Aim: Whole blood constitutes a robust and readily available resource for the identification and characterization of systemically circulating substances. Despite the relevant number of blood specimens archived in biobanks globally, their utilization in extracellular vesicle (EV) research remains considerably underexploited. In this study, we present an optimized PRotein Organic Solvent PRecipitation (PROSPR) workflow, specifically designed for the rapid and standardized enrichment of EVs directly from untreated blood samples.

Methods: PROSPR workflow was applied for EV enrichment directly from untreated blood. Comprehensive biophysical characterization was performed using dedicated flow cytometry, tunable resistive pulse sensing, and transmission electron microscopy. In parallel, discovery-driven lipidomics and high-depth proteomics were applied to assess the suitability of PROSPR-derived EVs for advanced systems biology and multi-omics integration.

Results: PROSPR consistently yielded highly purified EV preparations that preserved canonical vesicular morphology, structural integrity, and stable ζ-potentials, indicative of intact membrane characteristics. Multi-omics analyses revealed distinct and coordinated lipid-protein signatures uniquely enriched in EV populations derived from whole blood. These signatures predominantly reflected platelet- and mitochondria-associated cargo that was not detectable in corresponding plasma-derived EV fractions, highlighting key molecular features missed by plasma-based EV profiling approaches.

Conclusion: Consequently, the optimized PROSPR workflow presented here offers a robust and accessible methodological strategy to fully utilize dormant, archived blood resources within global and national biobanks, thereby enabling comprehensive mechanistic insights and facilitating the discovery of novel diagnostic and prognostic circulating biomarkers.

This conference report summarizes the Global Conference on Research and Application of Chinese herbal Medicine Derived Extracellular Vesicles-like Particles (CHM-EVLPs) held in Guangzhou. The conference gathered worldwide scholars to exchange cutting-edge advances in extracellular vesicle (EV) extraction, functional modification, disease treatment, and industrial transformation via six themed forums covering basic research, fruit EVLPs, laboratory testing, HIV intervention, and youth innovation. Experts shared standardized separation technologies, engineered modification strategies and multi-target therapeutic potentials of plant-derived extracellular vesicles (PDVs) against tumors, intestinal disorders, metabolic diseases and neurodegenerative injuries, while discussing key bottlenecks including large-scale production, quality control and regulatory guidelines. Cross-kingdom molecular communication, oral nano-delivery systems and interdisciplinary combinations with microneedles and artificial intelligence were highlighted as promising research directions. The event promoted international academic cooperation, clarified translational challenges of CHM-EVLPs products, and laid a foundation for advancing standardized, industrialized and globalized development of PDVs to facilitate worldwide medical care improvement.

Aim: Endometriosis is a chronic inflammatory disorder requiring clinical examination, imaging, and surgical intervention for accurate assessment. The aim of the present study was to provide a detailed biophysical and biochemical characterization of small extracellular vesicles (sEVs) derived from peritoneal fluid, and to explore disease-associated molecular differences that may inform future investigations.

Methods: sEVs were isolated from the peritoneal fluid of women with endometriosis (n = 7), women without endometriosis who underwent surgery for benign, noninflammatory conditions (n = 7), and follicular fluid samples (n = 3). The tetraspanin profile of the samples was determined using single-particle interferometric reflectance imaging sensing. After size exclusion chromatography and ultracentrifugation, atomic force microscopy (AFM) and Raman spectroscopy were employed to characterize the sEVs.

Results: Prominent Raman spectral variations in peritoneal sEVs between control women and those with endometriosis were observed at 891 cm^-1 (proteins) and 968 cm^-1 (lipids, CH wagging). Statistical analysis of molecular components influencing the Raman signature of sEVs suggest that their lipid composition appears to be influenced by the pathophysiology of endometriosis, reflecting changes similar to those observed in the eutopic endometrium in previous studies. sEVs isolated from peritoneal fluid and follicular fluid have markedly distinct biochemical compositions and morphometric properties. sEVs from follicular fluid were significantly smaller in size and exhibited a higher rigidity in this pilot cohort.

Conclusion: Distinct biophysical and biochemical differences in sEVs from endometriosis patients are documented in this pilot study, contributing to a deeper characterization of disease-associated extracellular vesicle (EV) alterations. Variations between follicular and peritoneal fluid sEVs suggest unique functional roles in reproductive health. These findings are hypothesis-generating and warrant validation in larger, well-controlled cohorts before any diagnostic application can be claimed.

Alzheimer’s disease (AD), a progressive neurodegenerative disorder, remains a major global health challenge owing to its complex pathogenesis and the presence of the blood-brain barrier (BBB), which substantially limits the delivery of effective therapeutics to the brain. Extracellular vesicles (EVs), which exhibit favorable biocompatibility, low immunogenicity, and the potential to cross or bypass the BBB under specific conditions, have emerged as promising therapeutic agents and delivery platforms for AD. This review focuses on the therapeutic potential of EV-based interventions in AD and summarizes recent advances in EV-mediated modulation of AD-related pathological processes, including amyloid-β (Aβ) clearance, tau protein regulation, neuroinflammation suppression, oxidative stress attenuation, and synaptic repair. Although EV-based therapies offer notable advantages, such as targeted BBB penetration and reduced immunogenic responses, their clinical translation remains constrained by safety concerns, including off-target effects, dose-dependent toxicity, and potential disturbances in neuroplasticity. In addition, this review discusses EV engineering strategies aimed at regulating the gut-brain axis (GBA), enhancing brain targeting, and advancing clinical translation. EV-based therapeutic interventions should therefore be developed within a safety-oriented framework supported by rigorous short- and long-term toxicological evaluation. Overall, this review highlights the therapeutic promise of EVs for AD while underscoring the need for rational engineering, standardized characterization, and safety-centered translational strategies to ensure clinical feasibility.

Aim: Olfactory ensheathing cells (OECs) are commonly utilized in cell-based therapies for neurodegenerative diseases and central nervous system injuries, due to their unique properties of suppressing gliosis and providing neuroprotection. Small extracellular vesicles (sEVs) have emerged as promising therapeutic agents. This study aimed to investigate OEC-sEVs’ effect on retinal degeneration.

Methods: Proteomics analysis identified differentially expressed proteins between OEC-sEVs and OECs. We conducted subretinal injection of OEC-sEVs and assessed their protective mechanisms in degenerative retina. Real-time quantitative PCR and Western blot were used to detect the expression of phagocytosis-related molecules in retinal pigment epithelium (RPE) cells.

Results: Proteomics analysis revealed that proteins enriched in OEC-sEVs were associated with phagocytosis. Following injection into the subretinal spaces of Royal College of Surgeons (RCS) rats, OEC-sEVs were internalized by RPE cells, leading to the transient upregulation of tyrosine-protein kinase Mer (MerTK), activation of the downstream phosphoinositide 3-kinase/Akt signaling pathway, and restoration of the phagocytic activity of RPE cells towards the photoreceptor outer segments. This delayed photoreceptor degeneration, thereby improving retinal function in RCS rats. In the ARPE-19 cell line treated with the specific MerTK inhibitor UNC2881, enhanced phagocytosis of fluorescent particles and porcine photoreceptor outer segments was observed following OEC-sEV internalization.

Conclusion: OEC-sEVs were abundant in proteins associated with phagocytosis, which may partially compensate for the phagocytic deficit of the RPE cells observed in a retinitis pigmentosa rat model via the MerTK-related signaling pathway.

The therapeutic landscape of probiotics is undergoing a paradigm shift from the live bacteria “bug” to the bioactive extracellular vesicles (EVs) “bubble” they secrete. Owing to their essential defense and metabolic functions, EVs derived from Lactobacillus, a genus of anaerobic or microaerophilic probiotics well known for their ability to ferment carbohydrates into lactic acid, have garnered particular interest for wound healing. As such, the presented mini-review began by elucidating why these Lactobacillus-derived EVs (L-EVs) attract upward attention by highlighting their superior safety, stability, multifunctionality, customizability, and scalability. This was followed by elaborating how these L-EVs accelerate wound healing by detailing their multifaceted mechanisms, including removing bacteria, reprogramming the microenvironment, repairing tissue, and remodeling the matrix during the wound-healing process. Lastly, it ended by envisioning where these L-EVs head for future therapy by addressing the translational hurdles, such as the lack of isolation and purification standardizations, the existing cargo and batch variabilities, as well as the ambiguous classification and regulation of L-EVs, which must be overcome to push them from bench to bedside. Moreover, it further proposed a specialized database based on artificial intelligence and multi-omics, the combined modification based on diverse engineering approaches, as well as the advanced formulation based on the material engineering strategies of L-EVs in the future to integrate the next generation of wound management modalities with optimal treatment efficacy.

Aim: Extracellular vesicles (EVs) are emerging as important mediators in myeloproliferative neoplasms (MPNs), but the characteristics of distinct EV subtypes remain unclear. This study aimed to characterize the molecular heterogeneity of MEG-01-derived large EVs (lEVs) and small EVs (sEVs).

Methods: lEVs and sEVs were isolated from MEG-01 cell culture medium by differential centrifugation followed by density gradient ultracentrifugation. EVs were characterized by transmission electron microscopy, nanoparticle tracking analysis, immunoblotting, and on-bead flow cytometry. Quantitative proteomic analysis was performed to compare the protein cargo of lEVs and sEVs, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses.

Results: Density gradient purification yielded high purity of lEV and sEV with distinct size, morphologies, and protein compositions. On-bead flow cytometry enabled efficient profiling of EV surface markers. Proteomic analysis revealed a shared core proteome between lEVs and sEVs, together with subtype specific protein signatures. Enrichment analyses indicated distinct molecular characteristics of the two EV populations, with sEVs showing enrichment of proteins associated with RNA-related processes and vesicle-mediated transport, whereas lEVs were enriched in proteins related to cytoskeletal organization and metabolism.

Conclusion: MEG-01 derived lEVs and sEVs comprise molecularly distinct subpopulations with shared and subtype specific protein cargo. The workflow established in this study provides a robust platform for high purity EV isolation and characterization, and offers insights into MEG-01 EV heterogeneity relevant to megakaryocyte biology and MPN research.

Aim: Abnormal skin pigmentation is not merely a common cosmetic concern but is also associated with pigmentation disorders and an elevated risk of malignant lesions such as melanoma. To overcome the adverse effects of existing antimelanogenic agents, we developed rose-derived extracellular vesicle-like particles (RS-EVLP) as a novel and biocompatible melanogenesis inhibitor.

Methods: RS-EVLP were isolated from roses and characterized by transmission electron microscopy and mass spectrometry. Their anti-melanogenic effects were evaluated in B16 cells via melanin content and tyrosinase activity assays. Molecular mechanisms were investigated using molecular docking, molecular dynamics simulations, and surface plasmon resonance (SPR) analysis. In vivo efficacy was assessed in a zebrafish model, followed by a clinical trial in human volunteers.

Results: RS-EVLP exhibited a bilayer nanostructure (50-150 nm) and contained natural anti-melanogenic components, including luteolin and procyanidin B3. They showed high binding affinity to microphthalmia-associated transcription factor (MITF) and tyrosinase-related protein 1 (TRP-1). In B16 cells, RS-EVLP dose-dependently reduced melanin content and tyrosinase activity by downregulating MITF and the transcription of tyrosinase (TYR)/TRP-1/TRP-2 enzyme system. In zebrafish, RS-EVLP reduced melanin spot area by 24% ± 17.5%. Clinical studies confirmed that a 5% RS-EVLP formulation progressively enhanced skin brightness as skin penetration efficiency increased.

Conclusion: RS-EVLP overcomes the limitations in bioavailability and toxicity associated with traditional synthetic whitening agents, offering a promising natural alternative for developing whitening cosmeceuticals.

Aim: Nidogen 1 (NID1) is a secreted pro-tumorigenic protein. While extracellular vesicle (EV)-NID1 is known to accelerate hepatocellular carcinoma (HCC) progression, its role in reshaping the HCC immune microenvironment remains undefined. This study investigated how EV-NID1 modulates dendritic cells (DCs) to establish an immunosuppressive niche that drives liver cancer growth.

Methods: The TIMER3.0 database was used to assess the correlations of NID1 with DCs and regulatory T cells (Tregs). A Transwell co-culture system using bone marrow-derived DCs was employed to evaluate the paracrine effects of HCC cells. EVs were isolated by differential ultracentrifugation, and their identity and purity were verified. Orthotopic and subcutaneous HCC mouse models were established to assess immune cell infiltration in vivo. DC adoptive transfer and Treg depletion assays were conducted to elucidate the underlying mechanism.

Results: High NID1 expression in HCC was associated with increased infiltration of myeloid DCs and Tregs, and their co-occurrence predicted poor patient survival. In vitro, EV-NID1 induced DC maturation and promoted their polarization toward mature DCs enriched in immunoregulatory molecules (mregDC), accompanied by elevated secretion of C-C chemokine ligand 17 (CCL17). These polarized DCs promoted CD4⁺ T cell proliferation and FOXP3⁺ Treg formation. In vivo, NID1-high tumors exhibited accelerated growth and increased infiltration of mregDCs and Tregs in tumors, lymph nodes, and spleens. NID1 knockdown reduced tumor growth, whereas adoptive transfer of EV-NID1-pulsed DCs partially rescued tumor progression. Furthermore, Treg depletion markedly suppressed the growth of NID1-high tumors.

Conclusions: This study suggests that NID1-enriched EVs derived from HCC promote tumor progression by inducing a regulatory phenotype in DCs and the expansion of Tregs.

Aim: We investigated whether the secretome, particularly small extracellular vesicles (sEVs), obtained from deferoxamine (DFO)-pretreated stem cells from human exfoliated deciduous teeth (SHED), could promote angiogenesis in human umbilical vein endothelial cells (HUVECs) and support vascularized tissue integration under preliminary ectopic implantation conditions.

Methods: SHED were treated with DFO (2.5, 5, 10 μM), and RNA sequencing was performed to analyze gene expression changes in DFO-treated SHED. The effects of the DFO-pretreated SHED secretome and the isolated sEVs (DFO-sEVs) on HUVEC migration were evaluated. The angiogenic capacity was assessed using wound healing, Transwell migration, and tube formation assays. DFO-sEVs were also tested for their ability to regulate macrophage polarization using immunofluorescence staining. Finally, DFO-sEVs were encapsulated in hydrogels and subcutaneously implanted into Sprague-Dawley rats, and their pro-angiogenic capacity in vivo was assessed via histological staining.

Results: RNA-seq analysis indicated that DFO pretreatment modulated SHED pathways related to hypoxia response, cell adhesion, and inflammation. The secretome from SHED pretreated with 5 μM DFO significantly promoted HUVEC migration and tube formation. DFO-sEVs, particularly from the 5 μM DFO group, demonstrated superior pro-angiogenic effects compared to sEVs from untreated SHED both in vitro and in vivo. Furthermore, DFO-sEVs drove macrophages to adopt an M2-like reparative polarization.

Conclusion: DFO pretreatment functionally potentiates the pro-angiogenic and immunomodulatory properties of SHED-derived sEVs. The findings suggest that DFO-sEVs may provide a bioactive component for promoting angiogenesis and facilitating tissue integration.

Drug resistance remains a critical obstacle in solid tumor therapy. While most studies have focused on chemical signaling within the tumor microenvironment, accumulating evidence has revealed that mechanical stress is a pivotal but underappreciated driver of drug resistance. Unlike previous reviews that have separately addressed mechanotransduction, exosome biology, and autophagy, this review proposes a vertically integrated framework - the “mechanical stress-cytoskeleton-exosomes-autophagy” axis - that connects these domains into a causal cascade from mechanical stress sensing to the dissemination of population-level resistance. Our central hypothesis is that cytoskeletal remodeling, driven by shear stress, matrix stiffness, and compressive stress, not only transduces mechanical signals intracellularly but also encodes these physical cues into exosomal cargo. These exosomes, in turn, transmit stress memory to recipient cells and trigger protective autophagy, thereby propagating drug-resistant phenotypes across heterogeneous tumor regions. We synthesize current evidence across biomechanics, vesicle trafficking, and stress biology and critically evaluate the intervention strategies that target each node of this axis. By reframing drug resistance as a mechanically propagated, exosome-mediated process, this review provides a new conceptual foundation and actionable directions for overcoming treatment failure.

Migrasomes are a newly identified class of extracellular vesicles (EVs) with a distinctive pomegranate-like morphology that are generated specifically from migrating cells. They are increasingly recognized as critically important mediators of intercellular communication involved in diverse physiological and pathological processes. However, their roles in cancers remain incompletely defined, and a consensus on migrasome-mediated malignant phenotypes is still lacking. In this review, we first summarize the current knowledge of migrasome biogenesis, distribution, and isolation methodologies. We then systematically discuss the multifaceted roles of migrasomes within the tumor microenvironment (TME), with particular emphasis on their contribution to cancer metastasis, progression and drug resistance. Furthermore, we explore the potential clinical applications of migrasomes in cancer diagnosis and therapy, and outline the key challenges and controversies that hinder their translation into clinical practice. Overall, this review provides an updated perspective on the roles of migrasomes in cancer biology and highlights the emerging opportunities for migrasome-informed targeted therapeutics.

Extracellular vesicles (EVs) are natural mediators of intercellular communication and hold promise for disease diagnosis and drug delivery, yet their intrinsic heterogeneity complicates isolation, molecular profiling, and targeted delivery. Aptamers, owing to their high affinity, specificity, chemical tunability, and scalable synthesis, offer a flexible molecular toolkit to address these challenges. In this review, we highlight recent progress in integrating aptamers into EV research across three interconnected domains: (1) the selection of aptamers against EV-associated surface markers; (2) the development of aptamer-based methods for EV capture and isolation; (3) the use of aptamer-functionalized EVs for targeted drug delivery and immunomodulation. We also discuss outstanding challenges, particularly the need for selection strategies that better preserve native EV epitope conformations and the rational design of multifunctional aptamer architectures. Looking forward, the convergence of aptamer technology with EV biology is expected to enable aptamer-drug conjugates and minimally invasive, dynamic liquid biopsy approaches for major diseases such as cancer, ultimately advancing early diagnosis and precision medicine.

Medicinal plants, as a core constituent of traditional medicine, provide an essential foundation for preventing and treating human diseases. Research on their pharmacologically active constituents has long focused on secondary metabolites, including alkaloids, terpenoids, flavonoids, and other compounds. However, with the rapid advancement of molecular biology techniques, nucleic acids have gradually become a research focus as a novel class of active components in medicinal plants that possess both nutritional and regulatory functions. This paper systematically reviews the types, characteristics, extraction techniques, pharmacological actions, and mechanisms of nucleic acids (including coding and non-coding RNAs) in medicinal plants. It also discusses current challenges and future directions for medicinal plant-derived nucleic acids in new drug development, providing a theoretical reference and technical basis for in-depth research and industrial application in this field.

Aim: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by excessive activation of pro-inflammatory macrophages and elevated levels of reactive oxygen species (ROS), resulting in synovial hyperplasia, cartilage destruction, and joint dysfunction. This study aimed to develop an engineered Tripterygium wilfordii-derived exosome-like nanoparticle system (TWELP@GlcN-HA) that integrates antioxidant, anti-inflammatory, immunomodulatory, and joint-targeting activities for synergistic RA therapy.

Methods:Tripterygium wilfordii-derived exosome-like nanoparticles (TWELP) were isolated from fresh Tripterygium wilfordii and sequentially surface-modified with glucosamine (GlcN) for cartilage protection and hyaluronic acid (HA) for targeted uptake by inflammatory cells. Their physicochemical properties, including morphology, particle size, structural integrity, and surface potential, were systematically characterized. In vitro assays were performed to evaluate ROS scavenging, inhibition of pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)], and macrophage polarization. Biodistribution and therapeutic efficacy were evaluated in a collagen-induced arthritis (CIA) mouse model by assessing synovial inflammation, cartilage damage, joint morphology, and joint function.

Results: TWELP@GlcN-HA exhibited a uniform spherical morphology, stable surface potential, and preserved structural integrity. In vitro, it efficiently scavenged ROS, suppressed TNF-α and IL-6 secretion, and promoted macrophage polarization from M1 to M2, thereby restoring immune homeostasis. In vivo, HA-mediated targeting promoted nanoparticle accumulation in inflamed joints. TWELP@GlcN-HA significantly alleviated synovial inflammation and cartilage destruction, improved joint morphology and function, and exhibited excellent biosafety with no systemic toxicity.

Conclusion: TWELP@GlcN-HA provides a multifunctional combinatorial strategy for RA treatment by synergistically regulating ROS, reprogramming pro-inflammatory macrophages, protecting cartilage, and achieving targeted joint delivery. Its enhanced therapeutic efficacy and favorable safety profile make it a promising candidate for future clinical translation in RA management.

Aim: Small RNA sequencing (small RNA-Seq) is a widely used and important method for biomarker discovery in extracellular vesicles (EVs), which carry small RNAs among their cargo and protect them from degradation in liquid biopsies. However, existing computational tools for small RNA-Seq data are not tailored to EV-RNA.

Methods: We present comprehensive and bias-aware analysis of small RNA gene expression (caRNAge), a biomarker discovery pipeline for small RNA-Seq data intended for users without extensive bioinformatic expertise. Ready to use as a Snakemake pipeline, caRNAge assists users in bias-aware normalization, differential gene expression analysis, and pathway over-representation analysis.

Results: Possible confounding factors can be identified and corrected to maximize data comparability between experiments and biomarker robustness. caRNAge is not limited to miRNAs; it also supports the detection of miRNA isomer (isomiRs), tRNA-derived fragments (tRFs), transfer RNA (tRNAs), and other small RNA species, thereby enhancing the specificity of biomarker signatures. Specific tissues can be selected for target prediction and pathway analysis to eliminate false-positive results. All results are provided in accessible and interactive HTML reports. We provide example data for each step of the pipeline to demonstrate caRNAge’s capabilities and advantages across three EV-miRNA biomarker discovery use cases. The pipeline is not limited to EV-miRNA data but can be used for data evaluation in any small RNA-Seq experiment.

Conclusion: By assisting users at every step of the discovery workflow, caRNAge fills an important gap in the field of EV-associated transcriptomic biomarker development. The pipeline is freely available at https://gitlab.lrz.de/kirchner/carnage.

Aim: The human heart contains two major mitochondrial subtypes, subsarcolemmal (SSL), fueling surface ion channels, and interfibrillar (IF), fueling interior contractile machinery. This study investigated whether the distinct extracellular vesicles (EVs) that we determined to be associated with these regions could influence cardiac energetics.

Methods: Following mitochondrial extraction, Subsarcolemmal Heart Extracellular Vesicles (SHEVs) and Interfibrillar Heart Extracellular Vesicles (IHEVs) were extracted from nonfailing human left ventricular lysates using precipitation-based methods. We characterized these EVs by size, proteomic profile, phospholipid content, and their ability to modulate mitochondrial function in primary neonatal rat ventricular myocytes (NRVMs).

Results: SHEVs (mean size: 148.9 ± 25.9 nm) and IHEVs (102.9 ± 31.0 nm) exhibited distinct proteomic signatures. SHEV and IHEV contained mostly structural proteins, but also contained 8% and 4% mitochondrial and glycolytic proteins, respectively. Lipidomic analysis revealed higher sphingomyelin and lower cardiolipin in EVs compared to whole tissue. SHEV treatment significantly increased mitochondrial ATP production, basal, and maximal respiration in NRVMs (P < 0.05). Conversely, IHEVs decreased maximal respiration without altering ATP production. Neither EV type exhibited intrinsic mitochondrial activity.

Conclusion: Regionally distinct EVs within the human heart have the capacity to modulate mitochondrial function. These findings suggest that cardiac EVs may act as localized regulators of cardiomyocyte bioenergetics.

Dental diseases, including dental caries, pulpitis, and periodontitis, can cause pain and tooth loss, compromising mastication, speech, aesthetics, and quality of life. These diseases have also been associated with systemic disorders, including Alzheimer’s disease. Conventional interventions, such as restorative procedures, root canal treatment, and periodontal scaling, have inherent limitations, underscoring the clinical need for novel therapeutic strategies. Plant-derived extracellular vesicles (PDEVs) have recently attracted considerable attention owing to their wide availability, cost-effectiveness, favourable biosafety profile, and capacity to mediate cross-kingdom communication. PDEVs exert diverse biological effects, including antimicrobial, anti-inflammatory, immunomodulatory, and tissue-regenerative activities. For example, ginger-derived PDEVs suppress the growth and virulence of Porphyromonas gingivalis through their lipid and miRNA cargoes, thereby alleviating periodontitis in mice. Given that effective dental therapy requires antimicrobial action, anti-inflammatory effects, immunomodulation, and tissue regeneration, PDEVs are promising therapeutic modalities. This review summarises the isolation, characterisation, composition, and biological functions of PDEVs, and discusses their potential applications in the management of dental diseases.

Extracellular vesicles (EVs) are promising platforms for next-generation drug delivery and regenerative therapeutics, yet biological heterogeneity, limited targeting specificity, variable cargo composition, and continued reliance on empirical optimization hinder clinical translation. Artificial intelligence (AI) offers new opportunities to make EV engineering more predictive and data-driven. Building on prior work in AI-assisted EV delivery and emerging virtual-EV modeling, this review presents an evidence-aware, multiscale framework that integrates parental-cell programming, vesicle design, recipient-system modeling, and experimental feedback. At the source level, AI virtual cells (AIVCs) may help predict how perturbations to parental cells influence EV biogenesis and composition. At the vesicle level, AI-guided approaches may support cargo prioritization, surface engineering, and loading optimization. At the recipient level, digital twin models could help simulate EV uptake, intracellular trafficking, microenvironmental interactions, and downstream therapeutic responses. These components may ultimately be integrated with experimental validation into iterative design-test-refinement workflows. Importantly, this review distinguishes direct EV-specific evidence from technologies transferable from adjacent fields and concepts that remain prospective. By defining these evidence boundaries and linking them to key challenges in data quality, model interpretability, manufacturing, and validation, this framework provides a practical roadmap for advancing AI-enabled EV therapeutics from conceptual modeling to testable, translationally relevant engineering strategies.