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BMP-2 Induced Signaling Pathways and Phenotypes: Comparisons Between Senescent and Non-senescent Bone Marrow Mesenchymal Stem Cells

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Abstract

The use of BMP-2 in orthopedic surgery is limited by uncertainty surrounding its effects on the differentiation of mesenchymal stem cells (MSCs) and how this is affected by cellular aging. This study compared the effects of recombinant human BMP-2 (rhBMP-2) on osteogenic and adipogenic differentiation between senescent and non-senescent MSCs. Senescent and non-senescent MSCs were cultured in osteogenic and adipogenic differentiation medium containing various concentrations of rhBMP-2. The phenotypes of these cells were compared by performing a calcium assay, adipogenesis assay, staining, real-time PCR, western blotting, and microarray analysis. rhBMP-2 induced osteogenic differentiation to a lesser extent (P < 0.001 and P = 0.005 for alkaline phosphatase activity and Ca2+ release) in senescent MSCs regardless of dose-dependent increase in both cells. However, the induction of adipogenic differentiation by rhBMP-2 was comparable between them. There was no difference between these two groups of cells in the adipogenesis assay (P = 0.279) and their expression levels of PPARγ were similar. Several genes such as CHRDL1, NOG, SMAD1, SMAD7, and FST encoding transcription factors were proposed to underlie the different responses of senescent and non-senescent MSCs to rhBMP-2 in microarray analyses. Furthermore, inflammatory, adipogenic, or cell death-related signaling pathways such as NF-kB or p38-MAPK pathways were upregulated by BMP-2 in senescent MSCs, whereas bone forming signaling pathways involving BMP, SMAD, and TGF- ß were upregulated in non-senescent MSCs as expected. This phenomenon explains bone forming dominance by non-senescent MSCs and possible frequent complications such as seroma, osteolysis, or neuritis in senescent MSCs during BMP-2 use in orthopedic surgery.

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Data Availability

All data generated or analyzed during this study are included in this article and could be available from the corresponding author on reasonable request.

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Acknowledgements

This study was supported by a Grant-in-Aid (No.06- 2011-218) from the SNUH Research Fund. We specially thank to Pf. In-Gyu Kim for his supervision of this study.

Funding

This study was supported by a Grant-in-Aid (No.06-2011-218) from the SNUH Research Fund.

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea

    Jae Hwan Cho

  2. Department of Orthopedic Surgery, Seoul National University College of Medicine, 103 Daehak-ro, Jongno-gu, Seoul, Republic of Korea

    Jae Hyup Lee, Kyung Mee Lee & Choon-Ki Lee

  3. Department of Orthopedic Surgery, SMG-SNU Boramae Medical Center, Boramae-ro 5-gil, Dongjak-Gu, Seoul, Republic of Korea

    Jae Hyup Lee

  4. Department of Biomedical Sciences, University of Ulsan College of Medicine, Seoul, Republic of Korea

    Dong-Myung Shin

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  1. Jae Hwan Cho
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Contributions

The concept was designed by JHC and JHL. Data acquisition was done by KML and DMS. Data were analyzed by JHL and DMS. Data interpretation was performed by JHC, JHL, CKL, and DMS. Initial manuscript was drafted by JHC. All authors contributed to revising the draft for intellectual content. All authors approved the final version of the manuscript.

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Correspondence to Jae Hyup Lee.

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Conflict of interest

All authors listed above (Jae Hwan Cho, Jae Hyup Lee, Kyung Mee Lee, Choon-Ki Lee, Dong-Myung Shin) have no conflicts of interest to declare.

Ethical Approval

All procedures performed in this study were under confirmation of Institutional Review Board of SMG-SNUBMC (Seoul Metropolitan Government Seoul National University Boramae Medical Center) (IRB No: 06-2011-218) with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Supplementary Information

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223_2021_923_MOESM1_ESM.docx

Supplementary file1 (DOCX 20 kb). Supplementary material 1. Specific methodology for comparisons between senescent and non-senescent MSCs induced by rhBMP-2

223_2021_923_MOESM2_ESM.pptx

Supplementary file2 (PPTX 259 kb). Supplementary material 2. GSEA leading edge subset analysis for P4 MSC (sample CD) vs senescent MSC (sample AB)

Supplementary file3 (XLSX 34 kb). Supplementary material 3. The raw data of real-time PCR

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Cho, J.H., Lee, J.H., Lee, K.M. et al. BMP-2 Induced Signaling Pathways and Phenotypes: Comparisons Between Senescent and Non-senescent Bone Marrow Mesenchymal Stem Cells. Calcif Tissue Int 110, 489–503 (2022). https://doi.org/10.1007/s00223-021-00923-3

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