Journal of Shanghai Jiao Tong University (Medical Science) >

2023 , Vol. 43 >Issue 4: 406 - 416

DOI: https://doi.org/10.3969/j.issn.1674-8115.2023.04.002

Basic research

Ameliorative effects on osteoporosis of small extracellular vesicles derived from bone marrow mesenchymal stem cells

  • Xuran LI ,
  • Shicong TAO ,
  • Shangchun GUO
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  • 1.Department of Orthopedic Surgery, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
    2.Clinical Medical College of Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
    3.Institute of Microsurgery on Extremities, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
TAO Shicong, E-mail: sctao@shsmu.edu.cn.
GUO Shangchun, E-mail: scguo@shsmu.edu.cn

Received date: 2022-12-23

  Accepted date: 2023-03-27

  Online published: 2023-04-28

Supported by

National Natural Science Foundation of China(81802226);Shanghai Pujiang Program(2019PJD038);Shanghai “Rising Stars of Medical Talent” Youth Development Program in 2020;Shanghai Sixth People's Hospital Funding(ynyq202101);"Two-hundred Talents" Program of Shanghai Jiao Tong University School of Medicine(2022-017)

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Abstract

Objective ·To investigate the effects of small extracellular vesicles (sEVs) derived from human bone marrow mesenchymal stem cells (BMSCs) on the regulation of osteoclast differentiation and macrophage polarization in mice, and mouse model of osteoporosis. Methods ·BMSCs were cultured and sEVs were isolated through differential centrifugation. The isolated sEVs were identified by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). RAW264.7 cells were cultured and stimulated with macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL) to differentiate the cells into osteoclasts. Tartrate-resistant acid phosphatase (TRAP) staining and phalloidin staining were performed to assess the effect of sEVs on osteoclast formation. The expression levels of osteoclast marker genes, i.e., cAMP-response element binding protein (CREB), cathepsin K (CTSK), and Jun proto-oncogene (c-Jun) were examined by real-time quantitative PCR. To polarize RAW264.7 cells to M1 phenotype, they were cultured with lipopolysaccharides; to polarize them to M2 phenotype, they were cultured with interleukin-4 (IL-4) and IL-13. Flow cytometry was performed to detect the effect of sEVs on macrophage polarization. Micro-computed tomography (micro-CT) and TRAP staining were performed to investigate the effect of sEVs on the bone tissues of lumbar vertebrae in osteoporosis mouse models. Results ·TEM and NTA demonstrated that the isolated sEVs had a typical globular structure with a diameter ranging from 30?150 nm. TRAP staining and phalloidin staining showed that BMSC-derived sEVs inhibited the fusion of RAW264.7 cells to form osteoblasts. PCR revealed that sEVs could decrease the expression of CREB, CTSK, and c-Jun (all P<0.05). Flow cytometry analysis indicated that BMSC-derived sEVs inhibited RAW264.7 macrophages polarization to M1 phenotype and induced RAW264.7 macrophages polarization to M2 phenotype. Micro-CT indicated that the number of trabeculae and the bone volume fraction of lumbar vertebrae were significantly higher in the sEV-intervened group than those in the control group (both P<0.05). TRAP staining revealed a reduction of osteoclast number in the lumbar vertebrae after intervention with sEVs. Conclusion ·The sEVs from human BMSCs can delay bone loss in osteoporosis mice, which may be related to its effects of inhibiting osteoclast differentiation and promoting the polarization of M2 type macrophages.

Key words： bone marrow mesenchymal stem cell (BMSC); small extracellular vesicle (sEV); osteoporosis; osteoclast; macrophage polarization

Cite this article

Xuran LI , Shicong TAO , Shangchun GUO . Ameliorative effects on osteoporosis of small extracellular vesicles derived from bone marrow mesenchymal stem cells[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023 , 43(4) : 406 -416 . DOI: 10.3969/j.issn.1674-8115.2023.04.002

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