【作者】 邹海蓉； 尚婷婷； 王惠； 钟毓； 张志君； 王志刚；

【Author】 Hairong Zou;Tingting Shang;Hui Wang;Yu Zhong;Zhijun Zhang;Zhigang Wang;Department of Ultrasound,the University-Town Hospital of Chongqing Medical University;Department of Ultrasound,the Second Affiliated Hospital of Chongqing Medical University;

【机构】 重庆医科大学附属大学城医院超声科； 重庆医科大学附属第二医院超声科；

【摘要】 目的自制一种同源肿瘤细胞膜包裹的普鲁士蓝纳米粒,探究其增效光热转换效应对乳腺癌治疗的影响。方法首先采用高效共沉淀法,以柠檬酸、FeCl3和K4[Fe(CN)6]为原料合成普鲁士蓝纳米粒(Prussian blue Nanoparticles,PB NPs);再利用Beyotime生物技术提取小鼠源性4T1乳腺癌细胞的细胞膜;最后通过声震法制备4T1乳腺癌细胞膜包裹PB的同源仿生M@PB纳米粒。通过透射电镜(TEM)、动态激光散射法(DLS)及紫外-可见光(UV-Vis)等体外表征方法验证其体外基本物化特征;观察M@PB在不同浓度及不同功率激光辐照下的光热性能;CCK8法和活死细胞染色法评估其对4T1的靶向性能以及激光对吞噬了M@PB的肿瘤细胞的增效杀伤作用;并探究M@PB在体内肿瘤部位的聚集情况及体内的生物代谢分布情况。结果成功制备纳米粒M@PB,呈现出均匀的立方形颗粒,其表面被覆一层薄薄的膜样结构。M@PB的流体动力学粒径和电位分别为85.7 nm和-6.3 mv,较PB粒径稍大电位稍低(PB粒径为77.6 nm,电位为-7.633 mv)。M@PB在体外可增强光声成像,光声信号随纳米粒浓度升高而增强。细胞吞噬实验显示4T1细胞对M@PB的吞噬大于对PB的吞噬。激光辐照后,活死细胞染色显示吞噬M@PB的4T1肿瘤细胞大量死亡,死亡量明显多于吞噬PB的4T1肿瘤细胞。体内光声(PA)成像结果显示,M@PB组在各个相同时间点的PA信号均比PB组强,随着注射时间的延长,PB组PA信号的强度呈先快后慢的持续上升趋势,而M@PB组呈现先快速上升后缓慢下降的趋势,在6h达峰值。在8～24 h内两组都处于一个稳定期,PA信号变化不大。结论研制的M@PB纳米粒具有特异靶向同源肿瘤细胞的能力,从而增效了纳米粒的光热转化。更多还原

【Abstract】 Objective Develop a homologous tumor cell membrane-encapsulated Prussian blue nanoparticle,and investigate its synergistic photothermal conversion effect on the treatment of breast cancer.Methods First,the efficient co-precipitation method was used to synthesize Prussian blue nanoparticles(PB NPs) using citric acid,FeCl3,and K4[Fe(CN)6] as raw materials.Then,the Beyotime biotechnology was used to extract the cell membrane from mouse-derived 4T1 breast cancer cells.Finally,the homologous mimic M@PB nanoparticles were prepared by sonication method with the 4T1 breast cancer cell membrane wrapped PB.The basic physical and chemical characteristics of M@PB were verified by TEM,DLS,and UV-Vis,etc.in vitro.The photothermal performance of M@PB under different concentrations and different power laser irradiation was observed.The targeting performance of M@PB towards 4T1 and the synergistic killing effect of laser on the tumor cells that ingested M@PB were evaluated by CCK8 assay and live/dead cell staining method.The aggregation of M@PB in the tumor site and the in vivo bio-metabolic distribution of M@PB were also explored.Results Successfully prepared nanoparticles M@PB,which showed uniform cubic particles with a thin film-like structure on their surface.The fluid dynamic diameter and potential of M@PB were 85.7 nm and-6.3 mv,slightly larger and lower in potential than PB(PB particle size was 77.6 nm,potential was-7.633 mv).M@PB enhanced in vivo photoacoustic imaging,and the photoacoustic signal increased with the increase of nanoparticle concentration.The cell phagocytosis experiment showed that 4T1 cells phagocytosed more M@PB than PB.After laser irradiation,live/dead cell staining showed that a large number of 4T1tumor cells that had phagocytosed M@PB died,with a significantly higher death rate than those that had phagocytosed PB.The in vivo photoacoustic(PA) imaging results showed that the PA signal of the M@PB group was stronger than that of the PB group at all the same time points,and the PA signal of the M@PB group showed a rapid increase followed by a slow decrease,reaching a peak at 6h.Both groups were in a stable period within8-24h,with little change in PA signal.Conclusion The M@PB nanoparticles developed have a specific targeting ability to homologous tumor cells,thus enhancing the photothermal conversion of the nanoparticles.更多还原