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Preparation of Nanoparticle-Loaded Extracellular Vesicles Using Direct Flow Filtration.

Shomit Mansur1, Shahriar Habib1, Mikayla Hawkins1

  • 1Chemical & Biological Engineering, The University of Alabama, Tuscaloosa, AL 35487, USA.

Pharmaceutics
|May 27, 2023
PubMed
Summary

Researchers developed a scalable method using direct flow filtration to produce quercetin-iron nanoparticle-loaded extracellular vesicles (EVs) from breast cancer cells. These engineered EVs show enhanced cellular uptake, suggesting improved drug delivery potential.

Keywords:
biomimetic nanocarrierscellular uptakedirect flow filtrationdrug deliveryextracellular vesicle protein markersnanoparticle-loaded EVs

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Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Cancer Therapeutics

Background:

  • Extracellular vesicles (EVs) show promise as cell-free therapeutics and drug delivery nanocarriers.
  • Scalable production and in vivo tracking of EVs remain significant challenges for their clinical application.

Purpose of the Study:

  • To develop a scalable method for producing nanoparticle-loaded EVs using direct flow filtration.
  • To characterize the nanoparticle-loaded EVs and evaluate their cellular uptake compared to free nanoparticles.

Main Methods:

  • Preparation of quercetin-iron complex nanoparticle-loaded EVs from MDA-MB-231br cancer cells via direct flow filtration.
  • Characterization using transmission electron microscopy, dynamic light scattering, SDS-PAGE, and antibody array for EV markers.
  • Comparison of cellular uptake of nanoparticle-loaded EVs versus free nanoparticles in MDA-MB-231br cells using iron staining.

Main Results:

  • Direct flow filtration yielded a significant increase in EV production compared to ultracentrifugation.
  • Characterization confirmed the morphology, size, protein markers (ALIX, TSG101, CD63, CD81), and successful loading of nanoparticles within EVs.
  • Nanoparticle-loaded EVs demonstrated enhanced and more uniform cellular uptake compared to free nanoparticles, suggesting improved delivery.

Conclusions:

  • Direct flow filtration is a feasible and scalable method for producing nanoparticle-loaded EVs from cancer cells.
  • The enhanced cellular uptake of nanoparticle-loaded EVs indicates their potential for deeper tissue penetration and improved therapeutic efficacy in drug delivery applications.