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In native tissues, stem cells reside within dynamic and mechanically heterogeneous microenvironments, where physical factors such as matrix stiffness, viscoelasticity, topography, and externally applied forces play essential roles in directing cellular behavior. Recent advances in mechanically active biomaterials have enabled precise control over the mechanical properties of stem cell microenvironments, providing powerful platforms to investigate mechanotransduction mechanisms and guide lineage specification. These materials\u2014including stiffness\u2010tunable and viscoelastic hydrogels, stimuli\u2010responsive scaffolds, and dynamic culture systems\u2014facilitate spatiotemporal modulation of mechanical cues and allow systematic interrogation of force\u2010mediated signaling pathways, such as integrin\u2010based adhesion, cytoskeletal remodeling, and YAP\/TAZ\u2010dependent transcriptional regulation. In this review, we summarize the fundamental mechanisms underlying mechanical regulation of stem cell differentiation and compare two\u2010 and three\u2010dimensional culture systems from a mechanobiological perspective. We further highlight how material design principles can enhance functional maturation and translational relevance. Finally, we discuss current challenges and future opportunities in the development of scalable, clinically compatible mechanically active biomaterials. By integrating insights from mechanobiology and materials science, this review aims to provide design\u2010oriented frameworks for the rational engineering of next\u2010generation stem cell\u2010based healthcare technologies.<\/jats:p>","DOI":"10.1002\/adhm.71724","type":"journal-article","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T04:44:52Z","timestamp":1789533892000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mechanically Active Biomaterials for Stem Cell Differentiation"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-1234-2427","authenticated-orcid":false,"given":"Lanxin","family":"Xu","sequence":"first","affiliation":[{"name":"Biomaterials and Tissue Engineering Research Center Shanghai Institute of Ceramics Chinese Academy of Sciences  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3963-0408","authenticated-orcid":false,"given":"Mingxia","family":"Lu","sequence":"additional","affiliation":[{"name":"Biomaterials and Tissue Engineering Research Center Shanghai Institute of Ceramics Chinese Academy of Sciences  Shanghai China"},{"name":"State Key Laboratory of High Performance Ceramics Shanghai Institute of Ceramics Chinese Academy of Sciences  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6932-1900","authenticated-orcid":false,"given":"Hongxu","family":"Lu","sequence":"additional","affiliation":[{"name":"Biomaterials and Tissue Engineering Research Center Shanghai Institute of Ceramics Chinese Academy of Sciences  Shanghai China"},{"name":"State Key Laboratory of High Performance Ceramics Shanghai Institute of Ceramics Chinese Academy of Sciences  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,15]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms26178255"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/adfm.202010626"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bea.2024.100143"},{"key":"e_1_2_12_5_1","doi-asserted-by":"crossref","DOI":"10.1002\/adbi.202000247","article-title":"Mechanical Properties of Materials for Stem Cell Differentiation","volume":"4","author":"Han S.\u2010B.","year":"2020","journal-title":"Advanced Biosystems"},{"key":"e_1_2_12_6_1","doi-asserted-by":"crossref","first-page":"9086","DOI":"10.1038\/s41598-019-45352-3","article-title":"Soft Culture Substrates Favor Stem\u2010Like Cellular Phenotype and Facilitate Reprogramming of Human Mesenchymal Stem\/Stromal Cells (hMSCs) Through Mechanotransduction","volume":"9","author":"Gerardo H.","year":"2019","journal-title":"Scientific Reports"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41580\u2010024\u201000721\u20103"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2020.08.007"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2015.10.034"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2020.120572"},{"key":"e_1_2_12_11_1","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1007\/s42791-024-00076-y","article-title":"3D Scaffolds\u2010Specific Cellular Mechanoresponse as a Pivotal Regulating Factor in Tissue Engineering","volume":"6","author":"Ahn Y.","year":"2024","journal-title":"JMST Advances"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41580\u2010023\u201000583\u20101"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1409802111"},{"key":"e_1_2_12_14_1","doi-asserted-by":"crossref","first-page":"1316","DOI":"10.1073\/pnas.1319569111","article-title":"TRPV4\u2010Mediated Mechanotransduction Regulates the Metabolic Response of Chondrocytes to Dynamic Loading","volume":"111","author":"O'Conor C. 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