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Related Experiment Video

Updated: Jul 17, 2026

A Magnetic Separation-Assisted High-Speed Homogenization Method for Large-Scale Production of Endosome-Derived Vesicles
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Published on: January 26, 2024

Development of Small Interfering RNA-Loaded Extracellular Vesicles via a High-Pressure Homogenization Approach.

Tatsuya Fukuta1, Masato Miyazaki1, Taiki Fujimoto2

  • 1Department of Physical Pharmaceutics, School of Pharmaceutical Sciences, Wakayama Medical University, 25-1 Shichiban-cho, Wakayama 640-8156, Japan.

ACS Applied Bio Materials
|July 16, 2026
PubMed
Summary

High-pressure homogenization effectively loads small interfering RNA (siRNA) into extracellular vesicles (EVs). This scalable method enhances targeted delivery and gene silencing for RNA therapeutics.

Keywords:
bovine milkcancerdrug deliveryextracellular vesicleshigh-pressure homogenizationsmall interfering RNA

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Drug Delivery Systems

Background:

  • Extracellular vesicles (EVs) show promise for RNA therapeutics delivery.
  • Current methods face challenges in RNA loading and scalability.
  • High-pressure homogenization (HPH) is explored for scalable EV preparation.

Purpose of the Study:

  • To investigate HPH for loading small interfering RNA (siRNA) into EVs.
  • To evaluate the efficiency of siRNA-loaded EVs for cytoplasmic delivery.
  • To assess the impact of tumor-targeting modification on delivery efficacy.

Main Methods:

  • Utilized high-pressure homogenization (HPH) for simultaneous loading of siRNA into milk-derived EVs (mEVs).
  • Modified mEVs with cyclo(Arg-Gly-Asp-d-Phe-Lys) (cRGD) for tumor targeting.
  • Assessed intracellular siRNA delivery and gene knockdown efficiency in a human glioma cell line.

Main Results:

  • Successfully demonstrated siRNA loading into mEVs using HPH.
  • cRGD-modified mEVs exhibited high affinity for glioma cells and enhanced intracellular delivery.
  • siRNA delivered via cRGD-mEVs showed superior gene knockdown efficiency compared to unmodified EVs.

Conclusions:

  • HPH is a viable method for scalable production of siRNA-loaded EVs.
  • cRGD modification improves targeted delivery and therapeutic efficacy of siRNA-loaded EVs.
  • This approach holds potential for developing novel EV-based therapeutics for cytoplasmic siRNA delivery.