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The Distinct Effects of Estrogen and Hydrostatic Pressure on Mesenchymal Stem Cells Differentiation: Involvement of Estrogen Receptor Signaling

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Abstract

This study aimed to investigate the differential and synergistic effects of mechanical stimulation and estrogen on the proliferation and osteogenic or chondrogenic differentiation potential of bone marrow mesenchymal stem cells (BMSCs) and the roles of estrogen receptor (ER) in them. BMSCs were isolated and cultured using the whole bone marrow adherence method, and flow cytometry was used to identify the surface marker molecules of BMSCs. Cells were pre-treated with 1 nM 17β-estradiol or 1 nM of the estrogen receptor antagonist tamoxifen. Then, the cells were stimulated with hydrostatic pressure. Assessment included flow cytometry analysis of the cell cycle; immunofluorescent staining for F-actin; protein quantification for MAPK protein; and mRNA analysis for Col I, OCN, OPN and BSP after osteogenic induction and Sox-9, Aggrecan and Col-II after chondrogenic induction. Hydrostatic pressure (90 kPa/1 h) and 1 nM 17β-estradiol enhanced the cellular proliferation ability and the cytoskeleton activity but without synergistic biological effects. Estrogen activated ERKs and JNKs simultaneously and promoted the osteogenic differentiation, whereas the pressure just caused JNK-1/2 activation and promoted the chondrogenic differentiation of BMSCs. Estrogen had antagonism effect on chondrogenic promotion of hydrostatic pressure. Mechanobiological effects of hydrostatic pressure are closely associated with ERα activity. MAPK molecules and F-actin were likely to be important mediator molecules in the ER-mediated mechanotransduction of BMSCs.

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Abbreviations

BMSCs:

Bone marrow mesenchymal stem cells

TAM:

Tamoxifen

ER:

Estrogen receptor

OA:

Osteoarthritis

Col-I:

Collagen I

OCN:

Osteocalcin

OPN:

Osteopontin

BSP:

Bone sialoprotein

Col-II:

Collagen II

MAPK:

Mitogen-activated protein kinase

JNK:

c-Jun N-terminal kinase

ERK:

External signal-regulated kinase

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (Nos. 81371188, 31570951).

Author contributions

YZ. and F-Z.Y. made the acquisition of data and most of the analysis of the data, and were also involved in the drafting of the manuscript. Y-H.Z. and Y-J.C. helped with the interpretation of the data. H.M. and M.Z. was involved in the conception and design of the study, and did the most of the manuscript drafting and revising. All authors read and approved the final manuscript.

Conflict of interest

The authors declare that they have no financial or non-financial competing interests related to this study.

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    Authors and Affiliations

    1. State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi’an, 710032, China

      Ying Zhao, Yin-Hua Zhao, Yong-Jin Chen & Min Zhang

    2. Department of Physiology, School of Basic Medicine, Fourth Military Medical University, No. 169 West Changle Road, Xi’an, 710032, China

      Ying Zhao & Heng Ma

    3. The 105th Hospital of PLA, No.424 West ChangJiang Road, Hefei, 230031, China

      Fei-Zhou Yi

    4. Department of Pathophysiology, School of Basic Medicine, Fourth Military Medical University, No. 169 West Changle Road, Xi’an, 710032, China

      Heng Ma

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    1. Ying Zhao
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    Correspondence to Heng Ma or Min Zhang.

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    Associate Editor Peter E. McHugh oversaw the review of this article.

    Ying Zhao and Fei-Zhou Yi have contributed equally to this article.

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    Zhao, Y., Yi, FZ., Zhao, YH. et al. The Distinct Effects of Estrogen and Hydrostatic Pressure on Mesenchymal Stem Cells Differentiation: Involvement of Estrogen Receptor Signaling. Ann Biomed Eng 44, 2971–2983 (2016). https://doi.org/10.1007/s10439-016-1631-5

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