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Extracellular vesicles (EVs) have shown great potential as cell-free therapeutics and biomimetic nanocarriers for drug delivery. However, the potential of EVs is limited by scalable, reproducible production and in vivo tracking after delivery. Here, we report the preparation of quercetin-iron complex nanoparticle-loaded EVs derived from a breast cancer cell line, MDA-MB-231br, using direct flow filtration. The morphology and size of the nanoparticle-loaded EVs were characterized using transmission electron microscopy and dynamic light scattering. The SDS-PAGE gel electrophoresis of those EVs showed several protein bands in the range of 20–100 kDa. The analysis of EV protein markers by a semi-quantitative antibody array confirmed the presence of several typical EV markers, such as ALIX, TSG101, CD63, and CD81. Our EV yield quantification suggested a significant yield increase in direct flow filtration compared with ultracentrifugation. Subsequently, we compared the cellular uptake behaviors of nanoparticle-loaded EVs with free nanoparticles using MDA-MB-231br cell line. Iron staining studies indicated that free nanoparticles were taken up by cells via endocytosis and localized at a certain area within the cells while uniform iron staining across cells was observed for cells treated with nanoparticle-loaded EVs. Our studies demonstrate the feasibility of using direct flow filtration for the production of nanoparticle-loaded EVs from cancer cells. The cellular uptake studies suggested the possibility of deeper penetration of the nanocarriers because the cancer cells readily took up the quercetin-iron complex nanoparticles, and then released nanoparticle-loaded EVs, which can be further delivered to regional cells.
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RAPID DIFFERENTIATION OF HOST AND PARASITE EXOSOME VESICLES USING PHOTONIC CRYSTAL BIOSENSOR
The analysis of membranous extracellular vesicles, such as exosomes vesicles (EV) opens a new direction for the rapid disease diagnosis because EVs can carry molecular constituents of their originating cells. Secreted by mammalian cells, the size of most membrane-bound phospholipid EVs ranges from 50 to 150 nm in diameter. Recent studies have demonstrated the potential of using EVs for cancer diagnosis and treatment monitoring. To diagnose infectious diseases using EVs, the ability to discriminate EVs from host cells and parasites is key. Here, we report a rapid EV analysis assay that can discriminate EVs based on a host-specific transmembrane protein (CD63 antigen) using a label-free optical biosensor.
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- Award ID(s):
- 1711839
- PAR ID:
- 10062942
- Date Published:
- Journal Name:
- Hilton Head Workshop 2018: A Solid-State Sensors, Actuators and Microsystems Workshop, June 3-7, 2018, Hilton Head Island, SC, USA.
- Page Range / eLocation ID:
- 303 - 305
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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