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Updated: Jun 5, 2025
Differentiation of Functional Osteoclasts from Human Peripheral Blood CD14+ Monocytes
Published on: January 27, 2023
Exosomes secreted from M2-polarized macrophages inhibit osteoclast differentiation via CYLD
Zi-Yan Guo1, Nan-Nan Yin2, Xiao-Fei Li2
1Department of Stomatology, Changhai Hospital Affiliated to Naval Medical University, 168 Changhai Road, Shanghai 200433, China.
Objective:
Bone resorption mediated by osteoclast differentiation induces the occurrence of bone-related diseases. Macrophages, an origin of osteoclasts, whose M2 type can reduce inflammation-induced bone damage. We aimed to investigate the effect of M2 macrophage-derived exosomes on osteoclast formation and elucidate its underlying mechanism.
Materials And Methods:
Exosomes were isolated from M2 macrophages (M2-exo) and were used to treat osteoclast-like cells. Osteoclast formation was evaluated using tartrate-resistant acid phosphatase, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting. The molecular mechanism of M2-exo function was analyzed by qRT-PCR, phosphor-kinase array analysis, and Western blotting.
Results:
M2-exo was internalized by osteoclasts and inhibited osteoclast differentiation in vitro. Moreover, CYLD was highly expressed in M2 macrophages and M2-exo-treated osteoclasts, and knockdown of it abrogated the inhibition of osteoclast differentiation caused by M2-exo. Additionally, CYLD suppressed the phosphorylation of STAT3, and STAT3 activator colivelin reversed the inhibition of osteoclast differentiation induced by CYLD overexpression.
Conclusion:
M2-exo inhibits osteoclast differentiation via delivering CYLD, which inactivates STAT3 signaling. These findings may provide a novel therapeutic option for bone diseases including periodontitis.
Insights
M2 macrophage-derived exosomes inhibit osteoclast formation by delivering CYLD, which inactivates STAT3 signaling. This offers a potential new treatment for bone diseases like periodontitis.
Area of Science:
- Cell Biology
- Immunology
- Bone Biology
Background:
- Osteoclast differentiation drives bone diseases.
- M2 macrophages can mitigate inflammation-induced bone damage.
- Investigating M2 macrophage-derived exosomes for therapeutic potential.
Purpose of the Study:
- To determine the effect of M2 macrophage-derived exosomes (M2-exo) on osteoclast formation.
- To elucidate the molecular mechanisms underlying M2-exo's function.
Main Methods:
- Exosomes isolated from M2 macrophages (M2-exo) were used to treat osteoclast-like cells.
- Osteoclast formation assessed via TRAP staining, qRT-PCR, and Western blotting.
- Molecular mechanism analyzed using qRT-PCR, phosphor-kinase array, and Western blotting.
Main Results:
- M2-exo inhibited osteoclast differentiation in vitro.
- CYLD, highly expressed in M2-exo, was crucial for inhibiting osteoclast differentiation.
- CYLD inactivated STAT3 signaling, reversing M2-exo's inhibitory effect.
Conclusions:
- M2-exo inhibits osteoclast differentiation by delivering CYLD, inactivating STAT3 signaling.
- This mechanism presents a novel therapeutic strategy for bone diseases, including periodontitis.
