The Hemostatic Effect Of Hydrophobically Modified Chitosan From Catharsius Molossus Linnaeus And Its Mechanism

BackgroundIt is reported that each type of injury occurs hundreds of millions of times every year.Excessive loss of blood is the biggest cause of death.Therefore,first aid is the first step and the key to treatment.Traditional first aid hemostasis materials such as ordinary gauze,tourniquet or bandages have limited hemostasis and they are difficult to adapt to complex bleeding conditions,thus requiring more flexible,simple and efficient hemostatic materials.The earliest reaction procedure for hemostasis after trauma is vasoconstriction,which slows blood flow,and promotes blood clotting and hemostasis.Hemostatic materials work,usually by aggregating platelets and activating the coagulation system.However,there is currently no condition that can fully satisfy the ideal first aid hemostatic agent.Chitosan is one of the most promising hemostatic agents in emergency due to its superior biocompatibility,biodegradability and safety,as well as its antibacterial properties and low cost.PurposeIn this study,experiments were carried out with three aspects: preparation and characterization of hydrophobically modified chitosan from Catharsius molossus Linnaeus(CHMC),investigation of hemostatic effect and mechanism investigation.The feasibility of its as a hemostatic material was revealed,and the factors affecting hemostasis were explored.The study will provide base for the application of hemostatic material.MethodsThe CHMC was prepared by the method of independent laboratory patent,and the product was characterized.The physical and chemical analysis was carried out by elemental analyzer and contact angle,and the structure was analyzed by infrared spectrometer and nuclear magnetic spectrometer.The biological evaluation was performed by CCK8 and flow cytometry.The hemostatic effect of CHMC was studied by coagulation and hemostatic experiments.Based on the above,the relationship between the main components of blood,such as red blood cells,nucleic acids,proteins and inorganic salts,was further studied to clarify the mechanism of coagulation.Results and ConclusionsThe yield of chitosan from Catharsius molossus Linnaeus(CCS)was about 17.27% and the yield of CHMC was about 91.54%.Compared with hydrophobic commercially available chitosan,CHMC has good solubility,low viscosity and high degree of substitution.Contact angle,infrared,and nuclear magnetic results confirmed the successful synthesis of hydrophobic products.The results of biological evaluation showed that the CHMC had no toxic effect on the cells.The mouse experiments and coagulation experiments showed that the CHMC had an excellent hemostatic effect.The investigation of hemostatic mechanism showed that CHMC had a certain effect on substances on the cell membrane.It could enrich cells,and change the permeability of cell membranes,and firmly link individual cells.There was no agglutination between nucleic acid and protein and CHMC.The inorganic salt ions could form a strong ionic bond with the CHMC,and form an ionic gel.Therefore,CHMCcould bind to the two most important substances in the blood,thus formed blood clots instead of relying on the waterfall coagulation mechanism.