可降解PLGA纳米粒包裹IR780和siRNA载药体系用于光热和光动力治疗膀胱癌的相关研究

发布时间：2018-04-02 22:17

  本文选题：膀胱癌　切入点：光热治疗　出处：《南京大学》2017年硕士论文

【摘要】：目的通过PLGA装载IR780及siRNA,增强光热和光动力治疗对小鼠膀胱癌MB49细胞的体外抑制作用。包括(1)制备IR780-siRNA-PLGA纳米粒并对其进行表征;(2)评价IR780-siRNA-PLGA纳米粒对抑制小鼠MB49细胞的体外毒性研究。方法(1)制备IR780-siRNA-PLGA NPs并对其进行表征:采用超声复乳法制备IR780-siRNA-PLGA纳米粒。内水相:survivin-siRNA溶于缓冲溶液;油相:PLGA 1mg/ml及IR780 0.2mg溶于二氯甲烷;外水相:0.5%PVA缓冲溶液。超声功率360w,初乳时间120s,复乳时间180s,激光粒度分析仪测定纳米粒径,电镜观察IR780-siRNA-PLGA纳米粒形态;(2)IR780-siRNA-PLGA纳米粒对抑制小鼠MB49细胞的体外细胞毒性研究:根据加入不同的纳米材料和剂量分组,并使用MTT法检测所构建的纳米粒体系对细胞的杀伤情况。结果(1)最优的制备工艺为PLGA浓度为1mg/ml,超声功率为360w,复乳时间180s。采用最优设计得到的IR780载药率为:1.8%,粒径为131nm。纳米粒在电子显微镜下呈典型的球形,形态圆润,分散性较好。(2)MTT试验证明,纳米粒包裹IR780+siRNA(IR780剂量为5 μg/ml)组予激光照射后所测OD值低于对照组(P0.05)。纳米粒包裹IR780+siRNA(IR780剂量为10μg/ml)组无论是否予激光照射OD值均低于对照组(P0.05),但予激光照射后OD值下降更明显(P0.01)。结论(1)我们成功构建了IR780-siRNA-PLGA NPs,采用优化后的超声复乳法制备的纳米粒粒径、形态较为理想,分散性好;(2)IR780-siRNA-PLGA NPs可以降低IR780毒性并提高光热和光动力治疗对小鼠MB49细胞的杀伤作用。
[Abstract]:Objective to enhance the inhibitory effect of photothermal and photodynamic therapy on mouse bladder cancer MB49 cells in vitro by loading IR780 and siRNAs with PLGA.IR780-siRNA-PLGA nanoparticles were prepared and characterized. The toxicity of IR780-siRNA-PLGA nanoparticles in inhibiting mouse MB49 cells was evaluated in vitro.Methods 1) IR780-siRNA-PLGA NPs was prepared and characterized: IR780-siRNA-PLGA nanoparticles were prepared by ultrasonic emulsion method.The inner water phase: survivvin-siRNA was dissolved in buffer solution; the oil phase, IR780 1mg/ml and IR780 0.2mg were dissolved in dichloromethane; and the outer water phase was: 0.5 PVA buffer solution.Ultrasonic power 360 wk, colostrum time 120 s, resuscitation time 180 s, laser particle size analyzer to measure nanometer particle size,The cytotoxicity of IR780-siRNA-PLGA nanoparticles to MB49 cells in vitro was observed by electron microscope. The cytotoxicity of the nanoparticles was determined by MTT method according to the addition of different nanomaterials and dosages.Results the optimal preparation conditions were as follows: the concentration of PLGA was 1 mg / ml, the ultrasonic power was 360 ws, and the resuscitation time was 180 s.The drug loading rate of IR780 obtained by the optimum design is 1: 1.8 and the particle size is 131 nm.The results showed that the OD values of the nanoparticles coated with IR780 siRNA(IR780 at 5 渭 g / ml were lower than those in the control group (P 0.05).The OD value of 10 渭 g / ml IR780 siRNA(IR780 group was lower than that of the control group, but the OD value decreased more significantly after laser irradiation than that in the control group.Conclusion (1) IR780-siRNA-PLGA NPs were successfully constructed. The particle size and morphology of the nanoparticles prepared by the optimized ultrasonic reemulsion method were ideal. The dispersible IR780-siRNA-PLGA NPs could reduce the toxicity of IR780 and enhance the killing effect of photothermal and photodynamic therapy on mouse MB49 cells.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14
，

本文编号：1702367