二氢卟吩和全氟己烷共载脂质体的制备及其体外性质

Preparation and in vitro characterizations of liposomes co-loading chlorin e6 and perfluorohexane

目的制备共载光敏剂二氢卟吩(Ce6)和氧载体全氟己烷(PFH)的长循环脂质体,对其进行表征,并进行小鼠黑色素瘤细胞摄取的初步研究。方法采用薄膜超声分散法制备Ce6和PFH共载的PEG化长循环脂质体(Ce6-PFH@PEG-Lip),运用单因素筛选法依次筛选探头超声功率、有机溶剂种类、卵磷脂与胆固醇及DSPE-PEG的投料比、PFH和Ce6用量、大豆油和e80磷脂用量,以及制备工艺;并对所制得纳米粒的粒度分布、Zeta电位、形态、包封率和载药量进行表征。同时应用流式细胞术研究细胞摄取情况。结果筛选出最优处方,成功制备了均一稳定的Ce6-PFH@PEGLip,其粒径为133.9±3.1 nm,PDI为0.211,Zeta电位为-39.1±2.3 m V,对Ce6的包封率为98.09%±1.6%。DSPEPEG的加入使得制剂更加稳定,提高了细胞对纳米粒的摄取效率。结论Ce6和PFH具有良好的相容性,能应用简单的处方和薄膜超声分散法制成共载长循环脂质体,在缺氧性肿瘤光动力治疗的应用具有广阔前景。

OBJECTIVE To prepare long-circulating liposomes co-loading photosensitizer chlorin e6(Ce6)and oxygen carrier perfluorohexane(PFH)(Ce6-PFH@PEG-Lip),and to characterize the liposomes and investigate them uptaken by mouse melanoma cells.METHODS Ce6-PFH@PEG-Lip were prepared by thin-film ultrasonic dispersion method.Preparation conditions were selected by single factor screening method,factors including ultrasonic power,organic solvent used and usage of lecithin,cholesterol and DSPE-PEG,as well as amount of PFH,Ce6,soybean oil and E80 phospholipid.Particle size,zeta potential,morphology,entrapment efficiency and drug loading of the liposomes were then characterized.Additionally,cell uptake was studied by flow cytometry.RESULTS The optimal formulation was selected and Ce6-PFH@PEG-Lip were successfully prepared.Particle size of Ce6-PFH@PEG-Lip was 133.9±3.1 nm with PDI of 0.211;Zeta potential was-39.1±2.3 m V;and entrapment efficiency of Ce6 was 98.09%±1.6%.The addition of DSPE-PEG stabilized the preparation,leading to enhanced cell uptake efficiency of the prepared liposomes.CONCLUSION Ce6 and PFH have good compatibility and can be used to prepare co-loading long-circulating liposomes by simple prescription and thin-film ultrasonic dispersion method,which processes large possibility in possible application in oxygen-deficient tumor photodynamic therapy for oxygen-deficient tumors.