RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层的血液相容性

Blood compatibility of RNAⅢ inhibiting peptide/bioinspired phosphorylcholine-based cytomembrane coating

背景:将葡萄球菌的全面抑制剂RNAⅢ抑制肽和稳定、无毒、具有良好血液相容性的磷酸胆碱结合,制备出磷酸胆碱基细胞膜仿生药物涂层,具有良好的药物传递性能,可减少生物材料表面细菌的黏附和生物被膜的形成,从而减少内置物的感染。目的:通过实验评价RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层的血液相容性。设计、时间及地点:随机对照动物实验,于2005-10/2007-10在解放军总医院临床药理研究所及医学动物实验中心完成。材料:采用有机化学固相合成Fmoc法合成RNAⅢ抑制肽,以甲基丙烯酸十八酯、甲基丙烯酸羟丙酯、甲基丙烯酸硅基丙酯和2-甲基丙烯酰氧基-2-三甲基氨基乙基磷酸酯为单体通过自由基溶液聚合合成可交联四元共聚物,将洁净的316L不锈钢片浸渍于5g/L的四元共聚物四氢呋喃和质量分数0.1的RNAⅢ抑制肽混合溶液中,制备聚合物涂层。方法:将聚合物涂层按1cm2/mL在37℃无菌条件下用生理盐水洗提72h,制得洗提液原液;加入同体积的无菌生理盐水制得50%洗提液。根据国家标准GB/T16886-1进行兔溶血实验、凝血实验、血小板聚集实验。主要观察指标:红细胞溶血率;凝血时间、凝血酶原时间、活化部分凝血酶时间;白细胞、红细胞和血小板数量;血小板聚集情况。结果:溶血实验结果显示RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层洗提液原液和50%洗提液的溶血率分别为4.88%和3.66%,两者的溶血率均<5%,符合医用生物材料的溶血实验要求。RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层洗提液原液和50%洗提液对兔全血凝固时间、凝血酶原时间和活化部分凝血酶时间与生理盐水阴性对照组比较差异无显著性(P>0.05);RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层洗提液原液组白细胞、红细胞和血小板数量及血小板聚集时间与50%洗提液组比较差异无显著性(P>0.05)。结论:RNAⅢ抑制肽/磷酸胆碱基细胞膜仿生药物涂层具有良好的血液相容性。

BACKGROUND： Combined RNAIII inhibiting peptide with bioinspired phosphorylcholine can prepare RNAⅢ inhibiting peptide/bioinspired phosphorylcholine-based cytomembrane coating （RIP/PC）, which has excellent drug transmissibility. RIP/PC can decrease the adhesion and formation of bacterial on the surface of biomaterial materials, accordingly, can decrease the infection of the implantation. OBJECTIVE： To evaluate blood compatibility of RIP/PC. DESIGN, TIME AND SETTING： Randomized control animal experiment was performed in the Experiment Animal Center and Clinical Institute of Pharmacologic Research of General Hospital of Chinese PLA from October 2005 to October 2007. MATERIALS： The solid phase Fmoc method was used to synthesize RIP. The cross link quaternionic polymer was synthesized from MPC, SMA, HPMA and TSMA. The stainless steel plate with 316 L was immersed into mixed solution of 5 g/L PC and 0.1 mass fraction of RIP to prepare coating and eluent. METHODS： Under aseptic condition, 1 cm2/mL coating was eluted with sodium chloride at 37 ~C for 72 hours to harvest 100% eluent, iso volume stroke-physiological saline solution was added into 100% eluent to get 50% eluent of RIP/PC. Then, haemolysis test, blood coagulation experiment, and platelet aggregation test was performed, respectively. MAIN OUTCOME MEASURES： The rate of haematolysis of erythrocyte, whole blood clotting time （WBCT）, prothrombin time （PT）, activation partial thrombin time （APTT）, counts of lencocyte, akaryocyte and plastocyte, as well as platelet aggregation. RESULTS： Haemolysis test showed that tile rate of haemolysis of 100% and 50% eluent of RIP/PC were 4.88% and 3.66%, respectively, which were coincidence with the criteria of biomaterial materials. There were no significantly differences between the 100% and 50% eluent of RIP/PC on WBCT, PT and APTT （P 〉 0.05）, the counts of lencocyte, akaryocyte and plastocyte, as well as and platelet aggregation （P 〉 0.05）. CONCLUSION： RIP/PC has good blood compatibility.