Porous chitosan microspheres for application as quick in vitro and in vivo hemostat

Controlling massive hemorrhage is of great importance to lower transfusional medical cost, and to reduce death and mobility rate in battlefield and civilian accidents. We reported the fabrication of porous chitosan microspheres (CSMS) with tunable surface pore size by microemulsion combined with thermally induced phase separation technique, and its application as a quick hemostat. Their hemostatic property was characterized by blood clotting kinetics, adherence interaction between red blood cells/platelets and CSMS, in vitro and in vivo hemostasis by rat tail amputation and liver laceration models, and histological analysis. Their density, surface area, porosity, water absorption ratio were 0.04-0.06g/cm³, 28.2-31.5m²/g, 98%, and 15.5-23.2g/g, respectively. The surface pore was controlled to be smaller than 2.0μm. The porous CSMS showed increasing hemostatic efficacy with decreasing surface pore size. Compared to the conventional compact chitosan particles (CCSP), the porous CSMS had much improved in vitro and in vivo hemostatic potential with respect to formation of blood clot, hemostatic time, and blood loss. For instance, the hemostatic time and blood loss of CSMS in the rat liver laceration model were down to respectively 70s and 0.026g from 175s and 0.28g of CCSP. Histological examination showed that application of porous CSMS to liver laceration caused no destruction of underlying hepatocytes, inflammatory reaction, and thermal injury to liver tissue. The porous CSMS is a biodegradable, quick and safe hemostat, which can be used in various wounds including complex and non-compressive ones.