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

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.

[1]  G. Palade,et al.  Temperature-sensitive steps in the transport of secretory proteins through the Golgi complex in exocrine pancreatic cells. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[2]  N. Roehm,et al.  An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. , 1991, Journal of immunological methods.

[3]  N. Singh,et al.  Modifications of alkaline microgel electrophoresis for sensitive detection of DNA damage. , 1994, International journal of radiation biology.

[4]  Paul Mulvaney,et al.  Synthesis of Nanosized Gold−Silica Core−Shell Particles , 1996 .

[5]  M. Fenech.  The in vitro micronucleus technique. , 2000, Mutation research.

[6]  Peng Huang,et al.  ROS stress in cancer cells and therapeutic implications. , 2004, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[7]  É. Duguet,et al.  Magnetic nanoparticle design for medical diagnosis and therapy , 2004 .

[8]  S. Franzen,et al.  Probing BSA binding to citrate-coated gold nanoparticles and surfaces. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[9]  Doina Bica,et al.  Antitumor effect of magnetite nanoparticles in cat mammary adenocarcinoma , 2005 .

[10]  Jun-Sung Kim,et al.  Cellular uptake of magnetic nanoparticle is mediated through energy-dependent endocytosis in A549 cells , 2006, Journal of veterinary science.

[11]  Thomas Kuhlbusch,et al.  Particle and Fibre Toxicology BioMed Central Review The potential risks of nanomaterials: a review carried out for ECETOC , 2006 .

[12]  Tzong-Ming Wu,et al.  Preparation and characterization of thermosensitive polymers grafted onto silica-coated iron oxide nanoparticles. , 2008, Journal of colloid and interface science.

[13]  Xu Ma,et al.  A novel method to prepare water-dispersible magnetic nanoparticles and their biomedical applications: magnetic capture probe and specific cellular uptake. , 2008, Journal of biomedical materials research. Part A.

[14]  S. Doak,et al.  NanoGenotoxicology: the DNA damaging potential of engineered nanomaterials. , 2009, Biomaterials.

[15]  Helinor Johnston,et al.  Development of in vitro systems for nanotoxicology: methodological considerations , 2009, Critical reviews in toxicology.

[16]  M. Marinovich,et al.  Risk Assessment of Products of Nanotechnologies , 2009 .

[17]  Bengt Fadeel,et al.  Better safe than sorry: Understanding the toxicological properties of inorganic nanoparticles manufactured for biomedical applications. , 2010, Advanced drug delivery reviews.

[18]  S. H. Lee,et al.  In vitro cytotoxicity screening of water-dispersible metal oxide nanoparticles in human cell lines , 2010, Bioprocess and biosystems engineering.

[19]  A. Sood,et al.  Nanomedicine based approaches for the delivery of siRNA in cancer , 2010, Journal of internal medicine.

[20]  Soonhag Kim,et al.  Gene Expression Profiles for Genotoxic Effects of Silica-Free and Silica-Coated Cobalt Ferrite Nanoparticles , 2012, The Journal of Nuclear Medicine.

[21]  A. Cuschieri,et al.  Dilemmas in the reliable estimation of the in-vitro cell viability in magnetic nanoparticle engineering: which tests and what protocols? , 2012, Nanoscale Research Letters.

[22]  Eleonore Fröhlich,et al.  The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles , 2012, International journal of nanomedicine.

[23]  Patrick Couvreur,et al.  Magnetic nanoparticles: design and characterization, toxicity and biocompatibility, pharmaceutical and biomedical applications. , 2012, Chemical reviews.

[24]  Sara A Love,et al.  Assessing nanoparticle toxicity. , 2012, Annual review of analytical chemistry.

[25]  M. Toprak,et al.  Uniform mesoporous silica coated iron oxide nanoparticles as a highly efficient, nontoxic MRI T(2) contrast agent with tunable proton relaxivities. , 2012, Contrast media & molecular imaging.

[26]  Warren C W Chan,et al.  The effect of nanoparticle size, shape, and surface chemistry on biological systems. , 2012, Annual review of biomedical engineering.

[27]  Mojca Pavlin,et al.  Visualization of internalization of functionalized cobalt ferrite nanoparticles and their intracellular fate , 2013, International journal of nanomedicine.

[28]  Tian Xia,et al.  Physicochemical properties determine nanomaterial cellular uptake, transport, and fate. , 2013, Accounts of chemical research.

[29]  H. Shokrollahi,et al.  The role of cobalt ferrite magnetic nanoparticles in medical science. , 2013, Materials science & engineering. C, Materials for biological applications.

[30]  Teófilo Rojo,et al.  The challenge to relate the physicochemical properties of colloidal nanoparticles to their cytotoxicity. , 2013, Accounts of chemical research.

[31]  Mikhail Y. Berezin,et al.  Nanotechnology for Biomedical Imaging and Diagnostics: From Nanoparticle Design to Clinical Applications , 2014 .

[32]  Andrew R Collins,et al.  Measuring oxidative damage to DNA and its repair with the comet assay. , 2014, Biochimica et biophysica acta.

[33]  A. Athanassiou,et al.  Toxicity Assessment of Silica Coated Iron Oxide Nanoparticles and Biocompatibility Improvement by Surface Engineering , 2014, PloS one.

[34]  M. Tourbin,et al.  Nanoparticles in wastewaters: hazards, fate and remediation , 2014 .

[35]  M. Çulha,et al.  Interaction of carbohydrate modified boron nitride nanotubes with living cells. , 2015, Colloids and surfaces. B, Biointerfaces.

[36]  Z. Su,et al.  Cancer Therapy: Facile and Scalable Synthesis of Novel Spherical Au Nanocluster Assemblies@Polyacrylic Acid/Calcium Phosphate Nanoparticles for Dual‐Modal Imaging‐Guided Cancer Chemotherapy (Small 26/2015) , 2015 .

[37]  Helmuth Möhwald,et al.  RGD peptide-modified dendrimer-entrapped gold nanoparticles enable highly efficient and specific gene delivery to stem cells. , 2015, ACS applied materials & interfaces.

[38]  Steven T. Wang,et al.  Nanoparticles for Bioimaging , 2015 .

[39]  H. Xiong,et al.  Serum albumin adsorbed on Au nanoparticles: structural changes over time induced by S-Au interaction. , 2015, Chemical communications.

[40]  Morteza Milani,et al.  Magnetic nanoparticles: Applications in gene delivery and gene therapy , 2015, Artificial cells, nanomedicine, and biotechnology.

[41]  E. Asik,et al.  2-Amino-2-deoxy-glucose conjugated cobalt ferrite magnetic nanoparticle (2DG-MNP) as a targeting agent for breast cancer cells. , 2016, Environmental toxicology and pharmacology.