可NIR-II荧光成像的多功能化疗栓塞微球PLGA@Re-DOX的制备及其在肝癌荧光手术导航中的应用

Preparation and Application of Multifunctional PLGA@Re-DOX Chemoembolization Microspheres with NIR-II Fluorescence Imaging for HCC Fluorescence-guided Surgery

目的:开发具备近红外二区(Near-Infrared II,NIR-II)荧光成像性能的多功能化疗栓塞微球,评估其在肝癌NIR-II荧光手术导航中的应用效果。方法:制备NaGdF_(4):Nd@NaGdF_(4)纳米颗粒(Re NPs)和聚乳酸-羟基乙酸共聚物(Polylactic-co-glycolic Acid,PLGA)基微球,利用透射电子显微镜(Transmission Electron Microscope,TEM)和荧光光谱仪分析Re NPs形貌和荧光性能。PLGA基微球的形貌、元素组成通过光学显微镜、扫描电子显微镜(Scanning Electron Microscope,SEM)、X射线能谱分析(Energy Dispersive Spectroscopy,EDS)进行表征。利用紫外光谱分仪测定PLGA微球的阿霉素(Doxorubicin,DOX)装载和DOX释放情况。细胞增殖-毒性实验(Cell Counting Kit-8,CCK-8)和溶血实验来评估其生物相容性。利用NIR-II成像系统分析PLGA@Re-DOX微球的成像效果和荧光稳定性。通过肝动脉插管将PLGA@Re-DOX微球递送至大鼠肝癌部位,观察其对NIR-II荧光成像效果。结果:Re NPs展现出良好的单分散性和均一球形结构,平均粒径为9.3±0.4 nm,并具备优异的NIR-II荧光成像性能。PLGA、PLGA@Re-DOX微球尺寸分别为294±7μm、288±13μm,具有高度的均一性。PLGA微球具有良好的生物相容性,其对细胞存活率和溶血率与对照组相比没有显著统计差异(P>0.05)。此外,DOX包封率可达2.5%,能缓慢释放药物。PLGA@Re-DOX微球具有良好的肝内外NIR-II荧光成像效果和优异的荧光稳定性,经肝动脉递送后进行NIR-II荧光成像可以清晰地显示肿瘤边缘。结论:制备的PLGA@Re-DOX微球可有望实现对肝癌的NIR-II荧光成像和荧光手术导航。

Objective:To prepare multifunctional chemoembolization microspheres featuring Near-Infrared II(NIR-II)fluorescence imaging capabilities,and to evaluate their effectiveness in NIR-II fluorescence-guided surgery for Hepatocellular Carcinoma(HCC).Methods:NaGdF_(4):Nd@NaGdF_(4)nanoparticles(Re NPs)and polylactic-co-glycolic acid(PLGA)-based microspheres.were prepared.The morphology and fluorescence properties of Re NPs were analyzed using Transmission Electron Microscopy(TEM)and fluorescence spectrometer.The morphology and elemental composition of PLGA-based microspheres were characterized by Optical Microscopy,Scanning Electron Microscopy(SEM),and Energy Dispersive Spectroscopy(EDS).The loading capacity and release profile of Doxorubicin(DOX)in PLGA microspheres were determined using UV spectrophotometry.Biocompatibility of the PLGA microspheres was evaluated through Cell Counting Kit-8(CCK-8)and hemolysis assays.The imaging effectiveness and fluorescence stability of PLGA@Re-DOX microspheres were evaluated using a NIR-II imaging system.PLGA@Re-DOX microspheres were delivered to hepatic artery of rats for in-situ tumor imaging.Results:Re NPs were characterized by good monodispersity and uniform spherical structure,with an average particle size of 9.3±0.4 nm,displaying excellent NIR-II fluorescence imaging capabilities.PLGA and PLGA@Re-DOX microspheres were noted for their high uniformity with diameters of 294±7?滋m and 288±13?滋m,respectively.Good biocompatibility was observed in the PLGA microspheres,as no significant statistical differences in cell viability and hemolysis rates were found compared to the control group(P>0.05).Additionally,a slow drug release was facilitated by a DOX encapsulation rate of up to 2.5%.The PLGA@Re-DOX microspheres demonstrated effective intrahepatic and extrahepatic NIR-II fluorescence imaging capabilities and excellent fluorescence stability,clearly delineating tumor margins post-delivery via hepatic artery.Conclusion:The synthesized PLGA@Re-DOX microspheres are promising for application in NIR-II fluorescence imaging and fluorescence-guided surgery of HCC.