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Yet, how atypical immune receptors contribute to this process remains unclear. Here, we identify Toll-like receptor 10 (TLR10) as a key regulator of osteogenic differentiation in human adipose-derived MSCs. Herein, ASC\/TERT1 MSCs were engineered to overexpress or silence TLR10 using lentiviral vectors, and osteogenic differentiation (0\u201314 days) was assessed by metabolic assays\u2014RT-qPCR of COL1A2, ALPL and BGLAP\u2014Alizarin Red S staining, and quantitative mass spectrometry. Enhancing TLR10 expression promoted osteogenic gene programs, extracellular matrix organization, metabolic adaptation, and robust matrix mineralization, whereas TLR10 suppression maintained proliferative states and impaired osteoblast maturation. Proteomic analyses revealed that TLR10 selectively activates osteogenic, ECM-remodeling, and vitamin D-responsive pathways, while restraining programs antagonistic to differentiation. Notably, active vitamin D induced TLR10 expression and partially restored osteogenesis in TLR10-deficient cells, indicating that TLR10 is associated with vitamin D-driven bone formation. Together, beyond its established role in innate immunity, TLR10 emerges as a vitamin D-responsive regulator of mesenchymal stem cell osteogenesis, highlighting a potential therapeutic axis to enhance bone regeneration and osteogenic outcomes.<\/jats:p>","DOI":"10.3390\/cells15080697","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T15:56:56Z","timestamp":1776268616000},"page":"697","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The TLR10\u2013Vitamin D Axis Facilitates Osteogenic Differentiation of Mesenchymal Stem Cells In Vitro"],"prefix":"10.3390","volume":"15","author":[{"given":"Anna","family":"Stierschneider","sequence":"first","affiliation":[{"name":"Institute Biotechnology, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7024-7493","authenticated-orcid":false,"given":"Benjamin","family":"Neuditschko","sequence":"additional","affiliation":[{"name":"Institute Krems Bioanalytics, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1288-6718","authenticated-orcid":false,"given":"Isabella","family":"Fischer","sequence":"additional","affiliation":[{"name":"Institute Biotechnology, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esther","family":"Hellmann","sequence":"additional","affiliation":[{"name":"Institute Biotechnology, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-7938-5914","authenticated-orcid":false,"given":"Daniel","family":"Zimmermann","sequence":"additional","affiliation":[{"name":"Institute Biotechnology, IMC Krems University of Applied Sciences, 3500 Krems, Austria"},{"name":"Institute of Biotechnology & Analytics, Biotech Campus Tulln, University of Applied Sciences Wiener Neustadt, 3400 Tulln, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5982-8521","authenticated-orcid":false,"given":"Katerina","family":"Prohaska","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology & Analytics, Biotech Campus Tulln, University of Applied Sciences Wiener Neustadt, 3400 Tulln, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Milchram","sequence":"additional","affiliation":[{"name":"Institute Krems Bioanalytics, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franz","family":"Herzog","sequence":"additional","affiliation":[{"name":"Institute Krems Bioanalytics, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3281-0695","authenticated-orcid":false,"given":"Christoph","family":"Wiesner","sequence":"additional","affiliation":[{"name":"Institute Biotechnology, IMC Krems University of Applied Sciences, 3500 Krems, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,15]]},"reference":[{"key":"ref_1","first-page":"1350","article-title":"Bone repair and key signalling pathways for cell-based bone regenerative therapy: A review","volume":"18","author":"Nasir","year":"2023","journal-title":"J. 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