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Here, we present a defined biomaterial platform in which maltose-binding protein\u2013fibroblast growth factor 2 (MBP-FGF2) is surface-immobilized to create a bioinstructive interface that directs in\u00a0vitro hMSC chondrogenesis without exogenous transforming growth factor-\u03b21 (TGF-\u03b21). The immobilized FGF2 interface promotes spontaneous, scaffold-free spheroid assembly via FGFR1\u2013heparan sulfate proteoglycan-mediated adhesion, providing localized and sustained signaling. Transcriptomic and molecular analyses revealed early activation of FGF- and TGF-\u03b2-related pathways, with up-regulation of SOX9, COL2A1, and aggrecan and suppression of COL1A1 and COL10A1. These effects were consistently observed in hMSCs derived from both human embryonic stem cells and human tonsil tissue. Upon subcutaneous implantation into immunodeficient mice, pre-differentiated spheroids generated ectopic cartilage tissue rich in type II collagen and aggrecan, even in the absence of TGF-\u03b21. Taken together, these findings establish FGF2-immobilized biointerfaces as bioinstructive matrices that enable exogenous TGF-\u03b2-independent chondrogenesis and ectopic cartilage tissue formation, providing a defined and reproducible route for generating chondrogenic constructs for regenerative medicine.<\/jats:p>","DOI":"10.34133\/bmr.0367","type":"journal-article","created":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T07:41:52Z","timestamp":1776670912000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark_01","source":"Crossref","is-referenced-by-count":0,"title":["Fibroblast Growth Factor 2-Immobilized Biointerfaces Drive Exogenous Transforming Growth Factor \u03b21-Independent Chondrogenesis of Human Mesenchymal Stem Cell and Ectopic Cartilage Tissue Formation"],"prefix":"10.34133","volume":"30","author":[{"given":"Ji Hoon","family":"Jeong","sequence":"first","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Chungnam-do, Republic of Korea."},{"name":"Department of Integrated Biomedical Science, \rSoonchunhyang University, Asan 31538, Chungnam-do, Republic of Korea."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae Hong","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology\u2013Head and Neck Surgery, College of Medicine, \rSoonchunhyang University Cheonan Hospital, Cheonan 31151, Republic of Korea."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myung Jin","family":"Ban","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology\u2013Head and Neck Surgery, College of Medicine, \rSoonchunhyang University Cheonan Hospital, Cheonan 31151, Republic of Korea."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeongrok","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology\u2013Head and Neck Surgery, College of Medicine, 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