Aim: Acute lung injury (ALI), marked by vigorous inflammatory reactions and elevated incidence, is a grave clinical issue for which efficacious pharmaceutical interventions are still lacking. This research delves into the anti-inflammatory actions and the associated pathways of nanovesicles originating from human umbilical cord mesenchymal stem cells (UCMSC-NVs) within a model of ALI induced by lipopolysaccharide (LPS).
Methods: UCMSC-NVs were prepared via serial extrusion and characterized using transmission electron microscopy and dynamic light scattering. Their effects on pulmonary inflammation and injury were evaluated in an LPS-induced ALI mouse model. Anti-inflammatory effects were analyzed using Western blot, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence, focusing on the role of hsa-let-7g-5p in regulating the nuclear factor κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) pathway.
Results: The UCMSC-NVs acquired through serial extrusion displayed bilayer vesicle structures, with a mean diameter of around 100 nm. Moreover, these vesicles exhibited elevated expression levels of CD9, CD63, and CD81. Administration of UCMSC-NVs significantly alleviated lung damage, accompanied by a reduction in alveolar leakage and neutrophil infiltration. Furthermore, it significantly downregulated the expression of pro-inflammatory cytokines, such as interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α). UCMSC-NVs reduced macrophage infiltration in the lungs of ALI mice by inhibiting the NF-κB/NLRP3 signaling, leading to a shift in macrophage polarization, with decreased M1 and increased M2 polarization. Our findings demonstrated a novel therapeutic mechanism wherein hsa-let-7g-5p encapsulated within UCMSC-NVs alleviates inflammation by inhibiting NF-κB/NLRP3 expression, thereby mitigating ALI.
Conclusion: These results provide a foundation for the development of novel cell-free therapies with clinical potential for treating inflammatory lung diseases such as ALI and acute respiratory distress syndrome (ARDS).
Aim: Extracellular vesicles (EVs) contribute to stroke rehabilitation by mediating intercellular signaling during inflammation and tissue repair. Here we report EV-associated surface proteins as potential biomarkers for predicting recovery of activities of daily living (ADL) during the subacute phase of ischemic stroke (IS).
Methods: IS patients and healthy controls (HCs) were recruited for this study, with serum samples analyzed across three study stages. In the discovery subset (10 IS, 6 HCs), serum proteomics was used to identify differentially expressed proteins (DEPros) and perform Gene Ontology (GO) enrichment analysis. In the exploration subset (7 IS, 12 HCs), a proximity-dependent barcoding assay (PBA) was employed to profile surface proteins on individual EVs and screen for biomarker candidates. In a validation cohort, patients were grouped by ADL improvement (little-effect recovery, LE, n = 30; obvious-effect recovery group, OE, n = 22) based on Longshi Scale and Barthel Index assessments at admission and at 3 months follow-up. Targeted biomarker validation was performed with enzyme-linked immunosorbent assay (ELISA) and receiver operating characteristic (ROC) analysis.
Results: A total of 113 DEPros were identified, with GO term enrichment in EV-related pathways. PBA profiling revealed matrix metalloproteinase 9 (MMP9), carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), melanoma cell adhesion molecule (MCAM), and gelsolin (GSN) as candidate biomarkers. In the validation cohort, MMP9 and CEACAM1 were significantly elevated in the LE group. ROC analysis showed area under the curve (AUC) of 0.726 for MMP9 and 0.700 for CEACAM1 in distinguishing LE from OE.
Conclusion: Elevated serum levels of EV-associated biomarkers MMP9 and CEACAM1 were associated with poor ADL recovery, supporting their potential as prognostic biomarkers for stroke rehabilitation outcomes.
Microbial extracellular vesicles (mEVs) are emerging as key mediators at the host-microbe interface in the lung, playing a remarkable dual role as both pathogenic drivers and therapeutic modulators. These nano-sized, membrane-bound structures (20-400 nm) secreted by bacteria, fungi, and other microorganisms carry diverse bioactive cargo including lipids, proteins, and nucleic acids that can profoundly influence host immune responses and tissue homeostasis. mEVs derived from probiotic bacteria demonstrate significant therapeutic potential as immunomodulatory agents capable of reducing pulmonary inflammation and enhancing epithelial barrier function. These probiotic-derived vesicles represent a novel class of postbiotics - bioactive microbial products that confer health benefits without requiring live microbial cells. Conversely, in pathogenic contexts, mEVs from bacteria such as Pseudomonas aeruginosa and Escherichia coli trigger robust inflammatory responses in the lung through pattern recognition receptor activation, particularly Toll-like receptors (TLRs)-2 and TLR-4, leading to upregulation of pro-inflammatory cytokines and contributing to chronic respiratory conditions including asthma and chronic obstructive pulmonary disease. Given its extensive surface area and highly specialized immune network, the lung represents a highly receptive site for mEV-mediated interactions. This review synthesizes evidence on pathogenic mechanisms of mEVs and explores their therapeutic potential in respiratory medicine, with specific focus on: (1) the role of environmentally-derived mEVs from dust and airborne sources in chronic respiratory inflammation; (2) recent experimental evidence of probiotic-derived mEVs therapeutic effects across diverse pulmonary conditions and (3) the concept of mEVs as both protective postbiotics and inflammatory triggers in the lung.
Extracellular vesicles (EVs) circulate in body fluids, carrying molecular cargo from their parent cells and exerting diverse biological functions. Consequently, they have attracted considerable attention as biomarkers for disease detection and pathophysiological understanding and have emerged as potential therapeutic targets. Although the number of clinical trials involving EVs is increasing, major challenges remain, including methodological transparency and the heterogeneity of EV subpopulations. Various EV isolation methods are commonly employed, and the primary approaches are summarized in the MISEV2023 guidelines. Each method has advantages and disadvantages; however, most conventional approaches require relatively large liquid volumes (e.g., hundreds of microliters or more) to obtain sufficient EV yields for analysis. In recent years, novel technologies have been developed to overcome these limitations by addressing constraints related to sample volume, simplicity, and accuracy. One such innovation is the cellulose nanofiber-EV sheet, which we developed in 2023. This method enables the capture and stable storage of EVs from microvolumes of body fluids (e.g., approximately 10 µL). Two application methods are available: the attaching method, in which the EV sheet is applied to moist tissue surfaces, and the soaking method, in which the sheet is soaked into body fluids. Each method offers distinct advantages. Given their unique properties, EV sheets may contribute to biomarker analysis and facilitate new research directions across diverse fields. Continued advances in EV isolation and analytical platforms will be essential to support the safe clinical implementation of EV-based diagnostics and therapeutics.
Diseases of the prostate gland, including benign prostatic hyperplasia (BPH) and prostate cancer (PCa), affect a significant proportion of men worldwide. The incidence of these diseases increases with advancing age decreased quality of life and mortality in cases of aggressive PCa. Advanced PCa shows a spectrum of disease states, including castration-resistant prostate cancer (CRPC) and therapy-resistant neuroendocrine prostate cancer (NEPC). NEPC is a highly aggressive, AR-independent state that evolves from CRPC by “lineage switching”. Although the underlying cellular mechanisms have been examined extensively, the role of extracellular-mediated intercellular communication is less well understood. Emerging evidence suggests an important role of extracellular vesicles (EVs) in the pathobiology of these diseases. EVs have been shown to be critical for imparting tumor aggressiveness via inducing epithelial to mesenchymal transition (EMT), stemness and immune evasion. Understanding EV-mediated signaling that governs the progression of prostatic diseases is crucial for developing effective targeted therapies and robust biomarkers for prognosis and risk stratification. Advances in EV engineering have enabled the development of targeted exosome-based therapeutics capable of delivering drugs, biologic payloads, or immune-stimulating signals. In this article, we review our current knowledge on the role of EVs in prostatic diseases, explore their diagnostic and therapeutic applications, and outline future directions aimed at translating EV-based technologies into tools for improved clinical management.
Aim: Oxidative stress is a key driver of cardiovascular disease, underscoring the need for safe and effective antioxidant therapies. This study aims to evaluate the cardioprotective potential of plant-derived nanovesicles (PDNVs) derived from Panax ginseng (Gin) and Rehmannia glutinosa (Glu) against hydrogen peroxide (H2O2)-induced oxidative injury in cardiac cells.
Methods: PDNVs were isolated from medicinal plants via differential ultracentrifugation and characterized for morphology, diameter, stability, and cellular uptake. The antioxidant and cytoprotective effects were assessed in H2O2-injured cardiomyoblasts through cell viability, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH·) radical scavenging, intracellular reactive oxygen species (ROS) and mitochondrial superoxide detection, and antioxidant enzyme (superoxide dismutase, SOD; glutathione peroxidase, GPx) activity assays. The composition of the PDNVs was determined through Coomassie Brilliant Blue staining for proteins, agarose gel electrophoresis for nucleic acids, and liquid chromatography-mass spectrometry system (LC-MS) for bioactive monomers.
Results: Gin-PDNVs and Glu-PDNVs significantly enhanced cardiomyoblast viability under oxidative stress. Glu-PDNVs demonstrated superior efficacy at lower concentrations, with stronger ROS scavenging capacity. Compositional analysis revealed that Glu-PDNVs carry proteins, nucleic acids, and antioxidant herbal compounds such as catalpol, rehmannioside D, and acteoside. Glu-PDNVs also dose-dependently scavenged DPPH· radicals, reduced mitochondrial superoxide accumulation, and significantly restored the H2O2-induced suppression of SOD and GPx activities.
Conclusion: This study provides the first evidence that Glu-PDNVs exert potent cardioprotection by regulating ROS and superoxide homeostasis, positioning them as a promising natural nanotherapeutic platform with translational potential.
Psoriasis is a chronic, immune-mediated inflammatory disease that affects approximately 2%-3% of the global population, and remains a major dermatologic and psychosocial burden. Despite advances in biologics targeting interleukin 17 (IL-17) and interleukin 23 (IL-23) pathways, effective and accessible treatment options for moderate psoriasis are lacking. Topical therapies and phototherapy are often inadequate, while systemic agents and biologics are limited by toxicity, high cost, and restricted reimbursement criteria, leaving patients with moderate disease without adequate therapeutic options. Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have emerged as a promising acellular therapeutic modality that harnesses the immunomodulatory and regenerative properties of parent MSCs. Unlike systemic biologics, MSC-EVs act locally and non-immunosuppressively. Topically applied MSC-EVs have demonstrated the ability to modulate cutaneous inflammation by attenuating complement activation [via CD59-mediated inhibition of complement terminal component 5b-9 (C5b-9) complex formation], reducing neutrophil infiltration, and subsequently lowering IL-17 and IL-23 expression in psoriatic lesions. Preclinical and early clinical studies suggest that MSC-EVs can restore local immune homeostasis through paracrine extracellular mechanisms, without systemic absorption or adverse effects. MSC-EVs represent a new class of cell-free nanotherapeutics inspired by biologics, capable of localized immunomodulation in psoriasis. By combining biologic-like efficacy with the safety and accessibility of topical therapy, MSC-EVs may bridge the long-standing therapeutic gap in moderate psoriasis. This review discusses current treatment limitations, the mechanistic rationale for MSC-EVs in psoriatic inflammation, and their potential to redefine dermatologic immunotherapy.