纳米磁颗粒Fe_(3)O_(4)@BSA@SeCyc2@FA用于海拉细胞的磁热和放疗增敏协同作用研究  

Synergistic magnetic hyperthermia/radiotherapy efficacy of magnetic nanoparticles Fe_(3)O_(4)@BSA@SeCyc2@FA for HeLa cells

作　　者：林海磊 陈北冬[2] 尹亮亮[1] 吉艳琴[1] Lin Hailei;Chen Beidong;Yin Liangliang;Ji Yanqin(Key Laboratory of Radiological Protection and Nuclear Emergency,China CDC,National Institute for Radiological Protection,Chinese Center for Disease Control and Prevention,Beijing 100088,China;Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所、辐射防护与核应急中国疾病预防控制中心重点实验室,北京100088 [2]北京医院、国家老年医学中心、中国医学科学院老年医学研究院,北京100730

出　　处：《中华老年医学杂志》2024年第7期882-888,共7页Chinese Journal of Geriatrics

摘　　要：目的构筑具有良好靶向性和生物相容性的新型磁性复合纳米颗粒,验证其对海拉(HeLa)细胞的磁热和放疗增敏协同作用效果。方法使用牛血清白蛋白(BSA)修饰四氧化三铁(Fe_(3)O_(4))纳米颗粒作为载体,利用碳二亚胺法将L-硒代胱氨酸(SeCyc2)和叶酸(FA)在BSA表面进行偶联,合成磁性复合纳米颗粒Fe_(3)O_(4)@BSA@SeCyc2@FA,并对其进行表征。通过评估颗粒的磁热特性,分析HeLa细胞对颗粒的内化作用及使用CellTiter和活性氧(ROS)试剂盒分别对不同实验组的细胞活力和ROS产生量进行检测,以此来评价在磁热联合高能X射线作用下,Fe_(3)O_(4)@BSA@SeCyc2@FA纳米颗粒对HeLa细胞的抑制或杀伤效果。结果Fe_(3)O_(4)@BSA@SeCyc2@FA纳米颗粒的平均粒径为(19.31±4.84)nm,Zeta电位为-25.4 mV(pH=7),其具备良好的分散性和稳定性,为后续生物实验奠定基础。通过BSA和FA特征吸收峰并结合纳米颗粒Se元素的含量为10.89μM,证实L-硒代胱氨酸和叶酸已经成功修饰在复合纳米颗粒表面。所制备颗粒的饱和磁化强度为47.2 emu/g,在518 kHz/16 kAm^(-1)交变磁场作用下,其比吸收率(SAR)为125.4 W/g,可在15 min内升温可达7℃,这表明该颗粒具有良好的发热特性。在0~200μg/ml浓度范围内,颗粒对HUVECs无明显毒性,但HeLa细胞对颗粒内化作用明显,并表现出一定的活力抑制敏感性。在磁热疗联合高能X射线后,HeLa细胞活力明显降低至54.7%,细胞内ROS的生成量较对照组增加了108.2%,实验结果表明,复合纳米颗粒明显增强了HeLa细胞的放射敏感性,磁热联合放疗协同作用对其抑制和杀伤更为有效。结论构筑的新型功能化磁性复合纳米颗粒具有作为一种协同磁热和放疗增敏作用纳米系统的潜力,其能够克服单一治疗模式的诸多不足,将为今后体内肿瘤综合治疗提供新的研究思路和基础。Objective To develop a novel magnetic composite nanoparticle with excellent targeting and biocompatibility and verify synergistic magnetic hyperthermia/radiotherapy efficacy in HeLa cells.Methods Bovine serum albumin(BSA)was used to modify ferroferric oxide(Fe_(3)O_(4))nanoparticles,and then L-selenocystine and folic acid(FA)were coupled on the surface of BSA by carbodiimide method to synthesize magnetic composite nanoparticles Fe_(3)O_(4)@BSA@[e-mail] as-prepared nanoparticles were characterized to determine the structure,composition,and properties of the nanoparticles to support their biological applications.By evaluating the magnetothermal properties of the as-prepared nanoparticles,analyzing the internalization effect of HeLa cells on the particles and using CellTiter and reactive oxygen species(ROS)kits to detect the cell viability and ROS production of different experimental groups,respectively,the killing and inhibiting effects on HeLa cells were evaluated when hyperthermia was combined with radiotherapy.Results The average particle size of the Fe_(3)O_(4)@BSA@SeCyc2@FA nanoparticles was(19.31±4.84)nm,and the zeta potential was -25.4 mV(pH=7),which indicated that the as-prepared nanoparticles maintained acceptable dispersion and stability for biological experiments.By BSA and FA characteristic absorption peaks combined with the measured content of Se(10.89μM),L-selenocystine and FA were successfully modified on the surface of the composite nanoparticles.The saturation magnetization value of the Fe_(3)O_(4)@BSA@SeCyc2@FA nanoparticles was 47.2 emu/g.The as-prepared nanoparticles maintained the specific absorption rate(SAR)value of 125.4 W/g at the alternating magnetic field of 518 kHz/16 kAm^(-1),indicating excellent heat generation efficiency(the temperature level ΔT=7℃ within 15 min).No significant toxicity was found for HUVEC cells even within the concentration range from 0 to 200μg/ml and inhibition of cell viability of the HeLa was shown.When magnetic hyperthermia combined with high-energy X

关 键 词：纳米粒子 硒代胱氨酸 热疗 辐射增敏剂 

分 类 号：R730.5[医药卫生—肿瘤]