Aim: Extracellular vesicles (EVs) hold great promise as emerging biomarkers for a variety of diseases. However, their clinical application is still hindered by complex and time-consuming isolation procedures. A clinically relevant scenario where improved biomarker-based diagnostics are urgently needed is prosthetic joint loosening, which may result from either aseptic inflammation or periprosthetic joint infection (PJI) - two conditions requiring fundamentally different therapeutic approaches. This study investigated whether imaging flow cytometry (IFCM) enables the discrimination between aseptic and septic loosening by profiling EVs directly in minimally processed synovial fluid.

Methods: We analyzed synovial fluid from 35 aseptic and 13 septic cases using IFCM to detect surface marker-defined EV subpopulations without prior isolation. Samples were classified based on clinical and microbiological findings. Marker abundance was quantified and analyzed using logistic regression.

Results: Septic loosening was associated with significantly increased levels of CD82⁺ EVs and decreased levels of CD9⁺ EVs. CD82⁺ EVs showed a sensitivity of 83.3% and specificity of 90.0%, while CD9⁺ EVs demonstrated 100% specificity but lower sensitivity (58.8%) for aseptic loosening. CD82⁺ EV abundance was identified as an independent predictor of septic loosening.

Conclusion: IFCM enables rapid and direct detection of diagnostically relevant EVs in native synovial fluid. CD9⁺ and CD82⁺ EVs serve as promising biomarkers for distinguishing aseptic from septic endoprosthesis loosening, offering a fast and robust diagnostic tool that may complement current clinical diagnostics and support timely treatment decisions.

In a recent study, Antounians and colleagues (2024) investigated the potential of amniotic fluid stem cell-derived extracellular vesicles (AFSC-EVs) as a therapeutic intervention for pulmonary hypoplasia resulting from congenital diaphragmatic hernia (CDH). They demonstrated the pronounced inflammatory signature of the fetal hypoplastic lungs, marked by heightened macrophage density in a CDH rat model. Intra-amniotic AFSC-EV administration mitigated the inflammatory process and enhanced fetal lung development by promoting branching morphogenesis and epithelial maturation. These findings add to a growing body of preclinical and clinical evidence supporting the therapeutic potential of AFSC-EVs for inflammation-driven pathologies.

Blebbisomes represent a newly identified class of large extracellular vesicles (EVs) that exhibit unique functional capabilities in intercellular communication. Beyond their motility, blebbisomes engage in complex vesicle trafficking functions. Strikingly, they are capable of both internalizing and secreting other EVs, essentially acting as intermediaries or “hubs” in intercellular communication networks.

Extracellular vesicles (EVs) are secreted by nearly all cell types and fulfil a crucial role in intercellular communication by transporting diverse cargo, including enzymes, mRNA, growth factors, chemokines, and cytokines. Although EVs were initially thought to primarily function in waste elimination, it is now clear that they can be diverted from degradation and instead actively secreted to mediate intercellular communication. While the processes of EV biogenesis, degradation, and release have been extensively studied, many aspects remain poorly understood. The involvement of molecular pathways shared by EV biogenesis and autophagy - a lysosome-mediated disposal mechanism - suggests the existence of common regulatory controls. Despite the partial overlap in molecular machineries involved in cargo sorting, the mechanisms that balance the degradation and secretory pathways of EVs, as well as their interplay with autophagy, remain elusive. This review discusses the molecular machinery that dictates the selective cargo loading into EVs. Additionally, it examines the coordination between degradation and secretory pathways in EV biology and situates these processes within the broader context of autophagy. The substantial overlap in molecular pathways, shared proteins, and complementary mechanisms suggests a high degree of coordination between these systems.

Aim: Gram-negative bacteria release outer membrane vesicles (OMVs) that fulfill many functions including survival during stress conditions, delivery of virulence factors, and nutrient acquisition. Additionally, they are increasingly used as an alternative for live bacteria in vaccine development and as a platform for bioengineering. Recently, OMVs have also been applied to express recombinant outer membrane proteins (OMPs) in their natural context as an alternative to the cumbersome reconstitution in liposomes. Here, we use an Escherichia coli strain that lacks four major OMPs for selective expression of the β-barrel assembly machinery (BAM) complex and PhoE in OMVs.

Methods: OMV production of Escherichia coli BL21(DE3) and its omp8 derivative upon overexpression of BAM and PhoE is compared and characterized.

Results: We find that overexpression of the BAM complex and PhoE causes a strong hypervesiculation phenotype, and the OMVs produced are intact and appear to recruit the BamA subunit of BAM and PhoE in their correctly folded and assembled conformations.

Conclusion: While the exact mechanism of hypervesiculation remains to be elucidated, it contributes to the suitability of the BL21(DE3)omp8 host strain to produce recombinant OMP-enriched OMVs that can be used for various purposes, including structural analysis.

Aim: Extracellular vesicles (EVs) have emerged as promising vehicles for the delivery of small non-coding RNAs (sncRNAs); however, their clinical translation is hindered by the lack of standardized manufacturing methods, RNA loading protocols, and dosing strategies in both preclinical and clinical settings. This review aims to analyze the current landscape of EV-based RNA therapeutics to identify key trends and discrepancies, providing insight for the clinical development of future sncRNA-loaded EVs.

Methods: PubMed and Google Scholar were used to identify 74 published articles using cell-derived EVs loaded with sncRNA. EV source, EV surface modifications, type of loaded RNA, loading methods, and dosages used in preclinical studies were quantitatively analyzed to identify trends and discrepancies.

Results: Most studies utilize naïve EVs derived from stem or immortalized cells, with electroporation and donor cell transfection being the predominant RNA loading strategies. EV loading and dosage schemes in preclinical studies are mainly based on protein content, while only a minority of studies use particle number. More generally, the variability in measurement units reflects the absence of standardized guidelines for both RNA loading and treatment dosing, generating variability and challenges in comparing results across studies.

Conclusion: Reliable dosing strategies are extremely important for determining the therapeutic potential of EVs in preclinical settings and ensuring clinical translatability. However, a standardized framework for EVs as robust platforms for RNA delivery remains to be established. We underscore the critical need for universal quantification methods, standardized measurement units, and reproducible protocols for EV production and application.

Exosomes, as key mediators of intercellular communication, have shown great potential in disease intervention and therapy in recent years. As natural nanocarriers, exosomes play a crucial role in transporting a wide array of cargo. Among these, miRNAs carried by exosomes are pivotal in gene regulation and the modulation of cellular signaling. Given that miRNAs are essential gene regulators, understanding how miRNAs are selectively loaded into exosomes is crucial for the development of novel diagnostic and therapeutic approaches. This review provides a detailed overview of the biogenesis and secretion mechanisms of exosomes, with a particular focus on the molecular mechanisms governing miRNA sorting into exosomes. Specifically, it highlights the miRNA motifs associated with exosomes enrichment (EXOmotifs), as well as those related to intracellular miRNA enrichment (CELLmotifs), along with RNA-binding proteins (RBPs) involved in sorting. We summarize the current progress in this field and discuss strategies for engineering Exosomes - such as gene editing, drug loading, and surface modification - to enhance their functionality and specificity. By exploring these mechanisms, this review offers a theoretical foundation for the application of engineered exosomes in disease treatment and outlines future research directions and potential applications.

The GISM Annual Meeting 2025 convened experts in regenerative medicine and nanomedicine to discuss recent advances in mesenchymal stromal/stem cell (MSC) research and extracellular vesicle (EV) technologies. The meeting emphasized novel strategies to enhance the therapeutic potential of MSCs and EVs, addressing both basic biological insights and translational challenges. Discussions highlighted the importance of standardizing production and characterization methods to improve scalability and reproducibility for clinical applications. Emerging therapeutic approaches, including cell engineering and targeted drug delivery, were showcased alongside preclinical and clinical studies. The conference provided a platform for interdisciplinary exchange, fostering collaboration and paving the way toward the clinical integration of EV and cell-based nanomedicine.

In December 2024, the United Kingdom Society for Extracellular Vesicles (UKEV) held its annual forum in Newcastle upon Tyne, marking 11 years since its founding. UKEV Forum 2024 brought together over 230 participants from the UK and abroad under the theme “Bridging Innovation and Impact”. The meeting emphasised translational science, regulatory foresight, methodological rigour, and cross-sector collaboration, reflecting the maturation of EV research towards clinical relevance. Hosted at Newcastle University, the event included plenary lectures, oral and lightning talks, poster sessions, and an Early Career Researcher (ECR) Day. Scientific discussions spanned EV biomarker discovery, mechanistic studies, tissue-derived EVs, and novel analytical tools such as cryo-TEM, electrochemical sensors, and DNA-PAINT microscopy. The forum showcased emerging topics in EV isolation, reproducibility, therapeutic development, and regulatory integration, drawing on diverse expertise across academia, biotech, and clinical sciences. Generous industry sponsorship and inclusive programming made UKEV 2024 a landmark event that reinforced the UK’s leadership in EV research.

Extracellular vesicles (EVs) are a type of cell-released phospholipid bilayer nanoscale carrier. However, research on EVs encounters several challenges, such as their heterogeneity, the complexities associated with their isolation and identification, the necessity for engineering optimization, and the limitations in exploring their mechanisms. The advancement of artificial intelligence (AI) technologies offers new opportunities for EV research. Here, the definition and brief history of AI, as well as types and common models of machine learning, are first introduced, and the interactions between AI, machine learning, and deep learning are explored. The article then discusses in detail a variety of applications of AI in EV research, including the use of AI for target identification and selective delivery of EVs, the design and optimization of drug delivery systems, the mapping of cellular communication networks, the analysis of multi-omics data, and synthetic biology-based research on EVs. These applications demonstrate the potential of AI in advancing EV research and applications. Finally, we offer an outlook on the major challenges and future prospects of AI. Overall, the introduction of AI technologies has provided new perspectives and tools for the study of EVs, which is expected to enhance the application of EVs in disease diagnosis and treatment.

In the past decade, extracellular vesicles (EVs) have gained increasing attention in biomedical research. These membrane-bound particles are naturally secreted by cells under both physiological and pathological conditions, and they exhibit a wide range of sizes and molecular compositions. EVs transport bioactive molecules - such as proteins, nucleic acids, and lipids - making them ideal candidates for biomarker discovery. Consequently, their accurate characterization and quantification are critical for understanding their roles in intercellular communication and evaluating their potential in diagnostics, prognostics, disease monitoring, and therapeutic applications. Microfluidic technologies offer promising solutions for EV analysis, addressing key limitations of conventional methods by enabling precise and sensitive measurements with small sample volumes. While microfluidic devices have been predominantly used for EV separation and isolation, their application in EV quantification remains underexplored. Compared to traditional techniques like nanoparticle tracking analysis or flow cytometry, microfluidic systems can provide faster, more accessible alternatives for EV quantification. This review summarizes recent advances in microfluidic technologies for EV quantification, discussing their advantages, current limitations, and future prospects.

Kidney disease, encompassing both acute kidney injury (AKI) and chronic kidney disease (CKD), represents a major global health challenge. A pivotal aspect of the pathogenesis of these conditions is damage to renal tubular epithelial cells (TECs), which contributes to maladaptive repair mechanisms and fibrosis. Due to their essential role, TECs are regarded as a promising target for innovative therapeutic strategies. Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have attracted increasing attention for their therapeutic potential in kidney disease, with extensive literature documenting their beneficial effects on TEC damage through targeted mechanisms. In this review, we critically examine the existing literature on the targeting of TECs by MSC-EVs in both in vitro and in vivo settings. Furthermore, we highlight the limitations and potential of MSC-EV-based strategies for TEC targeting, aiming to provide insights for future clinical trials and therapeutic applications.

Neuroaging is a complex biological process in which the brain undergoes progressive functional decline marked by synaptic loss, neuroinflammation, and cognitive decline. At the molecular and cellular level, aging is driven by multiple interconnected hallmarks, including genomic instability, telomere attrition, epigenetic alterations, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. Among these, cellular senescence, a state of irreversible cell cycle arrest, has emerged as a critical contributor to brain aging. Senescent cells accumulate with age, driven by the p53-p21 and p16-pRb pathways, and secrete pro-inflammatory factors via senescence-associated secretory phenotype (SASP), thereby exacerbating neurodegeneration, vascular dysfunction, and cognitive decline. Extracellular vesicles (EVs) are natural nanocarriers of proteins, lipids, and nucleic acids, and have emerged as key mediators of intercellular communication and therapeutics for aging and age-related conditions. EVs derived from various cell types, such as mesenchymal stem cells (MSCs), neural stem cells (NSCs), and induced pluripotent stem cells (iPSCs), can modulate senescence-related pathways, reduce inflammation, and promote tissue repair. Preclinical studies demonstrate that stem-cell-derived EVs can improve cognitive performance, enhance neurogenesis, reduce senescence phenotype, improve neuronal survival through neuroprotective miRNAs (miR-181a-2-3p), suppress neuroinflammation via inhibition of NLRP3 inflammasome, and support synaptic plasticity. Stem cell EVs possess natural biocompatibility, the ability to cross the blood-brain barrier (BBB), and targeted delivery mechanisms, making them promising candidates for anti-aging interventions. This review elaborates on the multifaceted role of stem cell EVs in mitigating brain aging, senescence, and age-associated chronic disease phenotype.

In recent years, the prevalence of ocular diseases has increased considerably. However, timely diagnosis and treatment are hampered by the challenge of early detection since symptoms often appear in advanced stages. Emerging research highlights extracellular vesicles (EVs) as potential biomarkers for ocular diseases, with tear-derived EVs offering a minimally invasive source for early diagnosis. Tears play a crucial role in maintaining eye health and reflect the physiological state of the eye; thus, abnormalities in tear composition can provide valuable insight into inflammatory eye diseases. Studies have demonstrated the utility of tear-derived EVs in identifying biomarkers not only for inflammatory eye diseases but also for neurodegenerative disorders, as they carry molecular signatures (including proteins and various RNA species) reflective of their cells of origin. In this review, we discuss the potential of tear-derived EVs as biomarkers for early detection and monitoring of ocular and neurodegenerative diseases and highlight the importance of standardizing tear collection and EV isolation protocols to ensure reproducibility.