An Investigation of the Simultaneous Effects of Bone Morphogenic Protein (BMP)-4 and Static Magnetic Field on Viability and Proliferation Rates of Rat Bone Marrow Stem Cells

Authors
F. Javani Jouni 1
P. Abdolmaleki 1
M. Movahedin 2
1 Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objectives: This study investigated the possible synergistic effect of simultaneous treatment of bone morphogenic protein (BMP)-4 as a chemical stimulator and static magnetic field (SMF) as a physical stimulator on viability percent and proliferation rate in rat bone marrow stem cells.
Methods: Passage 5 cells were trypsinized, and a cell suspension prepared after which the cells were counted and cultured in 25 cm2 flasks. Cells were incubated for one day and washed with phosphate-buffered saline. We added BMP-4 at the optimum concentration of 25 ng/ml at different times (24, 48 and 96 h) into the medium. The cells were exposed at an optimum intensity of 4 mT of the SMF at different exposure times (24, 48, and 96 h). Subsequently cells were washed with phosphate-buffered saline, trypsinized, and separate cell suspensions were prepared from each flask. We investigated the viability and proliferation rates of treated cells by staining them with Trypan blue and performed cell counts with an optical microscope. The mean numbers of whole cells and living cells were considered to be the proliferation and survival rates, respectively.
Results: Increased SMF exposure and BMP-4 increased the viability percent and change in proliferation rate in the treated groups compared with their corresponding controls. The maximum increased viability was observed in the group that was treated with BMP-4 for 96 h.
Conclusion: Our results have supported the hypothesis that SMF alters the viability and proliferation rate of treated BMSCs, which was enhanced when the cells were treated simultaneously with SMF and BMP-4.

Keywords

[1]     Aladjadjiyan A. Study of the influence of magnetic field on some biological characteristics of Zea mais. J Central Euro Agric 2002; 3 (2): 89 - 94.
[2]     Belyavskaya NA. Biological effects due to weak magnetic field on plants. Adv Space Res 2004; 34(7): 1566-74.
[3]     WHO. Environmental health criteria 69: magnetic fields, Geneva. 1987; Available at: http://www.inchem.org/documents/ehc/ehc/ehc69.htm
[4]     Tofani S, Barone D, Cintorino M, de Santi MM, Ferrara A, Orlassino R, Ossola P, Peroglio F, Rolfo K, Ronchetto F. Static and ELF magnetic fields induce tumor growth inhibition and apoptosis. Bioelectromagnetics 2001; 22(6): 419-28.
[5]     McLean M, Engström S, Holcomb R. Static magnetic fields for the treatment of pain. Epilepsy & Behavior 2001; 2(3): S74-S80.
[6]     Potenza L, Ubaldi L, De Sanctis R, De Bellis R, Cucchiarini L, Dachà M. Effects of a static magnetic field on cell growth and gene expression in Escherichia coli. Mutat Res 2004; 561(1-2): 53-62.
[7]     Pirozzoli MC, Marino C, Lovisolo GA, Laconi C, Mosiello L, Negroni A. Effects of 50 Hz electromagnetic field exposure on apoptosis and differentiation in a neuroblastoma cell line. Bioelectromagnetics 2003; 24(7): 510-6.
[8]     Tavasoli Z, Abdolmaleki P, Mowla SJ, Ghanati F, SarvestaniAS. Investigation of the effects of static magnetic field on apoptosis in bone marrow stem cells of rat. Environmentalist 2009; 29(2): 220-4.
[9]     Teodori L, Grabarek J, Smolewski P, Ghibelli L, Bergamaschi A, De Nicola M, Darzynkiewicz Z. Exposure of cells to static magnetic field accelerates loss of integrity of plasma membrane during apoptosis. Cytometry 2002; 49(3): 113-8.
[10]  Tofani S, Barone D, Berardelli M, Berno E, Cintorino M, Foglia L, Ossola P, Ronchetto F, Toso E, Eandi M. Static and ELF magnetic fields enhance the in vivo anti-tumor efficacy of cis-platin against lewis lung carcinoma, but not of cyclophosphamide against B16 melanotic melanoma. Pharmacol Res 2003; 48(1): 83-90.
[11]  Saito K, Suzuki H, Suzuki K. Teratogenic effects of static magnetic field on mouse fetuses. Reproductive Toxicology 2006; 22(1): 118-24.
[12]  Leszczynski D. Rapporteur report: cellular, animal and epidemiological studies of the effects of static magnetic fields relevant to human health. Prog Biophys Mol Biol 2005; 87(2-3): 247-53.
[13]  Aldinucci C, Palmi M, Sgaragli G, Benocci A, Meini A, Pessina F, Pessina GP. The effect of pulsed electromagnetic fields on the physiologic behaviour of a human astrocytoma cell line. Biochim Biophys Acta 2000; 1499(1-2): 101-108.
[14]  Liburdy RP, Callahan DE, Harland J, Dunham E, Sloma TR, Yaswen P. Experimental evidence for 60 Hz magnetic fields operating through the signal transduction cascade. Effects on calcium influx and c-MYC mRNA induction. FEBS Lett 1993; 334(3): 301-8.
[15]  Pipkin JL, Hinson WG, Young JF, Rowland KL, Shaddock JG, Tolleson WH, Duffy PH, Casciano DA. Induction of stress proteins by electromagnetic fields in cultured HL-60 cells. Bioelectromagnetics 1999; 20(6): 347-57.
[16]  Javani Jouni F, Abdolmaleki P, Ghanati F. Oxidative stress in broad bean (Vicia faba L.) induced by static magnetic field under natural radioactivity. Mutation Research 2012; 741(1): 116-21.
[17]  Sahebjamei H, Abdolmaleki P, Ghanati F. Effects of magnetic field on the antioxidant enzyme activities of suspension-cultured tobacco cells. Bioelectromagnetics 2007; 28(1): 42-7.
[18]  Holcomb RR, Worthington WB, McCullough BA, McLean MJ. Static magnetic field therapy for pain in the abdomen and genitals. Pediatr Neurol 2000; 23(3): 261-4.
[19]  Laurence JA, French PW, Lindner RA, Mckenzie DR. Biological effects of electromagnetic fields--mechanisms for the effects of pulsed microwave radiation on protein conformation. J Theor Biol 2000; 206(2): 291-8.
[20]  Owen M. Marrow stromal stem cells. J Cell Sci Suppl 1988; 10: 63-76.
[21]  Toma C, Pittenger MF, Cahill KS, Byrne BJ, Kessler PD. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation 2002; 105(1): 93-8.
[22]  Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007; 131(5): 861-72.
[23]  Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, Gromoll J, Engel W. Derivation of male germ cells from bone marrow stem cells. Lab Invest 2006; 86(7): 654-63.
[24]  Yuge L, Okubo A, Miyashita T, Kumagai T, Nikawa T, Takeda S, Kanno M, Urabe Y, Sugiyama M, Kataoka K. Physical stress by magnetic force accelerates differentiation of human osteoblasts. Biochem Biophys Res Commun 2003; 311(1): 32-8.
[25]  Makoolati Z, Movahedin M, Forouzandeh-Moghadam M. Effects of different doses of bone morphogenetic protein 4 on viability and proliferation rates of mouse embryonic stem cells. Yakhteh Med J 2009; 11(1): 29-34.
[26]  Makoolati Z, Movahedin M, Forouzandeh-Moghadam M. Assessment of Piwil2 gene expression pattern upon germ cell development from mouse embryonic stem cell. Iran Red Crescent Med J 2009;11(4):408-13.
[27]  Makoolati Z, Movahedin M, Forouzandeh-Moghadam M. effects of treatment with bone morphogenetic protein 4 and co-culture on expression of Piwil2 gene in mouse differentiated embryonic stem cells. IJFS 2009; 3(2): 78-83.
[28]  Makoolati Z, Movahedin M, Forouzandeh-Moghadam M. Bone morphogenetic protein 4 is an efficient inducer for mouse embryonic stem cell differentiation into primordial germ cell. In Vitro Cell Dev Biol Anim 2011; 47(5-6): 391-8.
[29]  Mazaheri Z, Movahedin M, Rahbarizadeh F, Amanpour S. Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells. In Vitro Cell Dev Biol Anim 2011; 47(8): 521-5.
[30]  Dini L, Abbro L. Bioeffects of moderate-intensity static magnetic fields on cell cultures. Micron 2005; 36(3): 195-217.
[31]  Coletti D, Teodori L, Albertini MC, Rocchi M, Pristerà A, Fini M, Molinaro M, Adamo S. Static magnetic fields enhance skeletal muscle differentiation in vitro by improving myoblast alignment. Cytometry A 2007; 71(10): 846-56.
[32]  Payne AG. Using immunomagnetic technology and other means to facilitate stem cell homing. Med Hypotheses 2004; 62(5): 718-20.
[33]  Chen CJ, Ou YC, Liao SL, Chen WY, Chen SY, Wu CW, Wang CC, Wang WY, Huang YS, Hsu SH. Transplantation of bone marrow stromal cells for peripheral nerve repair. Exp Neurol 2007; 204(1): 443-53.
[34]  Marques-Mari AI, Lacham-Kaplan O, Medrano JV, Pellicer A, Simón C. Differentiation of germ cells and gametes from stem cells. Hum Reprod Update 2009; 15(3): 379-90.
[35]  Human Experimentation: Code of Ethics of W.M.A. Br Med J 1964; 2(5402): 177. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/ PMC1816102/?page=1
[36]  Javani Jouni F, Abdolmaleki P, Movahedin M. Investigation on the effect of static magnetic field up to 15 mT on the viability and proliferation rate of rat bone marrow stem cells. In Vitro Cell Dev Biol Anim 2013; 49(3): 212-9.
[37]  Heng BC, Cao T, Haider HK, Wang DZ, Sim EK, Ng SC. An overview and synopsis of techniques for directing stem cell differentiation in vitro. Cell Tissue Res 2004; 315(3): 291-303.
[38]  Ankeny DP, McTigue DM, Jakeman LB. Bone marrow transplants provide tissue protection and directional guidance for axons after contusive spinal cord injury in rats. Exp Neurol 2004; 190(1): 17-31.
[39]  Chen Q, Long Y, Yuan X, Zou L, Sun J, Chen S, Perez-Polo JR, Yang K. Protective effects of bone marrow stromal cell transplantation in injured rodent brain: synthesis of neurotrophic factors. J Neurosci Res 2005; 80(5): 611-9.
[40]  Javani Jouni F, Abdolmaleki P, Movahedin M. Investigation on the effect of static magnetic field up to 15 mT on the viability and proliferation rate of rat bone marrow stem cells. In Vitro Cell Dev Biol Anim 2013: 49(3): 212-9.
[41]  Ahmadianpour MR, Abdolmaleki P, Mowla SJ, Hosseinkhani S. Static magnetic field of 6 mT induces apoptosis and alters cell cycle in p53 mutant Jurkat cells. Electromagn Biol Med 2013; 32(1): 9-19.
[42]  Dudley BM, Runyan C, Takeuchi Y, Schaible K, Molyneaux K. BMP signaling regulates PGC numbers and motility in organ culture. Mech Dev 2007; 124(1): 68-77.
[43]  Ying QL, Nichols J, Chambers I, Smith A. BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 2003; 115(3): 281-92.
Pathobiology Reserach
Volume 16, Issue 3
December 2013
Pages 95-108