Journal of Nanobiotechnology最新文献

Glioblastoma (GBM), the most aggressive adult primary brain tumor, faces lethal challenges due to its immunosuppressive microenvironment and blood-brain barrier (BBB) impedance. This study investigates how cRGD-functionalized macrophage-derived extracellular vesicles (cEV) loaded with the Liver X receptor (LXR) antagonist GSK2033 (cEV@GSK) enhance T cell-mediated antitumor immunity in GBM by targeting the LXR/ATP-binding cassette transporter A1 (Abca1) axis. Methods include isolating and cRGD-functionalizing RAW264.7 macrophage-derived extracellular vesicles, loading GSK2033 to form cEV@GSK, characterizing nanoparticles via TEM, size/zeta potential, and HPLC; evaluating BBB penetration, cellular uptake, and cytotoxicity in vitro; assessing in vivo distribution, antitumor efficacy, and biosafety using an orthotopic GBM mouse model; and analyzing mechanisms via proteomics and single-cell RNA sequencing (scRNA-seq), with T cell-LLM-GL261 co-cultures validating KLRB1 function. Results show cEV@GSK effectively crosses the BBB, exhibits biosafety, and significantly suppresses tumor growth. Mechanistically, it blocks the LXR/Abca1 axis, reducing myelin lipid transfer from lipid-laden macrophages (LLMs) and downregulating T cell KLRB1, thereby augmenting T cell activation and antitumor activity. Conclusion: cEV@GSK enhances T cell immunity by disrupting the LXR/Abca1 axis and LLM-mediated lipid transfer, offering a novel GBM immunotherapy strategy.

Diabetic wounds remain a major clinical challenge due to bacterial infection, persistent inflammation, and excessive accumulation of reactive oxygen species (ROS). This highlights the urgent need for wound dressings that provide sustained antibacterial effects, stable ROS scavenging, and effective immune modulation. To this end, we developed a multifunctional electrospun polyvinyl alcohol-chitosan (PVA-CS) nanofibrous dressing by integrating black phosphorus nanosheets (BPNSs) with polydopamine-coated zinc oxide nanoparticles (PDA@ZnO NPs). This dressing achieved synergistic photothermal-Zn²⁺ ion antibacterial activity, rapidly reducing the viability of MRSA, MSSA, and E. coli, along with their biofilms, to 0-2% in 3 min while enabling Zn²⁺ release to support a sustained antibacterial microenvironment. The dresssing also provided sustained ROS scavenging due to stabilized BPNSs, with intracellular ROS levels reduced to 13-20%, accompanied by an overall 50-70% improvement in antioxidant capacity, thereby alleviating oxidative stress and suppressing inflammation. In addition, ZnO NPs also promoted anti-inflammatory macrophage polarization, accelerated tissue healing, and supported natural repair processes. This work introduces a multifunctional wound dressing with rapid and sustained antibacterial, antioxidative, and anti-inflammatory properties, offering a promising solution for chronic diabetic wounds.

Myocardial infarction (MI) is one of the leading causes of death worldwide, characterized by irreversible loss of cardiomyocytes caused by ischemic injury. Despite the development of various therapeutic approaches in recent years, persistently high mortality underscores the urgent need for more advanced treatment strategies. Nanomaterials have attracted considerable attention in drug delivery due to their nanoscale dimensions, facile surface modification, and multifunctional properties. In particular, stimuli-responsive nanomaterials have demonstrated significant potential. These materials are capable of responding to endogenous cues (e.g., pH, ROS, and enzymes) or exogenous triggers (e.g., thermal, magnetic, and ultrasonic stimuli). This review systematically summarizes the recent advances and application prospects of stimuli-responsive nanomaterials for MI therapy. Through the summary and analysis of existing research, we examine the key pathological stimuli present in the infarcted microenvironment and classify nanocarriers according to their response mechanisms. We also discuss the diverse therapeutic and diagnostic applications of these stimuli-responsive nanomaterials in the context of MI. Overall, this article provides researchers and clinicians with an integrated perspective on harnessing stimuli-responsive nanomaterials for the precision diagnosis and treatment of MI.

Psoriasis is a chronic, immune-mediated skin disease characterised by epidermal hyperplasia and compromised barrier integrity, which significantly complicates effective drug delivery. Topical drug delivery (TDD) offers a promising, non-invasive, and patient-centric alternative therapy for its management. However, the efficacy of TDD is constrained by the markedly thickened stratum corneum in psoriatic lesion skin, which acts as a formidable barrier to effective drug penetration. In this work, we developed a high-drug-loading TDD system for the highly effective topical delivery of apremilast (APR), an FDA-approved oral treatment for psoriasis. Using a sequential nanoprecipitation method, lipid nanoparticles (LNPs) and polymer nanoparticles (PNPs) loaded with 40% APR, defined as the weight ratio of APR relative to the total nanoparticle formulation, were synthesized and embedded into Carbopol 940 gel for enhanced skin compatibility and topical application. Ex vivo studies revealed enhanced intradermal retention of LNPs Carbopol^® 940 gel (LNPG) and greater subdermal accumulation of PNPs Carbopol^® 940 gel (PNPG). In an imiquimod-induced psoriasis mouse model, treatment with both formulations resulted in marked clinical improvements, including reduced PASI scores, decreased epidermal thickness, and reduced spleen size. Furthermore, both LNPG and PNPG systems significantly downregulated psoriasis-associated cytokines (TNF-α, IL-1β, IL-6, CXCL8, and CCL20). These findings demonstrate the robust therapeutic potential of high-drug-loading NP gels and highlight their promise as a patient-friendly TDD platform for psoriasis and other dermatological conditions with a compromised barrier function.

Diabetic foot ulcers (DFUs), prone to infection and deterioration without timely intervention, significantly increase risks of amputation and mortality. Elevated local temperature in DFUs further impedes the healing process. Although microneedles (MNs) represent a promising strategy for DFUs treatment, conventional passive drug release systems suffer from slow release kinetics and low utilization efficiency. To address these limitations, we developed a dual-thermo-responsive microneedle patch (DTMN) that actively controls drug delivery through temperature-induced structural transitions. The inner layer consists of sodium alginate-poly (N-isopropylacrylamide) (SA-PNIPAM) loaded with sucrose octasulfate sodium salt (SOS), utilizing the volume phase transition of PNIPAM to accelerate drug expulsion in response to temperature change. The outer layer comprises a polyethylene glycol/polylactic acid-glycolic acid copolymer (PEG-PLGA) loaded with urea, which undergoes gel-sol transition to facilitate controlled urea release and wound cooling. An upper electrospun nanofiber membrane made of poly (ε-caprolactone)/chitosan (PCL/CS) incorporated with tetracycline hydrochloride (TH) and SOS provides enhanced antibacterial efficacy and increased drug loading capacity. The resulting DTMN exhibits efficient drug release (85.23% SOS and 35.44% urea-derived ammonia at 24 h), remarkable antioxidant activities, potent antibacterial performance, excellent biocompatibility, and significantly enhanced wound healing. This multifunctional system offers a novel and effective strategy for the management of DFUs.

Chronic non-healing wounds represent a severe complication of diabetes mellitus, which frequently progress to infection, limb amputation, and even mortality. The dysregulated wound microenvironment, marked by persistent inflammation and oxidative stress, severely impedes tissue repair, and the presence of MRSA biofilm infection further worsens these impairments and poses major clinical challenges. To address these challenges, we constructed a multifunctional injectable hydrogel (SOT) that integrates antibiofilm, antioxidant, and immunomodulatory properties. This hydrogel is formed through dynamic covalent crosslinking between thiolated hyaluronic acid (HA-SH) and dopamine-modified oxidized dextran (ODex-DA), which enables favorable injectability, self-healing, and in situ gelation. Tannic acid-silver nanoparticles (TA-Ag NPs) incorporated into the system impart antibiofilm and reactive oxygen species (ROS)-scavenging properties. In a diabetic MRSA biofilm infection model, the SOT hydrogel eradicated biofilms, reduced excessive ROS, and promoted wound closure. These findings suggest that this immuno-instructive hydrogel platform may offer a promising therapeutic approach for the treatment of MRSA biofilm-infected chronic diabetic wounds.

CRISPR-Cas9, an innovative genome-editing technique, holds immense promise in therapeutic applications; nevertheless, the lack of effective delivery methods for in vivo gene editing limits its utility in osteoarthritis (OA) treatment. Recently, exosomes, naturally derived nanosized vesicles secreted by cells, have attracted significant attention as potential vehicles for therapeutic cargo delivery. This study proposes a bioinspired engineered exosome-mediated CRISPR/Cas9 delivery platform for targeted editing of the Asporin (ASPN) gene as a potential precision therapy for OA. Specifically, chondrocyte affinity peptide (Cap)-modified MSC-derived exosomes were employed as natural, biocompatible carriers to deliver CRISPR/Cas9 components specifically to OA-affected chondrocytes, thereby achieving precise and efficient ASPN knockout. Flow cytometry analysis confirmed a modification efficiency of 79.1% for Cap, while the encapsulation efficiency of the ASPN-Cas9 plasmid into exosomes reached 9.5% ± 0.6%. Both in vivo and in vitro investigations revealed that this delivery approach markedly improved cellular uptake and gene-editing efficacy, achieving a substantial reduction of ASPN expression by 61.7%. This, in turn, alleviated ferroptosis, improved mitochondrial function, reduced chondrocyte senescence, inhibited inflammation, and enhanced the cartilage microenvironment. Altogether, these findings strongly suggest the promising therapeutic efficacy of this method in OA models, emphasizing its potential as a precise gene-targeting therapeutic intervention for OA.

Infectious wounds induce a cycle of bacterial proliferation, oxidative stress accumulation, and dysregulated macrophage polarization, collectively hindering tissue repair. Conventional wound dressings typically address these pathological factors in isolation, resulting in suboptimal therapeutic outcomes. Here, we report the design of a biocompatible multifunctional hydrogel (GAPC). This system integrates self-assembled Proanthocyanidin/Chlorhexidine nanoparticles into a dual-network GelMA/ADM scaffold. Consequently, the hydrogel exhibits simultaneous antibacterial activity, ROS scavenging, and immunomodulatory capacity. In vitro, GAPC hydrogel has superb antibacterial, antioxidant, and anti-inflammatory effects. On the other hand, GAPC hydrogel promotes M1-to-M2 macrophage transition and preserved cellular viability under oxidative stress. Furthermore, in vivo it accelerated infected burn wound closure, enhanced collagen remodeling, and stimulated neovascularization. Collectively, GAPC hydrogel interrupts the "infection-oxidative stress-inflammation" loop, offering a safe and promising option for managing infected wounds.

The CRISPR/Cas system has become an indispensable tool for programmable and accurate biosensing, with its performance critically dependent on precise activity control. While most regulatory strategies have focused on engineering Cas proteins or modifying CRISPR RNAs, relatively little attention has been given to the design of substrate probes. Here, we systematically characterize the trans-cleavage activity of split CRISPR/Cas12a on structured substrates and leverage this insight to engineer a tunable "Hooded" probe with switchable properties. This probe architecture confers protection against trans-cleavage, and its activity can be progressively modulated by varying the probe length. Utilizing this design, we constructed a multiplexed logic-gated detection platform for direct and simultaneous analysis of miRNA and PSA, which demonstrated high sensitivity and specificity. Furthermore, we validated the robust performance of this system for logic-operated imaging in diverse cellular models, confirming its reliability in complex biological settings. Overall, our Hooded probe strategy not only broadens the applicability of CRISPR/Cas12a in molecular diagnostics, but also provides a novel design principle for the multiplexed biosensing.

This study explored the molecular mechanisms by which T7 peptide-modified liposomal irisin (T7@Lipo@Irisin) alleviates perioperative neurocognitive disorders (PND) via regulation of the AMPK/PGC-1α metabolic pathway. T7@Lipo@Irisin nanoparticles were prepared by thin-film hydration and ultrasonic dispersion and showed favorable physicochemical performance, with an encapsulation efficiency of approximately 85%. Serum analysis of healthy donors (n = 10) and PND patients (n = 6) showed higher IL-6 and TNF-α and lower brain-derived neurotrophic factor (BDNF) in PND. In vitro, T7@Lipo@Irisin restored mitochondrial membrane potential, reduced reactive oxygen species (ROS) accumulation, enhanced Neuro-2a hippocampal neuron viability, and activated the AMPK/PGC-1α axis under oxidative stress. In a PND mouse model, it improved Garcia neurological scores, preserved neuronal morphology, and decreased apoptosis. Multi-omic integration of scATAC-seq/scRNA-seq and TMT-based proteomics demonstrated enhanced neuro-glial crosstalk, epigenetic activation of metabolic/antioxidant genes (e.g., Sirt1, Nfe2l2), and upregulated pathways (mitochondrial function, NAD-dependent metabolism, synaptic homeostasis). Proteomics confirmed upregulation of SIRT1, NDUFS2, and BDNF, forming a network linked to energy metabolism and neural repair. Collectively, T7@Lipo@Irisin mitigates PND by activating AMPK/PGC-1α to enhance mitochondrial function and stabilize the neuro-microenvironment.