| Background:After the absorbable hemostatic material comes into contact with human wounds,it can quickly stop bleeding by adsorbing platelets and activating the coagulation system.After being degraded and absorbed,it will enter the blood circulation and participate in body metabolism.If its biosafety is low and biocompatibility is poor,it will have many adverse effects on the body.Therefore,preclinical evaluations such as safety and effectiveness are indispensable before materials undergo clinical trials and then transformed into products.Preclinical evaluation of safety and effectiveness can be carried out through animal experiments and in vitro experiments.Animal experiments are divided into local evaluation and systemic evaluation.Local evaluation is to observe whether the wound using the material has inflammation,infection,ulceration and other reactions;systemic evaluation through the evaluation of blood compatibility,organ effects,etc.At present,the National Medical Products Administration has formulated guidelines for the registration of absorbable hemostatic materials,which clearly stipulates the requirements for absorbable hemostatic products.Before hemostatic materials conforming to the guidelines are transformed,the materials need to be evaluated according to this rule.Objective:Based on our preliminary study on the biosafety of the absorbable macroporous polysaccharides composite hemostatic materials(MPCHM),supplement the hemostatic efficacy in vivo and in vitro studies of the hemostatic materials,determine whether it causes an animal immunogenic reaction,explore its body Internal and external hemostatic performance and hemostatic mechanism,compared with the commercial compound microporous polysaccharide hemostatic powder(CMPHP),conpleted the preclinical biosafety and effectiveness evaluation of MPCHM,compile all the data into compliance with the National Medical Products Administration(NMPA)required product transformation registration information to assist in the registration of the materials.Methods:(1)Evaluation of in vitro hemostatic performance:The in vitro hemostatic properties of absorbable macroporous polysaccharides hemostatics material were determined,including water absorption,hemolysis rate coagulation index,four items of blood coagulation(APTT,TT,FIB,PT),blood cell aggregation,platelet adhesion and platelet factor fourth content.And compared with commercial compound microporous polysaccharide hemostatic powder to determine whether the safety of the material is suitable for hemostasis in vivo.(2)Evaluation of immunogenicity:The immunogenicity evaluation test was carried out in BLAB/c mice.Absorbable macroporous polysaccharides composite hemostasis material and commercial compound microporous polysaccharide hemostatic powder were implanted subcutaneously on the back of mice.The changes of organ coefficient,immunoglobulin content and spleen lymphocyte typing were detected at 2 and 4 weeks after implantation.(3)Evaluation of hemostatic effect of femoral artery dissection:Construct a rabbit femoral artery disconnection model,and the absorbable macroporous polysaccharides composite hemostasis material and commercial compound microporous polysaccharide hemostatic powder were used to stop the bleeding.Record the success rate of hemostasis,postoperative survival rate,hematological changes and changes of blood biochemical indexes.Results:(1)In vitro hemostasis properties:The water absorption rate of composite macroporous polysaccharides absorbable hemostatic materials was(4036.67±30.35)%,and that of CMPHP group was(2327.50±45.21)%.The water absorption rate of MPCHM group was higher than that of CMPHP group(P<0.05).The absorbance of blood cell aggregation in MPCHM group,CMPHP group and blank group was 0.37±0.05%,0.78 ±0.33%and 1.07 ±0.23%,respectively.The absorbance in blank group was higher than that in MPCHM group and CMPHP group(P<0.05).The absorbance in MPCHM group was significantly lower than that in CMPHP group(P<0.05).The platelet content of MPCHM group was 11.40±4.39,the platelet content of blank group was 16.80±2.49,which was lower than that of blank group(P<0.05),and lower than that of CMPHP group(P<0.05).The platelet content of CMPHP group was 15.67±2.07,and that of blank group was 16.80±2.49,blank group was significantly higher than that of CMPHP group(P<0.05).Compared with CMPHP,MPCHM has a stronger ability to absorb blood,it can promote blood cell aggregation and platelet adhesion.(2)Immunogenicity:Under these experimental conditions,the absorbable macroporous polysaccharides composite hemostatic materials did not cause abnormal organ coefficient,or increase of immunoglobulin IgG,IgM,and abnormal proportion of spleen lymphocytes during the experiment.At 2 weeks,the organ coefficient,immunoglobulin and lymphocytes of the CMPHP group were no different from those of the sham operation group,but at 4 weeks,the proportion of B lymphocytes of the CMPHP group was higher than that of the sham operation group(P<0.05,the difference was statistically significant).Therefore,the experimental observation period should be extended.The time when the contents of immunoglobulin and splenic lymphocytes in the CMPHP group returned to normal was determined,and it was confirmed that no immune response occurred in the MPCHM group at the later stage of the experiment.(3)Hemostasis effect of femoral artery dissection:The hemostatic success rate of absorbable macroporous polysaccharides composite hemostasis material was higher than that of commercial composite microporous polysaccharide hemostatic powder.The results of hematological and blood biochemical tests showed that the safety of composite microporous polysaccharide hemostatic powder was not lower than that of commercial compound microporous polysaccharide hemostatic powder,tissue section staining showed that the wound recovery of the absorbable macroporous polysaccharides composite hemostasis material was faster than that of the compound microporous polysaccharide hemostatic powder group.Conclusion:The hemostasis effectiveness in vivo and in vitro of absorbable macroporous polysaccharides hemostatics material is not lower than that of commercial compound microporous polysaccharide hemostatic powder,which has the function of promoting blood cell aggregation and platelet adhesion,fast blood absorption rate and large absorption volume,hemostasis is stronger in arteriolar bleeding.It can be used as a traumatic and emergency hemostatic product for body surface and visceral hemostasis. |
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