{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T17:54:09Z","timestamp":1760550849377,"version":"build-2065373602"},"reference-count":51,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2019,5,26]],"date-time":"2019-05-26T00:00:00Z","timestamp":1558828800000},"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":["MA 5158\/1-1"],"award-info":[{"award-number":["MA 5158\/1-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Co-culture studies investigating the role of periprosthetic fibroblasts (PPFs) in inflammatory osteoclastogenesis reveal contrary results, partly showing an osteoprotective function of fibroblasts and high OPG expression in monolayer. These data disagree with molecular analyses of original periosteolytic tissues. In order to find a more reliable model, PPFs were co-cultivated with peripheral blood mononuclear cells (PBMCs) in a transwell system and compared to conventional monolayer cultures. The gene expression of key regulators of osteoclastogenesis (macrophage colony-stimulating factor (MCSF), receptor activator of NF-\u03baB ligand (RANK-L), osteoprotegerin (OPG), and tumor necrosis factor alpha (TNF\u03b1)) as well as the ability of bone resorption were analyzed. In monolayer co-cultures, PPFs executed an osteoprotective function with high OPG-expression, low RANK-L\/OPG ratios, and a resulting inhibition of osteolysis even in the presence of MCSF and RANK-L. For transwell co-cultures, profound changes in gene expression, with a more than hundredfold decrease of OPG and a significant upregulation of TNF\u03b1 were observed. In conclusion, we were able to show that a change of culture conditions towards a transwell system resulted in a considerably more osteoclastogenic gene expression profile, being closer to findings in original periosteolytic tissues. This study therefore presents an interesting approach for a more reliable in vitro model to examine the role of fibroblasts in periprosthetic osteoclastogenesis in the future.<\/jats:p>","DOI":"10.3390\/ijms20102583","type":"journal-article","created":{"date-parts":[[2019,5,26]],"date-time":"2019-05-26T23:07:27Z","timestamp":1558912047000},"page":"2583","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Impact of Periprosthetic Fibroblast-Like Cells on Osteoclastogenesis in Co-Culture with Peripheral Blood Mononuclear Cells Varies Depending on Culture System"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8885-1624","authenticated-orcid":false,"given":"Miriam I.","family":"Koehler","sequence":"first","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"},{"name":"Department of Cardiology and Vascular Medicine, West-German Heart and Vascular Center, University Duisburg-Essen, Hufelandstr. 55, 45147 Essen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eliza S.","family":"Hartmann","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabine","family":"Schluessel","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Felicitas","family":"Beck","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia I.","family":"Redeker","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baerbel","family":"Schmitt","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina","family":"Unger","sequence":"additional","affiliation":[{"name":"Experimental Trauma Surgery, Department of Trauma Surgery, Klinikum Rechts der Isar, Technical University of Munich, Ismaninger Strasse 22, 81675 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5104-9881","authenticated-orcid":false,"given":"Martijn","family":"van Griensven","sequence":"additional","affiliation":[{"name":"Experimental Trauma Surgery, Department of Trauma Surgery, Klinikum Rechts der Isar, Technical University of Munich, Ismaninger Strasse 22, 81675 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Burkhard","family":"Summer","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Ludwig-Maximilians-University, Frauenlobstr. 9-11, 80337 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"Fottner","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6391-4086","authenticated-orcid":false,"given":"Susanne","family":"Mayer-Wagner","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistra\u00dfe 15, 81377 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,5,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"555","DOI":"10.3109\/08820139.2013.822766","article-title":"The role of cytokines in inflammatory bone loss","volume":"42","author":"Souza","year":"2013","journal-title":"Immunol. Invest."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1038\/nrrheum.2012.153","article-title":"Bone erosion in rheumatoid arthritis: Mechanisms, diagnosis and treatment","volume":"8","author":"Schett","year":"2012","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"511","DOI":"10.3389\/fimmu.2014.00511","article-title":"Biology of the RANKL-RANK-OPG System in Immunity, Bone, and Beyond","volume":"5","author":"Walsh","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1038\/nrrheum.2011.128","article-title":"Mediators of the inflammatory response to joint replacement devices","volume":"7","author":"Cobelli","year":"2011","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1001\/jama.292.4.490","article-title":"Clinical implications of the osteoprotegerin\/RANKL\/RANK system for bone and vascular diseases","volume":"292","author":"Hofbauer","year":"2004","journal-title":"JAMA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1359\/jbmr.2000.15.8.1459","article-title":"Fibroblastic stromal cells express receptor activator of NF-kappa B ligand and support osteoclast differentiation","volume":"15","author":"Quinn","year":"2000","journal-title":"J. Bone Miner. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3221","DOI":"10.1002\/art.22134","article-title":"The role of fibroblasts and fibroblast-derived factors in periprosthetic osteolysis","volume":"54","author":"Koreny","year":"2006","journal-title":"Arthritis Rheum."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1016\/j.orthres.2004.10.006","article-title":"Arthroplasty membrane-derived fibroblasts directly induce osteoclast formation and osteolysis in aseptic loosening","volume":"23","author":"Sabokbar","year":"2005","journal-title":"J. Orthop. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/j.arth.2007.06.010","article-title":"Substance P stimulates production of interleukin 1beta and tumor necrosis factor alpha in fibroblasts from hip periprosthetic membrane","volume":"23","author":"Qian","year":"2008","journal-title":"J. Arthroplasty"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1016\/j.arth.2013.01.012","article-title":"Revision surgery after total joint arthroplasty: A complication-based analysis using worldwide arthroplasty registers","volume":"28","author":"Sadoghi","year":"2013","journal-title":"J. Arthroplasty"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Saleh, K.J., Celebrezze, M., Kassim, R., Dykes, D.C., Gioe, T.J., Callaghan, J.J., and Salvati, E.A. (2003). Functional outcome after revision hip arthroplasty: A metaanalysis. Clin. Orthop. Relat. Res., 254\u2013264.","DOI":"10.1097\/01.blo.0000093006.90435.43"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1054\/arth.2002.35823","article-title":"Functional outcome after total knee arthroplasty revision: A meta-analysis","volume":"17","author":"Saleh","year":"2002","journal-title":"J. Arthroplasty"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1177\/1753425912451779","article-title":"Particle disease: Biologic mechanisms of periprosthetic osteolysis in total hip arthroplasty","volume":"19","author":"Gallo","year":"2013","journal-title":"Innate Immun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"8046","DOI":"10.1016\/j.actbio.2013.05.005","article-title":"Osteolysis around total knee arthroplasty: A review of pathogenetic mechanisms","volume":"9","author":"Gallo","year":"2013","journal-title":"Acta Biomater"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1002\/jor.22658","article-title":"Characterization of macrophage polarizing cytokines in the aseptic loosening of total hip replacements","volume":"32","author":"Jamsen","year":"2014","journal-title":"J. Orthop. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1196","DOI":"10.1302\/0301-620X.85B8.13311","article-title":"Imbalance of RANKL\/RANK\/OPG system in interface tissue in loosening of total hip replacement","volume":"85","author":"Mandelin","year":"2003","journal-title":"J. Bone Joint Surg. Br."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S0142-9612(03)00556-8","article-title":"Factors regulating osteoclast formation in human tissues adjacent to peri-implant bone loss: Expression of receptor activator NFkappaB, RANK ligand and osteoprotegerin","volume":"25","author":"Crotti","year":"2004","journal-title":"Biomaterials"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1002\/jor.23274","article-title":"Factors regulating bone remodeling processes in aseptic implant loosening","volume":"35","author":"Hartmann","year":"2017","journal-title":"J. Orthop. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1302\/0301-620X.80B3.0800531","article-title":"Cellular profile and cytokine production at prosthetic interfaces. Study of tissues retrieved from revised hip and knee replacements","volume":"80","author":"Goodman","year":"1998","journal-title":"J. Bone Joint Surg. Br."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1016\/j.joca.2014.08.004","article-title":"Comparison of periprosthetic tissues in knee and hip joints: Differential expression of CCL3 and DC-STAMP in total knee and hip arthroplasty and similar cytokine profiles in primary knee and hip osteoarthritis","volume":"22","author":"Tomankova","year":"2014","journal-title":"Osteoarthritis Cartilage"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1002\/jor.20486","article-title":"Expression profiling reveals alternative macrophage activation and impaired osteogenesis in periprosthetic osteolysis","volume":"26","author":"Koulouvaris","year":"2008","journal-title":"J. Orthop. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1002\/1529-0131(200002)43:2<259::AID-ANR4>3.0.CO;2-W","article-title":"Involvement of receptor activator of nuclear factor kappaB ligand\/osteoclast differentiation factor in osteoclastogenesis from synoviocytes in rheumatoid arthritis","volume":"43","author":"Takayanagi","year":"2000","journal-title":"Arthritis Rheum."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1002\/jcb.20795","article-title":"Gingival fibroblasts are better at inhibiting osteoclast formation than periodontal ligament fibroblasts","volume":"98","author":"Schoenmaker","year":"2006","journal-title":"J. Cell. Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/j.jaut.2012.06.001","article-title":"Rheumatoid and pyrophosphate arthritis synovial fibroblasts induce osteoclastogenesis independently of RANKL, TNF and IL-6","volume":"39","author":"Dickerson","year":"2012","journal-title":"J. Autoimmun."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1002\/jcp.21971","article-title":"Direct cell-cell contact between periodontal ligament fibroblasts and osteoclast precursors synergistically increases the expression of genes related to osteoclastogenesis","volume":"222","author":"Bloemen","year":"2010","journal-title":"J. Cell. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1111\/j.1365-2184.2011.00751.x","article-title":"Paracrine-mediated differentiation and activation of human haematopoietic osteoclast precursor cells by skin and gingival fibroblasts","volume":"44","author":"Fernandes","year":"2011","journal-title":"Cell Prolif."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5605","DOI":"10.1007\/s00216-013-6970-6","article-title":"The use of Transwells improves the rates of differentiation and growth of cultured 3T3L1 cells","volume":"405","author":"Sabater","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1038\/nature01658","article-title":"Osteoclast differentiation and activation","volume":"423","author":"Boyle","year":"2003","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0092-8674(00)81569-X","article-title":"Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation","volume":"93","author":"Lacey","year":"1998","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1016\/j.tem.2012.05.005","article-title":"New insights into osteoclastogenic signaling mechanisms","volume":"23","author":"Nakashima","year":"2012","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4424","DOI":"10.1210\/endo.139.10.6331","article-title":"A combination of osteoclast differentiation factor and macrophage-colony stimulating factor is sufficient for both human and mouse osteoclast formation in vitro","volume":"139","author":"Quinn","year":"1998","journal-title":"Endocrinology"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/S0092-8674(00)80209-3","article-title":"Osteoprotegerin: A novel secreted protein involved in the regulation of bone density","volume":"89","author":"Simonet","year":"1997","journal-title":"Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2021","DOI":"10.1016\/S0002-9440(10)63560-2","article-title":"Receptor activator of nuclear factor kappaB ligand (RANKL)\/osteoprotegerin (OPG) ratio is increased in severe osteolysis","volume":"163","author":"Grimaud","year":"2003","journal-title":"Am. J. Pathol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.2106\/JBJS.E.01038","article-title":"Serum levels of osteoprotegerin and receptor activator of nuclear factor-kappaB ligand as markers of periprosthetic osteolysis","volume":"88","author":"Granchi","year":"2006","journal-title":"J. Bone Joint Surg. Am."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10683","DOI":"10.3390\/ijms140610683","article-title":"RANK\/RANK-L\/OPG in patients with bone metastases treated with anticancer agents and zoledronic acid: A prospective study","volume":"14","author":"Mercatali","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2889","DOI":"10.1002\/art.39837","article-title":"RANK-Independent Osteoclast Formation and Bone Erosion in Inflammatory Arthritis","volume":"68","author":"Fissel","year":"2016","journal-title":"Arthritis Rheumatol"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1002\/art.38218","article-title":"Combination of tumor necrosis factor alpha and interleukin-6 induces mouse osteoclast-like cells with bone resorption activity both in vitro and in vivo","volume":"66","author":"Yokota","year":"2014","journal-title":"Arthritis Rheumatol"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4858","DOI":"10.1074\/jbc.275.7.4858","article-title":"Tumor necrosis factor-alpha induces differentiation of and bone resorption by osteoclasts","volume":"275","author":"Azuma","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1084\/jem.20050978","article-title":"Osteoclast differentiation independent of the TRANCE-RANK-TRAF6 axis","volume":"202","author":"Kim","year":"2005","journal-title":"J. Exp. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/S0736-0266(02)00106-7","article-title":"Two distinct cellular mechanisms of osteoclast formation and bone resorption in periprosthetic osteolysis","volume":"21","author":"Sabokbar","year":"2003","journal-title":"J. Orthop. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1111\/j.1600-0765.2008.01121.x","article-title":"Mechanical force augments the anti-osteoclastogenic potential of human gingival fibroblasts in vitro","volume":"44","author":"Kook","year":"2009","journal-title":"J. Periodontal Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1359\/jbmr.2002.17.2.210","article-title":"Periodontal ligament cells under mechanical stress induce osteoclastogenesis by receptor activator of nuclear factor kappaB ligand up-regulation via prostaglandin E2 synthesis","volume":"17","author":"Kanzaki","year":"2002","journal-title":"J. Bone Miner. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1177\/154405910608500812","article-title":"RANKL increase in compressed periodontal ligament cells from root resorption","volume":"85","author":"Yamaguchi","year":"2006","journal-title":"J. Dent. Res."},{"key":"ref_44","unstructured":"Hartmann, E.S., K\u00f6hler, M.I., Beck, F., Schl\u00fcssel, S., Redeker, J.I., Summer, B., Fottner, A., Jansson, V., and Mayer-Wagner, S. Fibroblast-like cells change gene expression of bone remodelling markers in 3D transwell cultures, in preparation."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/j.cell.2006.06.044","article-title":"Matrix elasticity directs stem cell lineage specification","volume":"126","author":"Engler","year":"2006","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1136\/ard.59.1.26","article-title":"Effect of osteoprotegerin and osteoprotegerin ligand on osteoclast formation by arthroplasty membrane derived macrophages","volume":"59","author":"Itonaga","year":"2000","journal-title":"Ann. Rheum. Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.2106\/00004623-199607000-00016","article-title":"Cellular biology of bone-resorbing cells","volume":"78","author":"Athanasou","year":"1996","journal-title":"J. Bone Joint Surg. Am."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"4029","DOI":"10.1182\/blood-2004-01-0041","article-title":"Immature dendritic cell transdifferentiation into osteoclasts: A novel pathway sustained by the rheumatoid arthritis microenvironment","volume":"104","author":"Rivollier","year":"2004","journal-title":"Blood"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1080\/08916930701694667","article-title":"Tartrate-resistant acid phosphatase (TRAP) and the osteoclast\/immune cell dichotomy","volume":"41","author":"Hayman","year":"2008","journal-title":"Autoimmunity"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1177\/002215540104900601","article-title":"Tartrate-resistant acid phosphatase (Acp 5): Identification in diverse human tissues and dendritic cells","volume":"49","author":"Hayman","year":"2001","journal-title":"J. Histochem. Cytochem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1006\/abio.2001.5369","article-title":"Identification and validation of endogenous reference genes for expression profiling of T helper cell differentiation by quantitative real-time RT-PCR","volume":"299","author":"Hamalainen","year":"2001","journal-title":"Anal. Biochem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/10\/2583\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:55:11Z","timestamp":1760187311000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/10\/2583"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,5,26]]},"references-count":51,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2019,5]]}},"alternative-id":["ijms20102583"],"URL":"https:\/\/doi.org\/10.3390\/ijms20102583","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2019,5,26]]}}}