Pulsed electromagnetic fields inhibit osteoclast differentiation in RAW264.7 macrophages via suppression of the protein kinase B/mammalian target of rapamycin signaling pathway

  • Authors:
    • Yutian Lei
    • Jinyu Su
    • Haixia Xu
    • Qiang Yu
    • Ming Zhao
    • Jing Tian
  • View Affiliations

  • Published online on: May 9, 2018     https://doi.org/10.3892/mmr.2018.8999
  • Pages: 447-454
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Abstract

When bone resorption, aided by the activity of osteoclasts, exceeds bone formation induced by osteoblasts, bone metabolism loses equilibration, which results in the development of bone diseases, including osteoporosis. Pulsed electromagnetic fields (PEMFs) are known to be involved in various biological processes, including cell proliferation, differentiation and apoptosis. However, the exact mechanism of action of osteoclasts remains poorly understood. In the present study, the effects of PEMFs on osteoclast differentiation and associated signaling pathways were systematically investigated in RAW264.7 macrophages. RAW264.7 cells were induced by receptor activator of nuclear factor‑κB ligand (RANKL) to obtain osteoclasts in vitro. The results of the present study demonstrated that PEMF exposure decreased osteoclast formation, limited tartrate‑resistant acid phosphatase activity, contracted bone resorption area and inhibited osteoclastic specific gene and protein expression. Furthermore, western blot analysis indicated that PEMFs distinctly abolished the upregulation of phosphorylated‑protein kinase B (Akt), ‑mammalian target of rapamycin (mTOR) and ‑ribosome S6 protein kinase (p70S6K) induced by RANKL, which was consistent with the effects of pharmacological inhibitor perifosine and rapamycin. Therefore, the present study suggested that PEMFs reduced osteoclast formation from RAW264.7 macrophages via inhibition of the Akt/mTOR signaling pathway. These findings provided novel insight into the mechanisms through which PEMFs suppress osteoclast differentiation.

References

1 

Liu W, Yang LH, Kong XC, An LK and Wang R: Meta-analysis of osteoporosis: Fracture risks, medication and treatment. Minerva Med. 106:203–214. 2015.PubMed/NCBI

2 

Musette P, Brandi ML, Cacoub P, Kaufman JM, Rizzoli R and Reginster JY: Treatment of osteoporosis: Recognizing and managing cutaneous adverse reactions and drug-induced hypersensitivity. Osteoporos Int. 21:723–732. 2010. View Article : Google Scholar : PubMed/NCBI

3 

Gartlehner G, Patel SV, Feltner C, Weber RP, Long R, Mullican K, Boland E, Lux L and Viswanathan M: Hormone therapy for the primary prevention of chronic conditions in postmenopausal women: Evidence report and systematic review for the US preventive services task force. JAMA. 318:2234–2249. 2017. View Article : Google Scholar : PubMed/NCBI

4 

Hadji P, Papaioannou N, Gielen E, Tepie Feudjo M, Zhang E, Frieling I, Geusens P, Makras P, Resch H, Möller G, et al: Persistence, adherence, and medication-taking behavior in women with postmenopausal osteoporosis receiving denosumab in routine practice in Germany, Austria, Greece and Belgium: 12-month results from a European non-interventional study. Osteoporosis Int. 26:2479–2489. 2015. View Article : Google Scholar

5 

Bone HG, Dempster DW, Eisman JA, Greenspan SL, McClung MR, Nakamura T, Papapoulos S, Shih WJ, Rybak-Feiglin A, Santora AC, et al: Odanacatib for the treatment of postmenopausal osteoporosis: Development history and design and participant characteristics of LOFT, the long-term Odanacatib fracture trial. Osteoporosis Int. 26:27212015. View Article : Google Scholar

6 

Hannon RA, Clack G, Rimmer M, Swaisland A, Lockton JA, Finkelman RD and Eastell R: Effects of the Src kinase inhibitor saracatinib (AZD0530) on bone turnover in healthy men: A randomized, double-blind, placebo-controlled, multiple-ascending-dose phase I trial. J Bone Miner Res. 25:463–471. 2013. View Article : Google Scholar

7 

Bonnick S, De Villiers T, Odio A, Palacios S, Chapurlat R, DaSilva C, Scott BB, Le Bailly De Tilleghem C, Leung AT and Gurner D: Effects of odanacatib on BMD and safety in the treatment of osteoporosis in postmenopausal women previously treated with alendronate: A randomized placebo-controlled trial. J Clin Endocrinol Metab. 98:4727–4735. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Bassett CA, Pawluk RJ and Pilla AA: Augmentation of bone repair by inductively coupled electromagnetic fields. Science. 184:575–577. 1974. View Article : Google Scholar : PubMed/NCBI

9 

Bassett CA, Pawluk RJ and Pilla AA: Acceleration of fracture repair by electromagnetic fields. A surgically noninvasive method. Ann N Y Acad Sci. 238:242–262. 1974. View Article : Google Scholar : PubMed/NCBI

10 

Wang T, Wang P, Cao Z, Wang X, Wang D, Shen Y, Jing D, Luo E and Tang W: Effects of BMP9 and pulsed electromagnetic fields on the proliferation and osteogenic differentiation of human periodontal ligament stem cells. Bioelectromagnetics. 38:63–77. 2017. View Article : Google Scholar : PubMed/NCBI

11 

Zhou J, He H, Yang L, Chen S, Guo H, Xia L, Liu H, Qin Y, Liu C, Wei X, et al: Effects of pulsed electromagnetic fields on bone mass and Wnt/β-catenin signaling pathway in ovariectomized rats. Arch Med Res. 274–282. 2012. View Article : Google Scholar : PubMed/NCBI

12 

Jing D, Zhai M, Tong S, Xu F, Cai J, Shen G, Wu Y, Li X, Xie K, Liu J, et al: Pulsed electromagnetic fields promote osteogenesis and osseointegration of porous titanium implants in bone defect repair through a Wnt/β-catenin signaling-associated mechanism. Sci Rep. 24:320452016. View Article : Google Scholar

13 

Urnukhsaikhan E, Cho H, Mishig-Ochir T, Seo YK and Park JK: Pulsed electromagnetic fields promote survival and neuronal differentiation of human BM-MSCs. Life Sci. 151:130–138. 2016. View Article : Google Scholar : PubMed/NCBI

14 

Kang KS, Hong JM, Seol YJ, Rhie JW, Jeong YH and Cho DW: Short-term evaluation of electromagnetic field pretreatment of adipose-derived stem cells to improve bone healing. J Tissue Eng Regen Med. 9:1161–1171. 2012. View Article : Google Scholar : PubMed/NCBI

15 

He J, Zhang Y, Chen J, Zheng S, Huang H and Dong X: Effects of pulsed electromagnetic fields on the expression of NFATc1 and CAII in mouse osteoclast-like cells. Aging Clin Exp Res. 27:13–19. 2015. View Article : Google Scholar : PubMed/NCBI

16 

Lei T, Liang Z, Li F, Tang C, Xie K, Wang P, Dong X, Shan S, Jiang M, Xu Q, et al: Pulsed electromagnetic fields (PEMF) attenuate changes in vertebral bone mass, architecture and strength in ovariectomized mice. Bone. 108:10–19. 2017. View Article : Google Scholar : PubMed/NCBI

17 

Lin HY and Lin YJ: In vitro effects of low frequency electromagnetic fields on osteoblast proliferation and maturation in an inflammatory environment. Bioelectromagnetics. 32:552–560. 2011. View Article : Google Scholar : PubMed/NCBI

18 

Zhang D, Huang Y, Huang Z, Zhang R, Wang H and Huang D: FTY-720P suppresses osteoclast formation by regulating expression of interleukin-6 (IL-6), interleukin-4 (IL-4) and matrix metalloproteinase 2 (MMP-2). Med Sci Monitor. 22:2187–2194. 2016. View Article : Google Scholar

19 

Yang J, Zhang J, Ding C, Dong D and Shang P: Regulation of osteoblast differentiation and iron content in MC3T3-E1 cells by static magnetic field with different intensities. Biol Trace Elem Res. Oct 19–2017.(Epub ahead of print). doi: 10.1007/s12011-017-1161-5.

20 

Barnaba SA, Ruzzini L, Di Martino A, Lanotte A, Sgambato A and Denaro V: Clinical significance of different effects of static and pulsed electromagnetic fields on human osteoclast cultures. Rheumatol Int. 32:1025–1031. 2012. View Article : Google Scholar : PubMed/NCBI

21 

Xie YF, Shi WG, Zhou J, Gao YH, Li SF, Fang QQ, Wang MG, Ma HP, Wang JF, Xian CJ, et al: Pulsed electromagnetic fields stimulate osteogenic differentiation and maturation of osteoblasts by upregulating the expression of BMPRII localized at the base of primary cilium. Bone. 93:22–32. 2016. View Article : Google Scholar : PubMed/NCBI

22 

Jing D, Li F, Jiang M, Cai J, Wu Y, Xie K, Wu X, Tang C, Liu J, Guo W, et al: Pulsed electromagnetic fields improve bone microstructure and strength in ovariectomized rats through a Wnt/Lrp5/β-catenin signaling-associated mechanism. PLoS One. 8:e793772013. View Article : Google Scholar : PubMed/NCBI

23 

Chang K, Chang WH, Huang S, Huang S and Shih C: Pulsed electromagnetic fields stimulation affects osteoclast formation by modulation of osteoprotegerin, RANK ligand and macrophage colony-stimulating factor. J Orthop Res. 23:1308–1314. 2005. View Article : Google Scholar : PubMed/NCBI

24 

Boyle WJ, Simonet WS and Lacey DL: Osteoclast differentiation and activation. Nature. 423:337–342. 2003. View Article : Google Scholar : PubMed/NCBI

25 

Xu H, Zhang J, Lei Y, Han Z, Rong D, Yu Q, Zhao M and Tian J: Low frequency pulsed electromagnetic field promotes C2C12 myoblasts proliferation via activation of MAPK/ERK pathway. Biochem Biophys Res Commun. 479:97–102. 2016. View Article : Google Scholar : PubMed/NCBI

26 

Park KH, Park B, Yoon DS, Kwon SH, Shin DM, Lee JW, Lee HG, Shim JH, Park JH and Lee JM: Zinc inhibits osteoclast differentiation by suppression of Ca2+-Calcineurin-NFATc1 signaling pathway. Cell Commun Signal. 11:742013. View Article : Google Scholar : PubMed/NCBI

27 

Kapetanakis NI, Uzan C, Jimenez-Pailhes AS, Gouy S, Bentivegna E, Morice P, Caron O, Gourzones-Dmitriev C, Le Teuff G and Busson P: Plasma miR-200b in ovarian carcinoma patients: Distinct pattern of pre/post-treatment variation compared to CA-125 and potential for prediction of progression-free survival. Oncotarget. 6:36815–36824. 2015. View Article : Google Scholar : PubMed/NCBI

28 

Halleen JM, Tiitinen SL, Ylipahkala H, Fagerlund KM and Vӓӓnӓnen HK: Tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of bone resorption. Clin Lab. 52:499–509. 2006.PubMed/NCBI

29 

Liu H, Li D, Liu S, Liu Z and Li M: Histochemical evidence of IGF2 mRNA-binding protein 2-mediated regulation of osteoclast function and adhesive ability. Histochem Cell Biol. 149:343–351. 2018. View Article : Google Scholar : PubMed/NCBI

30 

Rachner TD, Khosla S and Hofbauer LC: Osteoporosis: Now and the future. Lancet. 377:1276–1287. 2011. View Article : Google Scholar : PubMed/NCBI

31 

Liu HF, He HC, Yang L, Yang ZY, Yao K, Wu YC, Yang XB and He CQ: Pulsed electromagnetic fields for postmenopausal osteoporosis and concomitant lumbar osteoarthritis in southwest China using proximal femur bone mineral density as the primary endpoint: Study protocol for a randomized controlled trial. Trials. 16:2652015. View Article : Google Scholar : PubMed/NCBI

32 

Muramatsu Y, Matsui T, Deie M and Sato K: Pulsed electromagnetic field stimulation promotes anti-cell proliferative activity in doxorubicin-treated mouse osteosarcoma cells. In Vivo. 31:61–68. 2017. View Article : Google Scholar : PubMed/NCBI

33 

Visagie A, Kasonga A, Deepak V, Moosa S, Marais S, Kruger MC and Coetzee M: Commercial Honeybush (cyclopia spp.) tea extract inhibits osteoclast formation and bone resorption in RAW264.7 murine macrophages-an in vitro study. Int J Environ Res Public Health. 12:13779–13793. 2015. View Article : Google Scholar : PubMed/NCBI

34 

Collin-Osdoby P and Osdoby P: RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Biol. 816:187–202. 2012. View Article : Google Scholar : PubMed/NCBI

35 

Wong BR, Besser D, Kim N, Arron JR, Vologodskaia M, Hanafusa H and Choi Y: TRANCE, a TNF family member, activates Akt/PKB through a signaling complex involving TRAF6 and c-Src. Mol Cell. 4:1041–1049. 1999. View Article : Google Scholar : PubMed/NCBI

36 

Li L, Sapkota M, Gao M, Choi H and Soh Y: Macrolactin F inhibits RANKL-mediated osteoclastogenesis by suppressing Akt, MAPK and NFATc1 pathways and promotes osteoblastogenesis through a BMP-2/smad/Akt/Runx2 signaling pathway. Eur J Pharmacol. 815:202–209. 2017. View Article : Google Scholar : PubMed/NCBI

37 

Navé BT, Ouwens M, Withers DJ, Alessi DR and Shepherd PR: Mammalian target of rapamycin is a direct target for protein kinase B: Identification of a convergence point for opposing effects of insulin and amino-acid deficiency on protein translation. Biochem J. 344:427–431. 1999. View Article : Google Scholar : PubMed/NCBI

38 

Hsieh CJ, Kuo PL, Hou MF, Hung JY, Chang FR, Hsu YC, Huang YF, Tsai EM and Hsu YL: Wedelolactone inhibits breast cancer-induced osteoclastogenesis by decreasing Akt/mTOR signaling. Int J Oncol. 46:555–562. 2015. View Article : Google Scholar : PubMed/NCBI

39 

Takeshita S: SHIP-deficient mice are severely osteoporotic due to increased numbers of hyperresorptive osteoclasts. Nat Med. 9:943–949. 2002. View Article : Google Scholar

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APA
Lei, Y., Su, J., Xu, H., Yu, Q., Zhao, M., & Tian, J. (2018). Pulsed electromagnetic fields inhibit osteoclast differentiation in RAW264.7 macrophages via suppression of the protein kinase B/mammalian target of rapamycin signaling pathway. Molecular Medicine Reports, 18, 447-454. https://doi.org/10.3892/mmr.2018.8999
MLA
Lei, Y., Su, J., Xu, H., Yu, Q., Zhao, M., Tian, J."Pulsed electromagnetic fields inhibit osteoclast differentiation in RAW264.7 macrophages via suppression of the protein kinase B/mammalian target of rapamycin signaling pathway". Molecular Medicine Reports 18.1 (2018): 447-454.
Chicago
Lei, Y., Su, J., Xu, H., Yu, Q., Zhao, M., Tian, J."Pulsed electromagnetic fields inhibit osteoclast differentiation in RAW264.7 macrophages via suppression of the protein kinase B/mammalian target of rapamycin signaling pathway". Molecular Medicine Reports 18, no. 1 (2018): 447-454. https://doi.org/10.3892/mmr.2018.8999