Perfluorooctane sulfonate (PFOS) causes dysfunction of erythrocytes by calcium dysregulation, and a diabetic condition exacerbates PFOS-induced prothrombotic activation.

Microvesicles (MVs) are extracellular vesicles released from many cell types under physiological and pathological conditions, influencing viral transmission, immune regulation, and inflammation. This exploratory pilot study characterized and compared plasma MV profiles in patients infected with human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). Plasma samples (n = 125; HIV: 25, HBV: 50, HCV: 50) were analyzed using nanoparticle tracking analysis (NanoSight NS300) to assess MV size and concentration, classifying them as small (<300 nm) or large (>300 nm). Patients with HBV exhibited significantly larger mean MV size compared with both patients with HIV (131.5 ± 14.6 nm vs. 113.1 ± 14.0 nm, p < 0.0001) and HCV (131.5 ± 14.6 nm vs. 118.0 ± 18.5 nm, p = 0.0002). HCV infection showed higher concentrations of large MVs than HIV (p = 0.0022), while total and small MV levels did not differ. No sex-related differences were detected. Distinct MV size distributions appear linked to chronic viral infections, with HBV and HCV showing greater alterations than HIV. MVs may serve as potential biomarkers reflecting infection-associated biological processes; however, mechanistic, or functional roles were not assessed in this study and will require dedicated future investigations in larger controlled studies.

Signal transduction is essential for tumor progression and therapeutic response. Extracellular vesicles (EVs) have emerged as critical mediators of intercellular communication and therapeutic delivery. However, current EV-based strategies that rely on ex vivo manipulation encounter significant limitations, including nonspecific biodistribution, limited tumor accumulation, and diminished biological activity, underscoring the need for the localized production of therapeutic EVs within tumors. To address this, a rationally designed poly(D-amino acid) copolymer, methoxy poly(ethylene glycol)-block-poly(D-lysine10)-block-poly(O-phospho-D-tyrosine5-co-D-alanine25) (EG45-D-K-D-pYA), is developed, which undergoes alkaline phosphatase (ALP)-catalyzed self-assembly to form a tumor-localized, positively charged network. This assembly triggers the formation of localized microvesicles (MVs) and initiates a self-sustaining fission cascade, amplifying the cytotoxic signaling. Upon ALP catalysis, 88.3% of the phosphate group in EG45-D-K-D-pYA is hydrolyzed within 24 h, shifting zeta potential from -4.95 to +25.0 mV and increasing β-sheet content from 4.0% to 46.0%, promoting a morphological transition from nanoparticle to network. EG45-D-K-D-pYA disrupts tumor cell membranes, mitochondria, and nuclei, inducing potent tumor-selective cytotoxicity (IC50 = 0.58 μM at 24 h) and in situ generation of positively charged MVs (ζ = +32.9 mV). These MVs, enriched with membrane fragments, poly(amino acid)s, and mitochondrial and nuclear components, further mediate tumor-selective cell death and sustain cytotoxic propagation across four MV generations (P0-P4), with cytotoxicity rates of 53.13%, 28.71%, 36.35%, 24.69%, and 7.05%, respectively. EG45-D-K-D-pYA achieves 90.1% tumor growth inhibition and markedly suppresses metastases by forming network in vivo. This study presents a tumor-selective therapeutic approach that integrates enzyme-triggered self-assembly with MV-driven fission amplification to induce robust bystander cytotoxicity and sustained tumor suppression.

We previously showed that inhibition of the NF-κB signaling pathway in mesenchymal stromal cells (MSCs) (NKI-MSCs) induces quiescence in co-cultured hematopoietic stem cells (HSCs). This led us to investigate whether NKI-MSCs exert similar growth-inhibitory effects on leukemic cells. We found that both NKI-MSCs and their secretome induce cell cycle arrest in KG1a cells, a cell line of acute myeloid leukemia (AML) origin. Surprisingly, the extracellular vesicles (EVs) isolated from NKI-MSCs supported the proliferation of KG1a cells. This is perhaps the first report showing the opposite effects of MSCs and the EVs secreted by them. Further analysis revealed that microvesicles (MVs) from NKI-MSCs inhibited KG1a cell growth and induced apoptosis, whereas exosomes (Exos) supported proliferation. Our findings could have clinical implications. NKI-MVs, having apoptosis-inducing activity, could serve as an adjunct, off-the-shelf biologic to limit AML growth, enabling reduced-intensity chemotherapy in elderly patients and patients having co-morbidities. NF-κB inhibitors have been tried as chemotherapeutic agents for treating AML patients. However, systemic inhibition of NF-κB may also affect the bone marrow resident MSCs, which in turn could produce EVs supporting the proliferation of AML blasts. Our data could explain the inadequate clinical effectiveness of NF-κB inhibitors in treating AML, and also raise a concern for the systemic use of NF-κB inhibitors in the therapeutic regimen.

Sperm Selection through Microvesicle-Mediated ChemotaxisInduced by Follicular Fluid and Cumulus Cells.

Background: Myocardial infarction (MI) is a leading cause of heart failure. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show therapeutic potential for MI, largely attributed to paracrine effects. However, the role of microvesicles (MVs), a major class of extracellular vesicles, n this effect remains unclear. Therefore, it is imperative to ascertain whether hiPSC-CM derived MVs exhibit cardioprotective effects and to elucidate the underlying mechanisms in both murine and porcine models of MI. Methods: hiPSC-CM-MVs were isolated by differential ultracentrifugation and characterized via nanoparticle tracking, electron microscopy, flow cytometry, and western blot. Their therapeutic effects were evaluated in oxygen–glucose deprivation (OGD)-injured hiPSC-CMs and murine MI models. Proteomic profiling identified candidate effectors, which were further validated using cardiac-specific gene overexpression or knockout transgenic mouse models. A porcine MI model was also used to assess the safety and efficacy of MV through MDCT, hemodynamic measurements, and histological analysis. Results: In vitro, we initially isolated high-purity MVs from hiPSC-CMs, exhibiting a distinct enrichment of mitochondrial constituents.These MVs conferred robust cytoprotection against OGD-induced injury by suppressing cardiomyocyte apoptosis, augmenting mitochondrial membrane potential and calcium uptake capacity, and thereby sustaining calcium homeostasis. In vivo, the trentment of MV alleviated progressive cardiac dysfunction following MI in mice and concurrently suppressed a spectrum of pathological remodeling events, including myocardial hypertrophy, fibrosis, apoptosis, and calcium overload. The cardioprotective effects observed were likely the result of enhanced mitochondrial calcium uptake, which contributed to restore calcium homeostasis. To explore the underlying mechanism, proteomic profiling of MVs identified DExH-box helicase 9 (DHX9) as a key regulatory cargo. Cardiac-specific overexpression of DHX9 partially reproduced while knockout DHX9 abolished the effects of MVs. Finally, in a porcine MI model, MVs exhibited robust therapeutic efficacy and safety without systemic toxicity. Conclusion: This study identifies a novel mechanism by which hiPSC-CM-MVs restore calcium homeostasis and improve cardiac function post-MI via DHX9. These findings provide preclinical evidence supporting MVs as a safe and effective therapeutic strategy for MI.

Mechanistic role and prognostic value of CD20-positive microvesicles in rituximab-treated diffuse large B-cell lymphoma

Background: A comparative study was conducted to evaluate the potential of extracellular, tumor-derived microvesicles (MVs)s in promoting kidney regeneration. Methods: MVs were collected from L929 sarcoma, LLC, and B16 melanoma cells, and mesenchymal stem cells (MSCs). The regenerative activity of MVs was evaluated in an experimental murine model of chronic kidney injury (CKI). Results: Both tumor-derived MVs (T-MVs) and MSC-derived MVs (MSC-MVs) significantly improved kidney function and histological structure. Specifically, the height of collecting tubules in the middle third of the renal medulla returned to normal levels following MV treatment. Both T-MVs and MSC-MVs reduced the proportion of pro-inflammatory CD4+CD44+ T cells in renal cell infiltrates and spleens of CKI mice. Furthermore, treatment with these MVs increased the number of natural CD4+CD25+FoxP3+ regulatory T cells in the spleen, indicating their immunomodulatory effects. Conclusions: These findings suggest that T-MVs, similar to MSC-MVs, possess a universal capacity to promote kidney tissue regeneration and exert anti-inflammatory immunomodulatory effects.

ABSTRACTSickle cell disease (SCD) is a monogenic disorder caused by the presence of hemoglobin S (Hb S) and is associated with a wide range of clinical complications, including hemolytic anemia, vaso‐occlusive crises (VOC), and acute chest syndrome (ACS). This variability is largely driven by Hb S polymerisation and abnormal cell adhesion, which promote the release of circulating microvesicles (MVs). MVs are small vesicles (0.1–1.0 μm) released from various cell types in response to oxidative stress, cellular activation, or apoptosis. They possess pro‐coagulant and pro‐inflammatory properties and are increasingly recognised as potential modulators of disease processes. In this study, homozygous SCD patients and healthy controls were recruited to characterise their MVs profiles using flow cytometry and to explore associations with clinical and biological parameters. SCD patients exhibited significantly elevated levels of MVs compared to controls. Notably, red cell–derived MVs (RMVs) and phosphatidylserine‐positive MVs (AMVs) were strongly associated with elevated lactate dehydrogenase (LDH) levels and clinical severity. A negative correlation was observed between fetal hemoglobin (Hb F) and endothelial MVs (EMVs), suggesting a protective role of Hb F against endothelial injury. These findings support the potential of MVs as diagnostic and prognostic biomarkers for identifying SCD patients at higher risk of complications.

Certain invasive strains of the Gram-positive bacterium Streptococcus pyogenes exploit human plasminogen (hPg) to promote tissue invasion and pathogenesis. hPg is a single-chain multi-modular zymogen containing five kringle domains (K1-K5), four of which interact with lysine or pseudo-lysine residues on binding partners, positioning hPg for activation to plasmin and enhancing bacterial dissemination. The major hPg binding protein in S. pyogenes is the multicopy surface-resident M-protein, or other surface proteins, such as the homooctameric glycolytic enzyme, enolase (SEn). SEn lacks features for direct translocation from the cytoplasm to the bacterial surface, and it is unclear how Sen is translocated to the bacterial surface. Additionally, the mechanism by which SEn binds hPg is poorly understood. In this study, we show that SEn is exported via lipid microvesicles (MV), likely originating from the cytosolic membrane. Using cryogenic-electron microscopy, we provide a high-resolution (<3.4 Å) map of SEn reconstituted into dioleoyl phosphatidylglycerol (DOPG) liposomes, which serves as our MV model. The Sen-DOPG map reveals that two subunits of the SEn octamer are exposed to the extracellular medium, while six remain inserted within the membrane or vesicle interior. However, this interaction does not induce a conformational change in hPg, which remains in a closed conformation, thereby limiting the SEn stimulatory effect on many hPg activators, except for host tissue-type plasminogen activator (tPA). Instead, the ability of SEn to bind tPA is the primary factor driving enhanced hPg activation. These findings highlight a novel mechanism by which MV-associated SEn promotes hPg activation preferentially through tPA, independent of a hPg conformational rearrangement.

Lymph node metastasis (LNM) is a pivotal determinant of breast cancer (BC) patient prognosis and treatment efficacy. Cell surface heparan sulfate proteoglycans (HSPGs), namely, syndecan-1 (SDC1), SDC2, and SDC4, are involved in cancer progression, metastasis, and regulate extracellular vesicles (EVs) biogenesis, including the microvesicles (MVs). This study analyzed MV-enriched EVs isolated from blood plasma of BC patients with negative (n = 19) and positive (n = 20) LNM (nLNM and pLNM, respectively) using differential centrifugation. Western blot analysis revealed significantly elevated SDC2 levels in MV-enriched EVs from pLNM cases compared to nLNM. Additionally, fibronectin (FN), a SDC2-interacting protein identified through STRING analysis, was also upregulated in pLNM MV-enriched EVs. In contrast, qRT-PCR showed reduced SDC2 (P < 0.01) and FN (P < 0.05) mRNA levels in tumor tissues of pLNM patients compared to nLNM. ROC analysis highlighted the diagnostic value of SDC2 (AUC: 0.8376) and FN (AUC: 0.8803) mRNA in differentiating LNM status. Bioinformatics analyses further confirmed the association of SDC2 and FN expression with BC staging and prognosis. These findings underscore the potential of circulating MV-enriched EV-associated SDC2 and FN, along with their tumor tissue mRNA expression, as potential predictive biomarkers for LNM and chemotherapy response in chemotherapy-naïve obese BC patients.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-17638-2.

Mercury (Hg) exposure is a major environmental risk factor, closely linked to oxidative stress and cardiovascular disease. Red blood cells (RBC), due to their high oxygen exposure and lack of repair mechanisms, are particularly sensitive to oxidative injury and are key indicators of systemic redox imbalance. This study evaluates the protective effects of polyphenolic extracts from Annurca apple, specifically flesh and peel, from both ripe and unripe fruit, on HgCl2-exposed human RBCs. Key oxidative stress markers were measured, including ROS production, GSH levels, lipid peroxidation (TBARS), MetHb formation, SH group content, microvesicle (MV) generation, and morphological changes. Peel extracts, particularly those from ripe apples, consistently exhibited stronger antioxidant and cytoprotective effects than flesh extracts, effectively reversing Hg-induced oxidative damage and preserving RBC integrity. Notably, these extracts restored redox homeostasis and GSH levels, reduced ROS and TBARS accumulation, prevented MetHb formation, and mitigated MV release and morphological alterations. These protective effects appear to involve multifactorial mechanisms. These findings highlight the nutraceutical potential of Annurca apple extracts in counteracting heavy metal-induced oxidative stress and support their possible relevance for future studies aimed at health protection and waste valorization.

Potential use of serum microvesicle indicators to distinguish different types of drug-induced liver injury.

Microvesicles (MV) isolated from septic individuals were observed to impact systemic hemodynamics and cardiac function. The aim of this in vitro study was to analyze the effects of TNFα-induced endothelial MV (TMV) and MV from septic patients (SMV) on beating frequency and Ca2+ transient kinetics of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). MV were isolated from supernatants of TNFα-stimulated primary human pulmonary microvascular endothelial cells (HPMEC) and plasma from 20 sepsis patients by ultracentrifugation and quantified using flow cytometry. Spontaneous Ca2+ transients were measured in hiPSC-CM using the Ca2+-sensitive ratiometric indicator fura-2 at different time points of incubation with different MV concentrations. At 16 h of incubation, higher MV concentrations showed significant differences, especially regarding decay and beating frequency. Despite high variability, at 10 × 106 MV/mL and 16 h of incubation, TMV significantly decreased frequency compared to control MV (CMV). SMV from septic patients did not reveal any significant effects on Ca2+ transients under these experimental settings. MV isolated from control or TNFα-treated HPMEC affected Ca2+ handling and spontaneous activity of hiPSC-CM, however, the measured effects were not consistent throughout the different conditions. Further refinement of the experiment conditions is needed to specify the exact conditions for crosstalk between endothelium-derived MV and cardiomyocytes.

Cancer cells can release extracellular vesicles (EVs) of different sizes under stress conditions. Among the EVs, microvesicles (MVs), which have a size between 50 and 1000 nm, are bounded by a membrane lipid bilayer, exhibit proteins at their surface, and enclose some soluble proteins. MVs can interact with surrounding cells present in the tumor microenvironment to favor tumor resistance. Indeed, they can transport some oncoproteins such as epidermal growth factor receptor (EGFR) and modify phenotype of endothelial cells (ECs). Even if their role in cell communication is well established, the understanding of anticancer treatments on their release and their protein content change are of particular importance. In this work, we showed that head and neck squamous cell carcinoma (HNSCC) cells exposed to cetuximab, monoclonal antibody targeting EGFR, can modulate EGFR expression of MVs. Moreover, this work emphasizes the effect of cetuximab on the shedding and content of MVs by HNSCC cells as well as their interaction with ECs. Consequently, MVs can be used as surrogate markers for predicting the efficacy of anti‐EGFR therapies. Finally, the release of MVs after treatment must be envisaged as a resistance mechanism and must be considered in the future to evaluate the effect of therapy on the tumor microenvironment.

Microvesicles (MVs) have convenient clinical applications and play functional roles in cellular signal transduction. Although the clinical importance of MVs is being increasingly recognized, the current diversity of isolated protocols results in a heterogeneous population of their unknown origins, even expands to uncertain functions. Here, we systematically investigated the composition of MVs at different centrifugal speed intervals and discovered that centrifugation at 3000 g is critical in determining the composition of MVs. We observed that platelet-derived particles accounted for more than 80% of MVs under 3000 g, while only about 20% of MVs were obtained over 3000 g. The discovery that more than 80% of platelet-derived MVs sheds new light on their function, including procoagulation activity and clinical diagnosis, etc. Our work not only optimizes the method for MVs isolation but also clarifies the physiological functions and characteristics that should be attributed to platelets rather than MVs. Consequently, these findings will derive new conceptualizations regarding MVs’ composition and function.

Extracellular vesicles (EVs), which include exosomes (Exos) and microvesicles (MVs), play a crucial role in intercellular communication and exert various biological activities by delivering specific cargoes of functional molecules, such as RNAs and proteins, to target cells. EVs secreted by human mesenchymal stem cells (hMSCs) have demonstrated their capacity to replace intact MSCs in tissue repair and regeneration. Induced mesenchymal stem cells (iMSCs) derived from induced pluripotent stem cells (iPSCs) present a promising alternative to traditional MSCs for producing EVs. This study aimed to establish an alternative source of EVs from iMSCs and compare them with EVs from adipose-derived MSCs (ADMSCs). Both iMSCs and ADMSCs were expanded under xeno-free culture conditions, and conditioned media were collected for EV isolation and characterization. The effects of the isolated EVs on cellular viability, apoptosis, senescence, and cell migration were evaluated. Results indicated that iMSC-EVs had a larger particle size (~1.5-fold) with no significant differences in morphology or surface markers compared to ADMSC-EVs. Furthermore, both iMSC- and ADMSC-derived EVs significantly increased HDF viability at 48 and 72 hours (p ≤ 0.01, p ≤ 0.05). Both types of EVs significantly reduced apoptosis levels (p ≤ 0.01) in both HDFs and ADMSCs, while having no effect on senescence induction (p > 0.9999). Additionally, iMSC-EVs significantly enhanced ADMSC migration (p < 0.0001), whereas the effect was less pronounced with ADMSC-EVs. iMSC-EVs present a promising and a scalable option for regenerative applications, offering advantages over ADMSC-EVs. However, further investigation is needed to fully understand their effects and underlying mechanisms.

Abstract Triple‐negative breast cancer (TNBC) lacks estrogen receptor (ER), progesterone receptor (PR), and HER2 expression, rendering endocrine and HER2‐targeted therapies ineffective. As a result, chemotherapy remains the primary treatment, though with limited success. Micro‐vesicles, particularly exosomes, facilitate cell‐to‐cell communication and impact various physiological and pathological processes. Recent research has highlighted the therapeutic potential of plant‐derived micro‐vesicles (MVs), which contain bioactive molecules such as proteins, nucleic acids, lipids, and metabolites with anticancer, antioxidative, and anti‐inflammatory properties. This study characterized micro‐vesicles isolated from Elettaria cardamomum seeds (ECS MVs) using techniques such as SEM, TEM, DLS, AFM, NTA, and zeta potential analysis. Exosomes from 1 g of E. cardamom extract had an average size of 180 ± 98.1 nm, yielding 4.8 × 10⁷ particles/mL as per NTA. GC–MS identified key phytocompounds, whereas the Bradford assay and SDS‐PAGE confirmed protein quantification and molecular weight distribution. The anticancer effects of ECS MVs were examined on TNBC cell lines (MDA‐MB‐231 and 4T1) using colony formation, MTT, wound healing, and ROS assays. FACS analysis identified cell cycle arrest and trypan blue staining quantified cell viability. This is the first report demonstrating the anticancer potential of plant‐derived exosome‐like nanoparticles from E. cardamom.

Bioorthogonal catalytic microvesicle-mediated prodrug activation against liver cancer.

Microvesicles (MVs) are membrane-enclosed, plasma membrane-derived particles released by cells from all branches of life. MVs have utility as disease biomarkers and may participate in intercellular communication; however, physiological processes that induce their secretion are not known. Here, we isolate and characterize annexin-containing MVs and show that these vesicles are secreted in response to the calcium influx caused by membrane damage. The annexins in these vesicles are cleaved by calpains. After plasma membrane injury, cytoplasmic calcium-bound annexins are rapidly recruited to the plasma membrane and form a scab-like structure at the lesion. In a second phase, recruited annexins are cleaved by calpains-1/2, disabling membrane scabbing. Cleavage promotes annexin secretion within MVs. Our data support a new model of plasma membrane repair, where calpains relax annexin-membrane aggregates in the lesion repair scab, allowing secretion of damaged membrane and annexins as MVs. We anticipate that cells experiencing plasma membrane damage, including muscle and metastatic cancer cells, secrete these MVs at elevated levels.