Directing Neuronal Differentiation via Calcium‐Chirality Synergy in Supramolecular Nanofibers

ABSTRACT While chiral supramolecular nanofibrous networks (NFNs) show promise in regulating cell fate, precisely directing differentiation pathways remains challenging. This study systematically investigates the influence of various chiral supramolecular NFNs on neural cell differentiation. A series of chiral supramolecular NFNs was synthesized using L/D‐amino acids and their derivatives as building blocks, including L/D‐lysine (L/D‐Lys), L/D‐histidine (L/D‐His), L/D‐arginine (L/D‐Arg), and L/D‐glutamic acid derivatives (Fmoc‐L/D‐Glu). The regulatory effects of these NFNs on the NG108‐15 were evaluated during the differentiation process. Our findings demonstrate that the chiral microenvironment established by D‐Lys‐gel significantly enhances NG108‐15 differentiation and correlates with robust activation of the Ca 2+ signaling pathway. To further elucidate the relationship between supramolecular chirality and metal ions, we synthesized chiral supramolecular NFNs from L/D‐Lys‐Ca 2+ ‐gel and L/D‐Lys‐Mg 2+ ‐gel. Notably, the supramolecular NFNs derived from D‐Lys‐Ca 2+ ‐gel exhibited superior capacity to promote neurite outgrowth and maturation compared to their L‐enantiomer and Mg 2+ ‐incorporated counterparts. Proteomics analysis revealed that the chiral supramolecular NFNs obtained from D‐Lys‐Ca 2+ ‐gel enhance neuronal differentiation by upregulating key effectors involved in both axon guidance and calcium signaling pathways, particularly the Camkk2‐Tiam1 cascade. This study provides valuable insights into the design of chiral biomaterials and proposes novel strategies for neural regeneration and tissue engineering.