Accumulation of reactive oxygen species (ROS) following skin injury impairs tissue regeneration by inducing inflammation, oxidative damage, and cellular apoptosis. To address this challenge, we developed a multifunctional semi-interpenetrating polymer network hydrogel composed of silk fibroin methacrylate (SFMA) and Pluronic F127 (PF). The optimized SFMA9/PF1 hydrogel exhibits excellent mechanical compliance, electrical conductivity, antioxidant activity, and anti-freezing capacity. Incorporation of PF enhanced porosity, water retention, and ionic conductivity (∼0.42 S·cm−1), while preserving optical transparency and tunable viscoelasticity. Rheological and degradation analyses demonstrated outstanding elasticity, thermal stability, strain resistance, and sustained biodegradability under physiological conditions. Notably, the hydrogel maintains flexibility and structural integrity at subzero temperatures (−20°C and −80°C), indicating strong freeze resistance. In vitro assays confirmed superior cytocompatibility, with significant promotion of fibroblast adhesion, migration, and proliferation. The hydrogel also exhibited potent antioxidant capacity, effectively scavenging 1,1-diphenyl-2-picrylhydrazyl and H2O2 radicals and reducing intracellular ROS levels. In a full-thickness skin wound model in SD rats, SFMA9/PF1 hydrogel accelerated wound closure, enhanced re-epithelialization, and promoted organized collagen remodeling. Histological and immunofluorescence analyses revealed increased angiogenesis (CD31), reduced myofibroblast activation (α-SMA), and suppressed pro-inflammatory cytokine expression (IL-6), indicating the establishment of a regenerative and anti-inflammatory microenvironment. Overall, the SFMA9/PF1 hydrogel integrates mechanical adaptability, bioactivity, and oxidative stress modulation, representing a promising platform for advanced wound healing and skin tissue engineering.
Precise evaluation of cervical lymph node metastasis (CLNM) and genetic mutations (BRAFV600E/TERT promoter, TERTp) is pivotal for tailoring surgical and prognostic evaluation and adjuvant strategies in thyroid cancer (TC). Although current methods have limitations, we aim to develop deep learning (DL) models to predict CLNM and genetic mutations from TC frozen sections. We developed a DL framework using 2499 frozen-section whole-slide images from 2176 TC patients across five centers. The model was trained with a transfer learning-based feature extractor and an attention-based multiple instance learning (MIL) classifier, and validated on both internal and external cohorts. StyleGAN3-based data augmentation was employed to tackle class imbalance for TERTp prediction, while interpretability was assessed via attention heatmaps and Leiden clustering. The CLNM prediction model achieved a patient-level AUROC of 0.918 internally and 0.803–0.885 across three external validation datasets. For BRAFV600E prediction, AUROCs attained 0.814 internally and spanned 0.750––0.811 in external validation. In TERTp mutation prediction, GAN-based augmentation increased the AUROC to 0.804 (internal) and 0.732 (external), up from 0.782 and 0.724, respectively. Attention maps visualized CLNM correlations with invasive tumor margins, while mutations localized to specific cellular morphology features. Our DL models accurately predict CLNM and genetic mutations from TC frozen sections, potentially reducing unnecessary procedures and providing a rapid alternative to traditional molecular testing.
Owing to the rich repertoire of bioactive cargos inherited from parent cells, apoptotic extracellular vesicles (ApoEVs) have emerged as powerful biological agents in regenerative medicine. To augment ApoEVs regenerative potency, microenvironment-engineered ApoEVs (ME-ApoEVs) have been proposed, which mainly focused on embedding a single microenvironment factor within the vesicle cargo. Since the microenvironment operates as a system composed of multiple factors that collectively govern regeneration, we introduce a novel holistic microenvironment-engineered ApoEVs (hME-ApoEVs) for superior ME-ApoEVs. To validate the hME-ApoEV concept, we endowed ApoEV with a bone developing microenvironment, aiming to recapitulate developing features in adult bone repair. In the study, the developing bone microenvironment was first identified as an M2a-like macrophage driven immune microenvironment accounting for multidimensional developmental signatures. Based on these findings, we successfully fabricated hME-ApoEVs via stimulating the M2a-like developing microenvironment in vitro and confirmed its recapitulation of developmental signatures. Thirdly, we found that hME-ApoEVs recapitulated multidimensional developmental signatures in bone marrow-derived mesenchymal stem cell (BMSC) likely through activating Wnt/β-catenin signaling pathway. Finally, hME-ApoEVs were verified to retain development signatures in adult and senile cranial bone repair, improving bone regeneration. Instead of engineering a single microenvironment factor, the hME-ApoEV strategy effectively recapitulated the key features of the desired microenvironment in themselves, target cells and animal models via imprinting key features onto a simulated microenvironment generated by key cells in vitro. This strategy equips ApoEVs with complex microenvironment cues, amplifying their potency in regeneration medicine and providing a promising foundation for future microenvironment-inspired ApoEV engineering strategies.
Dental caries, a highly prevalent global health issue, is fundamentally driven by the acidogenic metabolism of cariogenic bacteria such as Streptococcus mutans. Conventional approaches for assessing acidogenic capacity remain limited by complex operational requirements and insufficient sensitivity for point-of-care use. Herein, we engineer a CRISPR/Cas12a-powered dendritic DNA nanostructure via hybridization chain reaction to construct a highly efficient signal amplification system. The designed activator strands self-assemble into dendritic nano-assemblies that enable multivalent immobilization of Cas12a-crRNA complexes through programmable base pairing, resulting in a confined microdomain with dramatically enhanced local enzyme density and accelerated trans-cleavage kinetics. The CRISPR/Cas dendritic aggregate enables high-density immobilization of Cas12a, resulting in increased local enzyme concentration and enhanced catalytic cleavage efficiency. By integrating this system with a smart pH-responsive molecular release mechanism, we further develop a paper-based origami biosensor (CDEOB) for the on-site and equipment-free quantification of acidogenic pathogens. The biosensor achieves an ultra-low detection limit of 98.1 CFU/mL in saliva and demonstrates excellent reliability across 101 clinical samples without pre-amplification. This work provides a novel functional nucleic acid material design strategy and a versatile biosensing platform, highlighting substantial potential for early diagnosis of dental caries and other infectious diseases at the point of care.
Multidrug-resistant hypervirulent Klebsiella pneumoniae (MDR-hvKP) infections present a therapeutic challenge owing to profound antibiotic resistance and the paucity of cross-serotype protective strategies against distinct serovars (e.g., K1/K2). Mice were subcutaneously immunized with the viable, hypovirulent, multidrug-resistant (MDR) strain Kp00198874 (serotype K64). Cross-reactive monoclonal antibodies (mAbs) were derived using hybridoma technology. Immunization elicited cross-serotype protective antibodies, ensuring 100% survival rates against lethal challenges posed by the K1/K2 serotype strains. In contrast to single neutralizing antibodies, mAbs 2D4 and 2E10 modulate phagocytic bacteria and immune responses. Prophylactic administration of mAbs conferred 100% survival rates against Kp-GZ8 and significantly prolonged the median survival time against ATCC43816 cells. Therapeutic intervention with mAbs mitigated post-infection pulmonary hemorrhage and inflammatory responses. mAbs 2D4 and 2E10 synergistically targeted capsular polysaccharide/protein epitopes to achieve cross-serotype-protective efficacy. This study established live Kp00198874 immunization as a novel platform for generating cross-protective mAbs against lethal MDR-hvKP, with 2D4 and 2E10 providing complementary mechanisms (rapid pathogen clearance and immunomodulation) through conserved epitope targeting, offering an antibody-based alternative for resistant infections.
Social isolation during adolescence is a critical risk factor for negative emotional symptoms, yet its neurobiological mechanisms remain incompletely understood. Here, we integrate human and mouse data to investigate the behavioral, functional, and molecular consequences of social isolation across development. In humans from the Human Connectome Project in Development, reduced social connectedness was robustly associated with greater depressive and anxiety symptoms across adolescence, with sex-specific patterns in effect distribution. At the neural level, social experience was preferentially associated with alterations in hippocampal functional connectivity (FC) rather than gross structure, and exhibited sex- and hemisphere-dependent coupling with large-scale cortical networks. In parallel, adolescent mice exposed to chronic social isolation exhibited marked reductions in hippocampal synaptic density and calcium signaling, accompanied by astrocyte-associated upregulation of complement components C3 and complement factor properdin and enhanced complement-mediated microglial synaptic pruning. Genetic deletion of C3 rescued synaptic deficits and alleviated depressive-like behaviors. Together, these findings identify a conserved neuroimmune pathway through which adolescent social isolation disrupts hippocampal FC, with sex- and species-specific phenotypic expression across behavioral and neural levels, highlighting a sex-specific mechanism linking social experience to neuropsychiatric risk.
The brain-gut axis shapes Crohn's disease (CD) pathogenesis, yet CD-associated neurophenotypes lack defined clinical and mechanistic significance. This work aimed to define these neurophenotypes, assess their prognostic impact, elucidate neurophenotype-driven progression mechanisms using multi-omics, and validate their therapeutic potential in vivo. 109 CD patients were prospectively recruited from two centers and underwent baseline multimodal neuro-MRI, MR enterography, ileocolonoscopy, and fecal/blood sample collection. The neurophenotypes were characterized using a multimodal neuro-MRI model developed from 13 of 13,870 features. 83 patients were followed up for disease progression, with repeated brain-gut assessments. Multi-omics (fecal microbiome/metabolomics, serum metabolomics/neurotransmitters) were used to decode the mechanisms underlying neurophenotype-driven progression. Dextran sulfate sodium (DSS)-induced colitis mice with different neurophenotypes were treated with repeated transcranial magnetic stimulation (rTMS) to explore its therapeutic potential. Multimodal neuro-MRI accurately mapped CD-specific neural signatures, stratifying patients into high-risk (neurophenotype score ≥ 0.45) and low-risk neurophenotypes with robust cross-center validity (training cohort AUC = 0.842, test cohort AUC = 0.824). High-risk neurophenotype was identified as an independent predictor of accelerated disease progression (Hazard ratio = 15.46, p = 0.030), independent of intestinal inflammation. Integrated multi-omics revealed that the high-risk neurophenotype contributed to CD progression through microbial-metabolic-neurotransmitter networks where tryptophan emerged as the central regulatory hub. Serum tryptophan levels were negatively correlated with neurophenotype severity and intestinal disease progression. rTMS targeting high-risk neurophenotype in DSS-induced colitis mice elevated tryptophan levels and ameliorated intestinal disease activity. This study redefines the high-risk neurophenotype as a sustained pathogenic driver rather than a mere phenomenon, proposing brain-gut axis modulation as a promising therapeutic strategy distinct from conventional anti-inflammatory approaches.
Sleep deprivation (SD) threatens human health and increases the prevalence of disease with abnormal glycolipid metabolism, including metabolic dysfunction-associated steatotic liver disease (MASLD) and insulin resistance (IR). However, the mechanism underlying SD induced MASLD and IR remains incompletely understood. Plasma samples were collected, and circulating exosomes were isolated. Differentially expressed miRNAs in exosomes were identified using an miRNA array and quantitative real-time PCR (qPCR). MiRNA mimics or inhibitors were transfected into the cells to mimic the change in miRNA expression, and the biological effects of differentially expressed miRNAs were detected. Western blot and dual-luciferase reporter assay were used to detect the specific target gene and molecular mechanism of miRNAs. Sleep deprived rats derived circulating exosomes strongly induced hepatic steatosis and IR. miRNA array combined with qPCR identified miR-3572-5p and miR-183-5p in exosomes that impacted abnormal glycolipid metabolism. Subsequently, miR-3572-5p inhibited fatty acid β-oxidation by targeting PGC-1α aggravated hepatic lipid accumulation. Meanwhile, both miR-3572-5p and miR-183-5p exerted an inhibitory effect on the PI3K/Protein Kinase B/GLUT4 insulin signaling pathway through distinct targeting of IRS1 and IGFBP7, which in turn contributed to the aggravation of IR. Notably, circulating exosomes enriched in miR-3572-5p were traced to peripheral blood monocytes, whereas those carrying miR-183-5p were pinpointed to adipose tissue. SD accelerates the progression of MASLD and IR by up-regulating the expression of miR-3572-5p and down-regulating the expression of miR-183-5p in circulating exosomes, thereby inhibiting fatty acid β-oxidation and disrupting insulin signaling pathways.
Fusobacterium nucleatum (F. nucleatum), recognized for its opportunistic pathogenicity, is a prevalent pathogen implicated in several diseases. Emerging evidence highlights the crucial role of F. nucleatum–derived extracellular vesicles (F. nucleatum EVs) in mediating virulence and serving as carriers of diverse bioactive components that facilitate host–pathogen interactions. F. nucleatum EVs can disseminate from the oral cavity to the gastrointestinal tract and joints, promoting the onset and progression of oral inflammatory diseases, head and neck squamous cell carcinoma, rheumatoid arthritis, inflammatory bowel disease, and colorectal cancer through various signaling pathways. A deeper understanding of F. nucleatum EVs will advance the understanding of the pathogenesis of related diseases and inform the development of effective infection control strategies. This review systematically summarizes recent progress in the biogenesis, molecular composition, and pathogenic roles of F. nucleatum EVs across various disease contexts and evaluates potential strategies for the diagnosis, treatment, and management of conditions involving F. nucleatum and its EVs.
Conventional drug delivery methods, such as oral administration and intravenous injection, are associated with several challenges including high systemic toxicity, difficulty in dosage control, and delayed therapeutic responses. These issues limit both the efficacy of treatments and patient compliance, thus becoming a bottleneck in the advancement of precision medicine. In recent years, implantable drug delivery systems (IDDS) have emerged as a promising frontier at the intersection of precision medicine and medical engineering. Through in vivo implantation coupled with signal response mechanisms, IDDS facilitates targeted drug delivery, dynamic regulation, and real-time responsiveness. These significantly enhance therapeutic efficacy while reducing toxicity levels, offering a new path to address the limitations inherent in traditional delivery methods. This article reviews the research progress of IDDS. The technical principles and module innovations are systematically elaborated based on three mechanisms: endogenous response, exogenous triggering, and closed-loop control. Furthermore, it explores the pivotal role that flexibilization and miniaturization of IDDS play in improving biocompatibility, tissue adhesion capabilities, and spatial adaptability. The article also summarizes application examples within typical disease models while analyzing core challenges related to clinical translation. Finally, it anticipates future development directions for integrating medical engineering into this evolving field.
Intracerebral hemorrhage (ICH) is a severe neurological condition characterized by high mortality and long-term disability. The pathophysiology of ICH involves not only primary hematoma formation but also secondary injury mechanisms such as blood–brain barrier disruption, oxidative stress, excitotoxicity, and neuroinflammation. Hematoma-derived factors like hemoglobin, iron, and thrombin, together with pro-inflammatory cytokines from activated glial and immune cells, exacerbate neurovascular dysfunction. Emerging research highlights the role of the bone-brain axis, where bone marrow-derived cells and osteokines influence systemic inflammation, angiogenesis, and tissue repair. Hydrogel-based drug delivery systems (HDDS) have gained attention for their potential to address these challenges, offering controlled release, biocompatibility, and targeted delivery. This review summarizes recent advancements in hydrogel-based therapeutic strategies for ICH treatment, focusing on structure-activity relationships and the integration of bone-brain axis modulation for enhanced neuroregeneration. We discuss key translational barriers, including biosafety, manufacturing scalability, and inter-patient heterogeneity. Future advances are expected to rely on the development of dynamic, microenvironment-responsive hydrogels that can synchronize drug release with stage-specific pathological cues, improving therapeutic precision. Moreover, combining HDDS with multimodal therapies and minimally invasive neurosurgical techniques may further enhance clinical outcomes. The potential for HDDS to target both local brain injury and systemic inter-organ crosstalk represents a promising direction for advancing clinical applications in ICH.
Artificial cells have emerged as a class of promising bioactive materials that recapitulate key structural and functional features of natural cells, serving as an effective alternative to address the inherent bottlenecks of conventional biomaterials in biomedical research, particularly in cancer theranostics. Fabricated primarily via bottom-up strategies, these bioactive constructs enable modular assembly of functional biomaterials to build hierarchical systems spanning artificial organelles, diverse artificial cells and prototissues, endowing artificial cells with tuneable mechanical properties, programmable environmental responsiveness and precise targeted delivery capabilities. Unlike traditional cancer diagnostic and therapeutic agents that suffer from poor tumour specificity, rapid systemic clearance and severe off-target effects, artificial cells leverage their biomimetic design to achieve prolonged circulatory retention, precise tumour site accumulation, and controlled release of diagnostic probes and therapeutic payloads. This review systematically explores the major types of artificial cells, modular bottom-up construction methods, and advanced biomimetic behaviours of artificial cell systems. Additionally, it highlights their cutting-edge applications in integrated cancer theranostics, including tumour biomarker sensing, targeted drug delivery, and immunomodulatory therapy. Finally, the review addresses key challenges and future directions to accelerate their clinical translation in oncology.
Inflammatory bowel disease (IBD) is characterized by gut microbial dysbiosis, dysregulated host immune responses and increased susceptibility to certain intestinal infections. These factors complicate the distinction between active infection, inflammatory flare, and mixed inflammatory infectious states. Conventional diagnostics remain essential in clinical practice, but a single test is often insufficient to separate colonization, true infection, and inflammation-related symptoms. This review summarizes current evidence on multi-omics biomarkers relevant to intestinal infection and inflammation in IBD, including metagenomics, metabolomics, proteomics, single-cell RNA sequencing, spatial transcriptomics and radiomics features. Biomarkers linked more closely to infection are considered alongside those that mainly reflect inflammatory activity, with attention to areas of overlap. The review also examines the degree of validation across studies, major sources of heterogeneity and confounding, and the strengths and limitations of emerging diagnostic platforms and artificial intelligence (AI) assisted integrative approaches. Overall, multi-omics strategies may improve diagnostic stratification and deepen understanding of disease mechanisms in IBD. However, most candidate biomarkers and related platforms remain insufficiently validated for routine clinical use.
Extracellular vesicles (EVs) are cell-derived particles delimited by a lipid bilayer and containing a wide variety of cargos that cannot replicate independently. Their key role as mediators of intercellular communication implicates them in a broad spectrum of physiological and pathological processes. In recent years, EVs have gained increasing attention as potential biomarkers, therapeutic targets and drugs delivery vehicles. However, the successful translation of EV-based strategies from bench to bedside requires a deeper understanding of EV composition and the biogenesis mechanisms. A particularly promising and actively investigated avenue in EV research is the targeted inhibition of EV biogenesis. Modulating these processes not only provides valuable tools for dissecting EV functions in vitro and in vivo, but also opens new therapeutic possibilities, especially in diseases where EVs contribute to pathogenesis, such as cancer, neurodegeneration, inflammation and infectious diseases. Identifying and characterizing compounds that interfere with EV biogenesis is therefore of high relevance, both for fundamental research and clinical applications. This review aims to contribute to the field by offering a comprehensive overview of the most studied EV inhibitors, highlighting their mechanisms of action, experimental utility, and potential for therapeutic exploitation. Particular attention is given to their advantages and limitations as modulators of EV biogenesis, with the goal of guiding future research toward more effective and selective strategies.
Hematologic malignancies (HMs) are aggressive neoplasms originating from lymphoid or myeloid lineages. They are characterized by heterogeneous clinical manifestations, high refractoriness, and frequent relapse. Recent advances in extracellular vesicle (EV) research have revealed their translational potential in improving the diagnosis, prognosis, and treatment of HMs. EVs are nanoscale membrane-bound particles secreted by cells. They carry bioactive cargo, including nucleic acids, proteins, and metabolites, and mediate intercellular communication. In HMs, EVs play essential roles in tumor initiation, progression, metastasis, angiogenesis, metabolic reprogramming, and drug resistance, positioning them as pivotal contributors to disease pathogenesis. Leveraging their functional versatility and disease-specific molecular signatures, EVs show promise as non-invasive biomarkers for early detection and disease monitoring. They also represent potential therapeutic targets to disrupt tumor-microenvironment crosstalk, and as engineered drug delivery systems for precision medicine. This review systematically examines the pathobiological functions of EVs in HMs and highlights emerging clinical applications, offering valuable insights to guide future research.