Extracellular vesicles derived from mesenchymal stromal cells mediate endogenous cell growth and migration via the CXCL5 and CXCL6/CXCR2 axes and repair menisci

Abstract

Background Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) are promising candidates for tissue regeneration therapy. However, the therapeutic efficacy of MSC-EVs for meniscus regeneration is uncertain, and the mechanisms underlying MSC-EV-mediated tissue regeneration have not been fully elucidated. The aims of this study were to evaluate the therapeutic efficacy of intra-articular MSC-EV injection in a meniscus defect model and elucidate the mechanism underlying MSC-EV-mediated tissue regeneration via combined bioinformatic analyses. Methods MSC-EVs were isolated from human synovial MSC culture supernatants via ultrafiltration. To evaluate the meniscus regeneration ability, MSC-EVs were injected intra-articularly in the mouse meniscus defect model immediately after meniscus resection and weekly thereafter. After 1 and 3 weeks, their knees were excised for histological and immunohistochemical evaluations. To investigate the mechanisms through which MSC-EVs accelerate meniscus regeneration, cell growth, migration, and chondrogenesis assays were performed using treated and untreated chondrocytes and synovial MSCs with or without MSC-EVs. RNA sequencing assessed the gene expression profile of chondrocytes stimulated by MSC-EVs. Antagonists of the human chemokine CXCR2 receptor (SB265610) were used to determine the role of CXCR2 on chondrocyte cell growth and migration induced by MSC-EVs. Results In the meniscus defect model, MSC-EV injection accelerated meniscus regeneration and normalized the morphology and composition of the repaired tissue. MSC-EVs stimulated chondrocyte and synovial MSC cell growth and migration. RNA sequencing revealed that MSC-EVs induced 168 differentially expressed genes in the chondrocytes and significantly upregulated CXCL5 and CXCL6 in chondrocytes and synovial MSCs. Suppression of CXCL5 and CXCL6 and antagonism of the CXCR2 receptor binding CXCL5 and CXCL6 negated the influence of MSC-EVs on chondrocyte cell growth and migration. Conclusions Intra-articular MSC-EV administration repaired meniscus defects and augmented chondrocyte and synovial MSC cell growth and migration. Comprehensive transcriptome/RNA sequencing data confirmed that MSC-EVs upregulated CXCL5 and CXCL6 in chondrocytes and mediated the cell growth and migration of these cells via the CXCR2 axis.

N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells

“…The ability of EV to stimulate cell migration has already been reported by several groups for different cell sources. The cancer cell-derived EV were shown to modulate the dissemination pattern of metastatic cells [ 43 ] and, in particular, MSC-EV have been seen to stimulate chondrocyte and synovial MSC migration via the CXCL5 and CXCL6/CXCR2 axes [ 12 ]. However, none of these studies referred to glycan-dependent mechanisms.…”

“…In this context, it is widely accepted that EV cargo (lipids, RNA and proteins) can be shuttled to the target cells by endocytosis (clathrin-dependent or caveolae-dependent) [ 3 , 4 , 5 , 6 ], phagocytosis [ 7 ], macropinocytosis [ 8 ] or by plasma/endosomal membrane fusion in acidic pH conditions, when EV and cells’ plasma membrane display the same fluidity [ 9 ]. Moreover, it has also been established that this transfer of vesicular content plays a crucial role in the intercellular communication in several biological processes, including angiogenesis [ 10 , 11 ], cell migration [ 12 ], inflammation regulation [ 13 , 14 ], bone formation [ 15 ] or interneuronal communication [ 16 ]. The transcriptomic, proteomic and lipidomic profiles of EV have been widely studied to find the molecules implicated in EV–cell interaction [ 17 ].…”

Research trends of mesenchymal stem cells application in orthopedics: A bibliometric analysis of the past 2 decades

“…At the same time, MSCs can also differentiate into ligament fibroblasts, contributing to the regeneration of cruciate ligament, and the differentiation process can be regulated ( 44 ). In addition, it can also promote the repair of meniscus injury ( 45 ). Platelet-rich plasma (PRP) is a product of platelet concentration obtained from peripheral blood after repeated centrifugation, which has been used in clinical treatment since the 1980s ( 46 ).…”

Source and hub of inflammation: The infrapatellar fat pad and its interactions with articular tissues during knee osteoarthritis

“…36 Extracellular vesicles (e.g., exosomes) secreted from MSCs have been reported to be able to enhance cartilage regeneration, maintain chondrocyte homeostasis and delay OA progression in preclinical in vitro and in vivo studies. [37][38][39] The chondrogenic properties of exosomes derived from IFP-MSCs were recently described in a preclinical study, which found that IFP-MSCs secrete a large amount of miR-100-5pabundant exosomes that protected chondrocytes from cell apoptosis and ameliorated OA progression in mice. 40 Cocultures of chondrocytes and IFP-MSCs can promote chondrogenic differentiation of IFP-MSCs, 41 and suppression of chondrocyte inflammation and MSC hypertrophy when compared to either of the cell populations cultured alone.…”

“…Moreover, human IFP‐MSCs show a higher chondrogenic differentiation potential than MSCs isolated from body fat tissue and bone marrow 36 . Extracellular vesicles (e.g., exosomes) secreted from MSCs have been reported to be able to enhance cartilage regeneration, maintain chondrocyte homeostasis and delay OA progression in preclinical in vitro and in vivo studies 37–39 . The chondrogenic properties of exosomes derived from IFP‐MSCs were recently described in a preclinical study, which found that IFP‐MSCs secrete a large amount of miR‐100‐5p‐abundant exosomes that protected chondrocytes from cell apoptosis and ameliorated OA progression in mice 40 …”