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

CGRP gene-modified rBMSCs show better osteogenic differentiation capacity in vitro 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Molecular Histology Springer Journals

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

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Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer Science+Business Media B.V., part of Springer Nature
Subject
Life Sciences; Cell Biology; Biomedicine, general; Developmental Biology
ISSN
1567-2379
eISSN
1567-2387
D.O.I.
10.1007/s10735-018-9775-2
Publisher site
See Article on Publisher Site

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.

Journal

Journal of Molecular HistologySpringer Journals

Published: May 30, 2018

References

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