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Citrate Capped Superparamagnetic Iron Oxide Nanoparticles Used for Hyperthermia Therapy

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Citrate Capped Superparamagnetic Iron Oxide Nanoparticles Used for Hyperthermia Therapy J. Biomedical Science and Engineering, 2012, 5, 715-719 JBiSE http://dx.doi.org/10.4236/jbise.2012.512089 Published Online December 2012 (http://www.SciRP.org/journal/jbise/) Citrate capped superparamagnetic iron oxide nanoparticles used for hyperthermia therapy Elham Cheraghipour1, Sirus Javadpour1, Ali Reza Mehdizadeh2 1Department of Materials Science and Engineering, Shiraz University, Shiraz, Iran 2Department of Medical Physics, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected] Received 13 October 2012; revised 15 November 2012; accepted 24 November 2012 ABSTRACT received increased attention during the last decade due to their characteristic, i.e. inducible magnetic moments in Superparamagnetic magnetite nanoparticles (MNP) of about 10 nm were designed with proper physico- the presence of an external magnetic field. Magnetite chemical characteristics by an economic, biocompati- nanoparticles have been widely used experimentally for ble chemical co-precipitation of Fe2+ and Fe3+ in an numerous biomedical and in vivo applications [1-3]. ammonia solution, for hyperthermia applications. Magnetic-based delivery strategies are based on bind- Synthetic methodology has been developed to get a ing drugs with magnetic fluids that concentrate the drug well dispersed and homogeneous aqueous suspension in the site of interest. The surface-bound drugs can be of MNPs. Citric acid was used to stabilize the mag- released from the drug carriers by changing the physio- netite particle suspension, it was anchored on the logical conditions, and are then taken up by the affected surface of freshly prepared MNPs by direct addition cells [4]. MNPs behave like magnets only in presence of method. Carboxylic acid terminal group not only an external magnetic field and do not retain any residual render the particles more water dispersible but also magnetism upon removal of the external magnetic field. provides a site for further surface modification. The This property makes it possible for their application as naked MNPs are often insufficient for their stability, magnetic resonance imaging (MRI) contrast agent or hydrophilicity and further functionalization. To over- their target ability under the influence of external mag- come these limitations, citric acid was conjugated on nets. However, MNPs are not stable at normal physio- the surface of the MNPs. The microstructure and logical conditions and show a tendency to aggregate be- morphology of the nanoparticles were characterized cause of the hydrophobic nature of these particles [5-7]. by X-ray diffraction (XRD) and transmission electron Challenges in fabricating and processing nanoparticles microscopy (TEM), and the interaction between citric in regard to the applications in biomedicine are the issues acid and MNPs were characterized by Fourier trans- of agglomeration, uniformity, hydrophilicity, and bio- form infrared spectroscopy (FTIR), whereas the ma- compatibility of nanoparticles. It is a technological chal- gnetic properties were investigated by vibrating sam- lenge to acquire control over the nanoparticle sizes and ple magnetometry (VSM). Magnetic measurement re- vealed that the saturation magnetization of the na- dispersibility in desired solvents. Because of the large noparticles was 74 emu/g and the nanoparticles were surface to volume ratio, nanoparticles possess high sur- superparamagnetic at room temperature. We also face energies. The particles tend to aggregate to mini- have analyzed the potential of these particles for hy- mize total surface energy. In the case of metal oxide sur- perthermia by determination of the specific absorp- faces, such energies are in excess of 0.1 N/m [8]. tion rate, the temperature increase (ΔT) of the parti- Suitable surface functionalization of the particles and cles was 37˚C. These ferrofluids with high self-heating choice of solvent are crucial to achieving sufficient re- capacity are a promising candidate for cancer hyper- pulsive interactions between particles to prevent aggre- thermia treatment. gation and obtain a thermodynamically stable colloidal solution. The surface of MNPs can be stabilized in an Keywords: Magnetite Nanoparticle; Citric Acid; aqueous dispersion by the adsorption of citric acid [9]. Hyperthermia This process, as described by Sahoo et al. [9], occurs by the citric acid being coordinated via one or two of the 1. INTRODUCTION carboxylate functionalities, depending upon steric neces- sity and the curvature of the surface. Carboxylates have Superparamagnetic magnetite nanoparticles (MNPs) have important effects on the growth of iron oxide nanoparti- OPEN ACCESS 716 E. Cheraghipour et al. / J. Biomedical Science and Engineering 5 (2012) 715-719 cles and their magnetic properties. Bee et al. demon- tion mixture to prevent the development of large poly- strated that the average diameter of citrate coated crystalline particles, turbulence was created by placing nanoparticles can be varied from 3 nm to 8 nm by de- the reaction flask in an ultrasonic bath (100 W) and con- creasing the amount of citrate ions [10]. Liu and Huang trolling the homogenization (overhead stirrer) rate (9000 have studied the effect of the presence of citric acid dur- rpm) during the initial two minutes of the reaction. ing iron oxide synthesis [11]. Increasing concentrations NH4OH (25 wt%) was added instantaneously to the reac- of citric acid caused significant decreases in the crystal- tion mixture and vigorous mechanical stirring was ap- linity of the iron oxides formed. Moreover, the presence plied to reach pH 10 - 11. of citrate led to changes in the surface geometry. The The surface of MNPs was stabilized with citric acid by stability range is strongly dependent upon pH and the a direct addition method, to obtain modified MNPs with concentration of adsorbed acids [12]. carboxylic groups. Briefly, the MNPs surface were treat- Here we successfully capped citrate on the surface of ed with citrate ions by incubating for 1 hour within 0.5 superparamagnetic iron oxide nanoparticles as a hyper- g/mL citric acid solution and reaction temperature was thermia device. The use of hyperthermia in the treatment raised up to 90˚C and the reaction was completed for 60 of malignant tumors is as old as medicine itself. Hippo- min with continuous stirring. The black precipitates were crates, the father of medicine, suggested that surface tu- acquired by cooling the reaction mixture to room tem- mors should be cauterized by application of hot iron. In perature. At last the suspensions were washed several modern times, more advanced methods (hot water bath, times with deionized water. The particles did not settle high-frequency radiation and magnetic fluid hyperther- down under the influence of magnet, as a result of stabi- mia) are employed to heat, and hopefully destroy tumors lity of solution. The as-formed reaction product con- [13]. tained an excess of citric acid and so, the nanoparticle Nowadays, there is a great interest for nano-ther- dispersion was centrifuged and washed several times. motherapy, i.e. killing tumor cells, not with chemothera- Magnetite nanoparticles stabilized with citric acid are ab- peutic agents, but with locally targeted nano-heaters that breviated as MNP-CA. would basically kill malignant cells [14]. This technique is an encouraging technique for cancer treatment because 2.3. Characterization of the Nanoparticles of ease of targeting the cancerous tissue, thus having The synthesized nanoparticles were characterized by va- fewer side effects than radiotherapy and chemotherapy. rious analytical techniques. The X-ray diffraction (XRD) Moreover, this method provides a straight forward way patterns were acquired from dried nanoparticle samples of treating cancer in combination with irradiation and with a BRUKER X-ray powder diffractometer by using chemotherapy. Cu-Kα1 (1.54060Å) radiation. The scans of selected dif- In this study we focused our interest on the physico- fraction peaks were carried out in step mode (step size chemical aspects of the particles. Finally, the heating 0.02˚, measurement time 2 s, measurement temperature properties of these nanoparticles were evaluated, show- 300 K and standard: Si powder). ing potential for hyperthermia. Transmission electron microscopy (TEM) images were acquired on a Phillips 400 TEM operating at 100 2. MATERIALS AND METHODS kV. TEM grids were prepared by depositing a drop of 2.1. Materials diluted nanoparticle suspension on 300 mesh silicon- monoxide support films and drying the grids under vac- Ferric chloride hexahydrate, Ferrous chloride tetrahy- uum for 2 h. drate, Citric acid (CA) and Ammonia solution (25%) The infrared spectra were recorded in the range 400 - were purchased from Merck (Germany). All chemicals 4000 cm−1 on a Fourier transform infrared spectrometer were of analytical grade and used as received. (FTIR-Shimadzu-8000). Samples of the surface modified nanoparticles were dried overnight using a Virtis Freeze- 2.2. Magnetite Nanoparticle Synthesis mobile freeze-drier. Nanoparticle powder (2 mg) was The preparation of magnetite nanoparticles was performed then milled with KBr and the mixture was pressed into a by a chemical coprecipitation of Fe2+ and Fe3+ ions (1:2 pellet for analysis. molar ratio) by the addition of NH4OH. Briefly, 50 ml of The saturation magnetization (Ms) and coercive force 2+ 3+ 1.0 mol/L Fe and 2.0 mol/L Fe solutions were pre- (Hc) of the samples were measured using a vibrating pared with deionized water in two beakers, and then sample magnetometer (VSM, Dexing, Model: 250) with transferred to a three necked flask and temperature was a sensitivity of 10−3 emu. The magnetic field was chang- slowly increased to 80˚C in refluxing condition under ed uniformly with a time rate of 66 Oe/s. nitrogen atmosphere. While vigorously stirring the reac- In order to test the heating capacity of the MNPs, we Copyright © 2012 SciRes. OPEN ACCESS E. Cheraghipour et al. / J. Biomedical Science and Engineering 5 (2012) 715-719 717 have prepared samples of the synthesized MNP-CA sus- 3.3. FTIR Analysis pended in distilled water with a magnetic composition of Figure 4 shows the FT-IR spectra of pure citric acid 1% w/v (g of Fe O /mL solution).
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