Secretive derived from hypoxia preconditioned mesenchymal stem cells promote cartilage regeneration and mitigate joint inflammation via extracellular vesicles

Yanmeng Yang; Yingnan Wu; Dahou Yang; Shu Hui Neo; Nurul Dinah Kadir; Doreen Goh; Jian Xiong Tan; Vinitha Denslin; Eng Hin Lee; Zheng Yang

doi:10.1016/j.bioactmat.2023.03.017

Secretive derived from hypoxia preconditioned mesenchymal stem cells promote cartilage regeneration and mitigate joint inflammation via extracellular vesicles

Bioact Mater. 2023 Mar 29:27:98-112. doi: 10.1016/j.bioactmat.2023.03.017. eCollection 2023 Sep.

Authors

Yanmeng Yang^{1

2

3}, Yingnan Wu^{1

2}, Dahou Yang³, Shu Hui Neo³, Nurul Dinah Kadir^{1

2}, Doreen Goh^{1

2}, Jian Xiong Tan¹, Vinitha Denslin², Eng Hin Lee^{1

2

3}, Zheng Yang^{1

2

3}

Affiliations

¹ Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
² NUS Tissue Engineering Program, Life Sciences Institute, National University of Singapore, Singapore.
³ Critical Analytics for Manufacturing Personalised-Medicine, Singapore-MIT Alliance for Research and Technology, Singapore.

Abstract

Secretome derived from mesenchymal stem cells (MSCs) have profound effects on tissue regeneration, which could become the basis of future MSCs therapies. Hypoxia, as the physiologic environment of MSCs, has great potential to enhance MSCs paracrine therapeutic effect. In our study, the paracrine effects of secretome derived from MSCs preconditioned in normoxia and hypoxia was compared through both in vitro functional assays and an in vivo rat osteochondral defect model. Specifically, the paracrine effect of total EVs were compared to that of soluble factors to characterize the predominant active components in the hypoxic secretome. We demonstrated that hypoxia conditioned medium, as well as the corresponding EVs, at a relatively low dosage, were efficient in promoting the repair of critical-sized osteochondral defects and mitigated the joint inflammation in a rat osteochondral defect model, relative to their normoxia counterpart. In vitro functional test shows enhancement through chondrocyte proliferation, migration, and matrix deposition, while inhibit IL-1β-induced chondrocytes senescence, inflammation, matrix degradation, and pro-inflammatory macrophage activity. Multiple functional proteins, as well as a change in EVs' size profile, with enrichment of specific EV-miRNAs were detected with hypoxia preconditioning, implicating complex molecular pathways involved in hypoxia pre-conditioned MSCs secretome generated cartilage regeneration.

Keywords: Cartilage regeneration; Extracellular vesicles; Hypoxia preconditioned MSCs; Joint inflammation; Secretome.