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Open Access Highly Accessed Research article

Influence of glucocorticoids on the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells

Da-An Zhou12, Hong-Xin Zheng3, Cheng-Wen Wang2, Dan Shi2 and Jian-Jun Li1*

Author Affiliations

1 Department of Spinal and Neural Function Reconstruction, School of Rehabilitation Medicine of Capital Medical University, China Rehabilitation Research Center, No. Jiaomeibei Road Fengtai District, Beijing 100068, China

2 Department of Rehabilitation, the 3rd Affiliated Hospital of Liaoning Medical University, Jinzhou 121000, China

3 Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China

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BMC Musculoskeletal Disorders 2014, 15:239  doi:10.1186/1471-2474-15-239

Published: 15 July 2014

Abstract

Background

Glucocorticoid has been used extensively in clinical applications, because of its several pharmacologic actions, which include immunosuppression, anti-inflammation, anti-shock, and relief of asthma. However, the long-term or high-dose application of glucocorticoid can induce adverse effects such as osteoporosis, which is known in this case as glucocorticoid-induced osteoporosis (GIOP). It is a secondary osteoporosis that results in easy fracturing, and even disability. Therefore it became a thorny issue.

Methods

The rat model of glucocorticoid-induced osteoporosis (GIOP) was replicated to isolate BMSCs. Rats were assigned into four groups: normal, normal induction, GIOP, and GIOP induction. The growth cycle was monitored by using flow cytometry. Osteogenic differentiation was compared by using alkaline phosphatase (ALP) staining with a modified calcium cobalt method. The quantitative detection of osteoprotegerin and the receptor activator of nuclear factor kappa-B ligand (RANKL) was conducted by using enzyme-linked immunoassay. Finally, renal Klotho mRNA expression was assessed by using RT-PCR.

Results

BMSC proliferation was reduced in GIOP rats. The ALP-positive expression of normal BMSCs to the osteogenic induction solution was stronger than that of BMSCs from GIOP rats (P < 0.01). Osteoprotegerin expression was significantly higher in the normal induction group than in the normal, GIOP (P < 0.01), and GIOP induction groups (P < 0.05). RANKL expression was significantly higher in the normal induction group than in the other groups (P < 0.01) and significantly higher in the normal group than in the GIOP and GIOP induction groups (P < 0.01). RT-PCR analysis showed that renal Klotho mRNA expression was significantly reduced in the GIOP group compared with the normal group (P < 0.01).

Conclusion

BMSC proliferation, osteogenic differentiation, and reactive activity to an osteogenic inductor were reduced in GIOP rats. Klotho mRNA expression decreased during GIOP induction.

Keywords:
Glucocorticoid; Osteogenic differentiation capacity; Osteoprotegerin; RANKL; Klotho gene