Small extracellular vesicles derived from four dimensional-culture of mesenchymal stem cells induce alternatively activated macrophages by upregulating IGFBP2/EGFR to attenuate inflammation in the spinal cord injury of rats

• Published in: Frontiers in bioengineering and biotechnology Vol. 11; p. 1146981
• Main Authors: Wang, Junhua, Wei, Qingshuai, Yang, Yue, Che, Mingtian, Ma, Yuanhuan, Peng, Lizhi, Yu, Haiyang, Shi, Huijuan, He, Guanheng, Wu, Rongjie, Zeng, Ting, Zeng, Xiang, Ma, Wenbin
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 28.04.2023
• Subjects: Bioengineering and Biotechnology; four dimensional culture; IGFBP2/EGFR; macrophage polarization; Small Extracellular Vesicles; spinal cord injury; spinal cord injury; four dimensional culture; Small Extracellular Vesicles; macrophage polarization; IGFBP2/EGFR
• ISSN: 2296-4185; 2296-4185
• DOI: 10.3389/fbioe.2023.1146981

Effectively reducing the inflammatory response after spinal cord injury (SCI) is a challenging clinical problem and the subject of active investigation. This study employed a porous scaffold-based three dimensional long-term culture technique to obtain human umbilical cord mesenchymal stem cell (hUC-MSC)-derived Small Extracellular Vesicles (sEVs) (three dimensional culture over time, the "4D-sEVs"). Moreover, the vesicle size, number, and inner protein concentrations of the MSC 4D-sEVs contained altered protein profiles compared with those derived from 2D culture conditions. A proteomics analysis suggested broad changes, especially significant upregulation of Epidermal Growth Factors Receptor (EGFR) and Insulin-like Growth Factor Binding Protein 2 (IGFBP2) in 4D-sEVs compared with 2D-sEVs. The endocytosis of 4D-sEVs allowed for the binding of EGFR and IGFBP2, leading to downstream STAT3 phosphorylation and IL-10 secretion and effective induction of macrophages/microglia polarization from the pro-inflammatory M1 to anti-inflammatory M2 phenotype, both and in the injured areas of rats with compressive/contusive SCI. The reduction in neuroinflammation after 4D-sEVs delivery to the injury site epicenter led to significant neuroprotection, as evidenced by the number of surviving spinal neurons. Therefore, applying this novel 4D culture-derived Small Extracellular Vesicles could effectively curb the inflammatory response and increase tissue repair after SCI.