Corneal neovascularization (CorNV) lacks effective therapies due to poor drug delivery and a complex inflammatory microenvironment, necessitating a deeper understanding of the underlying pathogenic mechanisms and the development of efficient delivery systems. Here we show that neutrophil extracellular traps (NETs) are pathologically activated in CorNV and identify peptidyl arginine deiminase 4 (PAD4) as a key therapeutic target. To achieve coordinated inhibition of NETosis and downstream vascular endothelial growth factor (VEGF) signaling, we engineer a reactive oxygen species (ROS) responsive copolymer encapsulating the PAD4 inhibitor Cl-amidine (Ca@PNH) that hierarchically targets neovascular endothelial cells and activated neutrophils via HRH and neutrophil elastase binding peptide (NEBP) peptides. Upon topical administration, Ca@PNH achieves prolonged ocular retention and lesion specific ROS triggered drug release, suppressing both NETs formation and angiogenesis. In a mouse CorNV model, this spatiotemporal strategy effectively treats neovascularization with excellent safety. Collectively, our work systematically establishes NETs as critical drivers of CorNV, identifies PAD4 as a druggable target, and offers a hierarchical delivery paradigm for inflammation driven ocular diseases.

Microbiota-targeted strategies, such as fecal microbiota transplantation (FMT), hold promise for enhancing immunotherapy but face clinical challenges including safety risks, batch-to-batch variability, and dependence on substantial quantities of donor fecal material. To overcome these limitations, we established a standardized in vitro expansion platform for responder patient-derived gut microbiota. Through optimization of culture conditions, we achieved stable viability and retention of key immunotherapy-enhancing strains over serial passages, enabling a scalable 6.4 × 10[7]-fold expansion. Leveraging this platform, we engineered hybrid nanovesicles derived from bacterial protoplasts, followed by silica surface coating (HNVs@Silica) for oral delivery. The silica shell significantly improved nanovesicle stability under gastrointestinal conditions and conferred superior antitumor efficacy compared with uncoated counterparts. Notably, HNVs@Silica produced from the 2nd, 4th, and 6th passages retained comparable potency in augmenting anti-PD-1 (αPD-1) therapy. Mechanistically, HNVs@Silica remodeled intestinal immunity, particularly within Peyer's patches, by activating and expanding effector CD8[+] T cells. This local response enhanced the priming and systemic dissemination of tumor-specific T cells, thereby improving the efficacy of αPD-1. Collectively, our findings establish a safe, scalable, and orally bioavailable alternative to FMT for enhancing cancer immunotherapy.

Frequent multidrug eyedrop instillation for ocular perioperative prophylaxis (e.g., cataract surgery) suffers from rapid nasolacrimal clearance, limited corneal penetration, and poor access to the posterior segment. To address these limitations, we engineered an interface-adaptive phenylboronate ester hydrogel for rapid in situ gelation and the co-entrapment of two frequently instilled drugs (moxifloxacin and dexamethasone) in their free or liposomal forms at physiological pH. This platform utilizes chemical crosslinking between hyaluronic acid-conjugated 2-aminophenylboronic acid and the diols of polyvinyl alcohol (HA-2APBA@PVA), forming a dynamic hydrogel network via phenylboronate ester bonds with a thermodynamically favorable formation energy (ΔG = -10.64 kcal/mol). This unique chemistry imparts shear-thinning injectability through fine ophthalmic needles, ensures mechanical stability against blink-induced shearing, and drives robust mucoadhesion with tear film mucins such as MUC5AC for prolonged ocular retention at the injection site. Architecturally, a "liposome-in-gel" system establishes hierarchical diffusion steps that suppress the initial burst effect and extend drug release duration by >2-fold compared to the free-drug solution control. The hydrogel demonstrates excellent ophthalmic compatibility, exhibiting physiological osmolality, >80% optical transmittance, and near 100% viability in human corneal and retinal pigment epithelial cells. Safety was further validated via Draize irritation testing and preserved ocular histology across topical and periocular administration routes. Unlike the rapid washout of standard eyedrops, this hydrogel depot ensures prolonged ocular surface retention. Pharmacokinetic analysis revealed route-dependent versatility: compared to standard eyedrops, subconjunctival administration enhanced anterior segment exposure of moxifloxacin and dexamethasone by 14.9- and 8.6-fold, respectively, while sub-Tenon dosing facilitated targeted delivery to the posterior segment. This single, dynamic covalent chemistry provides a scalable, clinically compatible alternative strategy to eyedrops and advances multidrug therapies across both ocular segments.

Cuproptosis-based in situ tumor vaccination provides a unique opportunity to transform tumor destruction into systemic antitumor immunity. However, the oxidative stress required for efficient tumor killing simultaneously disrupts immune competence through copper toxicity and immunosuppressive remodeling. Herein, we develop a spatial redox compartmentalized nanocomposite hydrogel platform to resolve this paradox by selectively amplifying intracellular cuproptosis while preserving the extracellular immune niche. Following peritumoral co-injection of gallic acid-grafted chitosan (CS-GA), dopamine-grafted oxidized hyaluronic acid (OHA-DA), and neocuproine-caffeic acid (NCP-CA) nanoparticles, the hydrogel establishes distinct redox microdomains for tumor eradication and immune protection. Specifically, NCP selectively exploits endogenous exchangeable Cu(I) pools and redirects copper toward mitochondria, where copper valence transformation and ferredoxin 1-mediated activation induce dihydrolipoamide S-acetyltransferase oligomerization, mitochondrial proteotoxic stress, and cuproptotic immunogenic cell death. Meanwhile, CA treatment was associated with increased AMPK phosphorylation and a 63.5% reduction in PD-L1 expression. Simultaneously, catechol-mediated redox cycling by CS-GA&OHA-DA continuously scavenges extracellular reactive oxygen species, alleviating oxidative immunosuppression and restoring T-cell viability to 94.4%. After 14 days of treatment, a substantial tumor regression was observed with a tumor volume reduction exceeding 64.2%. In a contralateral tumor rechallenge model, enhanced infiltration of CD8[+] T-cells (40.8%) and CD8[+] TEM-cells (26.4%) contributed to 75.6% inhibition of tumor recurrence and 93.3% suppression of metastasis. By exploiting endogenous copper resources rather than exogenous copper supplementation and spatially decoupling tumoricidal oxidation from immune-protective antioxidation, this hydrogel platform establishes a new approach for cuproptosis-based in situ vaccination, inducing durable antitumor immune memory and effectively preventing postoperative tumor recurrence and metastasis.

Myocardial ischemia-reperfusion injury remains a major cause of cardiomyocyte loss and adverse remodeling following acute myocardial infarction, yet effective therapeutic options remain limited. Although nicotinamide adenine dinucleotide (NAD[+]) plays a central role in mitochondrial function and redox homeostasis, its therapeutic application is limited by poor membrane permeability and inefficient restoration of intracellular NAD[+] pools. Here, we developed a cardiac-targeted lipid-polymer hybrid nanoparticle (LN-DPC) that enables efficient intracellular delivery of NAD[+] to ischemic myocardium. The nanoparticale integrates boronate-guanidine-mediated NAD[+] encapsulation with an ischemic myocardium-homing peptide (CSTSMLKAC), achieving stable loading, controlled release, and preferential accumulation in injured cardiac tissue. In vitro, LN-DPC enhanced cellular uptake and restored intracellular NAD[+] levels, leading to improved mitochondrial function, reduced reactive oxygen species generation, and alleviation of calcium overload under OGD/R conditions. In vivo, the nanoparticles preferentially accumulated in the injured myocardium, attenuated early inflammation, significantly reduced infarct size, improved cardiac function, and markedly limited chronic fibrosis. Mechanistically, NAD[+] delivery restored SIRT3 expression, improved mitochondrial homeostasis, and suppressed HMGB1-RAGE signaling to mitigate secondary myocardial injury; pharmacological SIRT3 inhibition attenuated the protective effects of LN-DPC both in vitro and in vivo, supporting a functional contribution of SIRT3 to LN-DPC-mediated cardioprotection. Collectively, this work establishes targeted NAD[+] delivery as an effective strategy to restore metabolic homeostasis and protect against ischemia-reperfusion injury, and provides a nanomedicine platform for metabolic intervention in cardiovascular diseases.

Penetration enhancers (PEs) are widely used in transdermal drug delivery systems (TDDS) due to their cost-effectiveness and excellent biocompatibility. However, the rapid selection of PEs for TDDS is hindered by two critical challenges: first, in vitro permeation tests (IVPT) are costly and time consumption; second, the datasets for drug-PE matching are relatively limited. To address these issues, this study established a rapid screening platform for PEs in transdermal formulations using machine learning (ML) approaches based on a self-constructed database and further explored the permeation process of "drug-PE" systems via molecular dynamics simulation (MD). Systematic evaluation of five ML algorithms showed that the Light Gradient Boosting Machine (LightGBM) model achieved the optimal predictive performance using 5-fold cross-validation based on random selection and unseen drug-excipient combination splitting, with R[2] values of 0.78 ± 0.03 (random split), 0.73 ± 0.01 (unseen PE split), and 0.72 ± 0.06 (unseen drug-PE combination split) on the test sets, respectively. In contrast, performance decreased substantially for the unseen-drug split (R[2] = -0.40), indicating that the model performs well primarily for interpolation within the chemical space represented in the dataset but does not reliably extrapolate to entirely unseen drugs. Feature importance analysis revealed a dose-dependent relationship between PE concentration and transdermal efficiency, while the log S of drugs exerted a non-singular effect on transdermal flux. Moreover, transdermal database analysis indicated that drugs with log P values of 2-3, when paired with PEs possessing log P values of 4-6, tended to achieve higher transdermal permeation efficiency. The accuracy of the prediction model was verified by IVPT using two model drugs, berberine and ferulic acid. Furthermore, MD simulations were employed to elucidate the penetration mechanism of the model drugs in "drug-PE" combinations at the atomic level. Finally, a rapid screening platform (https://permapp-online.streamlit.app) was developed, which enabled transdermal flux prediction through the input of molecular descriptors. This platform can provide valuable support for the future design of transdermal formulations.

Retinoblastoma is a rare pediatric intraocular malignancy for which intravitreal administration offers a clinically attractive route for localized therapy owing to its minimally invasive nature and direct access to intraocular tumors. However, efficient intravitreal delivery of nucleic acids remains challenging because the protein-poor vitreous microenvironment restricts nanoparticle transport and subsequent intracellular delivery. Conventional lipid nanoparticles (LNPs), which have been primarily developed for systemic administration, are therefore not necessarily suited to this unique biological environment. Here, we developed a C12-200-containing lipid nanoparticle (CLNP) engineered for efficient intravitreal gene delivery through systematic optimization of lipid composition. The optimized CLNP exhibited favorable diffusion within the vitreous, efficient intracellular mRNA delivery, and reduced dependence on extracellular protein-assisted uptake under vitreous-mimetic conditions. To demonstrate the therapeutic versatility of the platform, human RB transcriptional corepressor 1 mRNA and triptolide were co-delivered as representative gene and small-molecule therapeutics. CLNP-mediated co-delivery of hRB1 mRNA and triptolide produced greater antiproliferative activity than either monotherapy in vitro and markedly suppressed tumor progression in an orthotopic retinoblastoma model following a single intravitreal administration, with no detectable abnormalities in retinal function, structure, or apoptosis during the evaluation period. Collectively, this study establishes a vitreous-oriented lipid engineering strategy that simultaneously improves extracellular transport through the vitreous and intracellular nucleic acid delivery, providing a broadly applicable platform for localized gene delivery and combination gene-drug therapy for ocular diseases.

Glioblastoma (GBM) remains one of the most aggressive and treatment-resistant cancers, in part due to its highly immunosuppressive tumor microenvironment and the restricted therapeutic access imposed by the blood brain barrier (BBB). Focused ultrasound (FUS) has emerged as a promising noninvasive technology capable of transiently opening the BBB, enhancing localized therapeutic delivery, and directly modulating immune responses within the brain. Beyond its established role in drug delivery, accumulating evidence demonstrates that FUS can reshape both innate and adaptive immunity through mechanical, thermal, and cavitation-mediated bioeffects, leading to cytokine release, immune-cell recruitment, antigen presentation, and immunogenic cell death. In this review, we discuss the emerging role of FUS as an immunomodulatory platform in GBM. We first summarize the immunological consequences of different FUS modalities, including thermal ablation, histotripsy, sonodynamic therapy, and microbubble-mediated BBB opening, with emphasis on their effects on the tumor microenvironment. We then review preclinical studies combining FUS with chemotherapy, immune checkpoint blockade, CAR T-cell therapy, and nanoparticle-based therapeutics, highlighting how FUS can enhance both therapeutic delivery and antitumor immune activation. Subsequently, we examine the rapidly expanding clinical landscape of FUS in GBM, including trials focused on BBB opening, sonodynamic therapy, and sonobiopsy approaches for molecular profiling and treatment monitoring. Finally, we discuss current translational challenges, including treatment standardization, device heterogeneity, imaging requirements, and the development of next-generation ultrasound-responsive agents. Collectively, this review positions FUS not only as a delivery technology, but also as a versatile strategy for reshaping the immune microenvironment in GBM and enabling future combination immunotherapies.

Small molecule targeted therapies frequently fail to control brain metastases and primary brain tumors even when systemic exposure reaches toxicity-limiting levels. Systemic exposure-response relationships are often absent, which is well documented for tyrosine kinase inhibitors. Target-site information is therefore needed, since the concentration reached in the tumor is determined not by plasma exposure, but by passive and active transport across the blood-brain barrier and by intratumoral heterogeneity. Physiologically-based pharmacokinetic (PBPK) modeling provides the mechanistic framework to describe this. Models built solely on in vitro-to-in vivo extrapolation of permeability and transport have repeatedly required correction factors derived from rodent data, and their evaluation in humans still depends on invasive sampling or microdialysis. Microdialysis performs poorly for lipophilic, highly protein-bound drugs. We argue that quantitative imaging can supply the missing in vivo input. In a hybrid strategy, in vitro-derived transport parameters serve as the starting point, while blood-brain barrier influx and efflux are recalibrated on positron emission tomography (PET) microdose measurements in patients. This could lead to improved prediction of brain concentrations at therapeutic doses and allows simulated target-site exposure to be weighed against in vitro potency for compound and dose selection. Beyond whole-brain averages, magnetic resonance imaging (MRI) can provide permeability and perfusion parameters longitudinally and in larger cohorts, whereas mass spectrometry imaging (MSI) techniques can map intratumoral heterogeneity in drug distribution in the tumor microenvironment and identify potential sanctuary sites. We map each modality onto the specific PBPK parameters it can inform, and argue that imaging-informed, spatially resolved PBPK is a realistic route toward model-informed precision dosing in neuro-oncology.

Pseudomonas aeruginosa (PA)-induced bacterial keratitis represents one of the most severe corneal infections usually accompanied by prominent inflammatory infiltration. Clinical fluoroquinolone-based therapies yield unfavorable therapeutic outcomes due to the insufficient ocular bioavailability and compromised anti-inflammatory efficacy. In the present work, we report a molecularly engineering strategy to conjugate ciprofloxacin (Cip) with a self-immolative carbonyl sulfide (COS)/hydrogen sulfide (H2S) donating motif. The resulting prodrug (termed ASCip) could spontaneously self-assemble into nanosheets in aqueous solution, thereby improving ocular surface retention and drug bioavailability. Importantly, ASCip could be specifically activated by aminopeptidase N (APN), which is overexpressed at the inflammatory sites, enabling the controlled and simultaneous release of Cip and H2S, to realize antibacterial and anti-inflammatory effects. In vitro evaluations of this carrier-free nanoantibiotic revealed potent antibacterial activity against PA upon APN activation, favorable biocompatibility, and significant anti-inflammatory effects via modulating the inflammatory mediators and promoting macrophage polarization towards the anti-inflammatory M2 phenotype. In a murine model of PA-infected keratitis, the nanoantibiotic efficiently eradicated pathogens, attenuated ocular inflammation, and remarkably accelerated corneal repair, providing a promising strategy to improve the efficacy of fluoroquinolone-based therapies against bacterial keratitis. More importantly, the strategy developed provides a versatile way to co-deliver COS/H2S with other functional motifs.

Spermatogenic disorders are a major cause of male infertility, with oxidative stress-driven Leydig cell dysfunction serving as a key pathological contributor. However, effective antioxidant therapy remains limited by insufficient accumulation in injured testes and by the formulation complexity associated with conventional carrier-based delivery systems. Here, we investigated the pathological microenvironment of di(2-ethylhexyl) phthalate (DEHP)-injured testes and found increased inducible nitric oxide synthase (iNOS) expression together with upregulation of the cationic amino acid transporter 2 (CAT2) in the testicular interstitium, indicating altered arginine metabolism and transport. Based on this pathological feature, we constructed carrier-free maltol-arginine nanoparticles (MAS) through Mannich reaction-mediated covalent conjugation and non-covalent self-assembly. In this system, the arginine moiety functions as a CAT2-recognizable structural cue for lesion-oriented delivery, whereas maltol provides antioxidant activity for redox regulation. MAS preferentially accumulated in injured testes and showed enhanced uptake by Leydig cells in vitro. Mechanistically, MAS promoted Nrf2 nuclear translocation, improved redox homeostasis, and attenuated NF-κB/NLRP3/GSDMD-mediated pyroptotic signaling. In the murine injury model, MAS ameliorated testicular microenvironmental disruption, improved Leydig cell-associated endocrine function, and enhanced sperm quality. Overall, this study presents a functionally engineered, carrier-free biomaterial strategy that redirects pathology-related arginine transport remodeling into a metabolic delivery advantage, providing a viable strategy for targeted intervention in spermatogenic disorders and restoration of testicular homeostasis.

Epigenetic regulation plays a pivotal role in cancer development and progression, in which long non-coding RNAs (lncRNAs) have emerged as one of the most important epigenetic regulators because of their ability to promote immune evasion and tumor progression. Therefore, targeted regulation of lncRNA expression could be a promising epigenetic strategy for enhanced cancer therapy, which however remains challenged due to the localization of lncRNAs in various membrane-bound organelles (e.g., nucleus and mitochondrion). We herein developed a nucleus-targeted RNA interfering (RNAi) nanoplatform for specific silencing of epigenetically induced MYC interacting lncRNA 1 (EPIC1), a nucleus-localized oncogenic lncRNA that is negatively correlated with antitumor immune response and prognosis of breast cancer (BCa) patients. Using BCa orthotopic and metastatic tumor models, we demonstrated this RNAi nanoplatform could efficiently silence EPIC1 expression in BCa cells and significantly inhibit BCa tumor growth. Mechanism study shows this EPIC1 silencing could block the function of histone methyltransferase EZH2 to catalyze the methylation of lysine 27 on histone H3 (H3K27me3) at the promoter region of its target genes, which could not only directly inhibit BCa tumor growth by up-regulating tumor suppressor genes (e.g., BAX and TP53), but also enhance the expression of immunoactive genes (e.g., MHC-I, IFN-β, CXCL9, and CXCL10) to potentiate BCa immunotherapy. Our findings indicate that EPIC1 silencing with the nucleus-targeted RNAi nanoplatform could be a promising epigenetic strategy for effective cancer therapy.

The paradigm shift toward patient-centric precision medicine has established personalized cancer vaccines (PCVs) as a key strategy for eliciting tumor-selective immunity. However, traditional cell-based PCVs face significant translational barriers, including manufacturing complexity and safety concerns. While tumor-derived extracellular vesicles (TDEVs) offer a promising cell-free alternative that recapitulates the parental tumor's antigenic landscape, their therapeutic efficacy is frequently antagonized by intrinsic immunosuppressive cargo. Here, we report ChimeraVax, a chimeric vesicular vaccine platform engineered by covalently conjugating TDEVs with Lactobacillus acidophilus-derived membrane vesicles (LAMVs) via bio-orthogonal, copper-free click chemistry. In this hybrid assembly, the TDEVs serve as a comprehensive antigen reservoir, while the LAMVs provide potent natural adjuvanticity that bypasses TDEV-mediated immunosuppression. In vitro, ChimeraVax facilitated the synchronized delivery of tumor antigens and immunostimulatory molecules to dendritic cells, significantly enhancing antigen uptake and pro-inflammatory signaling. In a murine model of aggressive triple-negative breast cancer, subcutaneous administration of ChimeraVax coordinated both innate and adaptive immune responses, yielding robust antitumor efficacy. Notably, ChimeraVax showed strict antigen dependence, selectively suppressing matched lesions with minimal effects on mismatched tumors, highlighting its specificity and translational potential as a modular personalized cancer vaccine platform.

Multidrug-resistant bacterial infection has become increasingly severe due to the abuse of antibiotics. Recently, carbon monoxide (CO)-based gas therapy has stood out as a novel strategy to overcome antimicrobial resistance and clear infection. However, the application is hindered by the reactive gas nature and poor precise loading. Herein, we developed a peptide-based delivery strategy by covalently binding the photo-sensitive CO donor 3-hydroxylflavone (3-HF) to the N-terminus of antimicrobial peptides (AMPs), achieving controllable and quantitative loading of CO donors. Meanwhile, by altering the amino acid sequences of AMPs, constitutionally isomeric AMP-HF conjugates underwent distinct assemble behaviors and formed different morphologies, leading to distinct light-triggered CO-releasing patterns. In vitro experiments revealed that the sequence-dependent CO-releasing behavior could enhance the antibacterial and anti-inflammatory capacity of AMP-HF conjugates to different extents. In vivo results further confirmed that AMP-HF conjugates could differentially eliminate bacteria and reconstruct the inflammatory environment at the infected site to accelerate the wound healing process.