Preparation of therapy-grade extracellular vesicles from adipose tissue to promote diabetic wound healing

Chuqiao Pan; Peng Xu; Yi Zheng; Yikai Wang; Chuhsin Chen; Shibo Fu; Zibo Liu; Yahong Chen; Ke Xue; Qimin Zhou; Kai Liu

doi:10.3389/fbioe.2023.1129187

Preparation of therapy-grade extracellular vesicles from adipose tissue to promote diabetic wound healing

Front Bioeng Biotechnol. 2023 Mar 23:11:1129187. doi: 10.3389/fbioe.2023.1129187. eCollection 2023.

Authors

Chuqiao Pan¹, Peng Xu¹, Yi Zheng¹, Yikai Wang¹, Chuhsin Chen¹, Shibo Fu¹, Zibo Liu¹, Yahong Chen¹, Ke Xue¹, Qimin Zhou¹, Kai Liu¹

Affiliation

¹ Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Abstract

Background: Treatment of diabetic wounds is a major challenge in clinical practice. Extracellular vesicles (EVs) from adipose-derived stem cells have shown effectiveness in diabetic wound models. However, obtaining ADSC-EVs requires culturing vast numbers of cells, which is hampered by the need for expensive equipment and reagents, extended time cost, and complicated procedures before commercialization. Therefore, methods to extract EVs from discarded tissue need to be developed, for immediate application during surgery. For this reason, mechanical, collagenase-digestive, and constant in-vitro-collective methods were designed and compared for preparing therapy-grade EVs directly from adipose tissue. Methods: Characteristics and quantities of EVs were detected by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting firstly. To investigate the biological effects of EVs on diabetic wound healing, angiogenesis, proliferation, migration, and inflammation-regulation assays were then evaluated in vitro, along with a diabetic wound healing mouse model in vivo. To further explore the potential therapeutic mechanism of EVs, miRNA expression profile of EVs were also identified and analyzed. Results: The adipose tissue derived EVs (AT-EVs) were showed to qualify ISEV identification by nanoparticle tracking analysis and Western blotting and the AT-EVs yield from three methods was equal. EVs also showed promoting effects on biological processes related to diabetic wound healing, which depend on fibroblasts, keratinocytes, endothelial cells, and macrophages both in vitro and in vivo. We also observed enrichment of overlapping or unique miRNAs originate from different types of AT-EVs associated with diabetic wound healing for further investigation. Conclusion: After comparative analyses, a mechanical method was proposed for preparing immediate clinical applicable EVs from adipose tissue that would result in reduced preparation time and lower cost, which could have promising application potential in treating diabetic wounds.

Keywords: adipose tissue; diabetes; extracellular vesicles; therapy-grade; wound healing.

Grants and funding

This work was supported by the Industrial Support Fund of Huangpu District (XK2020003), Plastic surgery of Shanghai key clinical project (shslczdzk00901), Chuangke Fund of Shanghai Ninth hospital (CK2019009), Shanghai Sailing program (21YF1424300) the National Natural Science Foundation of China (82272290) and Hainan Province National Science Foundation of China (822CXTD537).