Yanan Li, Kai Xiang, Jun Ma, Qinao Wang, Sanwei Hao, Fangqing Wang, Ge Yang, Degang Li, Hailin Cong, Bing Yu
Current hemostatic materials often face challenges associated with delayed action and insufficient stability on wet bleeding surfaces. Here, we designed and screened three short self-assembling peptides by integrating amphiphilic motifs, β-sheet-associated assembly, and intrinsic antioxidant properties. Through systematic comparison of sequence variants, peptide P3 (PYFKWS) was identified as the optimized candidate, exhibiting enhanced hemostatic and wound-healing performance. Structural interrogation revealed that P3 adopts a β-sheet-stabilized amphiphilic architecture, enabling rapid in situ hydrogel formation within physiological milieu. Molecular simulations further indicated that the superior assembly behavior of P3 was associated with a more favorable intermolecular hydrogen-bonding organization rather than the total hydrogen-bond number alone. This rapid self-assembly facilitates hemostasis through hydrogel-mediated physical sealing, erythrocyte adsorption, and platelet recruitment. Augmenting this structural advantage, the indole moieties and phenolic hydroxyl constituents within P3 collectively confer notable radical-scavenging capacity, contributing to oxidative stress modulation. This effect significantly dampens pro-inflammatory responses (TNF-α: ↓63.2%; IL-6: ↓58.7% vs control) and promotes a microenvironment favorable for tissue repair. Validated in mouse tail bleeding, rat liver injury, and full-thickness wound models, P3 significantly reduced hemostasis time and blood loss while accelerating wound closure. This study demonstrates a minimal peptide design strategy that integrates rapid hemostasis with oxidative stress regulation and tissue repair, providing insights into the development of multifunctional peptide-based hemostatic materials.