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1.
Small ; 20(18): e2307240, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38100284

RESUMO

Extracellular vesicles (EVs) are nanosized biomolecular packages involved in intercellular communication. EVs are released by all cells, making them broadly applicable as therapeutic, diagnostic, and mechanistic components in (patho)physiology. Sample purity is critical for correctly attributing observed effects to EVs and for maximizing therapeutic and diagnostic performance. Lipoprotein contaminants represent a major challenge for sample purity. Lipoproteins are approximately six orders of magnitude more abundant in the blood circulation and overlap in size, shape, and density with EVs. This study represents the first example of an EV purification method based on the chemically-induced breakdown of lipoproteins. Specifically, a styrene-maleic acid (SMA) copolymer is used to selectively breakdown lipoproteins, enabling subsequent size-based separation of the breakdown products from plasma EVs. The use of the polymer followed by tangential flow filtration or size-exclusion chromatography results in improved EV yield, preservation of EV morphology, increased EV markers, and reduced contaminant markers. SMA-based EV purification enables improved fluorescent labeling, reduces interactions with macrophages, and enhances accuracy, sensitivity, and specificity to detect EV biomarkers, indicating benefits for various downstream applications. In conclusion, SMA is a simple and effective method to improve the purity and yield of plasma-derived EVs, which favorably impacts downstream applications.


Assuntos
Vesículas Extracelulares , Lipoproteínas , Maleatos , Poliestirenos , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Lipoproteínas/química , Lipoproteínas/metabolismo , Maleatos/química , Humanos , Animais , Cromatografia em Gel , Camundongos , Macrófagos/metabolismo
2.
Nat Nanotechnol ; 19(1): 13-20, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38110531

RESUMO

Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.


Assuntos
Vesículas Extracelulares , Vesículas Extracelulares/metabolismo , Comunicação Celular
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