Effect of Pulsed Electromagnetic Fields on Osteogenic Differentiation and Wnt/β-catenin Signaling Pathway in RatBone Marrow Mesenchymal Stem Cells

ZHOU Yu-jing, WANG Pu, CHEN Hong-ying. et al

Abstract

To investigate the effect of pulsed electromagnetic fields (PEMFs) on osteogenic differentiation and Wnt/β-catenin signaling pathway in rat bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were isolated and the passage 3 cells were divided into 3 groups. Cells were cultured in LG-DMEM complete medium for 1 d to ensure fully adherent. Then, change the medium. Cells were maintained in complete medium (Control group) or in osteo-induction medium (OM group). The cells in PEMFs group were cultured in complete medium and exposed to 8 Hz, 3.8 mT PEMF stimulation for 40 min/d. The intervention lasted for 21 d. Cell proliferation activity was determined by using MTT. The effects of PEMF onosteogenic differentiation were assessed by ALP and Alizarin Red S staining. Various osteoblast-relevant genes and genes of Wnt/β-catenin signaling were analyzed by quantitative real-time RT-PCR. Results We found that OM could significantly promote the proliferation of BMSC at 7 d, 14 d, 21 d (P<0.05), but the effect was not obviously found in PEMFs group. For osteogenic differentiation, the positive rates of ALP or Alizarin Red S staining were detected higher in PEMFs/OM group （P<0.05）. Quantitative RT-PCR revealed PEMFs or OM could increase mRNA levels of Wnt1, Wnt3a, LRP5, β-catenin, BMP-2, Runx2, ALP, OC at special time point （P<0.05）. Compared to OM group, PEMFs have a lower expression in each detection, but the trends were consistent.　　 Conclusion PEMFs (8 Hz, 3.8 mT) could induce the osteogenic differentiation of rat BMSCs via activating Wnt/β-catenin signaling pathway.

 

Keywords: Pulsed electromagnetic fields, Bonemarrow mesenchymal stem cells, Osteogenic differentiation, Wnt/β-catenin signaling pathway 

 

Full Text:

PDF


References


Cairoli E, Zhukouskaya VV, Eller-Vainicher C, et al. Perspectives on osteoporosis therapies. J Endocrinol Invest, 2015;38(3):303-311.

Gnecchi M. Melo LG. Bone marrow-derived mesenchymal stem cells; isolation, expansion, characterization, viral transduction, and production of conditioned medium. Methods Mol Biol, 2009; 482; 281-294. doi: 10. 1007/978-1-59745-060- 718.

Liu HF, Yang L, He HC, etal. Pulsed electromagnetic fields on postmenopausal osteoporosis in Southwest China: a randomized. active-controlled clinical trial. Bioelectro-magnetics, 20 13;34(4):323-332.

Liu H, Yang L, He H, et al. The hemorheological safety of pulsed electromagnetic fields in postmenopausal women with osteoporosis in southwest China; a randomized, placebo controlled clinical trial. Clin Hemorheol Microcirc, 2013; 55 (3):285-295.

Jansen JH. van der Jagt OP, Punt BJ, et al. Stimulation of osteogenic differentiation in human osteoprogenitor cells by pulsed electromagnetic fields; an in vitro study. BMC Musculoskelet Disord,2010; 11; 188. doi; 10. 1186/1471-2474- 11-188.

Kaivosoja E, Sariola V, Chen Y, et al. The effect of pulsed electromagnetic fields and dehydroepiandrosterone on viability and osteo-induction of human mesenchymal stem cells. J Tissue Eng Regen Med.2015;9( 1);31-40.

Song MY, Yu JZ, Zhao DM. et al. The time-dependent manner of sinusoidal electromagnetic fields on rat bone marrow mesenchymal stem cells proliferation, differentiation, and mineralization. Cell Biochem Biophys,2014 ;69(1) ;47-54.

Liu C, Yu J, Yang Y, et al. Effect of 1 mT sinusoidal electromagnetic fields on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stromal cells. Bioelectromagnetics,2013;34(6):453-464.

Sun LY, Hsieh DK. Yu TC, et al. Effect of pulsed electromagnetic field on the proliferation and differentiation potential of human bone marrow mesenchymal stem cells. Bioelectromagnetics,2009;30(4):251-260.

Wang Y, Li YP. Paulson C, et al. Wnt and the Wnt signaling pathway in bone development and disease. Front Biosci (Landmark Ed),2014; 19;379-407.

Gregory CA, Gunn WG, Reyes E, et al. How Wnt signaling affects bone repair by mesenchymal stem cells from the bone marrow. Ann N Y Acad Sci ,2005; 1049 ; 97-106.

Zhou J, He H. Yang L, et al. Effects of pulsed electromagnetic fields on bone mass and Wnt/beta-catenin signaling pathway in ovariectomized rats. Arch Med Res,2012; 43(4):274-282.

Grassel S. Stockl S. Jenei-Lanzl Z. Isolation, culture, and osteogenic/chondrogenic differentiation of bone marrow-derived mesenchymal stem cells. Methods Mol Biol. 2012; 879: 203- 267. doi: 10. 1007/978-1-61779-815-3_ 14.

Nardi NB. Camassola M. Isolation and culture of rodent bone marrow-derived multipotent mesenchymal stromal cells. Methods Mol Biol. 2011; 698: 151-160. doi: 10. 1007/978-1- 60761-999-4_12.

Zhu H. Guo ZK. Jiang XX. etal. A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone. Nat Protoc.2010;5(3):550-560.

Soleimani M. Nadri S. A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat Protoc, 2009;4(1) ; 102-106.

Tepper OM. Callaghan MJ. Chang El. etal. Electromagnetic fields increase in vitro and in vivo angiogenesis through endothelial release of FGF-2. FASEB J. 2004; 18(11): 1231- 1233.

Selvamurugan N. Kwok S. Vasilov A. etal. Effects of BMP-2 and pulsed electromagnetic field ( PEMF) on rat primary osteoblastic cell proliferation and gene expression. J Orthop Res.2007;25 < 9):1213-1220.

Ducy P. Zhang R. Geoffroy V, et al. Osf2/Cbfal : a transcriptional activator of osteoblast differentiation. Cell. 1997;89(5);747-754.

Orimo H. Shimada T. The role of tissue-nonspecific alkaline phosphatase in the phosphate-induced activation of alkaline phosphatase and mineralization in SaOS-2 human osteoblast-like cells. Mol Cell Biochem.2008;315( 1-2) ;51-60.

Stein GS, Lian JB. Stein JL. et al. Transcriptional control of osteoblast growth and differentiation. Physiol Rev, 1996; 76 (2):593-629.


Refbacks

  • There are currently no refbacks.