發(fā)布時間：2018-12-09 08:35

【摘要】：目的：制備穩(wěn)定的靶向性納米級脂質體超聲造影劑,為后期的靶向性超聲診斷和治療提供物質基礎。 方法： 1.納米級脂質體超聲造影劑的制備：通過改變蛋黃卵磷脂、二硬脂酰磷脂酰膽堿、棕櫚酸和膽固醇的比例,采用冷凍干燥-超聲法,探討制備納米級脂質體超聲造影劑的條件,觀察和記錄其物理特性參數(shù),并通過兔左心腔造影研究來評價不同脂質體的顯影效果。 2.靶向性納米級脂質體超聲造影劑的制備和鑒定：采用SPDP交聯(lián)法將IL-8單克隆抗體與自制納米級脂質體超聲造影劑偶聯(lián),并通過玻片凝集實驗和免疫熒光實驗證明偶聯(lián)是否成功。 結果： 1.通過單因素試驗優(yōu)化得到的納米級脂質體超聲造影劑的最佳膜材料組成為：卵磷脂(EPC)：二硬脂酰磷脂酰膽堿(DSPC):棕櫚酸：膽固醇(CH)=17:60:8:15(mol)。經(jīng)優(yōu)化得到的最佳制備工藝參數(shù)為：水化溫度55℃,100%功率超聲,脈沖(1s/1s)作用5mmin。在此條件下制備得到的脂質體平均粒徑為690+92nm、濃度4.12×109/ml、半衰期1.25h、Zeta電位-32.04mV、滲透壓范圍為278～310m0sm/kgH20。兔左心腔造影研究后經(jīng)Qlab8.0軟件分析得出自制納米級脂質體超聲造影劑能有效顯影,且在體內(nèi)顯影時間較長,脂質體凍干粉于常溫下放置1月后再復水仍具有反射性。造影過程及造影后1月內(nèi)兔子無死亡現(xiàn)象,證明自制脂質體超聲造影劑相對安全。 2.經(jīng)玻片凝集實驗和免疫熒光實驗證明SPDP交聯(lián)法能成功將IL-8偶聯(lián)于自制脂質體超聲造影劑上,偶聯(lián)IL-8單克隆抗體后的脂質體平均粒徑為692±81nm、濃度為3.05×109/ml、Zeta電位為-32.02mV、滲透壓為275～314mOsm/kgH2O,與偶聯(lián)前脂質體相比無明顯變化；同樣經(jīng)兔左心腔造影研究證明偶聯(lián)后的脂質體超聲造影劑仍能有效顯影,且相對安全。 結論：我們成功制備了形態(tài)規(guī)則、粒徑均一、穩(wěn)定性好,在動物體內(nèi)具有較好顯影效果和安全性的靶向性納米級脂質體超聲造影劑,為后期的靶向性超聲診斷和治療提供了物質基礎。
[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，

本文編號：2369098