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Controlled release of canine MSC-derived extracellular vesicles by cationized gelatin hydrogels

Yoshizaki, Karin Nishida, Hidetaka Tabata, Yasuhiko Jo, Jun-ichiro Nakase, Ikuhiko Akiyoshi, Hideo 京都大学 DOI:10.1016/j.reth.2022.11.009

2023.03

概要

INTRODUCTION: Canine mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) have emerged as a promising form of regenerative therapy. Therapeutic application of EVs remains difficult due to the short half-life of EVs in vivo and their rapid clearance from the body. We have developed cationized gelatin hydrogels that prolong the retention of EVs to overcome this problem. METHODS: Canine MSCs were isolated from bone marrow. MSC-derived EVs were isolated from the culture supernatant by ultracentrifugation. Gelatin was mixed with ethylene diamine anhydrate to cationized. Distinct cross-linked cationized gelatin hydrogels were created by thermal dehydration. Hydrogels were implanted into the back subcutis of mice in order to evaluate the degradation profiles. Hydrogels with collagenase were incubated at 37 °C in vitro to quantize the release of EVs from hydrogels. Lipopolysaccharide (LPS)-stimulated BV-2 cells were used to evaluate the immunomodulatory effect of EVs after release from the hydrogels. RESULTS: The cationized gelatin hydrogels suppressed EV release in PBS. More than 60% of immobilized EVs are not released from the hydrogels. The cationized hydrogels released EVs in a sustainable manner and prolonged the retention time of EVs depending on the intensity of cross-linking after degradation by collagenase. The expression of IL-1β in LPS-stimulated BV-2 cells was lower in EVs released from the hydrogels than in controls. CONCLUSIONS: Our results indicate that the controlled release of EVs can be achieved by cationized gelatin hydrogels. The released EVs experimentally confirmed to be effective in reducing proinflammatory response. The cationized gelatin hydrogels appear to be useful biomaterials for releasing canine MSC-derived EVs for regenerative therapy.

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