靶向性纳米级脂质体超声造影剂制备的方法学研究
发布时间：2018-12-09 08:35
【摘要】：目的：制备稳定的靶向性纳米级脂质体超声造影剂,为后期的靶向性超声诊断和治疗提供物质基础。 方法： 1.纳米级脂质体超声造影剂的制备：通过改变蛋黄卵磷脂、二硬脂酰磷脂酰胆碱、棕榈酸和胆固醇的比例,采用冷冻干燥-超声法,探讨制备纳米级脂质体超声造影剂的条件,观察和记录其物理特性参数,并通过兔左心腔造影研究来评价不同脂质体的显影效果。 2.靶向性纳米级脂质体超声造影剂的制备和鉴定：采用SPDP交联法将IL-8单克隆抗体与自制纳米级脂质体超声造影剂偶联,并通过玻片凝集实验和免疫荧光实验证明偶联是否成功。 结果： 1.通过单因素试验优化得到的纳米级脂质体超声造影剂的最佳膜材料组成为：卵磷脂(EPC)：二硬脂酰磷脂酰胆碱(DSPC):棕榈酸：胆固醇(CH)=17:60:8:15(mol)。经优化得到的最佳制备工艺参数为：水化温度55℃,100%功率超声,脉冲(1s/1s)作用5mmin。在此条件下制备得到的脂质体平均粒径为690+92nm、浓度4.12×109/ml、半衰期1.25h、Zeta电位-32.04mV、渗透压范围为278～310m0sm/kgH20。兔左心腔造影研究后经Qlab8.0软件分析得出自制纳米级脂质体超声造影剂能有效显影,且在体内显影时间较长,脂质体冻干粉于常温下放置1月后再复水仍具有反射性。造影过程及造影后1月内兔子无死亡现象,证明自制脂质体超声造影剂相对安全。 2.经玻片凝集实验和免疫荧光实验证明SPDP交联法能成功将IL-8偶联于自制脂质体超声造影剂上,偶联IL-8单克隆抗体后的脂质体平均粒径为692±81nm、浓度为3.05×109/ml、Zeta电位为-32.02mV、渗透压为275～314mOsm/kgH2O,与偶联前脂质体相比无明显变化；同样经兔左心腔造影研究证明偶联后的脂质体超声造影剂仍能有效显影,且相对安全。 结论：我们成功制备了形态规则、粒径均一、稳定性好,在动物体内具有较好显影效果和安全性的靶向性纳米级脂质体超声造影剂,为后期的靶向性超声诊断和治疗提供了物质基础。
[Abstract]:Aim: to prepare a stable ultrasound contrast agent for targeted nano-liposomes to provide material basis for the late diagnosis and treatment of targeted ultrasound. Methods: 1. Preparation of nanometer liposome ultrasound contrast agent: by changing the proportion of egg yolk lecithin, dicarbonyl phosphatidylcholine, palmitic acid and cholesterol, the conditions of preparing nanometer liposome ultrasound contrast agent were studied by freeze-drying ultrasonic method. The physical parameters were observed and recorded, and the development effects of different liposomes were evaluated by left ventricular angiography in rabbits. 2. Preparation and identification of targeted nano-liposome ultrasound contrast agent: IL-8 monoclonal antibody was coupled with self-made nano-liposome ultrasound contrast agent by SPDP crosslinking method. The results of slide agglutination and immunofluorescence test proved that the coupling was successful or not. Results: 1. The optimum membrane composition of nanoscale liposome ultrasound contrast agent optimized by single factor test is: lecithin (EPC): dicarbonyl phosphatidylcholine (DSPC): palmitic acid: cholesterol (CH) = 17: 60: 8: 15 (mol). The optimum preparation parameters were obtained as follows: hydration temperature 55 鈩，
本文编号：2369098
[Abstract]:Aim: to prepare a stable ultrasound contrast agent for targeted nano-liposomes to provide material basis for the late diagnosis and treatment of targeted ultrasound. Methods: 1. Preparation of nanometer liposome ultrasound contrast agent: by changing the proportion of egg yolk lecithin, dicarbonyl phosphatidylcholine, palmitic acid and cholesterol, the conditions of preparing nanometer liposome ultrasound contrast agent were studied by freeze-drying ultrasonic method. The physical parameters were observed and recorded, and the development effects of different liposomes were evaluated by left ventricular angiography in rabbits. 2. Preparation and identification of targeted nano-liposome ultrasound contrast agent: IL-8 monoclonal antibody was coupled with self-made nano-liposome ultrasound contrast agent by SPDP crosslinking method. The results of slide agglutination and immunofluorescence test proved that the coupling was successful or not. Results: 1. The optimum membrane composition of nanoscale liposome ultrasound contrast agent optimized by single factor test is: lecithin (EPC): dicarbonyl phosphatidylcholine (DSPC): palmitic acid: cholesterol (CH) = 17: 60: 8: 15 (mol). The optimum preparation parameters were obtained as follows: hydration temperature 55 鈩，
本文编号：2369098
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