Limin Song, Xinying Wang, Yiyao Pu, Xueyi Hu, Jun He, Rongrong Jin, Yu Nie
Abstract Cellular senescence significantly impairs tissue repair through disrupting tissue homeostasis and limiting regenerative capacity. Biomaterial based microenvironment modulation has emerged as a promising strategy to counteract senescence and promote tissue regeneration. While mechanical properties such as matrix stiffness are known to influence cell fate, whether different tissue-derived cells exhibit distinct mechanical requirements for senescence resistance remains poorly understood. Here, we used stiffness-tunable polyacrylamide hydrogels spanning physiologically relevant elastic moduli and systematically evaluated senescence responses in neuronal (SH-SY5Y), dermal fibroblast (NIH3T3), and osteoblast precursor (MC3T3-E1) cells. Our results revealed tissue-specific stiffness windows that maximally suppressed senescence, with optimal senescence resistance responses observed at 1 kPa, 10 kPa, and 250 kPa for SH-SY5Y, NIH3T3, and MC3T3-E1 cells, respectively. Mechanistically, integrin α1, a collagen-binding integrin, was identified as a key mechanosensor mediating these effects. Optimal stiffness conditions enhanced integrin α1-mediated mechanotransduction, cytoskeletal remodeling, and activation of PI3K–Akt and YAP signaling pathways, leading to the downregulation of senescence markers (p16, p21) and upregulation of proliferation related genes. These findings provide a mechanistic framework for the design of stiffness-matched biomaterials to counteract senescence and enhance tissue regeneration. Graphical Abstract