title: Bone marrow stromal/stem cell-derived extracellular vesicles regulate osteoblast activity and differentiation in vitro and promote bone regeneration in vivo.
abstract: Emerging evidence suggests that extracellular vesicles (EVs) are secreted by diverse tissues and play important roles in cell-cell communication, organ interactions and tissue homeostasis. Studies have reported the use of EVs to stimulate tissue regeneration, such as hepatic cell regeneration, and to treat diseases, such as pulmonary hypertension. However, little is known about the osteogenic effect of EVs. In this study, we explore the role of bone marrow stromal cell-derived EVs in the regulation of osteoblast activity and bone regeneration. We isolated bone marrow stromal/stem cell (BMSC)-derived EVs through gradient ultracentrifugation and ultrafiltration, and tested the influence of the EVs on osteogenesis both in vivo and in vitro. The results indicated that EVs positively regulated osteogenic genes and osteoblastic differentiation but did not inhibit proliferation in vitro. Furthermore, we constructed an EVs delivery system to stimulate bone formation in Sprague Dawley (SD) rats with calvarial defects. We found that BMSC-derived EVs led to more bone formation in the critical-size calvarial bone defects. Moreover, we found that miR-196a plays an essential role in the regulation of osteoblastic differentiation and the expression of osteogenic genes. We anticipate that our assay using bone marrow stromal cell-derived EVs will become a valuable tool for promoting bone regeneration.
authors.affiliation: Shanghai Jiaotong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.
authors.forename: Yunhao
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Shanghai Tongji University School of Medicine, Shanghai, 200029, China.
authors.forename: Lian
authors.initials: L
authors.lastname: Wang
authors.affiliation: Shanghai Tongji University School of Medicine, Shanghai, 200029, China.
authors.forename: Zhengliang
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Department of Anesthesiology, Affiliated Hospital of Guangdong Medical College, Yanjiang West Road, Guangzhou, 510120, China.
authors.forename: Genyin
authors.initials: G
authors.lastname: Chen
authors.affiliation: Shanghai Jiaotong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.
authors.forename: Changqing
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D001861:Bone Regeneration / Q000502:physiology*; D001842:Bone and Bones / Q000000981:diagnostic imaging / Q000502:physiology; D002454:Cell Differentiation; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000378:metabolism / Q000637:transplantation*; D005434:Flow Cytometry; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis; D053495:Osteopontin / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D055114:X-Ray Microtomography
chemical_list: D035683:MicroRNAs; D015675:Osteocalcin; D053495:Osteopontin