Chirality Regulates Stem Cell Fate and Promotes Corneal Epithelial Regeneration via Manipulating Notch Pathway

Abstract The differentiation and stemness maintenance of stem cells are the core topics in cell biology and regenerative medicine, involving cell fate determination, developmental regulation and tissue regeneration. Chirality is an essential factor influencing multiple biological processes, including protein interactions, stem cell development and disease pathogenesis. However, its roles in regulating stem cells fate, especially limbal epithelial stem cells (LESCs), remain elusive. Herein, it is first discovered that right‐handed chiral hydrogel (DH) enhanced LESCs proliferation, migration, and differentiation into corneal epithelial cells, while left‐handed chiral hydrogel (LH) can partially preserve LESCs stemness. Further in vivo experiments demonstrated that 3D DH effectively accelerated corneal wound healing process, inhibited both inflammation and vascularization in a partial limbal stem cell deficiency model. Mechanistically, DH activates Notch signaling by increasing its stereo‐affinity to Notch1, facilitating Notch intracellular domain release and HES1 transcription, thereby directing LESCs fate. Collectively, this work highlights the novel role of chirality in LESCs fate determination and confirms DH as a drug‐free, effective approach for corneal epithelial regeneration, offering a new direction for regenerative medicine and tissue engineering.