{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:06:18Z","timestamp":1790121978938,"version":"4.0.1"},"reference-count":56,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2022,10,18]],"date-time":"2022-10-18T00:00:00Z","timestamp":1666051200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea (NRF) grant funded by the Korean government (MSIT)","doi-asserted-by":"publisher","award":["NRF-2018R1D1A1B07048493"],"award-info":[{"award-number":["NRF-2018R1D1A1B07048493"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Electrical signals represent an essential form of cellular communication. For decades, electrical stimulation has been used effectively in clinical practice to enhance bone healing. However, the detailed mechanisms between electrical stimulation and bone healing are not well understood. In addition, there have been many difficulties in setting up a stable and efficient electrical stimulation system within the in vitro environment. Therefore, various conductive materials and electrical stimulation methods have been tested to establish an effective electrical stimulation system. Through these systems, many studies have been conducted on the effects of electrical stimulation on bone healing and osteogenic differentiation. However, previous studies were limited by the use of opaque conductive materials that obscure the cells; fluorescent observations and staining are known to be two of the critical methods to confirm the states of the cells. Indium tin oxide (ITO) glass is known to have excellent transparency and conductivity, but it is challenging to cultivate cells due to low cell adhesion characteristics. Therefore, we used O2 plasma treatment to increase the hydrophilicity and wettability of ITO glass. This enhanced cell affinity to the glass, providing a stable surface for the cells to attach. Then, electrical stimulation was applied with an amplitude range of 10 to 200 \u00b5A at a frequency of 10 Hz. Our results demonstrated that the osteogenic differentiation efficiency was maximized under the amplitude conditions of 10 \u00b5A and 50 \u00b5A. Accordingly, the results of our study suggest the development of an excellent platform in the field of biological research as a good tool to elucidate various mechanisms of cell bioactivity under electrical conditions.<\/jats:p>","DOI":"10.3390\/ijms232012490","type":"journal-article","created":{"date-parts":[[2022,10,19]],"date-time":"2022-10-19T00:58:51Z","timestamp":1666141131000},"page":"12490","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Electrical Stimulation of Human Adipose-Derived Mesenchymal Stem Cells on O2 Plasma-Treated ITO Glass Promotes Osteogenic Differentiation"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7000-6577","authenticated-orcid":false,"given":"Seungho","family":"Baek","sequence":"first","affiliation":[{"name":"PCL Inc., 128, Beobwon-ro, Songpa-gu, Seoul 08510, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7051-7644","authenticated-orcid":false,"given":"Heekyung","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, School of Integrative Engineering, Chung-Ang University, 221 Heukseok-Dong, Dongjak-gu, Seoul 06974, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatma Dilara","family":"Igci","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, School of Integrative Engineering, Chung-Ang University, 221 Heukseok-Dong, Dongjak-gu, Seoul 06974, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4554-5488","authenticated-orcid":false,"given":"Donghyun","family":"Lee","sequence":"additional","affiliation":[{"name":"PCL Inc., 128, Beobwon-ro, Songpa-gu, Seoul 08510, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1007\/s00198-009-0860-y","article-title":"Bone mineral: Update on chemical composition and structure","volume":"20","author":"Rey","year":"2009","journal-title":"Osteoporos. Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S7","DOI":"10.1097\/00003086-199810001-00003","article-title":"The cell and molecular biology of fracture healing","volume":"355","author":"Einhorn","year":"1998","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Dimitriou, R., Jones, E., McGonagle, D., and Giannoudis, P.V. (2011). Bone regeneration: Current concepts and future directions. BMC Med., 9.","DOI":"10.1186\/1741-7015-9-66"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1359\/jbmr.2002.17.3.513","article-title":"Differential temporal expression of members of the transforming growth factor \u03b2 superfamily during murine fracture healing","volume":"17","author":"Cho","year":"2002","journal-title":"J. Bone Miner. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"S193","DOI":"10.1038\/550S193a","article-title":"Non-union bone fracture: A quicker fix","volume":"550","author":"Holmes","year":"2017","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/S0925-4773(99)00142-2","article-title":"Does adult fracture repair recapitulate embryonic skeletal formation?","volume":"87","author":"Ferguson","year":"1999","journal-title":"Mech. Dev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1634\/theoncologist.9-4-422","article-title":"Biology and Therapeutic Advances for Pediatric Osteosarcoma","volume":"9","author":"Marina","year":"2004","journal-title":"Oncologist"},{"key":"ref_8","first-page":"S1","article-title":"Management of long-bone non-unions","volume":"38","author":"Giannoudis","year":"2007","journal-title":"Injury"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"S43","DOI":"10.1016\/S0020-1383(13)70010-X","article-title":"Treatment of non-unions with bone defects: Which option and why?","volume":"44","author":"Ashman","year":"2013","journal-title":"Injury"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1007\/s12257-019-0086-6","article-title":"Strategies towards Orthopaedic Tissue Engineered Graft Generation: Current Scenario and Application","volume":"24","author":"Mallick","year":"2019","journal-title":"Biotechnol. Bioprocess Eng."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1002\/jcp.25641","article-title":"Osteogenic Differentiation of Periosteal Cells During Fracture Healing","volume":"232","author":"Wang","year":"2016","journal-title":"J. Cell. Physiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"273","DOI":"10.5435\/JAAOS-20-05-273","article-title":"Assessment of Compromised Fracture Healing","volume":"20","author":"Bishop","year":"2012","journal-title":"J. Am. Acad. Orthop. Surg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1016\/j.injury.2011.04.016","article-title":"Enhancement of fracture healing with the diamond concept: The role of the biological chamber","volume":"42","author":"Calori","year":"2011","journal-title":"Injury"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S11","DOI":"10.1016\/S0020-1383(07)80004-0","article-title":"Risk factors contributing to fracture non-unions","volume":"38","author":"Calori","year":"2007","journal-title":"Injury"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"563","DOI":"10.5435\/00124635-201109000-00007","article-title":"Management of Aseptic Tibial Nonunion","volume":"19","author":"Hak","year":"2011","journal-title":"J. Am. Acad. Orthop. Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"17926","DOI":"10.1038\/s41598-017-17120-8","article-title":"Mediation of cellular osteogenic differentiation through daily stimulation time based on polypyrrole planar electrodes","volume":"7","author":"Liu","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"15020","DOI":"10.1038\/boneres.2015.20","article-title":"Dopaminergic effects on in vitro osteogenesis","volume":"3","author":"Lee","year":"2015","journal-title":"Bone Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1007\/BF02439451","article-title":"Dexamethasone enhances differentiation of human osteoblastic cells in vitro","volume":"15","author":"Yamanouchi","year":"1997","journal-title":"J. Bone Miner. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1263\/jbb.105.586","article-title":"Effect of ascorbic acid on bone marrow-derived mesenchymal stem cell proliferation and differentiation","volume":"105","author":"Choi","year":"2008","journal-title":"J. Biosci. Bioeng."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1186\/scrt165","article-title":"Effects of different serum conditions on osteogenic differentiation of human adipose stem cells in vitro","volume":"4","author":"Haimi","year":"2013","journal-title":"Stem Cell Res. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11814-017-0236-5","article-title":"Nanomaterials for bioelectronics and integrated medical systems","volume":"35","author":"Lee","year":"2017","journal-title":"Korean J. Chem. Eng."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1166\/sam.2016.2627","article-title":"Potentiostatically Stimulated Osteogenic Differentiation in a Mammalian Pre-Osteoblast Cell Line (MC3T3-E1)","volume":"8","author":"Kim","year":"2016","journal-title":"Sci. Adv. Mater."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"16503","DOI":"10.1039\/C8RA02072H","article-title":"Double-layered microsphere based dual growth factor delivery system for guided bone regeneration","volume":"8","author":"Xu","year":"2018","journal-title":"RSC Adv."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"23","DOI":"10.4161\/org.6048","article-title":"The use of mesenchymal stem cells in tissue engineering: A global assessment","volume":"4","author":"Rosenbaum","year":"2008","journal-title":"Organogenesis"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1007\/s10616-012-9428-3","article-title":"Mesenchymal stem cells from umbilical cord blood: Parameters for isolation, characterization and adipogenic differentiation","volume":"64","author":"Sibov","year":"2012","journal-title":"Cytotechnology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1007\/s12257-019-0332-y","article-title":"Exosome-mediated Bidirectional Signaling between Mesenchymal Stem Cells and Chondrocytes for Enhanced Chondrogenesis","volume":"24","author":"Kim","year":"2019","journal-title":"Biotechnol. Bioprocess Eng."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"113","DOI":"10.4252\/wjsc.v3.i12.113","article-title":"Hepatogenic differentiation of mesenchymal stem cells induced by insulin like growth factor-I","volume":"3","author":"Ayatollahi","year":"2011","journal-title":"World J. Stem Cells"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"82","DOI":"10.2174\/157488811795495486","article-title":"Mesenchymal Stem Cells Neuronal Differentiation Ability: A Real Perspective for Nervous System Repair?","volume":"6","author":"Scuteri","year":"2011","journal-title":"Curr. Stem Cell Res. Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1186\/s13287-018-0942-x","article-title":"In vitro osteoblastic differentiation of mesenchymal stem cells generates cell layers with distinct properties","volume":"9","author":"Hanna","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_30","first-page":"222","article-title":"Mesenchymal stem cell applications to tendon healing","volume":"2","author":"Chaudhury","year":"2012","journal-title":"Muscle Ligaments Tendons J."},{"key":"ref_31","first-page":"208","article-title":"Isolation, differentiation, and characterization of mesenchymal stem cells from human bone marrow","volume":"10","author":"Baghaei","year":"2017","journal-title":"Gastroenterol. Hepatol. Bed Bench"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"195","DOI":"10.4252\/wjsc.v6.i2.195","article-title":"Umbilical cord-derived mesenchymal stem cells: Their advantages and potential clinical utility","volume":"6","year":"2014","journal-title":"World J. Stem Cells"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1186\/s13287-015-0066-5","article-title":"Comparative analysis of human mesenchymal stem cells from bone marrow and adipose tissue under xeno-free conditions for cell therapy","volume":"6","author":"Li","year":"2015","journal-title":"Stem Cell Res. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/978-1-62703-330-5_21","article-title":"Skin-Derived Mesenchymal Stem Cells: Isolation, Culture, and Characterization","volume":"989","author":"Orciani","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"134","DOI":"10.4252\/wjsc.v6.i2.134","article-title":"Brain mesenchymal stem cells: The other stem cells of the brain?","volume":"6","author":"Appaix","year":"2014","journal-title":"World J. Stem Cells"},{"key":"ref_36","first-page":"2156478","article-title":"Impact of Tissue Harvesting Sites on the Cellular Behaviors of Adipose-Derived Stem Cells: Implication for Bone Tissue Engineering","volume":"2017","author":"Rad","year":"2017","journal-title":"Stem Cells Int."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1007\/s12257-019-0255-7","article-title":"Recent Clinical Trials in Adipose-derived Stem Cell Mediated Osteoarthritis Treatment","volume":"24","author":"Cho","year":"2019","journal-title":"Biotechnol. Bioprocess Eng."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1111\/j.1749-6632.1974.tb26824.x","article-title":"Panel discussion: To what extent can electrical stimulation be used in the treatment of human disorders?","volume":"238","author":"Bassett","year":"1974","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"31724","DOI":"10.1038\/srep31724","article-title":"Efficacy of Electrical Stimulators for Bone Healing: A Meta-Analysis of Randomized Sham-Controlled Trials","volume":"6","author":"Aleem","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1097\/00003086-198309000-00020","article-title":"Constant Direct-current Treatment for Established Nonunion of the Tibia","volume":"178","author":"Heppenstall","year":"1983","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.2106\/00004623-198264080-00011","article-title":"Treatment of therapeutically resistant non-unions with bone grafts and pulsing electromagnetic fields","volume":"64","author":"Bassett","year":"1982","journal-title":"J. Bone Jt. Surg."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1097\/00003086-198912000-00021","article-title":"Osteonecrosis of the femoral head. Results of core decompression and grafting with and without electrical stimulation","volume":"249","author":"Steinberg","year":"1989","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_43","first-page":"e34","article-title":"Electrical Stimulation in Bone Healing: Critical Analysis by Evaluating Levels of Evidence","volume":"11","author":"Griffin","year":"2011","journal-title":"Eplasty"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3369","DOI":"10.1002\/jbm.a.36181","article-title":"Time-dependent effect of electrical stimulation on osteogenic differentiation of bone mesenchymal stromal cells cultured on conductive nanofibers","volume":"105","author":"Zhu","year":"2017","journal-title":"J. Biomed. Mater. Res. Part A"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e2821","DOI":"10.7717\/peerj.2821","article-title":"In vitro effect of direct current electrical stimulation on rat mesenchymal stem cells","volume":"5","author":"Mobini","year":"2017","journal-title":"PeerJ"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Wang, Y., Cui, H., Wu, Z., Wu, N., Wang, Z., Chen, X., Wei, Y., and Zhang, P. (2016). Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0154924"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1097\/00003086-199212000-00035","article-title":"In vitro bone-cell response to a capacitively coupled electrical field. The role of field strength, pulse pattern, and duty cycle","volume":"285","author":"Brighton","year":"1992","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1615\/JLongTermEffMedImplants.2016016818","article-title":"Use of Combined Magnetic Field Treatment for Fracture Nonunion","volume":"26","author":"Phillips","year":"2016","journal-title":"J. Long-Term Eff. Med. Implants"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1097\/00003086-200402000-00005","article-title":"Treatment of Nonunions With Electric and Electromagnetic Fields","volume":"419","author":"Aaron","year":"2004","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1097\/00003086-200103000-00031","article-title":"Pulsed Electromagnetic Fields Increase Growth Factor Release by Nonunion Cells","volume":"384","author":"Guerkov","year":"2001","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_51","first-page":"395","article-title":"Surface Characterization of O2-Plasma-Treated Indium-Tin-Oxide (ITO) Anodes for Organic Light-Emitting-Device Applications","volume":"41","author":"Kim","year":"2002","journal-title":"J. Korean Phys. Soc."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1007\/BF02618296","article-title":"Surface treatments and cell attachment","volume":"20","author":"Ramsey","year":"1984","journal-title":"In Vitro"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2452","DOI":"10.1007\/s11814-018-0131-8","article-title":"Surface treatment of sol-gel bioglass using dielectric barrier discharge plasma to enhance growth of hydroxyapatite","volume":"35","author":"Soliman","year":"2018","journal-title":"Korean J. Chem. Eng."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1186\/ar2116","article-title":"Mesenchymal stromal cells. Biology of adult mesenchymal stem cells: Regulation of niche, self-renewal and differentiation","volume":"9","author":"Kolf","year":"2007","journal-title":"Arthritis Res. Ther."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1080\/14653240600855905","article-title":"Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement","volume":"8","author":"Dominici","year":"2006","journal-title":"Cytotherapy"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"65101","DOI":"10.1088\/0957-4484\/24\/6\/065101","article-title":"A multifunctional biphasic suspension of mesoporous silica encapsulated with YVO4:Eu3+ and Fe3O4 nanoparticles: Synergistic effect towards cancer therapy and imaging","volume":"24","author":"Singh","year":"2013","journal-title":"Nanotechnology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/20\/12490\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:56:44Z","timestamp":1760144204000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/20\/12490"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,18]]},"references-count":56,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["ijms232012490"],"URL":"https:\/\/doi.org\/10.3390\/ijms232012490","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,18]]}}}