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Open Access Research article

Osteogenic differentiation of amniotic epithelial cells: synergism of pulsed electromagnetic field and biochemical stimuli

Qian Wang1234, Wenchao Wu3, Xiaoyu Han3, Ai Zheng5, Song Lei6, Jiang Wu7, Huaiqing Chen37, Chengqi He124, Fengming Luo38 and Xiaojing Liu123*

Author Affiliations

1 Center of Rehabilitation Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China

2 Rehabilitation Key Laboratory of Sichuan Province, Chengdu, Sichuan, P. R. China

3 Laboratory of Cardiovascular Diseases, Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China

4 Sichuan University-The Hong Kong Polytechnic University Institute for Disaster Management and Reconstruction, Sichuan University, Chengdu 610041, China

5 Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu, Sichuan, P. R. China

6 Department of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China

7 Laboratory of Biomedical Engineering, West China Center of Medical Sciences, Sichuan University, Chengdu, Sichuan, P. R. China

8 Golden Card Ward, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China

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BMC Musculoskeletal Disorders 2014, 15:271  doi:10.1186/1471-2474-15-271

Published: 11 August 2014

Abstract

Background

Pulsed electromagnetic field (PEMF) is a non-invasive physical therapy used in the treatment of fracture nonunion or delayed healing. PEMF can facilitate the osteogenic differentiation of bone marrow mesenchymal stem cells in vitro. Amniotic epithelial cells (AECs) have been proposed as a potential source of stem cells for cell therapy. However, whether PEMF could modulate the osteogenic differentiation of AECs is unknown. In the present study, the effects of PEMF on the osteogenic differentiation of AECs were investigated.

Methods

AECs were isolated from amniotic membrane of human placenta by trypsin digestion and were induced by PEMF and/or osteo-induction medium. After 21 days we used real time RT-PCR and immunocytochemistry to study the expression of osteoblast markers. The signal transduction of osteogenesis was further investigated.

Results

The PEMF stimulation, or osteo-induction medium alone could induce osteogenic differentiation of AECs, as shown by expression of osteoblast specific genes and proteins including alkaline phosphatase and osteocalcin. Furthermore, a combination of PEMF and osteo-induction medium had synergy effects on osteogenic differentiation. In our study, the gene expression of BMP-2, Runx2, β-catenin, Nrf2, Keap1 and integrinβ1 were up-regulated in the osteogenic differentiation of AECs induced by PEMF and/or osteo-induction medium.

Conclusions

Combined application of PEMF and osteo-induction medium is synergistic for the osteogenic differentiation of AECs. It might be a novel approach in the bone regenerative medicine.

Keywords:
Amniotic epithelial cells; Osteogenic differentiation; Pulsed electromagnetic field; BMP-2; Wnt/β-catenin signaling; Reactive oxygen species; Integrinβ1