基于HA修饰的CoFe_(2)O_(4)-TiO_(2)纳米颗粒体外PDT灭活HL60细胞的试验研究

An Experimental Study Based on HA Modified CoFe_(2)O_(4)-TiO_(2)Nanoparticles by Photodynamic Therapy for HL60 Cells in vitro

通过水热法和表面修饰法制备了以CoFe_(2)O_(4)为内核、TiO_(2)为外壳、用透明质酸修饰的HA@CoFe_(2)O_(4)-TiO_(2)。利用场发射扫描电子显微镜(SEM)成像、能谱分析(EDS)、X射线衍射(XRD)、X射线光电子谱(XPS)、紫外-可见吸收光谱(UV-Vis)以及傅里叶变换红外光谱(FTIR)对复合纳米颗粒的理化性质进行表征。用细胞计数试剂(CCK-8)法研究HA@CoFe_(2)O_(4)-TiO_(2)复合纳米颗粒在暗室条件下对白血病HL60细胞的毒性,以及在光照条件下的光动力疗法(PDT)灭活效果。结合复合纳米颗粒的荧光光谱(FS)、细胞内活性氧(ROS)产量和摄取纳米颗粒水平分析,初步探究CoFe_(2)O_(4)和透明质酸与TiO_(2)结合影响PDT灭活HL60细胞的作用机制。试验结果表明,HA@CoFe_(2)O_(4)-TiO_(2)形貌规则呈球形,尺寸符合细胞摄取要求。CoFe_(2)O_(4)拓宽了TiO_(2)的可见光响应范围。暗毒性试验表明,HA@CoFe_(2)O_(4)-TiO_(2)复合纳米颗粒对HL60细胞的毒性较小。同时,与TiO_(2)和CoFe_(2)O_(4)-TiO_(2)纳米颗粒相比,HA@CoFe_(2)O_(4)-TiO_(2)对HL60细胞的光动力灭活效率高达82.6%,表明其有望成为应用于体外PDT治疗白血病的光敏剂,提高光动力效率及靶向性。

In this study,CoFe_(2)O_(4)-TiO_(2)was prepared by hydrothermal method,in which CoFe_(2)O_(4)was the core and TiO_(2)was the shell,and HA@CoFe_(2)O_(4)-TiO_(2)was obtained by hyaluronic acid modification.The physicochemical properties of the nanocomposites were characterized by scanning electron microscopy(SEM)imaging,energy dispersive spectroscopy(EDS),X-ray diffraction(XRD),X-ray photoelectron spectroscopy(XPS),ultraviolet-visible absorption spectroscopy(UV-Vis)and Fourier transform infrared spectroscopy(FTIR).CCK-8 method was used to investigate the toxicity of HA@CoFe_(2)O_(4)-TiO_(2)nanocomposites on HL60 cells in dark chamber,and the inactivation effect of photodymanic therapy(PDT).The mechanism of interaction between CoFe_(2)O_(4),hyaluronic acid and TiO_(2)on PDT inactivation of HL60 cells was preliminarily investigated by fluorescence spectra(FS),intracellular ROS production and uptake level analysis of nanocomposites.The experimental results showed that the morphology of HA@CoFe_(2)O_(4)-TiO_(2)was regular and spherical,and the size met the requirements of cell uptake.The visible response range of TiO_(2)was broadened with CoFe_(2)O_(4).Dark toxicity experiment proved that HA@CoFe_(2)O_(4)-TiO_(2)nanocomposites were less toxic to HL60 cells under darkroom conditions.Meanwhile,Compared with the groups treated with TiO_(2)and CoFe_(2)O_(4)-TiO_(2)nanoparticles,the photocatalytic inactivation efficiency of the group treated with HA@CoFe_(2)O_(4)-TiO_(2)was as high as 82.6%.Therefore,it has the potential to become a photosensitizer for the PDT treatment of leukemia in vitro,improving photodynamic efficiency and targeting.