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

Yub Raj Neupane; Harish K Handral; Syed Abdullah Alkaff; Wei Heng Chng; Gopalakrishnan Venkatesan; Chenyuan Huang; Choon Keong Lee; Jiong-Wei Wang; Gopu Sriram; Rhonnie Austria Dienzo; Wen Feng Lu; Yusuf Ali; Bertrand Czarny; Giorgia Pastorin

doi:10.1016/j.apsb.2022.10.022

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

Acta Pharm Sin B. 2023 May;13(5):1887-1902. doi: 10.1016/j.apsb.2022.10.022. Epub 2022 Nov 15.

Authors

Yub Raj Neupane^{1

2}, Harish K Handral³, Syed Abdullah Alkaff⁴, Wei Heng Chng^{1

5}, Gopalakrishnan Venkatesan^{1

6}, Chenyuan Huang^{7

8

9}, Choon Keong Lee¹, Jiong-Wei Wang^{7

8

9

10}, Gopu Sriram¹¹, Rhonnie Austria Dienzo¹², Wen Feng Lu³, Yusuf Ali^{12

13}, Bertrand Czarny^{4

12}, Giorgia Pastorin^{1

5

14}

Affiliations

¹ Department of Pharmacy, National University of Singapore, Singapore 117559, Singapore.
² Department of Pharmaceutical Sciences and Experimental Therapeutics, the University of Iowa College of Pharmacy, Iowa City, IA 52242, USA.
³ Department of Mechanical Engineering, National University of Singapore, Singapore 117575, Singapore.
⁴ School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798, Singapore.
⁵ Integrative Sciences and Engineering Programme, NUS Graduate School, National University of Singapore, Singapore 119077, Singapore.
⁶ Antimicrobial Resistance Interdisciplinary Research Group (AMR-IRG), Singapore-MIT Alliance for Research and Technology, Singapore 138602, Singapore.
⁷ Department of Surgery, National University of Singapore, Singapore 119228, Singapore.
⁸ Cardiovascular Research Institute, National University Heart Centre, Singapore 117599, Singapore.
⁹ Nanomedicine Translational Research Programme, Centre for NanoMedicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117609, Singapore.
¹⁰ Department of Physiology, National University of Singapore, Singapore 117593, Singapore.
¹¹ Faculty of Dentistry, National University of Singapore, Singapore 119085, Singapore.
¹² Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 639798, Singapore.
¹³ Singapore Eye Research Institute (SERI), Singapore General Hospital, Singapore 169856, Singapore.
¹⁴ NUSNNI-NanoCore, National University of Singapore, Singapore 117574, Singapore.

Abstract

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.

Keywords: Bionanotechnology; Cell migration; Cell proliferation; Cell-derived nanovesicles; ECM; Extracellular vesicles; Fibroblasts; Mesenchymal stem cells; Wound healing.