摘要
目的:制备一款由小胶质细胞膜(microglia cell membrane,MiCM)伪装和装载相变材料全氟己烷(perfluorohexane,PFH)的多功能脂质体(liposome,Lipid)纳米粒(MiCM@PFH@Lipid),并研究其基本性质,检测特异性靶向卒中后的损伤部位和实现栓塞部位血管再通的能力。方法:采用旋蒸法制备脂质体球载体,通过超声融合法将PFH和MiCM融合至脂质体形成MiCM@PFH@Lipid,透射电镜(transmission electron microscope,TEM)观察纳米粒的形样貌,马尔文粒径分析仪检测纳米粒第1天的粒径分布,和其在第1、3、5、7 d的电位分布,检测其稳定性。Western blot和考马斯亮蓝染色检测MiCM的包被情况,通过2,3,5-三苯基氯化四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)、动物行为学测试检测小鼠脑功能状态。结果:表征结果显示,MiCM@PFH@Lipid呈球形,粒径为(234.800±3.143)nm,平均电位为(-33.433±1.550)mV,具有良好的分散性,在7 d内具有较好的稳定性,MiCM成功包被在纳米粒上。小鼠离体荧光结果显示,纳米粒可以快速地在损伤部位蓄积,并且与非靶向组比较差异有统计学意义(P<0.05)。动物行为学实验和脑TTC染色结果显示,MiCM@PFH@Lipid纳米粒可以实现血管再通,恢复部分神经功能(P<0.05)。病理学检查显示,纳米粒治疗后没有对重要脏器造成明显损伤,而血常规检测与正常对照组并未见明显异常(P<0.05)。结论:本实验成功制备了多功能纳米粒MiCM@PFH@Lipid,其可以靶向至缺血性卒中后脑损伤区域并破坏血栓,用于缺血性卒中疾病的治疗。
Objective:To prepare MiCM@PFH@Lipid,a multifunctional liposome(Lipid)nanoparticles camouflaged with microglial cell membrane(MiCM)and loaded with the phase change material Perfluorohexane(PFH),to study its basic properties,and to test its ability to specifically target the site of injury after stroke and to achieve revascularization at the site of thrombus.Methods:Liposome sphere carriers were prepared by rotary evaporation method.MiCM and PFH were fused with the liposome to form MiCM@PFH@Lipid using the ultrasonic fusion method.Transmission electron microscopy was used to observe the morphology of the nanoparticles.The par⁃ticle size distribution of nanoparticles at day 1 and their potential distribution at days 1,3,5,and 7 were measured using a Malvern par⁃ticle size analyzer to detect their stability.Western blot and Coomassie brilliant blue staining were used to detect the encapsulation of nanoparticles with MiCM.The functional state of the mouse brain was detected using brain staining with 2,3,5-triphenyltetrazolium chloride and animal behavior test.Results:MiCM@PFH@Lipid nanoparticles were spherical in shape,with a particle size of(234.800±3.143)nm and an average potential of(-33.433±1.550)mV.The nanoparticles were well dispersed and stable within 7 days.MiCM was successfully encapsulated on the nanoparticles.Mouse ex vivo fluorescence image showed that the nanoparticles could rapidly accumulate at the site of injury,which was significantly different as compared with the non-targeted group(P<0.05).Animal behavior test and brain staining showed that MiCM@PFH@Lipid nanoparticles could achieve vascular recanalization and partly restore neurological functions(P<0.05).Pathological examination showed that there was no significant damage to important organs after nanoparticle treatment.Routine blood test showed no significant abnormality as compared with the normal control group(P<0.05).Conclusion:In this study,multifunctional nanoparticles(MiCM@PFH@Lipid)were successfully prepared.These nanoparticles can ta
作者
成睿琪
张融融
秦新月
Cheng Ruiqi;Zhang Rongrong;Qin Xinyue(Department of Neurology,The First Affiliated Hospital of Chongqing Medical University)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2024年第7期853-858,共6页
Journal of Chongqing Medical University
