Biostability of γ-Fe2O3 nano particles Evaluated using an in vitro cytotoxicity assays on various tumor cell lines

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Abstract

This study examined the cytotoxic effects of the γ-Fe2O3 magnetic nanoparticles (GMNs) on a range of tumor cell lines. GMNs, approximately 20 nm in diameter, were prepared using a chemical coprecipitation technique, and coated with two surfactants to obtain a water-based product. A 3-(4, 5-dimethylthiazol-2-yl) −2, 5-diphenyltetrazolium bromide assay revealed the GMNs to be non-toxic to human fibroblast cells and most of the tumor cell lines tested. However, the magnetic nanoparticles coated with the anti-cancer drugs exhibited cytotoxic activity against tumor cells. These results suggest that GMNs are not cytotoxic to the tumor cells and normal human cells tested in this study. In addition, the magnetic nanoparticles are biostable and magnetic nanoparticles coated with anti-cancer drugs are effective drug delivery vehicles. These results highlight the potential of drug-coated Fe2O3 magnetic nanoparticles as new anti-tumor agents.

Introduction

Nanotechniques have been touted as a new technology that may surpass the physical and chemical limitations of materials made from microparticles [1]. Recently, nanotechnology has attracted considerable interest in areas of biomedicine, such as cytotoxicity [2], [3], drug delivery [4], biosensors [5], and magnetic resonance imaging [6], catalyst supporters [7], and biomedical applications, such as magnetic carriers for bioseparation [8], enzyme and protein immobilization [9] and contrast-enhancing media. Kuckelhaus et al. performed an in vivo investigation of cobalt ferrite-based magnetic fluids using morphological tests [10]. Despite the presence of matrix metalloproteinase clusters, they did not observe any tissue damage or inflammatory cell infiltration. Macaroff et al. reported an in vitro evaluation of the toxicity of biocompatible magnetic fluids in an attempt to provide a better understanding of the interaction between the drug and protein [11]. Park et al demonstrated that a magnetic fluid coated with hematoporphyrin was stable without any phase transformations at body temperature [12]. Johannsen et al delivered target temperatures >70 °C using magnetic fluid hyperthermia, and reduced prostate cancer growth in an orthotopic Dunning R3327 rat model. However, the stability evaluation of magnetic nanoparticles as drug delivery was limited to the cells and tissues [13]. The toxicity of nanoparticles has not received as much attention as their applications. Recently, the organization for economic co-operation and development (OECD) was requested to sponsor a study of the toxicity of fourteen nanoparticles: fullerene, SWCNT, MWCNT, silver nanoparticle, iron nanoparticle, carbon black, titanium dioxide, aluminium oxide, cerium oxide, zinc oxide, silicon dioxide, polystyrene nanoparticle, dendrimers and nanoclay.

In particular, drug-loaded magnetic nanoparticles have been investigated for various biomedical applications, such as DNA separation, drug delivery, magnetic resonance imaging, hyperthermia and cell labeling [14]. To use these magnetic nanoparticles in biomedical applications, they often must be modified with biocompatible compounds [14]. Researchers have achieved this by either coating the magnetic nanoparticles with a layer of biodegradable polymers or evenly distributing a polymer matrix throughout the nanoparticles [15]. Using these approaches, magnetic drug targeting has been used to improve localized drug delivery and enhance drug-therapeutic efficiency in a variety of tumors [16].

This study examined the cytotoxic effects of GMNs on tumor cell lines as well as the precise drug effects of the GMNs. The cytotoxic effects of GMNs were investigated using tumor cells, DU145, PC3, LN3, C6, Siha and T98, and normal human fibroblast cells. The cytotoxicity of the GMNs loaded with anti-cancer drugs (Mitoxantrone and Hangamdan) on DU145 prostate cancer cells was then tested.

Section snippets

Synthesis of γ-Fe2O3 nanoparticles

The GMNs were made using FeCl3 6H2O (Junsei, Extra pure) and FeCl2 4H2O (Junsei, Extra pure) with NH4OH as a neutralizer. The Fe2+/Fe3+ ratio was 1. The solution was mixed well by shaking for 30 min, followed by vigorous shaking at 80 °C while adding NH4OH slowly until the pH reached 7. The addition of NH4OH was then stopped and the solution was stirred continuously for 2 h to produce red brown colored nanoparticles. The electrolytes remaining in solution were removed by washing the solution

Results and discussion

Fig. 1(a) shows the XRD pattern of the GMNs. All the peaks were indexed to the cubic γ-Fe2O3 phase according to the diffraction data reported in the 1997 Joint Committee on Powder Diffraction Standards (JCPDS). The particle size of the nanoparticles was 20 nm according to the Debye–Scherrer formula. The magnetic properties of the magnetic nanoparticles were examined using a vibration sample magnetometer at room temperature. The magnetic hysteresis loops of the magnetic nanoparticles showed zero

Conclusion

This study examined the cytotoxic effects of the γ-Fe2O3 magnetic nanoparticles (GMNs) on a range of tumor cell lines. GMNs, approximately 20 nm in diameter, were prepared using a chemical coprecipitation technique and coated with two surfactants to obtain a water-based product. The Raman spectra showed broad peaks at 213, 279, 389, 491, 593 cm−1. The HGMNs Raman spectra showed broad bands at near ∼362, ∼479, ∼695 and ∼1386 cm−1. The MGMNs Raman spectra of the showed six broad band at ∼375,

Acknowledgement

This study was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund) (KRF-2007-314-E00217) and by Basic Science Research Program Through the National Research Foundation of Korea (NRF) founded by the Ministry of Education, Science and Technology (2010-0011876).

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