{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T11:33:14Z","timestamp":1770550394932,"version":"3.49.0"},"reference-count":81,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,6,7]],"date-time":"2025-06-07T00:00:00Z","timestamp":1749254400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,6,7]],"date-time":"2025-06-07T00:00:00Z","timestamp":1749254400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["CICECO-Aveiro Institute of Materials, UIDB\/50011\/2020 project (doi: 10.54499\/UIDB\/50011\/2020), UIDP\/50011\/2020 (doi: 10.54499\/UIDP\/50011\/2020) and LA\/P\/0006\/2020 (doi:10.54499\/LA\/P\/0006\/2020)"],"award-info":[{"award-number":["CICECO-Aveiro Institute of Materials, UIDB\/50011\/2020 project (doi: 10.54499\/UIDB\/50011\/2020), UIDP\/50011\/2020 (doi: 10.54499\/UIDP\/50011\/2020) and LA\/P\/0006\/2020 (doi:10.54499\/LA\/P\/0006\/2020)"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["BetterBone project (2022.04286.PTDC, doi: 10.54499\/2022.04286.PTDC)"],"award-info":[{"award-number":["BetterBone project (2022.04286.PTDC, doi: 10.54499\/2022.04286.PTDC)"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["Portuguese National NMR Network (RNRMN), supported by Infrastructure Project N\u00ba 022161"],"award-info":[{"award-number":["Portuguese National NMR Network (RNRMN), supported by Infrastructure Project N\u00ba 022161"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/150655\/2020, doi: 10.54499\/SFRH\/BD\/150655\/2020"],"award-info":[{"award-number":["SFRH\/BD\/150655\/2020, doi: 10.54499\/SFRH\/BD\/150655\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["project \u201cCellFi\u201d, PTDC\/BTM-ORG\/3215\/2020 (DOI10.54499\/PTDC\/BTM-ORG\/3215\/2020)"],"award-info":[{"award-number":["project \u201cCellFi\u201d, PTDC\/BTM-ORG\/3215\/2020 (DOI10.54499\/PTDC\/BTM-ORG\/3215\/2020)"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Stem Cell Res Ther"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Mesenchymal stem cells (MSC) are pivotal bioengineering tools, offering significant promise for applications in bone regeneration. However, their therapeutic potential is limited by inter-donor variability and experimental issues. This study aimed to identify robust metabolic markers of osteodifferentiation applicable across multiple donors, while providing insight into the metabolic pathways actively involved in the process.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>Untargeted nuclear magnetic resonance (NMR) metabolomics was applied to characterize the intra- and extracellular metabolic adaptations of human adipose-derived MSC (hAMSC) undergoing osteogenic differentiation, compared to proliferation alone. Multivariate and univariate statistical analysis was carried out on data from three independent donors, and cross-validation was employed to evaluate the predictive capacity of the proposed markers.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Variations in the levels of selected (nine) intracellular and (seventeen) extracellular metabolites detect osteodifferentiation by day 7 (out of 21), with nearly 100% accuracy. These signatures suggest a metabolic shift from glycolysis\/OxPhos to lactic fermentation, fatty acid <jats:italic>\u03b2-<\/jats:italic>oxidation and phosphocreatine hydrolysis. Intracellular glucose, lactate, citrate and specific amino acids are redirected towards protein synthesis and glycosylation, with some of the secreted metabolites (<jats:italic>e.g.<\/jats:italic>, citrate) seemingly involved in biomineralization and other extracellular roles. Membrane metabolism, antioxidant mechanisms and adenosine metabolism are also impacted by osteodifferentiation.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>These findings reveal effective donor-independent markers of hAMSC osteodifferentiation, with a robust extracellular signature standing out for potential rapid and non-invasive detection of osteocommitted cells.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s13287-025-04419-x","type":"journal-article","created":{"date-parts":[[2025,6,7]],"date-time":"2025-06-07T13:51:22Z","timestamp":1749304282000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Metabolic markers detect early ostedifferentiation of mesenchymal stem cells from multiple donors"],"prefix":"10.1186","volume":"16","author":[{"given":"Daniela S. C.","family":"Bispo","sequence":"first","affiliation":[]},{"given":"In\u00eas C. R.","family":"Gra\u00e7a","sequence":"additional","affiliation":[]},{"given":"Catarina S. H.","family":"Jesus","sequence":"additional","affiliation":[]},{"given":"Jo\u00e3o E.","family":"Rodrigues","sequence":"additional","affiliation":[]},{"given":"Marlene C.","family":"Correia","sequence":"additional","affiliation":[]},{"given":"Sabrina","family":"Atella","sequence":"additional","affiliation":[]},{"given":"Iola F.","family":"Duarte","sequence":"additional","affiliation":[]},{"given":"Brian J.","family":"Goodfellow","sequence":"additional","affiliation":[]},{"given":"Mariana B.","family":"Oliveira","sequence":"additional","affiliation":[]},{"given":"Jo\u00e3o F.","family":"Mano","sequence":"additional","affiliation":[]},{"given":"Ana M.","family":"Gil","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2025,6,7]]},"reference":[{"key":"4419_CR1","doi-asserted-by":"publisher","first-page":"268","DOI":"10.3389\/fcell.2019.00268","volume":"7","author":"MR Iaquinta","year":"2019","unstructured":"Iaquinta MR, Mazzoni E, Bononi I, Rotondo JC, Mazziotta C, Montesi M, et al. Adult stem cells for bone regeneration and repair. Front Cell Dev Biol. 2019;7:268.","journal-title":"Front Cell Dev Biol"},{"key":"4419_CR2","doi-asserted-by":"publisher","first-page":"1400347","DOI":"10.3389\/fcell.2024.1400347","volume":"12","author":"AB \u010cesnik","year":"2024","unstructured":"\u010cesnik AB, \u0160vajger U. The issue of heterogeneity of MSC-based advanced therapy medicinal products\u2013a review. Front Cell Dev Biol. 2024;12:1400347.","journal-title":"Front Cell Dev Biol"},{"key":"4419_CR3","doi-asserted-by":"publisher","first-page":"782","DOI":"10.1016\/j.jcyt.2019.04.003","volume":"21","author":"DG Phinney","year":"2019","unstructured":"Phinney DG, Galipeau J. Manufacturing mesenchymal stromal cells for clinical applications: A survey of good manufacturing practices at U.S. Academic centers. Cytotherapy. 2019;21:782\u201392.","journal-title":"Cytotherapy"},{"key":"4419_CR4","doi-asserted-by":"publisher","first-page":"e0209060","DOI":"10.1371\/journal.pone.0209060","volume":"14","author":"F Feng","year":"2019","unstructured":"Feng F, Thompson MP, Thomas BE, Duffy ER, Kim J, Kurosawa S, et al. A computational solution to improve biomarker reproducibility during long-term projects. PLoS ONE. 2019;14:e0209060.","journal-title":"PLoS ONE"},{"key":"4419_CR5","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1677\/jme.0.0290023","volume":"29","author":"SA Bustin","year":"2002","unstructured":"Bustin SA. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol. 2002;29:23\u201339.","journal-title":"J Mol Endocrinol"},{"key":"4419_CR6","doi-asserted-by":"publisher","first-page":"632717","DOI":"10.3389\/fcell.2021.632717","volume":"9","author":"A Wright","year":"2021","unstructured":"Wright A, Arthaud-Day ML, Weiss ML. Therapeutic use of mesenchymal stromal cells: the need for inclusive characterization guidelines to accommodate all tissue sources and species. Front Cell Dev Biol. 2021;9:632717.","journal-title":"Front Cell Dev Biol"},{"key":"4419_CR7","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/s41232-021-00192-5","volume":"42","author":"Y Imai","year":"2022","unstructured":"Imai Y, Kanie K, Kato R. Morphological heterogeneity description enabled early and parallel non-invasive prediction of T-cell proliferation inhibitory potency and growth rate for facilitating donor selection of human mesenchymal stem cells. Inflamm Regen. 2022;42:8.","journal-title":"Inflamm Regen"},{"key":"4419_CR8","doi-asserted-by":"publisher","first-page":"10408","DOI":"10.1038\/s41598-020-67254-5","volume":"10","author":"S Payr","year":"2020","unstructured":"Payr S, Schuseil T, Unger M, Seeliger C, Tiefenboeck T, Balmayor ER, et al. Effect of donor age and 3D-cultivation on osteogenic differentiation capacity of adipose-derived mesenchymal stem cells. Sci Rep. 2020;10:10408.","journal-title":"Sci Rep"},{"key":"4419_CR9","doi-asserted-by":"publisher","first-page":"e55082","DOI":"10.1371\/journal.pone.0055082","volume":"8","author":"F Matsuoka","year":"2013","unstructured":"Matsuoka F, Takeuchi I, Agata H, Kagami H, Shiono H, Kiyota Y, et al. Morphology-Based prediction of osteogenic differentiation potential of human mesenchymal stem cells. PLoS ONE. 2013;8:e55082.","journal-title":"PLoS ONE"},{"key":"4419_CR10","doi-asserted-by":"publisher","first-page":"935","DOI":"10.1002\/stem.2322","volume":"34","author":"RA Marklein","year":"2016","unstructured":"Marklein RA, Lo Surdo JL, Bellayr IH, Godil SA, Puri RK, Bauer SR. High content imaging of early morphological signatures predicts long term mineralization capacity of human mesenchymal stem cells upon osteogenic induction. Stem Cells. 2016;34:935\u201347.","journal-title":"Stem Cells"},{"key":"4419_CR11","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1073\/pnas.0909597107","volume":"107","author":"MD Treiser","year":"2010","unstructured":"Treiser MD, Yang EH, Gordonov S, Cohen DM, Androulakis IP, Kohn J, et al. Cytoskeleton-based forecasting of stem cell lineage fates. Proc Natl Acad Sci U S A. 2010;107:610\u20135.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"4419_CR12","doi-asserted-by":"publisher","first-page":"1329840","DOI":"10.3389\/fcell.2023.1329840","volume":"11","author":"M Mai","year":"2023","unstructured":"Mai M, Luo S, Fasciano S, Oluwole TE, Ortiz J, Pang Y, et al. Morphology-based deep learning approach for predicting adipogenic and osteogenic differentiation of human mesenchymal stem cells (hMSCs). Front Cell Dev Biol. 2023;11:1329840.","journal-title":"Front Cell Dev Biol"},{"key":"4419_CR13","doi-asserted-by":"publisher","first-page":"84","DOI":"10.5966\/sctm.2014-0156","volume":"4","author":"A Bertolo","year":"2015","unstructured":"Bertolo A, Gemperli A, Gruber M, Gantenbein B, Baur M, P\u00f6tzel T, et al. In vitro cell motility as a potential mesenchymal stem cell marker for multipotency. Stem Cells Transl Med. 2015;4:84\u201390.","journal-title":"Stem Cells Transl Med"},{"key":"4419_CR14","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1016\/j.jbiotec.2010.01.007","volume":"148","author":"C Hildebrandt","year":"2010","unstructured":"Hildebrandt C, B\u00fcth H, Cho S, Impidjati, Thielecke H. Detection of the osteogenic differentiation of mesenchymal stem cells in 2D and 3D cultures by electrochemical impedance spectroscopy. J Biotechnol. 2010;148:83\u201390.","journal-title":"J Biotechnol"},{"key":"4419_CR15","doi-asserted-by":"publisher","first-page":"9913","DOI":"10.1039\/D0TB01968B","volume":"8","author":"Z Zhang","year":"2020","unstructured":"Zhang Z, Zheng T, Zhu R. Long-term and label-free monitoring for osteogenic differentiation of mesenchymal stem cells using force sensor and impedance measurement. J Mater Chem B. 2020;8:9913\u201320.","journal-title":"J Mater Chem B"},{"key":"4419_CR16","doi-asserted-by":"publisher","first-page":"e2210637","DOI":"10.1002\/adma.202210637","volume":"35","author":"Y Zhou","year":"2023","unstructured":"Zhou Y, Ping X, Guo Y, Heng BC, Wang Y, Meng Y, et al. Assessing Biomaterial-Induced stem cell lineage fate by machine Learning-Based artificial intelligence. Adv Mater. 2023;35:e2210637.","journal-title":"Adv Mater"},{"key":"4419_CR17","doi-asserted-by":"publisher","first-page":"1094","DOI":"10.3390\/ph15091094","volume":"15","author":"Z Feng","year":"2022","unstructured":"Feng Z, Su X, Wang T, Guo S. Identification of biomarkers that modulate osteogenic differentiation in mesenchymal stem cells related to inflammation and immunity: A Bioinformatics-Based comprehensive study. Pharmaceuticals. 2022;15:1094.","journal-title":"Pharmaceuticals"},{"key":"4419_CR18","doi-asserted-by":"publisher","first-page":"2157","DOI":"10.3390\/cimb43030150","volume":"43","author":"M Kanawa","year":"2021","unstructured":"Kanawa M, Igarashi A, Fujimoto K, Saskianti T, Nakashima A, Higashi Y, et al. The identification of marker genes for predicting the osteogenic differentiation potential of mesenchymal stromal cells. Curr Issues Mol Biol. 2021;43:2157\u201366.","journal-title":"Curr Issues Mol Biol"},{"key":"4419_CR19","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/j.scr.2013.09.009","volume":"12","author":"C Gran\u00e9li","year":"2014","unstructured":"Gran\u00e9li C, Thorfve A, Ruetschi U, Brisby H, Thomsen P, Lindahl A, et al. Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach. Stem Cell Res. 2014;12:153\u201365.","journal-title":"Stem Cell Res"},{"key":"4419_CR20","doi-asserted-by":"publisher","first-page":"2804","DOI":"10.1002\/stem.215","volume":"27","author":"MO Platt","year":"2009","unstructured":"Platt MO, Wilder CL, Wells A, Griffith LG, Lauffenburger DA. Multipathway kinase signatures of multipotent stromal cells are predictive for osteogenic differentiation: Tissue-specific stem cells. Stem Cells. 2009;27:2804\u201314.","journal-title":"Stem Cells"},{"key":"4419_CR21","doi-asserted-by":"publisher","first-page":"3745","DOI":"10.3390\/cells11233745","volume":"11","author":"DSC Bispo","year":"2022","unstructured":"Bispo DSC, Jesus CSH, Romek K, Marques IMC, Oliveira MB, Mano JF, et al. An intracellular metabolic signature as a potential Donor-Independent marker of the osteogenic differentiation of adipose tissue mesenchymal stem cells. Cells. 2022;11:3745.","journal-title":"Cells"},{"key":"4419_CR22","doi-asserted-by":"publisher","first-page":"642681","DOI":"10.3389\/fcell.2021.642681","volume":"9","author":"TB Sigmarsdottir","year":"2021","unstructured":"Sigmarsdottir TB, McGarrity S, Yurkovich JT, Rolfsson \u00d3, Sigurj\u00f3nsson \u00d3E. Analyzing metabolic States of adipogenic and osteogenic differentiation in human mesenchymal stem cells via genome scale metabolic model reconstruction. Front Cell Dev Biol. 2021;9:642681.","journal-title":"Front Cell Dev Biol"},{"key":"4419_CR23","doi-asserted-by":"publisher","first-page":"e0221378","DOI":"10.1371\/journal.pone.0221378","volume":"14","author":"AR Caseiro","year":"2019","unstructured":"Caseiro AR, Pedrosa SS, Ivanova G, Branquinho MV, Almeida A, Faria F, et al. Mesenchymal stem\/ stromal cells metabolomic and bioactive factors profiles: A comparative analysis on the umbilical cord and dental pulp derived stem\/ stromal cells secretome. PLoS ONE. 2019;14:e0221378.","journal-title":"PLoS ONE"},{"key":"4419_CR24","doi-asserted-by":"publisher","first-page":"2187","DOI":"10.1042\/BCJ20160241","volume":"473","author":"A Mastrangelo","year":"2016","unstructured":"Mastrangelo A, Panadero MI, Perez LM, Galvez BG, Garcia A, Barbas C, et al. New insight on obesity and adipose-derived stem cells using comprehensive metabolomics. Biochem J. 2016;473:2187\u2013203.","journal-title":"Biochem J"},{"key":"4419_CR25","doi-asserted-by":"publisher","first-page":"608","DOI":"10.1016\/j.jcyt.2021.12.009","volume":"24","author":"SA DeVeaux","year":"2022","unstructured":"DeVeaux SA, Ogle ME, Vyshnya S, Chiappa NF, Leitmann B, Rudy R, et al. Characterizing human mesenchymal stromal cells\u2019 immune-modulatory potency using targeted lipidomic profiling of sphingolipids. Cytotherapy. 2022;24:608\u201318.","journal-title":"Cytotherapy"},{"key":"4419_CR26","doi-asserted-by":"publisher","first-page":"622","DOI":"10.1194\/jlr.M030650","volume":"54","author":"L Kilpinen","year":"2013","unstructured":"Kilpinen L, Tigistu-Sahle F, Oja S, Greco D, Parmar A, Saavalainen P, et al. Aging bone marrow mesenchymal stromal cells have altered membrane glycerophospholipid composition and functionality. J Lipid Res. 2013;54:622\u201335.","journal-title":"J Lipid Res"},{"key":"4419_CR27","doi-asserted-by":"publisher","first-page":"654","DOI":"10.1021\/acs.jproteome.1c00832","volume":"21","author":"DSC Bispo","year":"2022","unstructured":"Bispo DSC, Jesus CSH, Correia M, Ferreira F, Bonifazio G, Goodfellow BJ, et al. NMR metabolomics assessment of osteogenic differentiation of Adipose-Tissue-Derived mesenchymal stem cells. J Proteome Res. 2022;21:654\u201370.","journal-title":"J Proteome Res"},{"key":"4419_CR28","doi-asserted-by":"publisher","first-page":"2003","DOI":"10.1007\/s12015-021-10193-z","volume":"17","author":"DSCC Bispo","year":"2021","unstructured":"Bispo DSCC, Jesus CSHH, Marques IMCC, Romek KM, Oliveira MB, Mano JF, et al. Metabolomic applications in stem cell research: a review. Stem Cell Rev Rep. 2021;17:2003\u201324.","journal-title":"Stem Cell Rev Rep"},{"key":"4419_CR29","doi-asserted-by":"publisher","first-page":"7908","DOI":"10.3390\/ijms22157908","volume":"22","author":"S Nov\u00e1kov\u00e1","year":"2021","unstructured":"Nov\u00e1kov\u00e1 S, Danchenko M, Okaj\u010dekov\u00e1 T, Baranovi\u010dov\u00e1 E, Kov\u00e1\u010d A, Grend\u00e1r M, et al. Comparative proteomic and metabolomic analysis of human osteoblasts, differentiated from dental pulp stem cells, hinted crucial signaling pathways promoting osteogenesis. Int J Mol Sci. 2021;22:7908.","journal-title":"Int J Mol Sci"},{"key":"4419_CR30","doi-asserted-by":"publisher","first-page":"3776","DOI":"10.1039\/C6AN00170J","volume":"141","author":"A Surrati","year":"2016","unstructured":"Surrati A, Linforth R, Fisk ID, Sottile V, Kim DH. Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting. Analyst. 2016;141:3776\u201387.","journal-title":"Analyst"},{"key":"4419_CR31","doi-asserted-by":"publisher","first-page":"1257","DOI":"10.3390\/cells11081257","volume":"11","author":"DSC Bispo","year":"2022","unstructured":"Bispo DSC, Mich\u00e1lkov\u00e1 L, Correia M, Jesus CSH, Duarte IF, Goodfellow BJ, et al. Endo- and exometabolome crosstalk in mesenchymal stem cells undergoing osteogenic differentiation. Cells. 2022;11:1257.","journal-title":"Cells"},{"key":"4419_CR32","doi-asserted-by":"publisher","first-page":"311","DOI":"10.33594\/000000377","volume":"55","author":"A Surrati","year":"2021","unstructured":"Surrati A, Evseev S, Jourdan F, Kim D-H, Sottile V. Osteogenic response of human mesenchymal stem cells analysed using combined intracellular and extracellular metabolomic monitoring. Cell Physiol Biochem. 2021;55:311\u201326.","journal-title":"Cell Physiol Biochem"},{"key":"4419_CR33","doi-asserted-by":"publisher","first-page":"117363","DOI":"10.1016\/j.bone.2024.117363","volume":"192","author":"AH Hoveidaei","year":"2025","unstructured":"Hoveidaei AH, Sadat-Shojai M, Nabavizadeh SS, Niakan R, Shirinezhad A, MosalamiAghili S, et al. Clinical challenges in bone tissue engineering - A narrative review. Bone. 2025;192:117363.","journal-title":"Bone"},{"key":"4419_CR34","doi-asserted-by":"publisher","first-page":"424","DOI":"10.1210\/edrv-14-4-424","volume":"14","author":"GS Stein","year":"1993","unstructured":"Stein GS, Lian JB. Molecular mechanisms mediating proliferation\/differentiation interrelationships during progressive development of the osteoblast phenotype. Endocr Rev. 1993;14:424\u201342.","journal-title":"Endocr Rev"},{"key":"4419_CR35","doi-asserted-by":"publisher","first-page":"1257","DOI":"10.1007\/s10529-011-0541-8","volume":"33","author":"A Shafiee","year":"2011","unstructured":"Shafiee A, Seyedjafari E, Soleimani M, Ahmadbeigi N, Dinarvand P, Ghaemi N. A comparison between osteogenic differentiation of human unrestricted somatic stem cells and mesenchymal stem cells from bone marrow and adipose tissue. Biotechnol Lett. 2011;33:1257\u201364.","journal-title":"Biotechnol Lett"},{"key":"4419_CR36","doi-asserted-by":"publisher","first-page":"1400","DOI":"10.3390\/biom11101400","volume":"11","author":"EC Torre","year":"2021","unstructured":"Torre EC, Bicer M, Cottrell GS, Widera D, Tamagnini F. Time-dependent reduction of calcium oscillations in adipose-derived stem cells differentiating towards adipogenic and osteogenic lineage. Biomolecules. 2021;11:1400.","journal-title":"Biomolecules"},{"key":"4419_CR37","doi-asserted-by":"publisher","first-page":"2562","DOI":"10.1002\/jcp.22605","volume":"226","author":"G Pattappa","year":"2011","unstructured":"Pattappa G, Heywood HK, de Bruijn JD, Lee DA. The metabolism of human mesenchymal stem cells during proliferation and differentiation. J Cell Physiol. 2011;226:2562\u201370.","journal-title":"J Cell Physiol"},{"key":"4419_CR38","doi-asserted-by":"publisher","first-page":"3368","DOI":"10.1002\/stem.2097","volume":"33","author":"Y Liu","year":"2015","unstructured":"Liu Y, Mu\u00f1oz N, Bunnell BA, Logan TM, Ma T. Density-Dependent metabolic heterogeneity in human mesenchymal stem cells. Stem Cells. 2015;33:3368\u201381.","journal-title":"Stem Cells"},{"key":"4419_CR39","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1186\/s13287-023-03549-4","volume":"14","author":"C Pradenas","year":"2023","unstructured":"Pradenas C, Luque-Campos N, Oyarce K, Contreras-Lopez R, Bustamante-Barrientos FA, Bustos A, et al. Lactate: an alternative pathway for the immunosuppressive properties of mesenchymal stem\/stromal cells. Stem Cell Res Ther. 2023;14:335.","journal-title":"Stem Cell Res Ther"},{"key":"4419_CR40","doi-asserted-by":"publisher","first-page":"654","DOI":"10.1089\/ten.tea.2011.0223","volume":"18","author":"GA Higuera","year":"2012","unstructured":"Higuera GA, Schop D, Spitters TWGM, Van Dijkhuizen-Radersma R, Bracke M, De Bruijn JD, et al. Patterns of amino acid metabolism by proliferating human mesenchymal stem cells. Tissue Eng Part A. 2012;18:654\u201364.","journal-title":"Tissue Eng Part A"},{"key":"4419_CR41","doi-asserted-by":"publisher","first-page":"110935","DOI":"10.1016\/j.nut.2020.110935","volume":"78","author":"T Sartori","year":"2020","unstructured":"Sartori T, Santos ACA, Oliveira da Silva R, Kodja G, Rogero MM, Borelli P, et al. Branched chain amino acids improve mesenchymal stem cell proliferation, reducing nuclear factor kappa B expression and modulating some inflammatory properties. Nutrition. 2020;78:110935.","journal-title":"Nutrition"},{"key":"4419_CR42","doi-asserted-by":"publisher","first-page":"402","DOI":"10.3390\/biom11030402","volume":"11","author":"J Mierziak","year":"2021","unstructured":"Mierziak J, Burgberger M, Wojtasik W. 3-Hydroxybutyrate as a metabolite and a signal molecule regulating processes of living organisms. Biomolecules. 2021;11:402.","journal-title":"Biomolecules"},{"key":"4419_CR43","doi-asserted-by":"publisher","first-page":"e1601273","DOI":"10.1126\/sciadv.1601273","volume":"2","author":"J Meiser","year":"2016","unstructured":"Meiser J, Tumanov S, Maddocks O, Labuschagne CF, Athineos D, Van Den Broek N, et al. Serine one-carbon catabolism with formate overflow. Sci Adv. 2016;2:e1601273.","journal-title":"Sci Adv"},{"key":"4419_CR44","doi-asserted-by":"publisher","first-page":"905","DOI":"10.1038\/sj.bjp.0701924","volume":"124","author":"G Yildiz","year":"1998","unstructured":"Yildiz G, Demiry\u00fcrek AT, Sahin-Erdemli I, Kanzik I. Comparison of antioxidant activities of Aminoguanidine, Methylguanidine and guanidine by luminol-enhanced chemiluminescence. Br J Pharmacol. 1998;124:905\u201310.","journal-title":"Br J Pharmacol"},{"key":"4419_CR45","doi-asserted-by":"publisher","first-page":"356","DOI":"10.1159\/000063301","volume":"92","author":"T Yokozawa","year":"2002","unstructured":"Yokozawa T, Muto Y, Wakaki K, Kashiwagi H. Site of Methylguanidine production and factors that influence production levels. Nephron. 2002;92:356\u201362.","journal-title":"Nephron"},{"key":"4419_CR46","doi-asserted-by":"publisher","first-page":"162","DOI":"10.5582\/ddt.2011.v5.4.162","volume":"5","author":"K Ienaga","year":"2011","unstructured":"Ienaga K, Yokozawa T, Creatinine. 5-hydroxy-1-methylhydantoin, NZ-419) as intrinsic hydroxyl radical scavengers. Drug Discov Ther. 2011;5:162\u201375.","journal-title":"Drug Discov Ther"},{"key":"4419_CR47","doi-asserted-by":"publisher","first-page":"1885","DOI":"10.1089\/scd.2010.0093","volume":"19","author":"A Valle-Prieto","year":"2010","unstructured":"Valle-Prieto A, Conget PA. Human mesenchymal stem cells efficiently manage oxidative stress. Stem Cells Dev. 2010;19:1885\u201393.","journal-title":"Stem Cells Dev"},{"key":"4419_CR48","doi-asserted-by":"publisher","first-page":"721","DOI":"10.1002\/jcp.21145","volume":"213","author":"M Iemata","year":"2007","unstructured":"Iemata M, Takarada T, Hinoi E, Taniura H, Yoneda Y. Suppression by glutamate of proliferative activity through glutathione depletion mediated by the cystine\/glutamate antiporter in mesenchymal C3H10T1\/2 stem cells. J Cell Physiol. 2007;213:721\u20139.","journal-title":"J Cell Physiol"},{"key":"4419_CR49","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1038\/s41580-022-00490-x","volume":"23","author":"Y Posor","year":"2022","unstructured":"Posor Y, Jang W, Haucke V. Phosphoinositides as membrane organizers. Nat Rev Mol Cell Biol. 2022;23:797\u2013816.","journal-title":"Nat Rev Mol Cell Biol"},{"key":"4419_CR50","doi-asserted-by":"publisher","first-page":"1172","DOI":"10.3389\/fphar.2019.01172","volume":"10","author":"DR Chhetri","year":"2019","unstructured":"Chhetri DR. Myo-inositol and its derivatives: their emerging role in the treatment of human diseases. Front Pharmacol. 2019;10:1172.","journal-title":"Front Pharmacol"},{"key":"4419_CR51","doi-asserted-by":"publisher","first-page":"2247","DOI":"10.1007\/s13577-023-00957-9","volume":"36","author":"BC Galgaro","year":"2023","unstructured":"Galgaro BC, Beckenkamp LR, Naasani LIS, Wink MR. Adenosine metabolism by mesenchymal stromal cells isolated from different human tissues. Hum Cell. 2023;36:2247\u201358.","journal-title":"Hum Cell"},{"key":"4419_CR52","doi-asserted-by":"publisher","first-page":"960","DOI":"10.1634\/stemcells.2007-0509","volume":"26","author":"C-T Chen","year":"2008","unstructured":"Chen C-T, Shih Y-RV, Kuo TK, Lee OK, Wei Y-H. Coordinated changes of mitochondrial biogenesis and antioxidant enzymes during osteogenic differentiation of human mesenchymal stem cells. Stem Cells. 2008;26:960\u20138.","journal-title":"Stem Cells"},{"key":"4419_CR53","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1089\/scd.2015.0193","volume":"25","author":"LC Shum","year":"2016","unstructured":"Shum LC, White NS, Mills BN, De Mesy Bentley KL, Eliseev RA. Energy metabolism in mesenchymal stem cells during osteogenic differentiation. Stem Cells Dev. 2016;25:114\u201322.","journal-title":"Stem Cells Dev"},{"key":"4419_CR54","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1007\/s11306-021-01829-9","volume":"17","author":"BB Misra","year":"2021","unstructured":"Misra BB, Jayapalan S, Richards AK, Helderman RCM, Rendina-Ruedy E. Untargeted metabolomics in primary murine bone marrow stromal cells reveals distinct profile throughout osteoblast differentiation. Metabolomics. 2021;17:86.","journal-title":"Metabolomics"},{"key":"4419_CR55","doi-asserted-by":"publisher","first-page":"4120","DOI":"10.3390\/ijms22084120","volume":"22","author":"A Donat","year":"2021","unstructured":"Donat A, Knapstein PR, Jiang S, Baranowsky A, Ballhause TM, Frosch KH, et al. Glucose metabolism in osteoblasts in healthy and pathophysiological conditions. Int J Mol Sci. 2021;22:4120.","journal-title":"Int J Mol Sci"},{"key":"4419_CR56","doi-asserted-by":"publisher","first-page":"2463","DOI":"10.3390\/ijms25052463","volume":"25","author":"A Cressman","year":"2024","unstructured":"Cressman A, Morales D, Zhang Z, Le B, Foley J, Murray-Stewart T, et al. Effects of spermine synthase deficiency in mesenchymal stromal cells are rescued by upstream Inhibition of ornithine decarboxylase. Int J Mol Sci. 2024;25:2463.","journal-title":"Int J Mol Sci"},{"key":"4419_CR57","doi-asserted-by":"publisher","first-page":"8","DOI":"10.22203\/eCM.v010a02","volume":"10","author":"I Gerber","year":"2005","unstructured":"Gerber I, Ap Gwynn I, Alini M, Wallimann T. Stimulatory effects of creatine on metabolic activity, differentiation and mineralization of primary osteoblast-like cells in monolayer and micromass cell cultures. Eur Cell Mater. 2005;10:8\u201322.","journal-title":"Eur Cell Mater"},{"key":"4419_CR58","doi-asserted-by":"publisher","first-page":"488","DOI":"10.3390\/jcm8040488","volume":"8","author":"DG Candow","year":"2019","unstructured":"Candow DG, Forbes SC, Chilibeck PD, Cornish SM, Antonio J, Kreider RB. Effectiveness of creatine supplementation on aging muscle and bone: focus on falls prevention and inflammation. J Clin Med. 2019;8:488.","journal-title":"J Clin Med"},{"key":"4419_CR59","doi-asserted-by":"publisher","first-page":"1432","DOI":"10.1016\/j.bbrc.2009.12.083","volume":"391","author":"A Sekrecka-Belniak","year":"2010","unstructured":"Sekrecka-Belniak A, Balcerzak M, Buchet R, Pikula S. Active creatine kinase is present in matrix vesicles isolated from femurs of chicken embryo: implications for bone mineralization. Biochem Biophys Res Commun. 2010;391:1432\u20136.","journal-title":"Biochem Biophys Res Commun"},{"key":"4419_CR60","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.bone.2017.08.024","volume":"115","author":"P Kushwaha","year":"2018","unstructured":"Kushwaha P, Wolfgang MJ, Riddle RC. Fatty acid metabolism by the osteoblast. Bone. 2018;115:8\u201314.","journal-title":"Bone"},{"key":"4419_CR61","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1038\/s41420-022-01077-3","volume":"8","author":"MB Gayatri","year":"2022","unstructured":"Gayatri MB, Gajula NN, Chava S, Reddy ABM. High glutamine suppresses osteogenesis through mTORC1-mediated Inhibition of the mTORC2\/AKT-473\/RUNX2 axis. Cell Death Discov. 2022;8:277.","journal-title":"Cell Death Discov"},{"key":"4419_CR62","doi-asserted-by":"publisher","first-page":"1261","DOI":"10.1016\/S0021-9258(18)91421-3","volume":"239","author":"B Flanagan","year":"1964","unstructured":"Flanagan B, Nichols G. Metabolic studies of bone in vitro. J Biol Chem. 1964;239:1261\u20135.","journal-title":"J Biol Chem"},{"key":"4419_CR63","doi-asserted-by":"publisher","first-page":"1034","DOI":"10.3390\/cells9041034","volume":"9","author":"C Morganti","year":"2020","unstructured":"Morganti C, Bonora M, Marchi S, Ferroni L, Gardin C, Wieckowski MR, et al. Citrate mediates crosstalk between mitochondria and the nucleus to promote human mesenchymal stem cell in vitro osteogenesis. Cells. 2020;9:1034.","journal-title":"Cells"},{"key":"4419_CR64","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.bbrc.2022.05.028","volume":"615","author":"F Nian","year":"2022","unstructured":"Nian F, Qian Y, Xu F, Yang M, Wang H, Zhang Z. LDHA promotes osteoblast differentiation through histone lactylation. Biochem Biophys Res Commun. 2022;615:31\u20135.","journal-title":"Biochem Biophys Res Commun"},{"key":"4419_CR65","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1016\/j.mce.2017.05.017","volume":"452","author":"Y Wu","year":"2017","unstructured":"Wu Y, Wang M, Feng H, Peng Y, Sun J, Qu X, et al. Lactate induces osteoblast differentiation by stabilization of HIF1\u03b1. Mol Cell Endocrinol. 2017;452:84\u201392.","journal-title":"Mol Cell Endocrinol"},{"key":"4419_CR66","doi-asserted-by":"publisher","first-page":"22425","DOI":"10.1073\/pnas.1009219107","volume":"107","author":"YY Hu","year":"2010","unstructured":"Hu YY, Rawal A, Schmidt-Rohr K. Strongly bound citrate stabilizes the apatite nanocrystals in bone. Proc Natl Acad Sci. 2010;107:22425\u20139.","journal-title":"Proc Natl Acad Sci"},{"key":"4419_CR67","doi-asserted-by":"publisher","first-page":"1097","DOI":"10.1089\/scd.2012.0432","volume":"22","author":"M Ciciarello","year":"2013","unstructured":"Ciciarello M, Zini R, Rossi L, Salvestrini V, Ferrari D, Manfredini R, et al. Extracellular purines promote the differentiation of human bone marrow-derived mesenchymal stem cells to the osteogenic and adipogenic lineages. Stem Cells Dev. 2013;22:1097\u2013111.","journal-title":"Stem Cells Dev"},{"key":"4419_CR68","doi-asserted-by":"publisher","first-page":"286","DOI":"10.1002\/jbmr.195","volume":"26","author":"MC Yadav","year":"2011","unstructured":"Yadav MC, Sim\u00e3o AMS, Narisawa S, Huesa C, McKee MD, Farquharson C, et al. Loss of skeletal mineralization by the simultaneous ablation of PHOSPHO1 and alkaline phosphatase function: A unified model of the mechanisms of initiation of skeletal calcification. J Bone Miner Res. 2011;26:286\u201397.","journal-title":"J Bone Miner Res"},{"key":"4419_CR69","doi-asserted-by":"publisher","first-page":"e166888","DOI":"10.1172\/jci.insight.166888","volume":"8","author":"G Hu","year":"2023","unstructured":"Hu G, Yu Y, Sharma D, Pruett-Miller SM, Ren Y, Zhang GF, et al. Glutathione limits RUNX2 oxidation and degradation to regulate bone formation. JCI Insight. 2023;8:e166888.","journal-title":"JCI Insight"},{"key":"4419_CR70","doi-asserted-by":"publisher","first-page":"786","DOI":"10.1111\/jcmm.13356","volume":"22","author":"C-H Lin","year":"2018","unstructured":"Lin C-H, Li N-T, Cheng H-S, Yen M-L. Oxidative stress induces imbalance of adipogenic\/osteoblastic lineage commitment in mesenchymal stem cells through decreasing SIRT1 functions. J Cell Mol Med. 2018;22:786\u201396.","journal-title":"J Cell Mol Med"},{"key":"4419_CR71","doi-asserted-by":"publisher","first-page":"966","DOI":"10.1016\/j.cmet.2019.01.016","volume":"29","author":"Y Yu","year":"2019","unstructured":"Yu Y, Newman H, Shen L, Sharma D, Hu G, Mirando AJ, et al. Glutamine metabolism regulates proliferation and lineage allocation in skeletal stem cells. Cell Metab. 2019;29:966\u201378.","journal-title":"Cell Metab"},{"key":"4419_CR72","doi-asserted-by":"publisher","first-page":"1673","DOI":"10.1007\/s00726-014-1729-8","volume":"46","author":"C Zhou","year":"2014","unstructured":"Zhou C, Zhang X, Xu L, Wu T, Cui L, Xu D. Taurine promotes human mesenchymal stem cells to differentiate into osteoblast through the ERK pathway. Amino Acids. 2014;46:1673\u201380.","journal-title":"Amino Acids"},{"key":"4419_CR73","doi-asserted-by":"publisher","first-page":"ziae006","DOI":"10.1093\/jbmrpl\/ziae006","volume":"8","author":"D Atanasova","year":"2024","unstructured":"Atanasova D, Mirgorodskaya E, Moparthi L, Koch S, Haarhaus M, Narisawa S, et al. Glycoproteomic profile of human tissue-nonspecific alkaline phosphatase expressed in osteoblasts. JBMR Plus. 2024;8:ziae006.","journal-title":"JBMR Plus"},{"key":"4419_CR74","doi-asserted-by":"publisher","first-page":"e85464","DOI":"10.7554\/eLife.85464","volume":"12","author":"Z Zhang","year":"2023","unstructured":"Zhang Z, Huang Z, Awad M, Elsalanty M, Cray J, Ball LE, et al. O-GlcNAc glycosylation orchestrates fate decision and niche function of bone marrow stromal progenitors. Elife. 2023;12:e85464.","journal-title":"Elife"},{"key":"4419_CR75","doi-asserted-by":"publisher","first-page":"113372","DOI":"10.1016\/j.yexcr.2022.113372","volume":"421","author":"CL Bartlett","year":"2022","unstructured":"Bartlett CL, Ralefatane MG, Cave EM, Crowther NJ, Ferris WF. Differential glycosylation of tissue non-specific alkaline phosphatase in mesenchymal stromal cells differentiated into either an osteoblastic or adipocytic phenotype. Exp Cell Res. 2022;421:113372.","journal-title":"Exp Cell Res"},{"key":"4419_CR76","doi-asserted-by":"publisher","first-page":"1208","DOI":"10.1074\/jbc.M109.035436","volume":"285","author":"HE Miwa","year":"2010","unstructured":"Miwa HE, Gerken TA, Jamison O, Tabak LA. Isoform-specific O-glycosylation of osteopontin and bone sialoprotein by polypeptide N-acetylgalactosaminyltransferase-1. J Biol Chem. 2010;285:1208\u201319.","journal-title":"J Biol Chem"},{"key":"4419_CR77","doi-asserted-by":"publisher","first-page":"jcs209452","DOI":"10.1242\/jcs.209452","volume":"131","author":"KM Wilson","year":"2018","unstructured":"Wilson KM, Jagger AM, Walker M, Seinkmane E, Fox JM, Kr\u00f6ger R, et al. Glycans modify mesenchymal stem cell differentiation to impact on the function of resulting osteoblasts. J Cell Sci. 2018;131:jcs209452.","journal-title":"J Cell Sci"},{"key":"4419_CR78","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1042\/BJ20040511","volume":"382","author":"SJ Roberts","year":"2004","unstructured":"Roberts SJ, Stewart AJ, Sadler PJ, Farquharson C. Human PHOSPHO1 exhibits high specific phosphoethanolamine and phosphocholine phosphatase activities. Biochem J. 2004;382:59\u201365.","journal-title":"Biochem J"},{"key":"4419_CR79","doi-asserted-by":"publisher","first-page":"123","DOI":"10.3109\/03008208909002412","volume":"23","author":"WT Butler","year":"1989","unstructured":"Butler WT. The nature and significance of osteopontin. Connect Tissue Res. 1989;23:123\u201336.","journal-title":"Connect Tissue Res"},{"key":"4419_CR80","doi-asserted-by":"publisher","first-page":"111669","DOI":"10.1016\/j.msec.2020.111669","volume":"120","author":"N Chauhan","year":"2021","unstructured":"Chauhan N, Singh Y. L-histidine controls the hydroxyapatite mineralization with plate-like morphology: effect of concentration and media. Mater Sci Engineering: C. 2021;120:111669.","journal-title":"Mater Sci Engineering: C"},{"key":"4419_CR81","first-page":"288","volume":"102","author":"A Kumar","year":"2012","unstructured":"Kumar A, Bachhawat AK. Pyroglutamic acid: throwing light on a lightly studied metabolite. Curr Sci. 2012;102:288\u201397.","journal-title":"Curr Sci"}],"container-title":["Stem Cell Research &amp; Therapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13287-025-04419-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13287-025-04419-x\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13287-025-04419-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,7]],"date-time":"2025-06-07T18:03:25Z","timestamp":1749319405000},"score":1,"resource":{"primary":{"URL":"https:\/\/stemcellres.biomedcentral.com\/articles\/10.1186\/s13287-025-04419-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,7]]},"references-count":81,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["4419"],"URL":"https:\/\/doi.org\/10.1186\/s13287-025-04419-x","relation":{},"ISSN":["1757-6512"],"issn-type":[{"value":"1757-6512","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,7]]},"assertion":[{"value":"19 February 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 May 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 June 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Adipose tissue from donor 1 was obtained under an agreement between the University of Aveiro and \u201cHospital da Luz\u201d, Aveiro, dated 17th February 2023 and upon written consent from the donor. The human adipose-derived stem cells (donors 2 and 3) were purchased from the American Type Culture Collection (Lot 70017032, Ref. ATCC PCS-500-011) and Lonza (Lot 22TL018258, Ref. LSLZPT-5006) having been accompanied by Certificates of Analysis stating that cells were isolated from donated human tissue after obtaining permission for their use in research applications by informed consent or legal authorization. Regarding osteoblasts, these were purchased from Lonza (CC-2538, Lot 19TL217387), certified to have been isolated from donated human tissue after obtaining permission for their use in research applications. All human cells were used within the framework of the BetterBone project (2022.04286.PTDC, doi: )","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Human ethics and consent to participate"}},{"value":"Not applicable.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"Authors declare that they have no competing interests.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"Not applicable.","order":6,"name":"Ethics","group":{"name":"EthicsHeading","label":"Clinical trial number"}}],"article-number":"294"}}