诊断超声与超声治疗仪联合载阿霉素超声纳米泡作用于鼻咽癌细胞的体外实验研究

In vitro experimental study on the action of diagnostic ultrasound and ultrasonic therapeutic apparatus combined with doxorubicin-loaded ultrasound nanobubbles on nasopharyngeal carcinoma cells

目的:制备载阿霉素超声纳米泡(Dox-NBs),评价其基本特性并对比Dox-NBs联合诊断超声(DUS)与超声治疗仪(TUS)作用于鼻咽癌细胞的体外实验研究。方法:通过薄膜水化法、机械振荡法制备Dox-NBs并检测其基本物理特性、载药量及包封率;体外药物释放实验分为Dox-NBs组、Dox-NBs+DUS组、Dox-NBs+TUS组3组,利用多功能酶标仪检测Dox累积释放量;培养鼻咽癌5-8F细胞,按照不同的处理方法分为4组:对照组、Dox-NBs组、Dox-NBs+DUS组、Dox-NBs+TUS组,各组处理3 h后完全培养基培养3 d,细胞计数试剂盒(CCK-8)法检测各组细胞活性。结果:Dox-NBs的平均粒径为(286.93±14.45)nm,Zeta电位(-13.3±2.17)mV,平均载药量(37.0±3.34)μg/mg,包封率(36.30±1.92)%;体外药物释放结果显示,Dox-NBs+DUS组与Dox-NBs+TUS组分别在2 h与24 h Dox的累积释放量比较,差异均有统计学意义(P<0.05);CCK-8检测细胞活性显示,Dox-NBs组的细胞存活率高于Dox-NBs+DUS组与Dox-NBs+TUS组(P<0.01),Dox-NBs+DUS组的细胞存活率最低,Dox-NBs+DUS组对鼻咽癌细胞活性抑制作用最大。结论:Dox-NBs联合辐照可以提高药物释放量及协同抑制鼻咽癌细胞活性,DUS联合Dox-NBs作用效果优于TUS联合Dox-NBs,DUS可作为更普及、简便工具介导载药纳米泡的药物释放。

Objective:An in vitro experimental study that aims to prepare doxorubicin-loaded ultrasound nanobubbles(Dox-NBs),evaluate their basic properties and compare Dox-NBs combined with diagnostic ultrasound(DUS)and therapeutic ultrasound(TUS)acting on nasopharyngeal carcinoma cells.Methods:Dox-NBs was prepared by film hydration and mechanical oscillation and their basic physical properties,drug loading and encapsulation rate were detected.The in vitro drug release experiments involved three groups:Dox-NBs group,Dox-NBs+DUS group and Dox-NBs+TUS group.The cumulative release of Dox was detected by multimode microplate reader;the nasopharyngeal carcinoma 5-8F cells were cultured and divided into 4 groups according to different treatment methods:control group,Dox-NBs group,Dox-NBs+DUS group,Dox-NBs+TUS group,and each group was treated with complete medium for 3 h and then cultured in complete medium for 3 days.The cell counting kit-8(CCK-8)was used to detect the cell activity of each group.Results:The mean particle size of Dox-NBs was(286.93±14.45)nm,zeta potential(-13.3±2.17)mV,mean drug loading(37.0±3.34)μg/mg,and encapsulation rate(36.30±1.92)%.The in vitro drug release results showed that the differences were statistically significant(P<0.05)when comparing the cumulative release of Dox in Dox-NBs+DUS group and Dox-NBs+TUS group at 2 h and 24 h,respectively.CCK-8 assay of cell activity showed that the cell survival rate of Dox-NBs group was higher than that of Dox-NBs+DUS group and Dox-NBs+TUS group(P<0.01).The cell survival rate of Dox-NBs+DUS group was the lowest,and the Dox-NBs+DUS group had the greatest inhibitory effect on nasopharyngeal carcinoma cell activity.Conclusion:Combined irradiation of doxorubicin-loaded nanobubbles with ultrasound can improve drug release and synergistically inhibit nasopharyngeal carcinoma cell activity.The effect of DUS combined with Dox-NBs is superior to that of TUS combined with Dox-NBs,and diagnostic ultrasound can be used as a more accessible and easier tool to mediate drug release from drug-laden nanobubbles.