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Dkk-1–TNF-α crosstalk regulates MC3T3E1 pre-osteoblast proliferation and differentiation under mechanical stress through the ERK signaling pathway

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Abstract

The study aims to explore the role of the ERK signaling pathway in the crosstalk between Dkk-1 and TNF-α in MC3T3E1 pre-osteoblasts under cyclic tensile/compressive stress. A forced four-point bending system was used to apply cyclic uniaxial tensile/compressive strain (2000 μ, 0.5 Hz) to MC3T3E1 cells. Dkk-1 and TNF-α expression were upregulated in MC3T3E1 cells under compressive strain. Cell proliferation, the cell cycle, osteogenesis-related gene (Wnt5a, Runx2, Osterix) expression, β-catenin expression, and the p-ERK/ERK ratio were significantly enhanced, whereas apoptosis, the RANKL/OPG ratio, and TNF-α expression were significantly attenuated, by Dkk-1 silencing. Dkk-1 expression increased and the effects of Dkk-1 silencing were reversed when exogenous TNF-α was added. Mechanically, TNF-α crosstalked with Dkk-1 through ERK signaling in MC3T3E1 cells. ERK signaling blockade impaired Dkk-1-induced TNF-α expression and TNF-α-mediated Dkk-1 expression. Dkk-1 and TNF-α crosstalked, partially through ERK signaling, in MC3T3E1 cells under compressive/tensile strain, synergistically modulating various biological behaviors of the cells. These findings not only provide mechanical insight into the cellular events and molecular regulation of orthodontic tooth movement (OTM), but also aid the development of novel strategies to accelerate OTM.

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Data Availability

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

Abbreviations

OTM:

Orthodontic tooth movement

Dkk-1:

Dickkopf-related protein 1

RANKL:

Nuclear factor-κB ligand

OPG:

Osteoprotegerin

TNF:

Tumor necrosis factor

ERK:

Extracellular-regulated protein kinase

MSCs:

Mesenchymal stem cells

ATCC:

American Typical Culture Collection

α-MEM:

α -Modified Eagle’s medium (

FBS:

Fetal bovine serum

siRNAs:

Small interfering RNAs

RT-PCR:

Reverse-transcription polymerase chain reaction

FITC:

Fluorescein isothiocyanate

DAPI:

4′,6-Diamidino-2-phenylindole

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Acknowledgements

This work was supported by grants from the National Nature Science Foundation of China (Nos. 81973684, 81500818, 81873334, and 82174358).

Funding

This work was supported by grants from the National Nature Science Foundation of China (Nos. 81973684, 82174358 and 81873334).

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Authors and Affiliations

  1. Department of Stomatology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China

    Yeke Wu & Yuling Zuo

  2. College of Stomatology, Chongqing Medical University, Chongqing, People’s Republic of China

    Zheng Jing

  3. Department of Gynaecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China

    Disi Deng, Jin Yan & Min Liu

  4. Department of Radiology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China

    Li Li

  5. Department of Geriatrics, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China

    Wenbin Wu

  6. Department of Laboratory Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, #39 Shierqiao Rd, Chengdu, 610072, People’s Republic of China

    Qiongying Hu

  7. Department of Nuclear Medicine, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, 610072, People’s Republic of China

    Yunfei Xie

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  1. Yeke Wu
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  2. Zheng Jing
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  4. Jin Yan
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  6. Li Li
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  8. Wenbin Wu
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Contributions

YW: conceptualization, investigation, methodology, and writing of original draft. ZJ: investigation, methodology, and writing of original draft. DD: investigation, methodology, and writing of original draft. JY: investigation, methodology, and writing of original draft. ML: conceptualization, project administration, and writing – review & editing. LL: conceptualization, project administration, and writing – review & editing. YZ: methodology and visualization. WW: visualization, funding acquisition, project administration, and supervision. QH: funding acquisition, project administration, and supervision. YX: funding acquisition, project administration, and supervision.

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Correspondence to Qiongying Hu or Yunfei Xie.

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Wu, Y., Jing, Z., Deng, D. et al. Dkk-1–TNF-α crosstalk regulates MC3T3E1 pre-osteoblast proliferation and differentiation under mechanical stress through the ERK signaling pathway. Mol Cell Biochem 478, 2191–2206 (2023). https://doi.org/10.1007/s11010-022-04645-4

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