聚乳酸乙醇酸/RNAⅢ抑制肽缓释微球的血液相容性(英文)

Blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide microspheres

目的:评价聚乳酸乙醇酸/RNAⅢ抑制肽缓释微球的血液相容性。方法:①聚乳酸乙醇酸/RNAⅢ抑制肽微球制备:采用Fmoc法由C端至N端先合成粗品肽;采用反相液相色谱法对RNAⅢ抑制肽粗品进行纯化分析,按紫外吸收峰收集组分,冷冻干燥,得到RNAⅢ抑制肽纯品。再采用液相复乳法制备直径50～70μm的聚乳酸乙醇酸/RNAⅢ抑制肽微球。②浸提液制备:聚乳酸乙醇酸/RNAⅢ抑制肽微球粉末按1g/L在37℃无菌条件下用生理盐水浸提72h,制得浸提液原液;加入同体积的无菌生理盐水制得0.5g/L的浸提液。以溶血实验、凝血实验、血小板聚集实验,测定凝血酶原时间和活化部分凝血酶时间、观察对兔白细胞、红细胞和血小板的影响,对聚乳酸乙醇酸/RNAⅢ抑制肽微球血液相容性进行初步评价。结果:①浸提液原液和0.5g/L浸提液的溶血率分别为3.24%和2.67%,两者的溶血率均<5%,符合医用生物材料的溶血实验要求。②浸提液原液和0.5g/L浸提液对兔凝血时间,各时间点凝血酶原时间和活化部分凝血酶时间,白细胞、红细胞和血小板数量,对血小板聚集无明显影响。结论:聚乳酸乙醇酸/RNAⅢ抑制肽缓释微球具有良好的血液相容性。

OBJECTIVE：To evaluate blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide （PLGA/RIP） delayed release microspheres.METHODS：① Preparation of PLGA/RIP microspheres：The solid-phase synthesis （Fmoc） method was used to synthesize RIP crude sample from C end to N end;the synthesized crude peptide was purified by the reverse phase high performance liquid chromatography.According to UV absorption peak,the components were collected and freeze-dried,to obtain RIP purifications.Then liquid-phase multiple emulsion method was used to prepare PLGA/RIP microspheres at the diameter of 50-70 μm.② Preparation of eluent：The PLGA/RIP microsphere powders were eluted with sterile physiological saline at 37 ℃,to prepare 1 g/L eluent;then 0.5 g/L eluent was obtained adding equal volume of sterile physiological saline.The hemolysis test,blood clotting test,and platelet aggregation test were conducted to measure prothrombin time and activated partial thromboplastin time,to observe the influence on rabbit leucocytes,erythrocytes and thrombocytes,and to preliminarily evaluate the blood compatibility of PLGA/RIP microspheres.RESULTS：①The haemolysis rates of eluent stock solution and 0.5 g/L eluents were 3.24% and 2.67% respectively,which were in coincidence with the criteria of medical biomaterials,less than 5%.② The eluent stock solution and 0.5 g/L eluents of PLGA/RIP microspheres had no significant effect on rabbit clotting time,prothrombin time and activated partial thromboplastin time,the number of rabbit leucocytes,erythrocytes and thrombocytes,as well as platelet aggregation.CONCLUSION：PLGA/RIP delayed release microspheres have a good blood compatibility.