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Cellular uptake genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells

机译:人体乳腺细胞中钴铁氧体磁性纳米颗粒的细胞吸收遗传毒性和细胞毒性

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摘要

Magnetic nanoparticles (MNPs) have been increasingly used for many years as MRI agents and for gene delivery and hyperthermia therapy, although there have been conflicting results on their safety. In this study, cobalt ferrite magnetic nanoparticles (CoFe-MNPs) were prepared by the co-precipitation method and their surfaces were modified with silica by the sol–gel method. The particle and hydrodynamic sizes, morphology and crystal structure of the bare and silica-coated CoFe-MNPs were evaluated by transmission electron microscopy (TEM), dynamic light scattering (DLS), X-ray diffraction spectroscopy (XRD) and Fourier transform infrared spectroscopy (FTIR). The size of the bare CoFe-MNPs was in the range 8–20 nm and they were homogeneously coated with 3–4 nm silica shells. The bare and silica-coated CoFe-MNPs were agglomerated at physiological pH. However, the sizes of the agglomerates were below 200 nm both in water and complete medium. The cytotoxic and genotoxic potentials of the bare and silica-coated CoFe-MNPs were evaluated in a metastatic breast cancer cell line, MDA-MB-231, as well as a noncancerous mammary epithelial cell line, MCF-10A, by using XTT cytotoxicity, single-cell gel electrophoresis (comet), and cytokinesis-blocked (CB) micronucleus (CBMN) assays. Characterization studies with TEM, inductively coupled plasma optical emission spectroscopy (ICP-OES) and Prussian blue staining indicated that the CoFe-MNPs were internalized into the cells by energy-dependent endocytosis. The highest amount of uptake was observed in the cancer cells and the uptake of the silica-coated CoFe-MNPs was higher than that of the bare ones in both cell lines. The bare CoFe-MNPs showed higher levels of both cytotoxicity and genotoxicity than the silica-coated CoFe-MNPs. Moreover, the cancer cells seemed to be more susceptible to the CoFe-MNPs’ toxicity compared to the noncancerous cells. There was a concentration and time-dependent increase in DNA damage and the micronucleus (MN) frequency, which was statistically significant starting with the lowest concentration of bare CoFe-MNPs (p < 0.05), while no significance was observed below the concentration of 250 μg mL–1 for the silica-coated MNPs. Also, the extent of both DNA damage and MN frequency was much higher in the cancer cells compared to the noncancerous cells. According to our results, the silica coating ameliorated both the cytotoxicity and genotoxicity as well the internalization of the CoFe-MNPs.
机译:磁性纳米颗粒(MNP)多年来已越来越多地用作MRI试剂以及用于基因传递和热疗治疗,尽管在安全性方面存在矛盾的结果。在这项研究中,通过共沉淀法制备了钴铁氧体磁性纳米粒子(CoFe-MNPs),并通过溶胶-凝胶法对其表面进行了二氧化硅改性。通过透射电子显微镜(TEM),动态光散射(DLS),X射线衍射光谱(XRD)和傅里叶变换红外光谱对裸露的和涂覆二氧化硅的CoFe-MNP的颗粒和流体动力学尺寸,形态和晶体结构进行了评估。 (FTIR)。裸露的CoFe-MNP的大小在8–20 nm范围内,并且均匀地涂有3–4 nm的二氧化硅壳。裸露的和涂覆二氧化硅的CoFe-MNP在生理pH值下团聚。然而,在水和完全培养基中,附聚物的尺寸均低于200nm。通过使用XTT细胞毒性,评估了转移性乳腺癌细胞MDA-MB-231以及非癌性乳腺上皮细胞系MCF-10A中裸露的和涂有二氧化硅的CoFe-MNP的细胞毒性和遗传毒性,单细胞凝胶电泳(彗星)和胞质阻滞(CB)微核(CBMN)测定。用TEM,电感耦合等离子体发射光谱(ICP-OES)和普鲁士蓝染色进行的表征研究表明,CoFe-MNPs通过能量依赖性内吞作用被内化到细胞中。在癌细胞中观察到最高的摄取量,并且在两种细胞系中,二氧化硅包覆的CoFe-MNP的摄取均高于裸细胞。裸露的CoFe-MNPs比二氧化硅包被的CoFe-MNPs表现出更高的细胞毒性和遗传毒性。而且,与非癌细胞相比,癌细胞似乎更容易受到CoFe-MNPs毒性的影响。 DNA损伤和微核(MN)频率呈浓度和时间依赖性增加,从裸露的CoFe-MNP的最低浓度开始(p <0.05),这具有统计学意义,而低于250的浓度则无显着性二氧化硅包被的MNP为μgmL –1 。同样,与非癌细胞相比,癌细胞中DNA损伤的程度和MN频率都高得多。根据我们的结果,二氧化硅涂层改善了CoFe-MNP的细胞毒性和基因毒性以及内在化。

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