critical comparative analysis contrasting Operational Validity against traditional clinical evaluation dimensions (analytical validity, clinical validity, and clinical utility) and post-market metrics. While existing literature outlines isolated usability principles, the significance of this study lies in its synthesis of these fragmented concepts into a formalized, lifecycle-based “Operational Validity” framework that explicitly maps the causal mechanisms connecting initial user interaction directly to downstream clinical outcomes. By synthesizing international standards (IEC 62366-1) alongside the newly finalized May 2026 U.S. Food and Drug Administration (FDA) guidance on the Content of Human Factors Information in Medical Device Marketing Submissions, we examine how human factors engineering (HFE) and usability engineering serve as the methodological foundation for operational validity. We analyze the specific complexities of molecular workflows, identify key parameters of use-related failure modes in pre-analytical and interpretation stages, and detail the mandatory role of iterative formative and final summative usability testing in mitigating these risks. Finally, we propose a lifecycle-based approach to HFE that integrates design, simulated-use validation, and post-market surveillance. Establishing operational validity is essential to ensure that the high analytical sensitivity of molecular POC platforms translates into consistent clinical utility across the full spectrum of decentralized care. Full article

Background: 5-(3′,4′-Dihydroxyphenyl)-γ-valerolactone (DHPV) is a postbiotic gut microbiota-derived flavanol metabolite with reported anti-inflammatory activity. Despite growing interest in its potential dermatological applications, its pharmaceutical properties and suitability for topical delivery have not been systematically investigated. This study aimed to perform the first comprehensive preformulation and formulation-oriented evaluation of DHPV and to develop stable topical ointment formulations suitable for further dermatological research. Methods: The physicochemical properties of DHPV were characterized using powder X-ray diffraction (PXRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), quantitative solubility assessment, and excipient compatibility studies. Based on the obtained preformulation data, two anhydrous ointment formulations containing DHPV were developed. The formulations were evaluated for homogeneity, rheological behavior, chemical stability under accelerated storage conditions, and in vitro drug release performance. Results: DHPV was identified as a crystalline compound with heterogeneous particle morphology and limited aqueous solubility. Quantitative solubility studies demonstrated the highest solubility in PEG 300 and glycol-based solvents. Compatibility testing revealed increased impurity formation in hydrophilic environments, whereas lipophilic excipients provided improved chemical stability. Both ointment formulations exhibited acceptable physical characteristics and maintained DHPV stability throughout accelerated storage. However, marked differences in release behavior were observed. The lipid–wax formulation showed significantly higher release rates, lower variability, and more reproducible release profiles than the petrolatum-based reference formulation, indicating more efficient diffusion of DHPV from the semisolid matrix. Conclusions: The physicochemical characteristics of DHPV strongly influence formulation design and performance. Anhydrous lipid-based systems provide a favorable environment for maintaining DHPV stability, while formulation composition significantly affects drug release. The developed lipid–wax formulation represents a promising platform for future skin permeation, pharmacodynamic, and efficacy studies. Full article

Chemical contaminants in food pose a serious threat to public health, driving the need for sensitive, rapid, and on-site screening methods. Lateral flow immunoassay (LFIA) is rapid and portable but suffers from single-signal readout and insufficient label stability. Nanozymes, nanomaterials with enzyme-like catalytic activity and excellent stability, have emerged as promising signal labels to address these limitations. Moreover, their diverse physiochemical properties enable multiplex signal readout, where two or more complementary signals (e.g., colorimetric, fluorescent, chemiluminescent, photothermal, and surface-enhanced Raman scattering) are generated simultaneously from a single test line. This multiplex strategy significantly enhances detection sensitivity, accuracy, and reliability through signal amplification and self-calibration. This review provides a systematic overview of the catalytic properties and their major types used in multiplex signal LFIA. The signal combination strategies employed in nanozyme-based multiplex signal LFIA were also summarized, and their applications in detecting veterinary drugs, mycotoxins, pesticides, and other food chemical contaminants are highlighted. Ultimately, current challenges and future prospectives in this field are discussed. This review offers guidance for designing high-performance, nanozyme-based multiplex signal LFIA platforms for food safety monitoring. Full article

The vitreous body is not only a transparent filling material of the posterior segment; it is a soft, hydrated, and biologically active matrix that supports structural, optical, and biochemical homeostasis. Vitrectomy therefore leaves a functional deficit that current substitutes only partly address. Intraocular gases, silicone oils, and perfluorocarbon liquids remain essential surgical tools, but they mainly provide mechanical tamponade and do not reproduce native viscoelasticity, diffusion control, or protection against oxidative and inflammatory stress. This review considers vitreous replacement as a functional biomaterials challenge. We discuss native vitreous physiology, the limitations of present tamponade agents, and emerging bioengineered substitutes designed to create a more physiological intravitreal environment. Particular attention is given to hydrogel and polymer-based systems, especially hyaluronic acid-based and in situ crosslinked platforms, which are being developed to combine optical clarity, injectability, soft mechanical support, controlled degradation, and favorable tissue interaction. We also emphasize the need for standardized preclinical testing of swelling, enzymatic stability, drug diffusion, rheology, and long-term biocompatibility. Although next-generation materials may move the field beyond passive space filling, manufacturing reproducibility, regulatory validation, chronic safety, and cautious early-phase trials remain major translational barriers. Full article

Melanoma is the most aggressive type of skin cancer, driven by early invasion, phenotypic plasticity, and frequent resistance to targeted therapies. Although genomic profiling informs treatment selection, genotype alone often fails to predict therapeutic response, underscoring the need for rapid and physiologically relevant functional testing platforms. Here, we present a three-dimensional melanoma–skin organoid (mSO) model that integrates primary skin cells with melanoma cell lines in a self-assembling, high-throughput format. The spherical mSOs recapitulate native human skin architecture, including a stratified epidermis and a dermal–hypodermal core, while supporting melanoma growth within an appropriate tissue microenvironment. In this niche, melanoma cells display epidermal spreading in radial growth-like patterns, outward invasion, and transcriptional shifts toward a pro-invasive phenotype. Using live confocal imaging coupled with a custom automated image analysis pipeline, we quantitatively measured tumor growth, migration beyond the organoid boundary, and interactions between melanoma cells and normal melanocytes. The mSOs also captured genotype-specific drug responses: BRAF-mutant melanoma cells were sensitive to BRAF and MEK inhibition, whereas NRAS-mutant, BRAF–wild-type cells were resistant to BRAF inhibition but remained responsive to MEK inhibition. Altogether, our mSO platform combines architectural and functional complexity with experimental scalability, providing a robust framework for modeling melanoma progression and evaluating targeted therapeutic responses within a relevant skin microenvironment. In the future, adaptation of this system to include patient-derived tumor cells could support personalized therapeutic decision-making in melanoma. Full article

The myocardium possesses one of the highest vascular densities in the body. The outermost wall layer of large and medium-sized vessels, the adventitia, forms a critical interface between the vasculature and the myocardium and serves as a reservoir for stem and progenitor cells capable of differentiating into all vascular wall lineages as well as innate immune cells, including macrophages. Current cardiac organoid models intrinsically develop networks of endothelial cords and small capillary-like structures that resemble cardiac microvessels. However, these microvessels mostly lack an adventitial compartment in vivo. Here, we present a potential alternative assembloid strategy that combines vascular segments from mouse and human origin with either cardiomyocytes or cardiac spheroids derived from human induced pluripotent stem cells, thereby incorporating large diameter vessels and the vascular adventitia into a cardiac tissue model. Within the assembloids, the myocardial component remained contractile and connected to the vascular adventitia, which displayed cellular sprouting toward the hiPSC-derived cardiac tissue. Immunostaining for vascular and immune markers revealed that the adventitia gave rise to endothelial sprouts and macrophage-like cells which integrated into the myocardial tissue. In summary, we present proof of concept for complex assembloids composed of vessel segments and human iPSC-derived cardiomyocytes which contain and maintain an in vivo-like adventitial compartment. We suggest this model may serve as a platform for investigating myocardial–stromal interactions, cardiac tissue repair, and functional remodeling under both physiological and pathological conditions. Furthermore, the incorporation of large-lumen vessel segments may enable future experimental perfusion, rendering the model particularly suitable for drug testing via intravascular delivery. Full article

Phase-inversion in situ implants (PIISIs) represent a versatile polymer platform in which the rational choice of matrix former and solvent system directly governs the macroscopic properties of the resulting depot. This study applied a Quality by Design (QbD) approach to rationalize a bleached shellac–based PIISI, with particular focus on the physicochemical interactions between the polymer and the injection vehicle. Bleached shellac—a natural, low-cost, biodegradable oligomeric resin bearing –COOH, –OH, and ester functional groups—was selected as the matrix former and screened in seven neat solvents and five 1:1 binary combinations at 25% (m/m). Twelve formulations were evaluated against a predefined set of critical quality attributes, including injectability, phase-inversion kinetics, solvent diffusion volume, and implant structure (n = 5 per formulation; mean ± standard deviation (SD); one-way analysis of variance (ANOVA) with Tukey’s post hoc test, p < 0.05). Three lead solvent systems—propylene glycol/N-methylpyrrolidone (PG+NMP), PG/dimethyl sulfoxide (PG+DMSO), and DMSO/benzyl alcohol (DMSO+BA)—were identified as those providing an optimal balance between hydrogen-bond donor/acceptor solvation and controlled solvent extraction. In the second stage, shellac concentration (20–35%) was optimized, with 30% shellac in PG+NMP yielding the fastest phase inversion (~50 s), a structurally uniform matrix, and the lowest swelling (22%). A working mechanistic framework consistent with all observed critical quality attribute (CQA) trends in which solvent hydrogen-bond donor/acceptor balance and water miscibility govern implant architecture is proposed, and it is intended as a hypothesis-generating basis for the rational design of PIISI formulations; direct validation by spectroscopic, thermal-analytical, and biological methods is identified as the next step. The developed formulations are presented as a preliminary physicochemical platform; biological validation (in vitro cytocompatibility and inflammatory response assessment) is required before the system can be considered a validated formulation for dental drug delivery. Full article

The rapid expansion of individualized treatment strategies necessitates advanced patient-specific screening platforms recapitulating tumor complexity. Here, we present such a platform preserving heterotypic cellular interactions and three-dimensional architecture which are both critical for predicting therapeutic responses. This cost-effective and versatile platform allows the rapid generation and functional testing of spheroids derived from primary human glioblastoma specimens. To maximize accessibility and ease of integration, we took advantage of commercially available kits to optimize the protocol of spheroid generation we had previously established. This enabled the robust, adaptable, and reproducible formation of homogeneous, multicellular spheroids within a few days. Spheroids generated from one patient specimen were structurally stable and showed a high degree of homogeneity over time. Immunohistochemical and flow cytometric analyses further revealed patient-specific cellular heterogeneity, underscoring the platform’s ability to preserve clinically relevant tumor features. Functionally, we demonstrate the applicability of this system for drug response profiling by assessing invasion dynamics following treatment with clinically relevant compounds. Collectively, our results establish a scalable and adaptable 3D screening platform that enables rapid, patient-specific phenotypic and functional analyses. This approach provides a powerful tool to complement existing clinical workflows and holds promise for improving the prediction of therapeutic responses in glioblastoma. Full article

Background/Objectives: Providing optimized and accurate treatment to cancer patients remains a major challenge in oncology care. The emergence of precision medicine tools to match the correct therapy to the patient has significantly advanced treatment modalities in the last few years. While genomics has been shown to be critical in selecting targeted therapies for a specific somatic mutation, the overall clinical benefit of broad genomic sequencing has been found lacking. Here, we evaluate the utility of our previously clinically validated ex vivo functional assay across different treatment scenarios, demonstrating its ability to transform predicted non-responders into predicted responders, rule out ineffective treatments, provide multiple treatment options, and validate physician choices. Methods: The evaluation was performed on a post-market surveillance study analyzing 312 patients, from which 278 patients had successful test reports (an 89.1% test success rate), with clinical outcomes available from 45 of those patients. Results: We show that in the group of patients with clinical response data, the tests yield a PPV of 91.18% and NPV of 90.91% with clinical utility impacting physician decision in 51.1% of cases. Further analysis of the entire cohort showed the potential of clinical utility to reach up to 59.3% on a large group of patients. Conclusions: The accurate prediction of patient response using the test suggests the potential for the platform to improve patient treatment in clinical practice by reducing ineffective drug use and optimizing personalized patient drug regiments. Full article

An isolate of the diatom Staurosirella pinnata is a promising platform for drug discovery due to its ability to produce bioactive metabolites. As previously shown, S. pinnata extracts exhibit bioactivities, with hydrophilic fractions showing selective cytotoxicity against human melanoma cells and lipidic fractions promoting thermogenesis in murine white adipocytes. In this work, we focused on the interaction between S. pinnata metabolism and light irradiance exposure to evaluate bioactivity targeting medulloblastoma cells. Cultures under standard, control, irradiance (80 µmol photons m⁻² s⁻¹) were exposed in the stationary phase to increased light intensities (200 and 600 µmol photons m⁻² s⁻¹) for 126 h. Growth, photosynthetic performance and metabolic profile were monitored, while the bioactivity of small-molecule fractions was assessed at the end. Exposure to 200 µmol photons m⁻² s⁻¹ significantly enhanced growth (92.6% increase in absorbances compared to the control), whereas 600 µmol photons m⁻² s⁻¹ induced growth inhibition (41.3% decrease in absorbances with respect to the control culture) and impaired photosynthesis. Metabolomic analysis revealed a shift from carbohydrate to lipid metabolism. Bioactivity assays showed that extracts from the highest irradiance exhibited cytotoxic effects on medulloblastoma cells, similar to the 80 µmol photons m⁻² s⁻¹ cultures on DAOY (68% vs. 82% of cell death induction levels, respectively), while intermediate irradiance did not show a significant effect in any of the tested cell lines. The results showed that different light intensities impact S. pinnata metabolism, demonstrating effects exploitable for drug discovery and the importance of investigating the impact of cultivation parameters in modulating S. pinnata bioactivity potential. Full article

Background: A transdermal drug delivery system has significant benefits over conventional routes; however, its effectiveness is limited by the barrier properties of the stratum corneum. Dissolving microneedles (DMNs) have emerged as a minimally invasive strategy to enhance drug permeation while improving patient compliance. The integration of advanced fabrication techniques such as 3D printing enables precise control over microneedle geometry and reproducibility. Objective: This study aimed to fabricate and characterize pregabalin-loaded PVA/PVP dissolving microneedle arrays using a 3D-printing-assisted mold fabrication approach for efficient transdermal drug delivery. Methods: Microneedle master molds were fabricated using 3D printing, followed by replication using polydimethylsiloxane (PDMS) to obtain negative molds. Pregabalin-loaded bilayer microneedles were prepared using a micromolding technique with PVA/PVP polymers. The formulation was evaluated through rheological analysis, scanning electron microscopy (SEM), mechanical strength testing, insertion studies, swelling behavior, drug loading efficiency, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRD), and in vitro drug release studies. Results: The fabricated microneedles exhibited uniform geometry with sharp tips and no structural defects. Rheological analysis confirmed shear-thinning behavior suitable for mold filling. The microneedles demonstrated adequate mechanical strength (~3.3 N/needle) and efficient insertion into the parafilm model. Drug loading efficiency was high (92.4%), indicating effective encapsulation. FTIR analysis confirmed compatibility between drug and polymers, while DSC and XRD results indicated partial amorphization of pregabalin within the polymer matrix. The formulation showed a biphasic drug release profile with an initial burst followed by sustained release, achieving ~96.8% cumulative release over 24 h. Conclusions: The study successfully demonstrates a robust and reproducible 3D-printing-assisted approach for fabricating pregabalin-loaded dissolving microneedles. The developed system exhibited desirable mechanical, physicochemical, and drug release properties, highlighting its potential as an effective transdermal drug delivery platform. Full article

Background/Objectives: Cannabis sativa is the source of cannabidiol (CBD), a non-intoxicating phytocannabinoid with analgesic and anti-inflammatory qualities that has demonstrated therapeutic potential in inflammatory skin conditions like dermatitis. However, low bioavailability and poor water solubility restrict its topical application. This study attempted to improve CBD solubility and transdermal delivery using an amorphous solid dispersion (ASD)-based hydrogel system. Methods: CBD was stabilized in its amorphous form using an ASD strategy and incorporated into a hydrogel matrix. The CBD-ASD hydrogel was characterized by particle size analysis, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), rheological assessment, swelling studies, and diffusion experiments using Franz cells. Biological evaluations included cytotoxicity testing in human dermal fibroblast (HDF) cells, wound-healing assays, RT-qPCR-based anti-inflammatory analysis, antioxidant activity (DPPH assay), and antibacterial testing against Staphylococcus aureus. Results: Physicochemical analyses confirmed successful amorphous dispersion of CBD within a stable hydrogel network. The formulation exhibited sustained drug release over 144 h, achieving 86.32% cumulative release with diffusion-controlled kinetics. Rheological and swelling properties demonstrated mechanical stability and hydration suitability for long-term topical application, while Franz diffusion studies confirmed effective transdermal permeation. The CBD-ASD hydrogel showed no cytotoxicity in HDF cells and significantly enhanced wound closure. It also downregulated pro-inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Additionally, the formulation demonstrated 65.63 ± 10.00% DPPH radical scavenging activity and over 99% antibacterial inhibition. Conclusions: The CBD-ASD hydrogel represents a stable, multifunctional delivery platform that overcomes CBD solubility limitations and enhances therapeutic efficacy for inflammatory skin diseases. Full article

Adverse drug reactions (ADRs) remain a major public health issue, and genetic factors contribute importantly to interindividual variability in drug response. Pharmacogenetic testing helps reduce ADR risk by optimizing drug selection and dosage, particularly in monogenic disorders. Whole-exome sequencing of 6739 samples from the Russian population was performed on the DNBSEQ-G400 platform (MGI). Variants in 48 genes were examined, focusing on inherited arrhythmias, enzyme deficiencies (Glucose-6-Phosphate Dehydrogenase Deficiency [G6PDD], Porphyrias), Dravet Syndrome (DS) and Malignant Hyperthermia (MH). Variants reported as pathogenic (P), likely pathogenic (LP), or variants of uncertain significance (VUS) in ClinVar were manually re-evaluated using ACMG criteria. A total of 75 unique variants in 18 genes were observed in 119 individuals (1.77%), including 21 carriers and 13 women with a G6PD mutation. Of these, 44 variants were classified as P, 24 as LP, and 7 as VUS. Missense variants accounted for the largest proportion (73.33%). The most affected genes were KCNQ1 (24/119), which exhibited the highest number of unique variants (18), G6PD (20/119), SCN1A (15/119), and RYR1 (14/119). Regarding associated conditions, mutations linked to arrhythmias were found in 51 individuals, MH in 27, G6PDD in 20, DS in 15, and Porphyrias in 6. Integrating common and rare clinically actionable genetic variants with attention to penetrance and clinical validity may improve medication safety, reduce preventable ADRs, and enhance personalized pharmacotherapy. Full article

Background/Objectives: Insomnia severely impairs quality of life. Oral melatonin (MEL) suffers from poor brain delivery. Intranasal administration bypasses the blood–brain barrier, but rapid mucociliary clearance shortens drug retention, and MEL poor water solubility limits its nasal dissolution. Traditional in situ gels have “gelation-first, spreading-second” defects, causing uneven distribution. Herein, we developed a two-step sequential ion-triggered in situ gel combined with MEL liposomes (MEL-Lips-Gel) to enhance solubility, achieve instant uniform coating, and prolong retention for efficient nose-to-brain delivery. Methods: MEL-Lips were dispersed in alginate (first component) and calcium gluconate served as the second component. After sequential spray, the two components mix and form an ion-crosslinked gel. Rheology, in vivo fluorescence imaging, in vitro release, open-field/sucrose preference tests, and H&E staining were performed. Results: MEL-Lips showed uniform size and good encapsulation. The sequential system achieved instant widespread spreading and rapid gelation, significantly prolonged nasal retention, enabled sustained brain delivery, and reversed insomnia-induced hyperactivity and anxiety-like behaviors more effectively than oral MEL, intranasal MEL solution, liposomes alone, or non-liposomal gel, with good nasal safety. Conclusions: This sequential ion-triggered liposome-in-gel strategy synergistically overcomes rapid clearance (via gel) and poor solubility (via liposomes), enhancing nose-to-brain delivery of melatonin and providing a promising platform for insomnia therapy. Full article

Background/Objectives: Andrographolide (ADG) is a plant-derived compound with promising anticancer properties, but its medical use is limited due to poor water solubility and low bioavailability. This study proposes developing a gold-based nanocomposite drug delivery system, using a simplified synthesis method, to improve ADG’s hydrophilicity and enhance its delivery efficiency. Methods: A one-step method was used to synthesize gold nanocomposites with carbon quantum dots (CBQDs) and doped CBQDs acting as reducing and stabilizing agents. These nanocomposites were then conjugated with ADG and thoroughly characterized using multiple structural and spectroscopic techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), ultraviolet–visible spectroscopy (UV–Vis), transmission electron microscopy (TEM), Raman spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Hydrophilicity enhancement was evaluated using NMR-based log P measurements. Biological assessment involved cell viability assays and confocal microscopy studies in PC3 prostate cancer cells, along with the morphological evaluation of human red blood cells. Results: XRD confirmed the formation of crystalline, face-centered cubic gold nanoparticles, while spectroscopic analyses verified successful nanocomposite formation and ADG conjugation. NMR results showed enhanced hydrophilicity of ADG. Biological tests demonstrated that the nanocomposites were compatible with cells. Conclusions: This study presents a straightforward strategy for synthesizing gold-based nanocomposites that enhance the hydrophilicity and delivery potential of andrographolide, supporting their applicability as nanocarrier platforms for anticancer drug delivery. Full article