Generalizability of Enzyme-Based Isolation Approach for Extracellular Vesicles From Traditional Medicinal Plants

Area of Science:

• Plant biotechnology
• Nanotechnology
• Pharmacology

Background:

• Plant-derived extracellular vesicles (EVs) show therapeutic potential.
• Effective manufacturing protocols for plant EVs are needed for clinical use.
• Current extraction methods may not be optimal for diverse plant sources and conditions.

Purpose of the Study:

• To optimize an enzyme-based method for extracting plant EVs.
• To demonstrate the generalizability of the method across various medicinal plants and storage conditions.
• To evaluate the bioactivity and therapeutic potential of extracted EVs.

Main Methods:

• Systematic optimization of an enzyme-based pulping process (Phyto-EVpure).
• Application of Phyto-EVpure to seventeen medicinal plants (leaves, rhizomes, roots), including fresh, frozen, and dried samples.
• In vitro and in vivo assessment of ginger-derived EVs for hepatoprotection.
• Comparative analysis of phloem and xylem EVs from dried vine samples for anti-tumor activity.

Main Results:

• Phyto-EVpure improved EV yield and purity compared to grinding for ginger, Morus alba leaves, and Isatis indigotica roots.
• Ginger EVs extracted via Phyto-EVpure maintained bioactivities and showed hepatoprotective effects.
• The method efficiently isolated EVs from various plant parts regardless of storage (fresh, frozen, dried).
• EVs from the phloem of dried vines demonstrated superior anti-tumor activity compared to xylem EVs.

Conclusions:

• Phyto-EVpure offers a manufacturing-friendly and generalizable protocol for isolating medicinal plant EVs.
• The method is effective across a wide range of traditional medicinal plants and storage conditions.
• Plant EVs isolated using Phyto-EVpure retain bioactivity and exhibit therapeutic potential, such as hepatoprotection and anti-tumor effects.