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p-Smad3 differentially regulates the cytological behavior of osteoclasts before and after osteoblasts maturation

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Abstract

Background

A series of previous investigations have revealed that p-Smad3 plays a facilitative role in the differentiation and maturation of osteoblasts, while also regulating the expression of certain intercellular communication factors. However, the effects of p-Smad3 in osteoblasts before and after maturation on the proliferation, migration, differentiation, apoptosis and other cellular behaviors of osteoclasts have not been reported.

Methods

MC3T3-E1 cells were cultured in osteogenic induction medium for varying durations, After that, the corresponding conditioned medium was collected and the osteoclast lineage cells were treated. To elucidate the regulatory role of p-Smad3 within osteoblasts, we applied the activator TGF-β1 and inhibitor SIS3 to immature and mature osteoblasts and collected corresponding conditioned media for osteoclast intervention.

Results

We observed an elevation of p-Smad3 and Smad3 during the early stage of osteoblast differentiation, followed by a decline in the later stage. we discovered that as osteoblasts mature, their conditioned media inhibit osteoclasts differentiation and the osteoclast-coupled osteogenic effect. However, it promotes apoptosis in osteoclasts and the angiogenesis coupled with osteoclasts. p-Smad3 in immature osteoblasts, through paracrine effects, promotes the migration, differentiation, and osteoclast-coupled osteogenic effects of osteoclast lineage cells. For mature osteoblasts, p-Smad3 facilitates osteoclast apoptosis and the angiogenesis coupled with osteoclasts.

Conclusions

As pre-osteoblasts undergo maturation, p-Smad3 mediated a paracrine effect that transitions osteoclast cellular behaviors from inducing differentiation and stimulating bone formation to promoting apoptosis and coupling angiogenesis.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by National Natural Science Foundation of China (grant number 81873320, 82274546, 81973878 and 82205142); The Scientific research project of Wuxi Municipal Health Commission (grant number Q202232); 2020 Jiangsu Province Traditional Chinese Medicine Science and Technology Development Plan Project (grant number YB2020042) and Graduate Research.and Innovation Projects of Jiangsu Province (grant number NO. KYCX22_2061).

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Author notes

  1. Jiapeng Ye, Zhen Hua and Jirimutu Xiao contributed equally to this work.

Authors and Affiliations

  1. Wuxi Hospital Affiliated to Nanjing University of Chinese Medicine, Wuxi, Jiangsu, 214071, China

    Jiapeng Ye, Zhen Hua, Yang Shao, Shaoshuo Li, Heng Yin, Mao Wu, Yi Rong, Bowen Hong & Jianwei Wang

  2. Laboratory of New Techniques of Restoration & Reconstruction of Orthopedics and Traumatology, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, 210023, China

    Jirimutu Xiao, Yang Guo & Yong Ma

  3. Mongolian Medicine College, Inner Mongolia Medical University, Hohhot, Inner Mongolia, 010159, China

    Jirimutu Xiao

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  1. Jiapeng Ye
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Contributions

Design of study were performed by J. Y. and J. W. Implementation of experiment, and data collection were performed by J. Y. and J. X. Data analysis and drafting manuscript were performed by J. Y. and Z. H.Drawing and revising manuscript were performed by Y. S., S. L., H.Y, M. W., Y. R., B. H., Y. G. and Y. M. All authors read and approved the final manuscript.

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Correspondence to Jianwei Wang.

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Ye, J., Hua, Z., Xiao, J. et al. p-Smad3 differentially regulates the cytological behavior of osteoclasts before and after osteoblasts maturation. Mol Biol Rep 51, 525 (2024). https://doi.org/10.1007/s11033-024-09400-0

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