本文選題：膀胱癌　切入點(diǎn)：光熱治療　出處：《南京大學(xué)》2017年碩士論文

【摘要】：目的通過PLGA裝載IR780及siRNA,增強(qiáng)光熱和光動(dòng)力治療對(duì)小鼠膀胱癌MB49細(xì)胞的體外抑制作用。包括(1)制備IR780-siRNA-PLGA納米粒并對(duì)其進(jìn)行表征;(2)評(píng)價(jià)IR780-siRNA-PLGA納米粒對(duì)抑制小鼠MB49細(xì)胞的體外毒性研究。方法(1)制備IR780-siRNA-PLGA NPs并對(duì)其進(jìn)行表征:采用超聲復(fù)乳法制備IR780-siRNA-PLGA納米粒。內(nèi)水相:survivin-siRNA溶于緩沖溶液;油相:PLGA 1mg/ml及IR780 0.2mg溶于二氯甲烷;外水相:0.5%PVA緩沖溶液。超聲功率360w,初乳時(shí)間120s,復(fù)乳時(shí)間180s,激光粒度分析儀測(cè)定納米粒徑,電鏡觀察IR780-siRNA-PLGA納米粒形態(tài);(2)IR780-siRNA-PLGA納米粒對(duì)抑制小鼠MB49細(xì)胞的體外細(xì)胞毒性研究:根據(jù)加入不同的納米材料和劑量分組,并使用MTT法檢測(cè)所構(gòu)建的納米粒體系對(duì)細(xì)胞的殺傷情況。結(jié)果(1)最優(yōu)的制備工藝為PLGA濃度為1mg/ml,超聲功率為360w,復(fù)乳時(shí)間180s。采用最優(yōu)設(shè)計(jì)得到的IR780載藥率為:1.8%,粒徑為131nm。納米粒在電子顯微鏡下呈典型的球形,形態(tài)圓潤(rùn),分散性較好。(2)MTT試驗(yàn)證明,納米粒包裹IR780+siRNA(IR780劑量為5 μg/ml)組予激光照射后所測(cè)OD值低于對(duì)照組(P0.05)。納米粒包裹IR780+siRNA(IR780劑量為10μg/ml)組無論是否予激光照射OD值均低于對(duì)照組(P0.05),但予激光照射后OD值下降更明顯(P0.01)。結(jié)論(1)我們成功構(gòu)建了IR780-siRNA-PLGA NPs,采用優(yōu)化后的超聲復(fù)乳法制備的納米粒粒徑、形態(tài)較為理想,分散性好;(2)IR780-siRNA-PLGA NPs可以降低IR780毒性并提高光熱和光動(dòng)力治療對(duì)小鼠MB49細(xì)胞的殺傷作用。
[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.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.14
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本文編號(hào)：1702367