Injectable platelet-mimicking silk protein-peptide conjugate microspheres for hemostasis modulation and targeted treatment of internal bleeding.

Abstract

Uncontrolled deep bleeding, commonly encountered in surgical procedures, combat injuries, and trauma, poses a significant threat to patient survival and recovery. The development of effective hemostatic agents capable of precisely targeting trauma sites in deep tissues and rapidly halt bleeding remains a considerable challenge. Drawing inspiration from the natural hemostatic cascade, we present platelet-like microspheres composed of silk fibroin (SF) and thrombus-targeting peptides, engineered to mimic natural platelets for rapid hemostasis in vivo. These peptide/SF hemostatic microspheres, formulated using a freezing self-assembly technology, closely resemble natural platelets in terms of size, shape, and zeta potential. Moreover, they exhibit favorable cytocompatibility, hemocompatibility, and anti-cell adhesion. Assessment of fibrin polymerization revealed that these hemostatic microspheres possessed enzymatic physiological functions, similar to activated platelets, facilitating platelet adhesion, fibrin binding, and wound-triggered hemostasis. Notably, these hemostatic microspheres rapidly target the bleeding site in vivo within 5 min, with minimal dispersion elsewhere, persisting after blood clot formation. Furthermore, these microspheres exhibit favorable metabolic kinetics, with 71% degradation occurring within one-day post-subcutaneous injection. Histological assessment revealed well-preserved organ structures and minimal inflammatory responses at 14 d post-injection, supporting their long-term biocompatibility. Importantly, they can be injected and targeted into damaged blood vessels, selectively binding to fibrin and forming blood clots within 2 min, resulting in a 74% reduction in bleeding volume compared to SF microspheres alone. Therefore, these injectable SF-based hemostatic microspheres emerge as promising candidates for future rapid hemostasis in tissue injuries.