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Journal of Molecular Histology

, Volume 49, Issue 4, pp 357–367 | Cite as

CGRP gene-modified rBMSCs show better osteogenic differentiation capacity in vitro

  • Xijiao Yu
  • Shuang Liu
  • Hui Chen
  • Xinyu Zhao
  • Xue Chen
  • Yi Du
  • Shu Li
Original Paper

Abstract

Calcitonin gene-related peptide (CGRP) is a marked and important neuropeptide expressed in nerve fibers during bone repair. This study investigated the role of CGRP overexpression on osteogenic differentiation of rat bone mesenchymal stem cells (rBMSCs). rBMSCs were infected with viral stocks of pLenO-DCE-CGRP (CGRP group) or pLenO-DCE (Vector group), while normal rBMSCs were used as a control. Transfection efficiency of rBMSCs was analyzed by flow cytometry. Cell proliferation was examined using a Cell Counting Kit-8 and flow cytometry. Expressions of alkaline phosphatase(ALP), bone sialoprotein (BSP) and Runt-related transcription factor 2(Runx2) in rBMSCs were detected at 1 and 2 weeks after mineral induction by real-time PCR and western blotting. Alizarin Red staining was applied at 28 days. The ratio of osteoprotegerin (OPG) to receptor activator of nuclear factor kappa B ligand (RANKL) was also detected to determine the underlying mechanism. pLenO-DCE-CGRP-induced rBMSCs stably overexpressing CGRP were successfully established. Overexpression of the CGRP gene significantly promoted rBMSC proliferation (p < 0.05). In addition, expressions of osteogenesis-related indexes were upregulated in the CGRP group (p < 0.05) compared with vector and control groups, and more mineralization nodules were observed in the CGRP group (p < 0.05). CGRP gene increased OPG and reduced RANKL in rBMSCs. Hence, the OPG/ RANKL ratio was increased in the CGRP group compared with the other two groups. CGRP gene-modified rBMSCs show better osteogenic differentiation capacity compared with rBMSCs in vitro.

Keywords

Calcitonin gene-related peptide Bone mesenchymal stem cells Transfection OPG/RANKL Tissue engineering 

Notes

Acknowledgements

National Natural Science Foundation of China (81271138). Natural Science Foundation of Shandong Province (ZR2017QH007). Open Foundation of Shandong Provincial Key Laboratory of Oral Tissue Regeneration (SDKQ201704).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Xijiao Yu
    • 1
    • 2
  • Shuang Liu
    • 1
  • Hui Chen
    • 2
  • Xinyu Zhao
    • 1
  • Xue Chen
    • 1
  • Yi Du
    • 2
  • Shu Li
    • 1
  1. 1.Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Department of Periodontology, School and Hospital of StomatologyShandong UniversityJinanPeople’s Republic of China
  2. 2.Department of EndodonticsJinan Stomatological HospitalJinanPeople’s Republic of China

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