Immunoregulatory role of exosomes derived from differentiating mesenchymal stromal cells on inflammation and osteogenesis
Fei Wei
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Search for more papers by this authorZhengmao Li
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Search for more papers by this authorRoss Crawford
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Yin Xiao
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Correspondence
Yin Xiao, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Yinghong Zhou, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Yinghong Zhou
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Correspondence
Yin Xiao, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Yinghong Zhou, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Search for more papers by this authorFei Wei
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Search for more papers by this authorZhengmao Li
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Search for more papers by this authorRoss Crawford
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Yin Xiao
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Correspondence
Yin Xiao, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Yinghong Zhou, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Yinghong Zhou
The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia
The Australia-China Centre for Tissue Engineering and Regenerative Medicine (ACCTERM), Queensland University of Technology, Brisbane, Australia
Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, China
Correspondence
Yin Xiao, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Yinghong Zhou, The Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, 60 Musk Avenue, Kelvin Grove, Brisbane, Queensland 4059, Australia.
Email: [email protected]
Search for more papers by this authorAbstract
Bone marrow-derived mesenchymal stem/stromal cells (BMSCs) can differentiate into bone-forming osteoblasts, playing a crucial role in bone regeneration. Exosomes are naturally cell-secreted nanovesicles and are lately regraded as an emerging mediator of cellular communication in physiological and pathological conditions. The present study aimed at investigating the complex cellular communications, especially those among the differentiating BMSCs, immune cells (e.g., macrophages), and newly recruited BMSCs via exosome-mediated pathways. Exosomes were first isolated from osteogenically differentiating BMSCs at various stages (Day 0, Day 3, Day 7, and Day 14, respectively). The cellular uptake of isolated exosomes was examined in macrophages and human BMSCs (hBMSCs). The exosomes collected at various osteogenic differentiation stages (0d-exo, 3d-exo, 7d-exo, and 14d-exo) had no effect on the viability of hBMSCs. The uptake of exosomes (0d-exo, 3d-exo, and 7d-exo) significantly decreased proinflammatory-gene expression and the level of an M1 phenotypic marker. Our results then revealed that 3d-exo, 7d-exo, and 14d-exo led to a remarkable increase in mesenchymal stem/stromal cell migration. In addition, 0d-exo significantly promoted the expression of early osteogenic markers, such as alkaline phosphatase and bone morphogenetic protein 2, indicating a pro-osteogenic role of hBMSC-derived exosomes. Collectively, these results suggest that exosomes derived from differentiating mesenchymal stem/stromal cells play a unique osteoimmunomodulatory role in the regulation of bone dynamics.
CONFLICT OF INTEREST
The authors have declared that there is no conflict of interest.
Supporting Information
| Filename | Description |
|---|---|
| TERM_2947-Supp_0001_Figures.docxWord 2007 document , 15 MB |
Figure S1. Alizarin red S staining for mineralization in hBMSCs. Cells were cultured in a 24-well plate for 3, 7, or 14 days with a standard osteogenic medium. Mineralization was measured by Alizarin red S staining, where the calcified nodules appear bright red in color. Scale bar: 100 μm. Figure S2. Alkaline phosphatase (ALP) activity of hBMSCs. Cells were cultured in a 24-well plate for 3, 7, or 14 days in a standard osteogenic medium (OM). ALP activity was measured with the Alkaline Phosphatase Assay Kit. Values are expressed as the mean ± SD. Significance was found to be ****p < .0001. Figure S3. Characterization of exosome uptake by RAW 264.7 cells. Exosomes (10 μg by protein) were pre-labeled with exosome-labeling marker PKH67, and then incubated with RAW 264.7 cells for 24 hr at 37°C. The uptake of exosomes by RAW 264.7 cells was observed with a confocal microscopy. Low-magnification scale bar: 100 μm, high-magnification scale bar: 5 μm. Figure S4. Characterization of exosome uptake by hBMSCs at 24 hr. Exosomes (10 μg by protein) were pre-labeled with PKH67, and then incubated with hBMSCs for 24 hr at 37°C. The uptake of the fluorescently labeled exosomes by hBMSCs was observed with a confocal microscopy. Low-magnification scale bar: 100 μm, high-magnification scale bar: 5 μm. Figure S5. BCA assay for protein OD values. hBMSCs were cultured with the osteogenic medium supplemented with exosomes (10 μg by protein) for 7 days. Protein quantification was conducted using the Pierce BCA Protein Assay Kit. Figure S6. A schematic diagram illustrating the proposed mechanism of action of exosomes derived from differentiating MSCs. During osteogenic differentiation, a differentiating MSC secretes important nanocarriers to recruit additional naïve MSCs to the site of injury. This activity may generate a positive feedback loop to magnify naïve-MSC exosome signals, thereby reducing subsequent inflammation and promoting bone regeneration. Therefore, exosomes derived from osteogenically differentiating MSCs are extracellular vesicles important for control over inflammation and recruitment of naïve MSCs for bone regeneration. |
| TERM_2947-Supp_0002_Table.docxWord 2007 document , 19.9 KB | Table S1. Primers used in qRT-PCR |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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