Int J Med Sci 2021; 18(7):1628-1638. doi:10.7150/ijms.52316

Research Paper

Co-culture with Endothelial Progenitor Cells promotes the Osteogenesis of Bone Mesenchymal Stem Cells via the VEGF-YAP axis in high-glucose environments

Peilian Wu1,2,3*, Xia Zhang1,4*, Yun Hu1,2,3, Dongrong Liu1,2,3, Jinlin Song1,2,3, Wenjie Xu1,2,3, Hao Tan1,2,3, Rui Lu1,2,3, Leilei Zheng1,2,3✉

1. The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, 401147, China.
2. Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, 401147, China.
3. Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, 401147, China.
4. West china dental hospital of Chongqing, Chongqing, 401147, China.
*These authors contributed equally to this work.

This is an open access article distributed under the terms of the Creative Commons Attribution License ( See for full terms and conditions.
Wu P, Zhang X, Hu Y, Liu D, Song J, Xu W, Tan H, Lu R, Zheng L. Co-culture with Endothelial Progenitor Cells promotes the Osteogenesis of Bone Mesenchymal Stem Cells via the VEGF-YAP axis in high-glucose environments. Int J Med Sci 2021; 18(7):1628-1638. doi:10.7150/ijms.52316. Available from

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Patients with type 2 diabetes mellitus (T2DM) have a high risk of fracture and experience poor bone healing. In recent years, bone mesenchymal stem cells (BMSCs) and endothelial progenitor cells (EPCs) have become the most commonly used cells in cell therapy and tissue engineering. In this study, we found that high glucose levels had a negative effect on the differentiation of BMSCs and EPCs. Considering that EPCs-BMSCs sheets can provide endothelial cells and osteoblastic cells, we transplanted cell sheets into T2DM rats with bilateral skull defects. The outcomes of the in vivo study revealed that EPCs-BMSCs sheets promoted ossification, which was verified by micro-CT and immunohistochemistry (IHC) analyses. Furthermore, we detected the VEGF content in the culture supernatant using an enzyme-linked immunosorbent assay (ELISA). The results showed that the BMSCs co-cultured with EPCs presented a higher level of VEGF than other cells. To assess the differentiation and migration of BMSCs exposed to VEGF, ALP staining, scratch assay and qRT-PCR analysis were performed. In addition, we used immunofluorescence and western blotting analysis to further explore the related mechanisms. The results showed that cells cultured with VEGF had a stronger actin cytoskeleton and a greater amount of nuclear and total YAP than cells cultured without VEGF. Taken together, our results indicate that co-culture with EPCs could promote the osteogenesis of BMSCs partially via VEGF. Furthermore, YAP and F-actin play important roles in this process.

Keywords: type 2 diabetes mellitus, High glucose, BMSCs, EPCs, VEGF, YAP