Extracellular vesicles (EVs) are a unique and heterogeneous class of lipid bilayer nanoparticles secreted by most cells. EVs are regarded as important mediators of intercellular communication in both prokaryotic and eukaryotic cells due to their ability to transfer proteins, lipids and nucleic acids to recipient cells. In addition to their physiological role, EVs are recognized as modulators in pathological processes such as cancer, infectious diseases, and neurodegenerative disorders, providing new potential targets for diagnosis and therapeutic intervention. For a complete understanding of EVs as a universal cellular biological system and its translational applications, optimal techniques for their isolation and characterization are required. Here, we review recent progress in those techniques, from isolation methods to characterization techniques. With interest in therapeutic applications of EVs growing, we address fundamental points of EV‐related cell biology, such as cellular uptake mechanisms and their biodistribution in tissues as well as challenges to their application as drug carriers or biomarkers for less invasive diagnosis or as immunogens.This article is categorized under: Diagnostic Tools > Biosensing Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease

This study sought to measure medium‐sized extracellular vesicles (mEVs) in plasma, when patients have low Plasmodium falciparum early in infection. We aimed to define the relationship between plasma mEVs and: (i) parasitaemia, (ii) period from onset of malaria symptoms until seeking medical care (patient delay, PD), (iii) age and (iv) gender. In this cross‐sectional study, n = 434 patients were analysed and Nanosight Tracking Analysis (NTA) used to quantify mEVs (vesicles of 150–500 nm diameter, isolated at 15,000 × g, β‐tubulin‐positive and staining for annexin V, but weak or negative for CD81). Overall plasma mEV levels (1.69 × 10 ¹⁰ mEVs mL ⁻¹ ) were 2.3‐fold higher than for uninfected controls (0.51 × 10 ¹⁰ mEVs mL ⁻¹ ). Divided into four age groups, we found a bimodal distribution with 2.5‐ and 2.1‐fold higher mEVs in infected children (45 yo) (median:1.92 × 10 ¹⁰ mEVs mL ⁻¹ ), respectively, compared to uninfected controls; parasite density varied similarly with age groups. There was a positive association between mEVs and parasite density (r = 0.587, p < 0.0001) and mEVs were strongly associated with PD (r = 0.919, p < 0.0001), but gender had no effect on plasma mEV levels ( p = 0.667). Parasite density was also exponentially related to patient delay. Gender ( p = 0.667) had no effect on plasma mEV levels. During periods of low parasitaemia (PD = 72h), mEVs were 0.93‐fold greater than in uninfected controls. As 75% (49/65) of patients had low parasitaemia levels (20–500 parasites µL ⁻¹ ), close to the detection limits of microscopy of Giemsa‐stained thick blood films (5–150 parasites µL ⁻¹ ), mEV quantification by NTA could potentially have early diagnostic value, and raises the potential of Pf markers in mEVs as early diagnostic targets.

Extra Low-frequency Magnetic Fields (ELF-MFs) significantly enhance cellular uptake of methotrexate by inducing transient plasma membrane pores/damage. This enhanced 'dose loading' of methotrexate via the electromagnetically induced membrane pores leads to similar outcomes as the normal control while using significantly smaller therapeutic doses in vitro when compared to non-ELF-MF treated control. Approximately 10% of the typical therapeutic dose yielded similar results when used with ELF-MF. ELFMFs increase PC12, THP-1 and HeLa proliferation in vitro (120% of the control). Analysis of adherent cells demonstrate significantly less migration towards an induced scratch injury (20 mm in 24 h when compared to a control). Our results suggest an important role for the use of ELF-MFs in the treatment of tumours that opens some new and exciting possibilities including using smaller therapeutic doses of chemotherapeutic agents and disrupting tumour metastasis.(c) 2022 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Cancer progression in skeletal muscle (SkM) is very rare, and mechanisms remain unclear. This study assessed the potential of SkM (myocyte)-derived EVs (C2C12-EVs) as anti-cancer agents. Using murine in vitro models, we showed that following treatment with C2C12-EVs, lung carcinoma cells failed to colonise SkM cells, and that C2C12-EVs selectively exerted apoptosis on cancer cells. Uptake of C2C12-EVs by carcinoma cells caused changes in lysosomal function and mitochondrial membrane properties inducing cell death with elevated caspase 3 and 9. The C2C12-EVs also inhibited cell proliferation, affecting cell cycle arrest at S phase and inhibited cell migration. Proteomic analysis of C2C12-EV cargoes highlighted functional enrichment pathways involved in lysozyme function, HIF-1 and PI3K-Akt signalling, regulation of actin cytoskeleton, pyruvate metabolism, platelet activation, and protein processing in ER. Decorin, a muscle cell-specific cytokine released from myocytes in response to stress, was significantly enriched in C2C12-EVs and may contribute to C2C12-EVs’ inhibitory activity on cancer cells. C2C12-EVs may suppress cancer and potentially be used as therapeutic agents for cancer metastasis.