Cell-derived nanovesicles from mesenchymal stem cells as extracellular vesicle-mimetics in wound healing

Wound healing is a dynamic process that involves a series of molecular and cellular events aimed at replacing devitalized and missing cellular components and/or tissue layers. Recently, extracellular vesicles (EVs), naturally cell-secreted lipid membrane-bound vesicles laden with biological cargos including proteins, lipids, and nucleic acids, have drawn wide attention due to their ability to promote wound healing and tissue regeneration. However, current exploitation of EVs as therapeutic agents is limited by their low isolation yields and tedious isolation processes. To circumvent these challenges, bioinspired cell-derived nanovesicles (CDNs) that mimic EVs were obtained by shearing mesenchymal stem cells (MSCs) through membranes with different pore sizes. Physical characterisations and high-throughput proteomics confirmed that MSC-CDNs mimicked MSC-EVs. Moreover, these MSC-CDNs were efficiently uptaken by human dermal fibroblasts and demonstrated a dose-dependent activation of MAPK signalling pathway, resulting in enhancement of cell proliferation, cell migration, secretion of growth factors and extracellular matrix proteins, which all promoted tissue regeneration. Of note, MSC-CDNs enhanced angiogenesis in human dermal microvascular endothelial cells in a 3D PEG-fibrin scaffold and animal model, accelerating wound healing in vitro and in vivo. These findings suggest that MSC-CDNs could replace both whole cells and EVs in promoting wound healing and tissue regeneration. Agency for Science, Technology and Research (A*STAR) Ministry of Education (MOE) Nanyang Technological University Published version This work was supported by the National University of Singapore (NanoNash Program A-0004336-00-00 & A-0008504- 00-00, Singapore), and Nanyang Technological University, Singapore (grant number 001487-00001). Giorgia Pastorin. would also like to thank the Industry Alignment FunddPre-Positioning (IAF-PP) grant (A20G1a0046 and R-148-000-307-305/A0004345-00-00). This work was also supported by the Singapore Ministry of Education, under its Singapore Ministry of Education Academic Research Fund Tier 1 (10051 - MOE AcRF Tier 1: Thematic Call 2020) from Bertrand Czarny. Jiong-Wei Wang would like to thank the National University of Singapore NanoNASH Program (NUHSRO/2020/002/NanoNash/LOA) and the National University of Singapore Yong Loo Lin School of Medicine Nanomedicine Translational Research Program (NUHSRO/ 2021/034/TRP/09/Nanomedicine). Authors would also like to thank for the financial supports from Agency for Science, Technology, and Research (A*STAR, Singapore) Advanced Manufacturing and Engineering Individual Research Grant (AME IRG) (Project ID: A1883c0013, Singapore).