An enzyme-based system for the extraction of small extracellular vesicles from plants

Abstract

Plant-derived nanovesicles (NVs) and extracellular vesicles (EVs) are considered to be the next generation of nanocarrier platforms for biotherapeutics and drug delivery. However, EVs exist not only in the extracellular space, but also within the cell wall. Due to the limitation of isolation methods, the extraction efficiency is low, resulting in the waste of a large number of plants, especially rare and expensive medicinal plants.There are few studies comparing EVs and NVs. To overcome these challenges, we proposed and validated a novel method for the isolation of plant EVs by degrading the plant cell wall with enzymes to release the EVs in the cell wall, making it easier for EVs to break the cell wall barrier and be collected. We extracted EVs from the roots of Morinda officinalis by enzymatic degradation(MOEVs) and nanoparticles by grinding method (MONVs) as a comparison group. The results showed smaller diameter and higher yield of MOEVs.Both MOEVs and MONVs were readily absorbed by endothelial cells without cytotoxicity and promoted the expression of miR-155. The difference is that the promotion of miR-155 by MOEVs is dose-effective. More importantly, MOEVs and MONVs are naturally characterized by bone enrichment. These results support that EVs in plants can be efficiently extracted by enzymatic cell wall digestion and also confirm the potential of MOEVs as therapeutic agents and drug carriers.