{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T19:27:51Z","timestamp":1791228471493,"version":"4.1.0"},"reference-count":56,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,21]],"date-time":"2022-01-21T00:00:00Z","timestamp":1642723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003710","name":"Korea Health Industry Development Institute","doi-asserted-by":"publisher","award":["HI19C0757"],"award-info":[{"award-number":["HI19C0757"]}],"id":[{"id":"10.13039\/501100003710","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001321","name":"National Research Foundation","doi-asserted-by":"publisher","award":["NRF-2019R111A2A01063305"],"award-info":[{"award-number":["NRF-2019R111A2A01063305"]}],"id":[{"id":"10.13039\/501100001321","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Stroke is among the leading causes of death worldwide, and stroke patients are more likely to live with permanent disabilities even after treatment. Several treatments are being developed to improve the quality of life of patients; however, these treatments still have important limitations. Our study thus sought to evaluate the neural differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) at various pulsed electromagnetic field (PEMF) frequencies. Furthermore, the effects of selected frequencies in vivo were also evaluated using a mouse ischemia stroke model. Cell proliferation decreased by 20% in the PEMF group, as demonstrated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, and lactate dehydrogenase (LDH) secretion increased by approximately 10% in an LDH release assay. Fluorescence-activated cell sorting (FACS) analysis demonstrated that CD73 and CD105 were downregulated in the PEMF group at 60 Hz. Moreover, microtubule-associated protein 2 (MAP-2) and neurofilament light chain (NF-L) were upregulated in cell cultures at 60 and 75 Hz. To assess the effects of PEMF in vivo, cerebral ischemia mice were exposed to a PEMF at 60 Hz. Neural-related proteins were significantly upregulated in the PEMF groups compared with the control and cell group. Upon conducting rotarod tests, the cell\/PEMF group exhibited significant differences in motor coordination at 13 days post-treatment when compared with the control and stem-cell-treated group. Furthermore, the cell and cell\/PEMF group exhibited a significant reduction in the expression of matrix metalloproteinase-9 (MMP-9), tumor necrosis factor-alpha (TNF-\u03b1) and interferon-gamma (IFN-\u03b3) in the induced ischemic area compared with the control. Collectively, our findings demonstrated that PEMFs at 60 and 75 Hz could stimulate hBM-MSCs neural differentiation in vitro, in addition to promoting neurogenesis to enhance the functional recovery process by reducing the post-stroke inflammatory reaction.<\/jats:p>","DOI":"10.3390\/ijms23031177","type":"journal-article","created":{"date-parts":[[2022,1,21]],"date-time":"2022-01-21T08:37:18Z","timestamp":1642754238000},"page":"1177","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Induced Neurodifferentiation of hBM-MSCs through Activation of the ERK\/CREB Pathway via Pulsed Electromagnetic Fields and Physical Stimulation Promotes Neurogenesis in Cerebral Ischemic Models"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9910-9991","authenticated-orcid":false,"given":"Hee-Jung","family":"Park","sequence":"first","affiliation":[{"name":"Department of Medical Biotechnology, Dongguk University, Goyang-si 10326, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ju-Hye","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, Dongguk University, Goyang-si 10326, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myeong-Hyun","family":"Nam","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, Dongguk University, Goyang-si 10326, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7533-9605","authenticated-orcid":false,"given":"Young-Kwon","family":"Seo","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, Dongguk University, Goyang-si 10326, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Avan, A., Digaleh, H., Di Napoli, M., Stranges, S., Behrouz, R., Shojaeianbabaei, G., Amiri, A., Tabrizi, R., Mokhber, N., and Spence, J.D. (2019). Socioeconomic status and stroke incidence, prevalence, mortality, and worldwide burden: An ecological analysis from the Global Burden of Disease Study 2017. BMC Med., 17.","DOI":"10.1186\/s12916-019-1397-3"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1612","DOI":"10.1016\/S0140-6736(08)60694-7","article-title":"Stroke","volume":"371","author":"Donnan","year":"2008","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e139","DOI":"10.1161\/CIR.0000000000000757","article-title":"Heart Disease and Stroke Statistics-2020 Update: A Report From the American Heart Association","volume":"141","author":"Virani","year":"2020","journal-title":"Circulation"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1517\/14712598.2013.804507","article-title":"Cell-based therapy for ischemic stroke","volume":"13","author":"Zhang","year":"2013","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1002\/stem.269","article-title":"Mesenchymal stromal cells: Current understanding and clinical status","volume":"28","author":"Salem","year":"2010","journal-title":"Stem Cells"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1126\/science.284.5411.143","article-title":"Multilineage potential of adult human mesenchymal stem cells","volume":"284","author":"Pittenger","year":"1999","journal-title":"Science"},{"key":"ref_7","first-page":"2899","article-title":"Bone marrow cells differentiation into organ cells using stem cell therapy","volume":"20","author":"Yang","year":"2016","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1006\/bcmd.2001.0423","article-title":"Adult rat and human bone marrow stromal stem cells differentiate into neurons","volume":"27","author":"Black","year":"2001","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1097\/00001756-200207020-00023","article-title":"Differentiation of adult bone marrow stem cells into neuroprogenitor cells in vitro","volume":"13","author":"Kim","year":"2002","journal-title":"Neuroreport"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1089\/scd.2005.14.65","article-title":"Neural cell differentiation in vitro from adult human bone marrow mesenchymal stem cells","volume":"14","author":"Long","year":"2005","journal-title":"Stem Cells Dev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1186\/s13287-018-0913-2","article-title":"Neural stem cell therapy for subacute and chronic ischemic stroke","volume":"9","author":"Boese","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"908","DOI":"10.3389\/fneur.2019.00908","article-title":"Adult Stem Cells and Induced Pluripotent Stem Cells for Stroke Treatment","volume":"10","author":"Castillo","year":"2019","journal-title":"Front. Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"382","DOI":"10.3389\/fnhum.2014.00382","article-title":"Neurorestorative therapy for stroke","volume":"8","author":"Chen","year":"2014","journal-title":"Front. Hum. Neurosci."},{"key":"ref_14","first-page":"739","article-title":"Effects of electromagnetic field (PEMF) exposure at different frequency and duration on the peripheral nerve regeneration: In vitro and in vivo study","volume":"126","author":"Hei","year":"2016","journal-title":"Int. J. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1002\/bem.21832","article-title":"Pulsed electromagnetic fields (PEMF) promote early wound healing and myofibroblast proliferation in diabetic rats","volume":"35","author":"Cheing","year":"2014","journal-title":"Bioelectromagnetics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1472","DOI":"10.1007\/s12035-014-8650-8","article-title":"Epigenetic modulation of adult hippocampal neurogenesis by extremely low-frequency electromagnetic fields","volume":"49","author":"Leone","year":"2014","journal-title":"Mol. Neurobiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1007\/s12975-014-0334-1","article-title":"Effect of pulsed electromagnetic field (PEMF) on infarct size and inflammation after cerebral ischemia in mice","volume":"5","author":"Yang","year":"2014","journal-title":"Transl. Stroke Res."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Liu, X., Zuo, H., Wang, D., Peng, R., Song, T., Wang, S., Xu, X., Gao, Y., Li, Y., and Wang, S. (2015). Improvement of spatial memory disorder and hippocampal damage by exposure to electromagnetic fields in an Alzheimer\u2019s disease rat model. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0126963"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Jang, S., Cho, H.H., Cho, Y.B., Park, J.S., and Jeong, H.S. (2010). Functional neural differentiation of human adipose tissue-derived stem cells using bFGF and forskolin. BMC Cell Biol., 11.","DOI":"10.1186\/1471-2121-11-25"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/S0006-291X(02)00469-2","article-title":"Neurogenic differentiation of murine and human adipose-derived stromal cells","volume":"294","author":"Safford","year":"2002","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1007\/s11064-009-0101-y","article-title":"Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells","volume":"35","author":"Qian","year":"2010","journal-title":"Neurochem. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1634\/stemcells.21-1-50","article-title":"Matrix cells from Wharton\u2019s jelly form neurons and glia","volume":"21","author":"Mitchell","year":"2003","journal-title":"Stem Cells"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"507301","DOI":"10.1155\/2013\/507301","article-title":"Prospective isolation of murine and human bone marrow mesenchymal stem cells based on surface markers","volume":"2013","author":"Mabuchi","year":"2013","journal-title":"Stem Cells Int."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"53","DOI":"10.4252\/wjsc.v5.i2.53","article-title":"Comparison of phenotypic markers and neural differentiation potential of multipotent adult progenitor cells and mesenchymal stem cells","volume":"5","author":"Singh","year":"2013","journal-title":"World J. Stem Cells"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.ceb.2004.08.006","article-title":"Wnt signaling and the regulation of stem cell function","volume":"16","author":"Sommer","year":"2004","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Br\u00e4utigam, C., Raggioli, A., and Winter, J. (2013). The Wnt\/\u03b2-catenin pathway regulates the expression of the miR-302 cluster in mouse ESCs and P19 cells. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0075315"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.3892\/mmr.2013.1713","article-title":"Canonical Wnt signaling activates miR-34 expression during osteoblastic differentiation","volume":"8","author":"Tamura","year":"2013","journal-title":"Mol. Med. Rep."},{"key":"ref_28","first-page":"5978","article-title":"Parathyroid hormone induces epithelial-to-mesenchymal transition via the Wnt\/\u03b2-catenin signaling pathway in human renal proximal tubular cells","volume":"7","author":"Guo","year":"2014","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1002\/stem.589","article-title":"Wnt signaling regulates symmetry of division of neural stem cells in the adult brain and in response to injury","volume":"29","author":"Piccin","year":"2011","journal-title":"Stem Cells"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/S0074-7742(09)85026-5","article-title":"Role of Akt and ERK signaling in the neurogenesis following brain ischemia","volume":"85","author":"Shioda","year":"2009","journal-title":"Int. Rev. Neurobiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"299","DOI":"10.4077\/CJP.2010.AMK063","article-title":"Role of ERK signaling in the neuroprotective efficacy of magnesium sulfate treatment during focal cerebral ischemia in the gerbil cortex","volume":"53","author":"Huang","year":"2010","journal-title":"Chin. J. Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Tzeng, H.H., Hsu, C.H., Chung, T.H., Lee, W.C., Lin, C.H., Wang, W.C., Hsiao, C.Y., Leu, Y.W., and Wang, T.H. (2015). Cell Signaling and Differential Protein Expression in Neuronal Differentiation of Bone Marrow Mesenchymal Stem Cells with Hypermethylated Salvador\/Warts\/Hippo (SWH) Pathway Genes. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0145542"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1002\/(SICI)1097-4695(199901)38:1<65::AID-NEU5>3.0.CO;2-Q","article-title":"Retinoic acid and neurotrophins collaborate to regulate neurogenesis in adult-derived neural stem cell cultures","volume":"38","author":"Takahashi","year":"1999","journal-title":"J. Neurobiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1006\/mcne.1996.0595","article-title":"The adult rat hippocampus contains primordial neural stem cells","volume":"8","author":"Palmer","year":"1997","journal-title":"Mol. Cell. Neurosci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/S0896-6273(02)00828-0","article-title":"Function and regulation of CREB family transcription factors in the nervous system","volume":"35","author":"Lonze","year":"2002","journal-title":"Neuron"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/j.neuint.2013.02.002","article-title":"Electromagnetic fields induce neural differentiation of human bone marrow derived mesenchymal stem cells via ROS mediated EGFR activation","volume":"62","author":"Park","year":"2013","journal-title":"Neurochem. Int."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"U\u0142amek-Kozio\u0142, M., Czuczwar, S.J., Januszewski, S., and Pluta, R. (2020). Proteomic and Genomic Changes in Tau Protein, Which Are Associated with Alzheimer\u2019s Disease after Ischemia-Reperfusion Brain Injury. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21030892"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"12827","DOI":"10.18632\/aging.102547","article-title":"Tau as a potential therapeutic target for ischemic stroke","volume":"11","author":"Chen","year":"2019","journal-title":"Aging"},{"key":"ref_39","first-page":"665","article-title":"Nestin structure and predicted function in cellular cytoskeletal organisation","volume":"20","author":"Michalczyk","year":"2005","journal-title":"Histol. Histopathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1090","DOI":"10.1038\/nn.2385","article-title":"Neurod1 is essential for the survival and maturation of adult-born neurons","volume":"12","author":"Gao","year":"2009","journal-title":"Nat. Neurosci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1038\/nn.2360","article-title":"Wnt-mediated activation of NeuroD1 and retro-elements during adult neurogenesis","volume":"12","author":"Kuwabara","year":"2009","journal-title":"Nat. Neurosci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1073\/pnas.0909015107","article-title":"NeuroD1 induces terminal neuronal differentiation in olfactory neurogenesis","volume":"107","author":"Boutin","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"104","DOI":"10.5115\/acb.2015.48.2.104","article-title":"The effects of repetitive transcranial magnetic stimulation on proliferation and differentiation of neural stem cells","volume":"48","author":"Abbasnia","year":"2015","journal-title":"Anat. Cell Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1002\/glia.20126","article-title":"Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells","volume":"49","author":"Li","year":"2005","journal-title":"Glia"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.neuroscience.2005.08.092","article-title":"Intracarotid transplantation of bone marrow stromal cells increases axon-myelin remodeling after stroke","volume":"137","author":"Shen","year":"2006","journal-title":"Neuroscience"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.brainres.2007.02.047","article-title":"Axonal sprouting into the denervated spinal cord and synaptic and postsynaptic protein expression in the spinal cord after transplantation of bone marrow stromal cell in stroke rats","volume":"1149","author":"Liu","year":"2007","journal-title":"Brain Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1684\/ecn.2013.0332","article-title":"Extremely low frequency electromagnetic field and wound healing: Implication of cytokines as biological mediators","volume":"24","author":"Pesce","year":"2013","journal-title":"Eur. Cytokine Netw."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.pmbts.2017.02.005","article-title":"Biochemical and Biological Attributes of Matrix Metalloproteinases","volume":"147","author":"Cui","year":"2017","journal-title":"Prog. Mol. Biol. Transl. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Cabral-Pacheco, G.A., Garza-Veloz, I., Castruita-De la Rosa, C., Ramirez-Acu\u00f1a, J.M., Perez-Romero, B.A., Guerrero-Rodriguez, J.F., Martinez-Avila, N., and Martinez-Fierro, M.L. (2020). The Roles of Matrix Metalloproteinases and Their Inhibitors in Human Diseases. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21249739"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1007\/s12010-016-2289-z","article-title":"Neuroprotective Effect of Low Frequency-Pulsed Electromagnetic Fields in Ischemic Stroke","volume":"181","author":"Urnukhsaikhan","year":"2017","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7043","DOI":"10.4049\/jimmunol.172.11.7043","article-title":"Effects of TNF-alpha and IFN-gamma on nitric oxide-induced neurotoxicity in the mouse brain","volume":"172","author":"Blais","year":"2004","journal-title":"J. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Batsaikhan, B., Wang, J.Y., Scerba, M.T., Tweedie, D., Greig, N.H., Miller, J.P., Hoffer, B.J., Lin, C.T., and Wang, J.Y. (2019). Post-Injury Neuroprotective Effects of the Thalidomide Analog 3,6\u2032-Dithiothalidomide on Traumatic Brain Injury. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20030502"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1007\/s11481-014-9560-2","article-title":"Pro-inflammatory interferon gamma signaling is directly associated with stroke induced neurodegeneration","volume":"9","author":"Seifert","year":"2014","journal-title":"J. Neuroimmune Pharmacol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1002\/ana.410170513","article-title":"Induction of reproducible brain infarction by photochemically initiated thrombosis","volume":"17","author":"Watson","year":"1985","journal-title":"Ann. Neurol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1002\/jnr.22715","article-title":"Comparison of matrix metalloproteinase activation after focal cortical ischemia in young adult and aged mice","volume":"90","author":"Aleksy","year":"2012","journal-title":"J. Neurosci. Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1397","DOI":"10.1002\/biot.201600057","article-title":"Effect of human mesenchymal stem cell transplantation on cerebral ischemic volume-controlled photothrombotic mouse model","volume":"11","author":"Choi","year":"2016","journal-title":"Biotechnol. J."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1177\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:05:27Z","timestamp":1760133927000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1177"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,21]]},"references-count":56,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23031177"],"URL":"https:\/\/doi.org\/10.3390\/ijms23031177","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,21]]}}}