{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T23:07:05Z","timestamp":1790377625800,"version":"4.1.0"},"reference-count":54,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,30]],"date-time":"2020-08-30T00:00:00Z","timestamp":1598745600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["JO 1483\/1-1"],"award-info":[{"award-number":["JO 1483\/1-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["CRC 1270 ELAINE"],"award-info":[{"award-number":["CRC 1270 ELAINE"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011970","name":"International Team for Implantology","doi-asserted-by":"publisher","award":["1107_2015"],"award-info":[{"award-number":["1107_2015"]}],"id":[{"id":"10.13039\/501100011970","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>While several studies investigated the effects of mechanical or electrical stimulation on osseointegration and bone fracture healing, little is known about the molecular and cellular impact of combined biophysical stimulation on peri-implant osseointegration. Therefore, we established an in vitro system, capable of applying shear stress and electric fields simultaneously. Capacitively coupled electric fields were used for electrical stimulation, while roughened Ti6Al4V bodies conducted harmonically oscillating micromotions on collagen scaffolds seeded with human osteoblasts. Different variations of single and combined stimulation were applied for three days, while samples loaded with Ti6Al4V bodies and untreated samples served as control. Metabolic activity, expression of osteogenic markers and bone remodeling markers were investigated. While combined stimulation showed no substantial benefit compared to sole mechanical stimulation, we observed that 25 \u00b5m micromotions applied by roughened Ti6Al4V bodies led to a significant increase in gene expression of osteocalcin and tissue inhibitor of metalloprotease 1. Additionally, we found an increase in metabolic activity and expression of bone remodeling markers with reduced procollagen type 1 synthesis after 100 mVRMS electrical stimulation. We were able to trigger specific cellular behaviors using different biophysical stimuli. In future studies, different variations of electrical stimulation will be combined with interfacial micromotions.<\/jats:p>","DOI":"10.3390\/cells9091995","type":"journal-article","created":{"date-parts":[[2020,8,30]],"date-time":"2020-08-30T06:06:22Z","timestamp":1598767582000},"page":"1995","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Establishment and Evaluation of an In Vitro System for Biophysical Stimulation of Human Osteoblasts"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1049-7335","authenticated-orcid":false,"given":"Martin","family":"Stephan","sequence":"first","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics, Rostock University Medical Centre, 18057 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1486-0426","authenticated-orcid":false,"given":"Julius","family":"Zimmermann","sequence":"additional","affiliation":[{"name":"Institute of General Electrical Engineering, University of Rostock, 18059 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annett","family":"Klinder","sequence":"additional","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics, Rostock University Medical Centre, 18057 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franziska","family":"Sahm","sequence":"additional","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics, Rostock University Medical Centre, 18057 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1042-2058","authenticated-orcid":false,"given":"Ursula","family":"van Rienen","sequence":"additional","affiliation":[{"name":"Institute of General Electrical Engineering, University of Rostock, 18059 Rostock, Germany"},{"name":"Department Life, Light &amp; Matter, University of Rostock, 18051 Rostock, Germany"},{"name":"Department of Ageing of Individuals and Society, University of Rostock, 18051 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1671-3764","authenticated-orcid":false,"given":"Peer W.","family":"K\u00e4mmerer","sequence":"additional","affiliation":[{"name":"Department of Oral and Maxillofacial Surgery, University Medical Centre Mainz, 55131 Mainz, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rainer","family":"Bader","sequence":"additional","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics, Rostock University Medical Centre, 18057 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anika","family":"Jonitz-Heincke","sequence":"additional","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics, Rostock University Medical Centre, 18057 Rostock, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,30]]},"reference":[{"key":"ref_1","unstructured":"Department of Economic and Social Affairs, and Population Division (2019). World Population Prospects, United Nations."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1080\/17453674.2018.1446463","article-title":"Projections of primary hip arthroplasty in Germany until 2040","volume":"89","author":"Pilz","year":"2018","journal-title":"Acta Orthop."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1424","DOI":"10.1177\/0022034518792567","article-title":"Trends in Dental Implant Use in the U.S., 1999\u20132016, and Projections to 2026","volume":"97","author":"Elani","year":"2018","journal-title":"J. Dent. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/S0022-3913(83)80101-2","article-title":"Osseointegration and its experimental background","volume":"50","author":"Branemark","year":"1983","journal-title":"J. Prosthet. Dent."},{"key":"ref_5","first-page":"61","article-title":"Biology of implant osseointegration","volume":"9","author":"Mavrogenis","year":"2009","journal-title":"J. Musculoskelet. Neuronal Interact."},{"key":"ref_6","unstructured":"Betts, J.G., DeSaix, P., Johnson, E., Johnson, J.E., Korol, O., Kruse, D.H., Poe, B., Wise, J.A., Womble, M., and Young, K.A. (2015). Anatomy and Physiology, Rice University."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1016\/j.micron.2005.07.008","article-title":"Peri-implant osteogenesis in health and osteoporosis","volume":"36","author":"Marco","year":"2005","journal-title":"Micron"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1161\/CIRCRESAHA.111.246116","article-title":"Effects of aging on angiogenesis","volume":"110","author":"Rosenzweig","year":"2012","journal-title":"Circ. Res."},{"key":"ref_9","unstructured":"Herold, G. (2018). Innere Medizin 2018. Eine Vorlesungsorientierte Darstellung: Unter Ber\u00fccksichtigung des Gegenstandskataloges f\u00fcr die \u00c4rztliche Pr\u00fcfung: Mit ICD 10-Schl\u00fcssel im Text und Stichwortverzeichnis, Gerd Herold."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2104","DOI":"10.1177\/0300060517732697","article-title":"Current methods of preventing aseptic loosening and improving osseointegration of titanium implants in cementless total hip arthroplasty: A review","volume":"46","author":"Apostu","year":"2018","journal-title":"J. Int. Med. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1002\/jbm.820070404","article-title":"The effect of movement on the bonding of porous metal to bone","volume":"7","author":"Cameron","year":"1973","journal-title":"J. Biomed. Mater. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1002\/jor.1100100216","article-title":"Tissue ingrowth into titanium and hydroxyapatite-coated implants during stable and unstable mechanical conditions","volume":"10","author":"Hansen","year":"1992","journal-title":"J. Orthop. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1097\/01.ID.0000034394.75768.E3","article-title":"Osseointegration under immediate loading: Biomechanical stress-strain and bone formation\u2014Resorption","volume":"12","author":"Kawahara","year":"2003","journal-title":"Implant Dent."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"707","DOI":"10.2106\/00004623-199705000-00010","article-title":"In vivo skeletal responses to porous-surfaced implants subjected to small induced motions","volume":"79","author":"Jasty","year":"1997","journal-title":"J. Bone Joint Surg. Am."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1016\/S0883-5403(96)80136-7","article-title":"Differences in stiffness of the interface between a cementless porous implant and cancellous bone in vivo in dogs due to varying amounts of implant motion","volume":"11","author":"Bragdon","year":"1996","journal-title":"J. Arthroplast."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jonitz-Heincke, A., Sellin, M.-L., Seyfarth, A., Peters, K., Mueller-Hilke, B., Fiedler, T., Bader, R., and Klinder, A. (2019). Analysis of Cellular Activity and Induction of Inflammation in Response to Short-Term Exposure to Cobalt and Chromium Ions in Mature Human Osteoblasts. Materials (Basel), 12.","DOI":"10.3390\/ma12172771"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Jonitz-Heincke, A., Tillmann, J., Klinder, A., Krueger, S., Kretzer, J.P., H\u00f8l, P.J., Paulus, A.C., and Bader, R. (2017). The Impact of Metal Ion Exposure on the Cellular Behavior of Human Osteoblasts and PBMCs: In Vitro Analyses of Osteolytic Processes. Materials (Basel), 10.","DOI":"10.3390\/ma10070734"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Markhoff, J., Krogull, M., Schulze, C., Rotsch, C., Hunger, S., and Bader, R. (2017). Biocompatibility and Inflammatory Potential of Titanium Alloys Cultivated with Human Osteoblasts, Fibroblasts and Macrophages. Materials (Basel), 10.","DOI":"10.3390\/ma10010052"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5490","DOI":"10.3390\/ma8085259","article-title":"Influence of Different Three-Dimensional Open Porous Titanium Scaffold Designs on Human Osteoblasts Behavior in Static and Dynamic Cell Investigations","volume":"8","author":"Markhoff","year":"2015","journal-title":"Materials (Basel)"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1302\/2046-3758.72.BJR-2017-0228.R1","article-title":"Effects of interfacial micromotions on vitality and differentiation of human osteoblasts","volume":"7","author":"Ziebart","year":"2018","journal-title":"Bone Joint Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0021-9290(02)00301-9","article-title":"Effect of micromechanical stimulations on osteoblasts: Development of a device simulating the mechanical situation at the bone\u2013implant interface","volume":"36","author":"Pioletti","year":"2003","journal-title":"J. Biomech."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/j.bone.2007.09.055","article-title":"Microstimulation at the bone-implant interface upregulates osteoclast activation pathways","volume":"42","author":"Stadelmann","year":"2008","journal-title":"Bone"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"5178640","DOI":"10.1155\/2016\/5178640","article-title":"A Novel In Vitro System for Comparative Analyses of Bone Cells and Bacteria under Electrical Stimulation","volume":"2016","author":"Dauben","year":"2016","journal-title":"Biomed Res. Int."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Wang, Q., Zhong, S., Ouyang, J., Jiang, L., Zhang, Z., Xie, Y., and Luo, S. (1998). Osteogenesis of electrically stimulated bone cells mediated in part by calcium ions. Clin. Orthop. Relat. Res., 259\u2013268.","DOI":"10.1097\/00003086-199803000-00037"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1514","DOI":"10.2106\/00004623-200110000-00009","article-title":"Signal Transduction in Electrically Stimulated Bone Cells","volume":"83","author":"Brighton","year":"2001","journal-title":"J. Bone Jt. Surg. Am. Vol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1007\/s12013-014-9984-6","article-title":"Establishment of a novel in vitro test setup for electric and magnetic stimulation of human osteoblasts","volume":"70","author":"Grunert","year":"2014","journal-title":"Cell Biochem. Biophys."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"57","DOI":"10.3892\/ijmm.2016.2590","article-title":"Magnetically induced electrostimulation of human osteoblasts results in enhanced cell viability and osteogenic differentiation","volume":"38","author":"Hiemer","year":"2016","journal-title":"Int. J. Mol. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1002\/jor.22595","article-title":"Up-regulation of expression of selected genes in human bone cells with specific capacitively coupled electric fields","volume":"32","author":"Clark","year":"2014","journal-title":"J. Orthop. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1007\/s002490000100","article-title":"Capacitively coupled electric fields accelerate proliferation of osteoblast-like primary cells and increase bone extracellular matrix formation in vitro","volume":"29","author":"Hartig","year":"2000","journal-title":"Eur. Biophys. J."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Brighton, C.T., Okereke, E., Pollack, S.R., and Clark, C.C. (1992). In vitro bone-cell response to a capacitively coupled electrical field. The role of field strength, pulse pattern, and duty cycle. Clin. Orthop. Relat. Res., 255\u2013262.","DOI":"10.1097\/00003086-199212000-00035"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/S0167-4889(00)00135-X","article-title":"Electrical stimulation influences mineral formation of osteoblast-like cells in vitro","volume":"1538","author":"Wiesmann","year":"2001","journal-title":"Biochim. Biophys. Acta (BBA) Mol. Cell Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1006\/bbrc.1997.7118","article-title":"Electrical stimulation induces the level of TGF-beta1 mRNA in osteoblastic cells by a mechanism involving calcium\/calmodulin pathway","volume":"237","author":"Zhuang","year":"1997","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Krueger, S., Achilles, S., Zimmermann, J., Tischer, T., Bader, R., and Jonitz-Heincke, A. (2019). Re-Differentiation Capacity of Human Chondrocytes in Vitro Following Electrical Stimulation with Capacitively Coupled Fields. J. Clin. Med., 8.","DOI":"10.3390\/jcm8111771"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1016\/j.msec.2016.01.032","article-title":"Commercially pure titanium (cp-Ti) versus titanium alloy (Ti6Al4V) materials as bone anchored implants\u2014Is one truly better than the other?","volume":"62","author":"Shah","year":"2016","journal-title":"Mater. Sci. Eng. C Mater. Biol. Appl."},{"key":"ref_35","first-page":"1055","article-title":"The Potential Role of Human Osteoblasts for Periprosthetic Osteolysis Following Exposure to Wear Particles","volume":"28","author":"Lochner","year":"2011","journal-title":"Int. J. Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.dental.2012.10.003","article-title":"In vitro evaluation of elastin-like polypeptide-collagen composite scaffold for bone tissue engineering","volume":"29","author":"Amruthwar","year":"2013","journal-title":"Dent. Mater."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s00393-016-0083-5","article-title":"Biochemische Marker des Knochenstoffwechsels und ihre Bedeutung","volume":"75","author":"Schwetz","year":"2016","journal-title":"Z. Rheumatol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Heinrich, P.C., M\u00fcller, M., Graeve, L., and L\u00f6ffler, G. (2014). L\u00f6ffler\/Petrides Biochemie und Pathobiochemie, Springer.","DOI":"10.1007\/978-3-642-17972-3"},{"key":"ref_39","unstructured":"Neumeister, B., B\u00f6hm, B.O., and Beneke, H. (2015). Klinikleitfaden Labordiagnostik, Elsevier Urban & Fischer. [5th ed.]."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1166\/jbn.2019.2749","article-title":"Promoting Proliferation and Differentiation of Pre-Osteoblasts MC3T3-E1 Cells Under Combined Mechanical and Electrical Stimulation","volume":"15","author":"Li","year":"2019","journal-title":"J. Biomed. Nanotechnol."},{"key":"ref_41","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_42","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1002\/jor.1100030310","article-title":"Fracture healing in the rabbit fibula when subjected to various capacitively coupled electrical fields","volume":"3","author":"Brighton","year":"1985","journal-title":"J. Orthop. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1","DOI":"10.22203\/eCM.v024a01","article-title":"In search of an osteoblast cell model for in vitro research","volume":"24","author":"Czekanska","year":"2012","journal-title":"Eur. Cell. Mater."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"9622","DOI":"10.1016\/j.biomaterials.2011.09.009","article-title":"Biomimetic nanofibrous scaffolds for bone tissue engineering","volume":"32","author":"Holzwarth","year":"2011","journal-title":"Biomaterials"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/0304-4165(86)90054-1","article-title":"Embryonic bone matrix formation is increased after exposure to a low-amplitude capacitively coupled electric field, in vitro","volume":"882","author":"Fitzsimmons","year":"1986","journal-title":"Biochim. Biophys. Acta (BBA) Gen. Subj."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"S573","DOI":"10.1002\/jbmr.5650081327","article-title":"Optimization of electric field parameters for the control of bone remodeling: Exploitation of an indigenous mechanism for the prevention of osteopenia","volume":"8","author":"Rubin","year":"1993","journal-title":"J. Bone Miner. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"230805","DOI":"10.1155\/2013\/230805","article-title":"Extracellular matrix degradation and tissue remodeling in periprosthetic loosening and osteolysis: Focus on matrix metalloproteinases, their endogenous tissue inhibitors and the proteasome","volume":"2013","author":"Syggelos","year":"2013","journal-title":"Biomed. Res. Int."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"6663","DOI":"10.1016\/j.actbio.2013.01.014","article-title":"Micromotion-induced strain fields influence early stages of repair at bone-implant interfaces","volume":"9","author":"Wazen","year":"2013","journal-title":"Acta Biomater."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0169-6009(08)80016-5","article-title":"Cyclic stretch increases alkaline phosphatase activity of osteoblast-like cells: A role for prostaglandin E2","volume":"21","author":"Nishioka","year":"1993","journal-title":"Bone Miner."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1767","DOI":"10.1089\/ten.tea.2013.0157","article-title":"Combined effect of three types of biophysical stimuli for bone regeneration","volume":"20","author":"Kang","year":"2014","journal-title":"Tissue Eng. Part A"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s41232-018-0059-8","article-title":"Piezoelectric smart biomaterials for bone and cartilage tissue engineering","volume":"38","author":"Jacob","year":"2018","journal-title":"Inflamm. Regen."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"502B","DOI":"10.1143\/JJAP.3.502B","article-title":"Piezoelectric Effects in Collagen","volume":"3","author":"Fukada","year":"1964","journal-title":"Jpn. J. Appl. Phys."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1158","DOI":"10.1143\/JPSJ.12.1158","article-title":"On the Piezoelectric Effect of Bone","volume":"12","author":"Fukada","year":"1957","journal-title":"J. Phys. Soc. Jpn."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.abb.2008.03.018","article-title":"Functions of RANKL\/RANK\/OPG in bone modeling and remodeling","volume":"473","author":"Boyce","year":"2008","journal-title":"Arch. Biochem. Biophys."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/1995\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:04:46Z","timestamp":1760177086000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/1995"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,30]]},"references-count":54,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["cells9091995"],"URL":"https:\/\/doi.org\/10.3390\/cells9091995","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,30]]}}}