Protonated-chitosan sponge with procoagulation activity for hemostasis in coagulopathy.

Hemostatic materials are essential for managing acute bleeding in medical settings. Chitosan (CS) shows promise in hemostasis but its underlying mechanism remains incompletely understood. We unexpectedly discovered that certain protonated-chitosan (PCS) rapidly assembled plasma proteins to form protein membrane (PM) upon contact with platelet-poor plasma (PPP). We hypothesized that the novel observation was intricately related to the procoagulant effect of chitosan. Herein, the study aimed to elucidate the conditions necessary and mechanism for PM formation, identify the proteins within the PM and PCS's procoagulant action at the molecule levels. We confirmed that the amount of -NH3 + groups (>4.9 mmol/g) on PCS molecules played a crucial role in promoting coagulation. The -NH3 + group interacted with blood's multiple active components to exert hemostatic effects: assembling plasma proteins including coagulation factors such as FII, FV, FX, activating blood cells and promoting the secretion of coagulation-related substances (FV, ADP, etc) by platelets. Notably, the hemostatic mechanism can be extended to protonated-chitosan derivatives like quaternized, alkylated, and catechol-chitosan. In the blood clotting index (BCI) experiment, compared to other groups, PCS95 achieved the lowest BCI value (∼6 %) within 30 s. Protonated-chitosan exhibited excellent biocompatibility and antibacterial properties, with PCS95 demonstrating inhibition effectiveness of over 95 % against Escherichia coli (E.coil) and Staphylococcus aureus (S. aureus). Moreover, PCS performed enhanced hemostatic effectiveness over chitosan-based commercially agents (Celox™ and ChitoGauze®XR) in diverse bleeding models. In particular, PCS95 reduced bleeding time by 70 % in rabbit models of coagulopathy. Overall, this study investigated the coagulation mechanism of materials at the molecular level, paving the way for innovative approaches in designing new hemostatic materials.

Calcium Ion-Coupled Polyphosphates with Different Degrees of Polymerization for Bleeding Control.

The development of efficient hemostatic materials is crucial for achieving rapid hemorrhage control and effective wound healing. Inorganic polyphosphate (polyP) is recognized as an effective modulator of the blood coagulation process. However, the specific effect of polyP chain length on coagulation is not yet fully understood. Furthermore, calcium ions (Ca2+) are essential for the coagulation process, promoting multiple enzyme-catalyzed reactions within the coagulation cascade. Hence, calcium ion-coupled polyphosphate powders with three different degrees of polymerization (CaPP-n, n = 20, 50, and 1500) are synthesized by an ion-exchange reaction. CaPP exhibits a crystalline phase at a low polymerization degree and transitions to an amorphous phase as the polymerization degree increases. Notably, the addition of Ca2+ enhances the wettability of polyP, and CaPP promotes hemostasis, with varying degrees of effectiveness related to chain length. CaPP-50 exhibits the most promising hemostatic performance, with the lowest blood clotting index (BCI, 12.1 ± 0.7%) and the shortest clotting time (302.0 ± 10.5 s). By combining Ca2+ with polyP of medium-chain length, CaPP-50 demonstrates an enhanced ability to accelerate the adhesion and activation of blood cells, initiate the intrinsic coagulation cascade, and form a stable blood clot, outperforming both CaPP-20 and CaPP-1500. The hemostatic efficacy of CaPP-50 is further validated using rat liver bleeding and femoral artery puncture models. CaPP-50 is proven to possess hemostatic properties comparable to those of commercial calcium-based zeolite hemostatic powder and superior to kaolin. In addition, CaPP-50 exhibits excellent biocompatibility and long-term storage stability. These results suggest that CaPP-50 has significant clinical and commercial potential as an active inorganic hemostatic agent for rapid control of bleeding.