Cyclic mechanical stretch enhances BMP9-induced osteogenic differentiation of mesenchymal stem cells
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The purpose of this study was to investigate whether mechanical stretch can enhance the bone morphogenetic protein 9 (BMP9)-induced osteogenic differentiation in MSCs.
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.
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.
Our results suggest that mechanical stretch enhances BMP9-induced osteoblastic lineage specification in C3H10T1/2 MSCs.
KeywordsMechanical stretch BMP9 Osteogenic differentiation Mesenchymal stem cells
Yang Song and Yinhong Tang contributed equally to this paper.
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.
This article does not contain any studies with human participants or animals performed by any of the authors.
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