Exosomes derived from miR‐375‐overexpressing human adipose mesenchymal stem cells promote bone regeneration

Abstract

Objectives:The present study aimed to investigate whether exosomes derived from miR-375-overexpressing human adipose mesenchymal stem cells (hASCs) could enhance bone regeneration. Materials and Methods:Exosomes enriched with miR-375 (Exo [miR-375]) were generated from hASCs stably overexpressing miR-375 after lentiviral transfection and identified with transmission electron microscopy, nanosight and western blotting.The construction efficiency of Exo (miR-375) was evaluated with qRT-PCR and incubated with human bone marrow mesenchymal stem cells (hBMSCs) to optimize the effective dosage. Then, the osteogenic capability of Exo (miR-375) was investigated with ALP and ARS assays. Furthermore, dual-luciferase reporter assay and western blotting were conducted to reveal the underlying mechanism of miR-375 in osteogenic regulation. Finally, Exo (miR-375) were embedded with hydrogel and applied to a rat model of calvarial defect, and μ-CT analysis and histological examination were conducted to evaluate the therapeutic effects of Exo (miR-375) in bone regeneration. Results: miR-375 could be enriched in exosomes by overexpressing in the parent cells. Administration of Exo (miR-375) at 50 μg/mL improved the osteogenic differentiation of hBMSCs. With miR-375 absorbed by hBMSCs, insulin-like growth factor binding protein 3 (IGFBP3) was inhibited by binding to its 3′UTR, and recombinant IGFBP3 protein reduced the osteogenic effects triggered by Exo (miR-375). After incorporated with hydrogel, Exo (miR-375) displayed a slow and controlled release, and further in vivo analysis demonstrated that Exo (miR-375) enhanced the bone regenerative capacity in a rat model of calvarial defect. Conclusions: Taken together, our study demonstrated that exosomes derived from miR-375-overexpressing hASCs promoted bone regeneration. S U PP O RTI N G I N FO R M ATI O N Additional supporting information may be found online in the Supporting Information section at the end of the article. How to cite this article: Chen S, Tang Y, Liu Y, et al. Exosomes derived from miR-375-overexpressing human adipose mesenchymal stem cells promote bone regeneration. Cell

Therapeutic miRNA-Enriched Extracellular Vesicles: Current Approaches and Future Prospects

“…miR-92-a-3p in MSC-derived exosomes may increase chondrogenesis and suppress cartilage degeneration via wnt5A, thus, can serve as treatment for OA [ 70 ]. Moreover, ADSC-derived exosomes carrying miR-375 target 3′UTR of insulin like growth factor binding protein 3 in bone marrow-derived stem cells in vitro to promote bone formation which may serve as a good therapeutic option for calvarial defects [ 71 ]. However, in-depth investigation is required to apply these therapies in in-vivo models and humans.…”

“…Although GMP ensures obtaining clinical-grade EVs, which can be loaded with RNAs, the problem of ultra-purification for human interventional studies still persist. Moreover, batch-to-batch variation pose an obstacle for human studies [ 71 ]. Future studies are required to develop appropriate protocols for miRNA-enriched EVs.…”

“…A detailed criterion for characterization of EV is required, specifying the features of EVs suitable and safe for clinical usage. The choice of parent cell as a source of EVs has to be determined for engineering therapies best suited for physiological environment [ 71 ]. Moreover, the route of administration of EVs is also important, taking in consideration how long EVs can remain functionally stable in body fluids.…”

Current Knowledge and Future Perspectives on Mesenchymal Stem Cell-Derived Exosomes as a New Therapeutic Agent

“…Other research showed that exosomes from miR-92a-3p-overexpressing MSCs promoted chondrogenesis in vitro and suppressed cartilage degradation in an OA mouse model by repressing WNT5A expression [157]. Recent studies showed that exosomes from miR-375-overexpressing MSCs promoted osteogenic differentiation in vitro by inhibiting the expression of insulin-like growth factor binding protein 3 and improved bone regeneration in a calvarial defect rat model [158].…”

Umbilical Cord Mesenchymal Stem Cell-Derived Nanovesicles Potentiate the Bone-Formation Efficacy of Bone Morphogenetic Protein 2

“…Released exosomes are internalized in target cells and subsequently affect their biological behaviors and promote the tissue regenerative process [ 18 , 19 , 20 ]. Exosomes secreted from mesenchymal stem cells (MSCs) can promote the proliferation, migration, and tube formation of endothelial cells in vitro, angiogenesis in vivo [ 21 , 22 , 23 , 24 ], and in vivo bone regeneration [ 25 , 26 , 27 ]. The microRNAs (miRNAs) in exosomes play the role of promoting endothelial cell proliferation and vessel formation, and regulating the osteogenic differentiation of targeted MSCs into mature osteoblasts and the subsequent mineralization process [ 18 , 28 ].…”