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International Orthopaedics

, Volume 42, Issue 4, pp 947–955 | Cite as

Cyclic mechanical stretch enhances BMP9-induced osteogenic differentiation of mesenchymal stem cells

  • Yang Song
  • Yinhong Tang
  • Jinlin Song
  • Mingxing Lei
  • Panpan Liang
  • Tiwei Fu
  • Xudong Su
  • Pengfei Zhou
  • Li Yang
  • Enyi Huang
Original Paper

Abstract

Purpose

The purpose of this study was to investigate whether mechanical stretch can enhance the bone morphogenetic protein 9 (BMP9)-induced osteogenic differentiation in MSCs.

Methods

Recombinant adenoviruses were used to overexpress the BMP9 in C3H10T1/2 MSCs. Cells were seeded onto six-well BioFlex collagen I-coated plates and subjected to cyclic mechanical stretch [6% elongation at 60 cycles/minute (1 Hz)] in a Flexercell FX-4000 strain unit for up to 12 hours. Immunostaining and confocal microscope were used to detect cytoskeleton organization. Cell cycle progression was checked by flow cytometry. Alkaline phosphatase activity was measured with a Chemiluminescence Assay Kit and was quantified with a histochemical staining assay. Matrix mineralization was examined by Alizarin Red S Staining.

Results

Mechanical stretch induces cytoskeleton reorganization and inhibits cell proliferation by preventing cells entry into S phase of the cell cycle. Although mechanical stretch alone does not induce the osteogenic differentiation of C3H10T1/2 MSCs, co-stimulation with mechanical stretch and BMP9 enhances alkaline phosphatase activity. The expression of key lineage-specific regulators (e.g., osteocalcin (OCN), SRY-related HMG-box 9, and runt-related transcription factor 2) is also increased after the co-stimulation, compared to the mechanical stretch stimulation along. Furthermore, mechanical stretch augments the BMP9-mediated bone matrix mineralization of C3H10T1/2 MSCs.

Conclusions

Our results suggest that mechanical stretch enhances BMP9-induced osteoblastic lineage specification in C3H10T1/2 MSCs.

Keywords

Mechanical stretch BMP9 Osteogenic differentiation Mesenchymal stem cells 

Notes

Author contributions

Yang Song and Yinhong Tang contributed equally to this paper.

Funding information

This work was supported by the National Natural Science Foundation of China (81301551), the Chongqing Research Program of Basic Research and Frontier Technology (cstc2013jcyjA10022), the Visiting Scholar Foundation of Key Laboratory of Biorheological Science and Technology (Chongqing University), the Ministry of Education (CQKLBST-2012-004), the Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJ1702024), the Scientific and Technological Research Program of Chongqing Yubei district (2017 nongshe 42), and the Program for Innovation Team Building at Institutions of Higher Education in Chongqing in 2016 (CXTDG201602006). Mingxing Lei is supported by projects funded by China Postdoctoral Science Foundation (2016M590866) and Special Funding for Postdoctoral Research Projects in Chongqing (Xm2015093).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© SICOT aisbl 2018

Authors and Affiliations

  1. 1.Key Laboratory of Biorheological Science and Technology (Chongqing University)Ministry of EducationChongqingPeople’s Republic of China
  2. 2.Chongqing Key Laboratory of Oral Diseases and Biomedical SciencesStomatological Hospital of Chongqing Medical UniversityChongqingPeople’s Republic of China
  3. 3.Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, College of StomatologyChongqing Medical UniversityChongqingPeople’s Republic of China
  4. 4.Integrative Stem Cell CenterChina Medical University Hospital, China Medical UniversityTaichungTaiwan
  5. 5.Institute of New Drug Development, College of Biopharmaceutical and Food SciencesChina Medical UniversityTaichungTaiwan

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