Recent advances in enzymatic polyester depolymerization have highlighted the potential of biocatalysis for sustainable plastic recycling. In contrast, efficient enzymatic strategies for polyamide (PA) degradation remain limited. Here, we investigate how complementary amidases influence product formation during the enzymatic treatment of bulk nylon-6 (PA6) and nylon-6,6 (PA66). Guided by previously established substrate preferences, enzyme combinations are designed to couple NylC-mediated oligomer cleavage with downstream conversion by NylB or cyclic oligomer ring-opening by NylA. Sequential enzyme treatments demonstrate efficient conversion of characteristic PA6- and PA66-derived intermediates, while one-pot systems enable coordinated downstream conversion at 34°C-40°C. For PA6, NylB efficiently converts the NylC-generated 6-aminohexanoic acid dimer to 6-aminohexanoic acid, whereas for PA66, NylA selectively redirects accessible cyclic PA66-derived species toward the corresponding linear repeat-unit product. These changes in product distribution are not accompanied by a substantial increase in the overall amount of soluble polymer-derived material. The detected soluble products represent less than 1% of the initial polymer mass under the investigated conditions, demonstrating that overall soluble product release remains limited. Together, these results show that complementary amidases can control downstream intermediate conversion and product selectivity during enzymatic nylon depolymerization.

Transcription factors organize into liquid-like condensates to facilitate gene expression, yet the physical mechanisms governing their formation and properties remain poorly understood. We study the size statistics of transcriptional condensates in human HAP1 cells using widefield and super-resolution microscopy tagging the epigenetic reader BRD4. We find that hubs that appear monolithic in widefield resolve into clusters of smaller droplets that resist coarsening. We link this size control to Active Model B+, a non-equilibrium field theory that captures a regime of reverse Ostwald ripening out of thermal equilibrium. In this regime, chemically driven currents cause larger droplets to transfer mass back to smaller ones, stabilizing a state of microphase segregation. The observed exponential size distribution of BRD4 foci quantitatively matches our numerical simulations, suggesting a universal physical picture for the non-equilibrium self-limitation of cellular condensates.

3D in vitro cancer models provide a more physiologically relevant representation of tumor architecture and cell-matrix interactions than conventional monolayer cultures. This study evaluated bacterial nanocellulose (BNC) as a scaffold for the 3D culture of colorectal cancer cells. BNC scaffolds were produced using paraffin spheres as porogens and subsequently characterized in terms of morphology, chemical structure, thermal stability, and mechanical properties. Human colorectal cancer cell lines SW480 and SW620, along with CHO-K1 cells, were exposed to BNC leachates and cultured on 2D and 3D BNC matrices. Cells grown on 2D BNC exhibited typical monolayer proliferation, whereas 3D matrices allowed cell infiltration and volumetric distribution, supporting multicellular organization and the formation of spheroid-like aggregates. Cell viability remained in both leachate and direct scaffold cultures. Genotoxicity assays showed low levels of DNA damage, with most cells displaying no detectable genotoxic effects, and mutagenicity tests revealed no significant differences compared to controls (p > 0.05). Overall, the results demonstrate that 3D BNC supports cell viability, multicellular organization, and genomic safety, highlighting its suitability as a biocompatible scaffold for in vitro 3D colon cancer models.

Delivering hydrophobic active pharmaceutical ingredients remains a major challenge due to poor aqueous solubility, limited bioavailability, and chemical instability. Drug-in-cyclodextrin-in-liposome (DCL) technology combines cyclodextrin inclusion complexation with liposomal encapsulation to address these limitations. In this hybrid approach, the hydrophobic drug is first incorporated into the apolar cyclodextrin cavity, and the resulting complex is subsequently encapsulated within the aqueous compartment of liposomes. This review provides a comprehensive, data-driven analysis of DCL technology, covering its conceptual development, cyclodextrin properties, lipid composition, vesicle architecture, and preparation methods. Quantitative parameters, including encapsulation efficiency, particle size, zeta potential, and drug release, are compared with conventional liposomes and cyclodextrin complexes. DCL systems demonstrated 2 to 11 fold higher encapsulation efficiency than conventional liposomes, together with sustained release, improved vesicle stability, and enhanced protection of labile drugs against degradation. Preclinical evidence indicates potential therapeutic benefits, although in vivo validation remains limited. Molecular docking and molecular dynamics further provide mechanistic insights into drug-cyclodextrin and cyclodextrin-membrane interactions. Emerging strategies include surface modification, stimuli-responsive systems, dual-drug loading, green manufacturing, and computational design, supporting the future clinical translation of DCL technology.

Silk nanofibrils (SNF) demonstrate substantial potential as biomedical materials and biomineralization templates as a result of their biocompatibility, biodegradability, and resemblance to ECM structural proteins. However, a typical drawback is that the domesticated silk nanofibrils lack sufficient biological functional sites such as cationic groups that are beneficial for increasing cell adhesion, inhibiting bacterial growth, and promoting mineralized deposition. In this study, we developed a cationic SNF with dual functionalities of antibacterial properties and enhanced biomineralization using natural polysaccharide chitosan (CS) and synthetic polymer polyethyleneimine (PEI). The successful cationization of SNF was demonstrated through zeta potential measurement, XPS analysis, and antibacterial performance testing. Compared with polyethyleneimine modification, the CS-modified SNF exhibited superior cytocompatibility, highlighting the advantages of functionalizing proteins with natural cationized polysaccharides. Furthermore, the CS-modified SNF significantly accelerated the biomineralization process, enabling the formation of a uniform flower-like hydroxyapatite structure within 3 days in a 1.5 times simulated body fluid. This study offers a facile strategy for the surface functionalization of natural silk nanofibrils, thereby expanding the application of silks as functional biomaterials.

Pancreatic ductal adenocarcinoma (PDAC) accumulates vast amounts of hyaluronic acid (HA), generating a rigid, high-pressure stroma that blocks drug delivery and infiltration. Although hyaluronidase (HAase) can dismantle this barrier, systemic administration of HAase depletes HA from healthy tissues and causes dose-limiting toxicities. Here we exploited the inherent hypoxia-driven tumor-homing capacity of Escherichia coli BL21 to create an autonomous, self-amplifying delivery system (SHEc) that synthesized and secreted the highly active HA-specific HAase (SHAase) from Streptomyces koganeiensis exclusively within the tumor. Guided by the pelB signal peptide, SHAase could be translocated to the periplasm and released into the extracellular space only where SHEc accumulates. In vitro, SHEc selectively colonized the necrotic core of HA-rich multicellular tumor spheroids (MCTS) formed by Panc02 cells and TGF-β-activated NIH/3T3 cancer-associated fibroblasts, degrading 44.43% of HA within 24 h. This on-site HA digestion increased drug penetration depth by 2.92-fold, decreased hypoxic fraction by 54.81%, and enhanced gemcitabine-induced apoptosis. Thus, SHEc converts a systemically toxic protein into a tumor-restricted stromal remodeling agent, offering a safe and potent strategy to enhance chemotherapy against PDAC.

Poor penetration into tumor tissue remains a major barrier to effective nanomedicine delivery, particularly in glioblastoma multiforme (GBM). Here, nanoparticle transport was computationally studied to rationally design an ultrasmall chemotactic nanomotor that improves penetration into dense tumor tissue. The nanomotor comprises a targeting human heavy-chain ferritin (HFn) nanocage and a catalytic cerium oxide (CeO2) component. Leveraging transferrin receptor‑1 overexpression on brain microvascular endothelial cells and GBM cells, HFn enables transport across an in vitro blood-brain barrier (BBB) model and accumulation in GBM spheroids. The CeO2 domain catalyzes hydrogen peroxide decomposition in the tumor‑mimicking microenvironment, generating oxygen and imparting directional propulsion along H2O2 gradients. In chemotaxis assays, the nanomotors actively migrated toward localized H2O2 sources and GBM cells embedded in Matrigel. In large U87 spheroids (>400 µm), HFn@CeO2 nanomotors penetrated deeply and distributed throughout the spheroid core, whereas non‑propelled HFn nanocarriers remained confined mainly to the periphery. Doxorubicin‑loaded nanomotors (HFn@CeO2-DOX) achieved substantially enhanced intratumoral DOX distribution and reduced IC50 (0.33 µM) compared with HFn-DOX (1.13 µM) and free DOX (1.75 µM) in GBM spheroids. These results provide a promising basis for future in vivo evaluation in brain tumor therapy.

The use of hot melt extrusion combined with fused deposition modeling 3D printing was explored to enhance the solubility and bioavailability of poorly water-soluble compounds intended for Alzheimer's disease (AD) therapy. AD, a neurodegenerative disorder with no definitive treatment, affects millions worldwide, underscoring the urgent need for effective and personalized therapeutic strategies. Four sulfonamide-derived compounds, identified as potential AD therapeutics, were synthesized and successfully incorporated (loads between 10 and 50 wt.%) into customized, 3D-printed oral dosage forms. These 3D-printed tablets were evaluated for properties such as hardness, thermo-analytical characterization, dissolution, cytotoxicity, and stability. Notably, successful amorphization of the incorporated APIs was observed in most 3D-printed tablets, which may be critical for improving solubility. In vitro dissolution testing revealed multiple favorable release profiles, with several formulations demonstrating sustained or enhanced drug release suitable for the therapeutic requirements of AD. When compared to the dissolution data of the corresponding conventional matrix tablets, the present 3D-printed tablets offered superior or comparable dissolution characteristics, demonstrating the potential of personalized medicine and remote digital control for optimizing patient-specific treatments.

Conventional blood-based diagnostics provide essential clinical information but can be less suitable for frequent or continuous biomarker monitoring because they generally require repeated invasive sampling and laboratory-based analysis. Microneedle platforms provide minimally invasive access to dermal interstitial fluid (ISF) and can couple transdermal sampling with on-needle or in situ sensing. This review critically summarizes the microneedle diagnostic systems by comprehensively considering the microneedle architecture, material and polymer properties, mechanical design, sensing modality, disease-specific biomarker requirements and translational performance. Solid, hollow, coated, dissolving, and hydrogel microneedles are discussed together with recent advances in polymer structure-property engineering and 3D-printed microneedle platforms. Electrical/electrochemical, optical, and Raman-based sensing strategies are comparatively evaluated, followed by their applications in metabolic, oncological, inflammatory, allergic, stress-related, and renal disorders. Particular attention is given to the trade-offs among mechanical reliability, ISF transport, analytical sensitivity, biofouling, signal stability, manufacturing reproducibility, and clinical validation. Finally, scalable manufacturing, regulatory translation, wearable integration, and artificial-intelligence-assisted analysis are discussed as key requirements for moving microneedle diagnostics from proof-of-concept devices toward reliable personalized and decentralized healthcare.

Chronic diabetic wounds are difficult to heal due to impaired neovascularization, dysregulated tissue repair, and prolonged inflammation. To address these challenges, PLA/nanoclay/Nd2O3 nanocomposite fiber mats were fabricated by electrospinning. Structural and morphological analyses confirmed successful incorporation and uniform dispersion of nanoclay and Nd2O3 within the PLA matrix. Nanoclay modulated the scaffold surface, while Nd2O3 enhanced mechanical strength. The intercalated nanoclay network further improved load transfer, reduced fiber diameter, and enhanced surface wettability, supporting cellular interactions. The scaffold showed more than 90% cell viability across all tested samples, demonstrating good cytocompatibility. The optimized scaffold exhibited significant antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). The enhanced vascular density of the scaffolds in the chick chorioallantoic membrane (CAM) assay confirms their angiogenic potential. The scaffold also demonstrated controlled degradation, with 8.1 ± 0.5% weight loss after 28 days in simulated body fluid. In vivo evaluation using a diabetic rat wound model showed accelerated wound contraction, complete reepithelialization, neovascularization and minimal inflammatory response by day 21. The findings of this study suggest that PLA/Clay/Nd2O3 bioactive nanofibrous scaffolds have considerable potential for diabetic wound-healing applications.

Effective bleeding control and accelerated wound healing are crucial for patient survival. Conventional dressings often have structural limitations that hinder their effectiveness in controlling hemorrhages and promoting skin tissue regeneration. Here, a gelatin-based nanofibrous sponge incorporating mesoporous silica nanoparticles (MSNs) was developed by transforming two-dimensional nanofiber membranes into three-dimensional structures via electrospinning, gas foaming, and freeze-drying techniques. The sponges were characterized using SEM, EDS, and FTIR. Their swelling, degradation behavior, and mechanical properties showed improved permeability and fluid absorption. Incorporating MSNs significantly enhanced the compressive strength of sponges, making them suitable for deep wounds and creating a favorable microenvironment for cellular activity, despite reduced elastic recovery. In vitro tests confirmed good cytocompatibility and low hemolysis ratios. In vivo experiments on a mouse model of full-thickness skin defects showed that the sponge with a combination of (6%) MSNs effectively accelerated wound healing, with a markedly higher re-epithelialization than the control group, achieving nearly complete healing by Day 14. Additionally, this composite sponge facilitated rapid hemostasis with minimal blood loss. These results highlight the potential of the gelatin-based porous nanofibrous sponge combined with MSNs as a promising candidate for clinical applications, serving as an effective wound dressing and an absorbable hemostatic agent.

Fluorescent organic nanoparticles (FONPs) with intrinsic fluorescence and self-assembled vesicles (VES) are receiving widespread interest in biomedicine. Both FONPs and VES have individual drawbacks like cytotoxicity of FONPs while VES gets eliminated through reticuloendothelial system impeding their applications. Development of nanoconjugates through amalgamation of individual nanocarrier with complementing features would find greater significance by overcoming individual hurdles. Herein, we report design and synthesis of FONP-VES nanoconjugate by combining naphthalimide-based pH-responsive hydrazone moiety containing FONP (NIHYDH) and cholesterol-based vesicle (CBAV) via electrostatic interaction. FONP, VES and FONP-VES conjugate were characterized by spectroscopy and microscopy. Orange emitting NIHYDH-CBAV nanoconjugate was utilised in bioimaging and as delivery vehicle which demonstrated targeted transport of tamoxifen (TAM) to (ER+) MCF7 cells through pH-responsive dissipation of nanoconjugate owing to cleavage of hydrazone bond under acidic pH gradient. TAM has demonstrated clinical drug resistance along with limited bioavailability. Upon encapsulation within nanoconjugate, NIHYDH-CBAV-TAM offered ∼2.8-fold higher cytotoxicity toward MCF7 cells compared to normal NIH3T3 cells alongside demonstrating ∼2.1-fold reduction in the size of MCF7 3D tumour spheroids. TAM loaded nanoconjugate was ∼2.2-3.4-fold more cytotoxic than TAM loaded individual nanocarrier. Cytotoxicity of NIHYDH-CBAV-TAM got enhanced due to increased intracellular oxidative stress that triggered p53 upregulated cell apoptotic pathway.

Nanomedicine has broad prospects in tumor diagnosis and treatment, but its clinical translation has long been limited by targeted delivery efficiency. The heterogeneity of the tumor microenvironment is like a "sea fog" that hinders precision medicine. This makes it difficult for traditional active targeting that relies on natural targets to achieve precise delivery. For this difficulty, this review summarizes the constraints of tumor microenvironment heterogeneity on nanomaterials in terms of time and space. Pre-targeting strategies enhance delivery efficiency by constructing an "artificial lighthouse." According to the source of the target and the mode of specific binding, this review divides pre-targeting strategies into two categories: biomolecular recognition-mediated and bioorthogonal click chemistry-mediated. On this basis, this review comprehensively introduces recent progress in pre-targeting-enabled nanomedicine in body fluid-based detection, in vivo imaging, and multi-therapies. Furthermore, this review is the first to summarize the synergistic mechanisms of pre-targeting with prodrugs, insitu self-assembly, and dual-targeting strategies. This review systematically summarizes the complete framework of pre-targeting strategies, from mechanisms and classifications to applications and expansions. It aims to provide inspiration for the development of nanomedicine in the field of oncology for effective cancer diagnosis and treatment.

Colorectal cancer (CRC) exhibits intrinsic resistance to conventional therapeutic regimens. By integrating single-cell and bulk transcriptomic data from CRC cohorts, we demonstrate that proliferative capacity alone constitutes an inadequate prognostic predictor. Instead, an immunosuppressive tumor microenvironment (TME) emerges as the dominant driver of disease progression. Galectin-1 (LGALS1), highly upregulated in metastatic CRC, correlates with robust infiltration of regulatory T cells (Tregs) and M2 macrophages to facilitate immune evasion and poor clinical outcomes, thereby serving as a promising therapeutic target for anti-tumor immunotherapy. The stromal barrier mediated by cancer-associated fibroblasts (CAFs) restricts intratumoral penetration of antitumor agents and diminishes therapeutic efficacy. To overcome this physical barrier, we constructed a CAF extracellular vesicle-based co-delivery platform termed CEV@Comb, encapsulating the LGALS1 inhibitor OTX008 and 5-fluorouracil (5-FU). CEV@Comb selectively accumulates in tumors, suppresses growth in vitro and in vivo, and remodels the immunosuppressive TME by boosting M1 macrophages and cytotoxic CD8[+] T cells while reducing intratumoral Tregs. Combination therapy with CEV@Comb and anti‑programmed death‑1 antibody (αPD-1) further amplifies anti-tumor immunity in the Apc[-/-], Kras[G12D], Trp53[-/-], Smad4[-/-](AKPS) CRC mouse model. This study identifies LGALS1 as a core immunosuppressive mediator and provides an innovative chemoimmunotherapeutic strategy to improve clinical outcomes for patients with advanced CRC.

Acute full‑thickness skin wounds represent a significant clinical challenge due to tissue loss, risk of bacterial contamination, and the need for rapid and effective regeneration. Injectable thermosensitive hydrogels incorporating antibacterial nanomaterials represent a promising strategy for advanced wound care. In this study, a multifunctional hydrogel composed of Graphene oxide (GO), branched polyethyleneimine (BPEI), and silver nanoparticles (AgNPs) dispersed in Pluronic F127 was developed and evaluated for wound healing applications. The GO-BPEI-AgNP nanocomposite was synthesized via amidation and microwave-assisted methods and characterized using DLS, UV-vis, FTIR, XRD, and SEM-EDX analyses. Biocompatibility was assessed in HaCaT keratinocytes using the Alamar Blue assay, while scratch assays and antibacterial activity against. In vivo wound healing efficacy and histopathological changes were evaluated in a rat full-thickness skin wound model, together with qRT-PCR analysis of wound-healing-related genes. The hydrogel exhibited favorable injectability, nanoscale morphology, and high biocompatibility (IC50: 246.7 µg/mL). It significantly enhanced keratinocyte migration and demonstrated antibacterial activity. In vivo findings revealed accelerated wound closure, enhanced epidermal regeneration, and modulation of PDGFA, VEGFC, COL1A1, TIMP1, and MMP9 expression. Collectively, the GO‑BPEI‑AgNP hydrogel shows strong potential as an injectable antibacterial biomaterial for acute full‑thickness wound repair, with potential relevance to infection‑associated wound environments.