Iron oxide nanoparticles doped with alkali metals or alkali earth metals capable of gigantic AC magnetic self-heating in biocompatible AC magnetic field and method of preparing the same

Abstract

Disclosed herein are iron oxide nanoparticles prepared through high-temperature thermal decomposition of an Fe.sup.3+ precursor and an M.sup.+ or M.sup.2+ (M=Li, Na, K, Mg, and Ca) precursor in an oxygen atmosphere. The iron oxide nanoparticles are nanoparticles, in which an alkali metal or alkali earth metal is doped into an Fe vacancy site of -Fe.sub.2O.sub.3, and generate explosive heat even in a biocompatible low AC magnetic field. Through both in vitro and in vivo tests, it was proven that cancer cells could be killed by performing low-frequency hyperthermia using the iron oxide nanoparticles set forth above.

Claims

1. Iron oxide nanoparticles in which -Fe.sub.2O.sub.3 is doped with an alkali metal or alkali earth metal, wherein: an Fe vacancy site of -Fe.sub.2O.sub.3 is doped with the alkali metal or alkali earth metal; the iron oxide nanoparticles generate heat in a biocompatible AC magnetic field of f.sub.appl.H.sub.appl of 3.010.sup.9 Am.sup.1s.sup.1 or less; the iron oxide nanoparticles are represented by M.sub.x-Fe.sub.2-xO.sub.3, where M is selected from the group consisting of Li, Na, K, Mg, and Ca, and x satisfies 0.05x0.15; and the iron oxide nanoparticles are superparamagnetic.

2. The iron oxide nanoparticles according to claim 1, wherein M is an alkali metal selected from the group consisting of lithium (Li), sodium (Na), and potassium (K).

3. The iron oxide nanoparticles according to claim 1, wherein M is an alkali earth metal selected from the group consisting of magnesium (Mg) and calcium (Ca).

4. The iron oxide nanoparticles according to claim 1, wherein the iron oxide nanoparticles generate heat in a biocompatible AC magnetic field of f.sub.appl.H.sub.appl of 1.810.sup.9 Am.sup.1s.sup.1 (f.sub.appl<120 kHz, H.sub.appl<15.12 kA/m) or less.

5. The iron oxide nanoparticles according to claim 1, wherein the iron oxide nanoparticles have an intrinsic loss power (ILP) of 13.5 nHm.sup.2/Kg to 14.5 nHm.sup.2/Kg in an AC magnetic field of f.sub.appl.H.sub.appl of 1.810.sup.9 Am.sup.1s.sup.1 (f.sub.appl<120 kHz, H.sub.appl<15.12 kA/m) or less.

6. The iron oxide nanoparticles according to claim 1, which generate heat sufficient to reach a temperature of at least 50 C. when placed in the biocompatible AC magnetic field.

7. A method of preparing iron oxide nanoparticles capable of heating in a biocompatible low AC magnetic field, the method comprising: preparing iron oxide nanoparticles in which alkali metal or alkali earth metal is doped with -Fe.sub.2O.sub.3 by mixing an Fe.sup.3+ precursor, an M.sup.+ or M.sup.2+ precursor where M is selected from the group consisting of Li, Na, K, Mg, and Ca, a surfactant, and a solvent in an oxygen atmosphere to form a mixture, and thermally decomposing the mixture at high temperature, wherein: an Fe vacancy site of -Fe.sub.2O.sub.3 is doped with the alkali metal or alkali earth metal; the iron oxide nanoparticles generate heat in a biocompatible AC magnetic field of fa.sub.ppl.H.sub.appl of 3.010.sup.9 Am.sup.1s.sup.1 or less; and the iron oxide nanoparticles are represented by M.sub.x-yFe.sub.2-xO.sub.3, where M is selected from the group consisting of Li, Na, K, Mg, and Ca, and x satisfies 0.05<x<0.15, wherein the iron oxide nanoparticles are superparamagnetic.

8. The method according to claim 7, wherein the Fe.sup.3+ precursor and the M.sup.+ or M.sup.2+ precursor comprises at least one member selected from the group consisting of metal nitrate, metal sulfate, metal acetylacetonate, metal fluoroacetoacetate, metal halide, metal perchlorate, metal alkyl oxide, metal sulfamate, metal stearate, and organic metal compounds.

9. The method according to claim 7, wherein the surfactant comprises at least one of organic acids with the chemical formula C.sub.nCOOH wherein 7<n<30.

10. The method according to claim 7, comprising: (a) heating a mixed solution of an Fe.sup.3+ precursor, an M.sup.+ or M.sup.2+ precursor where M is selected from the group consisting of Li, Na, K, Mg, and Ca, a surfactant, and a solvent to a temperature less than a boiling point of the solvent in a mixed atmosphere of oxygen and argon, followed by maintaining the mixed solution at the temperature for a certain period of time; (b) heating the mixed solution again to the boiling point of the solvent in a mixed atmosphere of oxygen and argon, followed by maintaining the mixed solution at the boiling point for a certain period of time; (c) removing a heat source and cooling the mixed solution to room temperature; and (d) performing precipitation and separation of nanoparticle powder by adding a polar solvent to the mixed solution and then performing centrifugation.

11. The method according to claim 7, wherein a doping level is adjusted by adjusting an amount of the Fe.sup.3+ precursor or the M.sup.+ or M.sup.2+ precursor.

12. The method according to claim 7, wherein the iron oxide nanoparticles generate heat sufficient to reach a temperature of at least 50 C. when placed in the biocompatible AC magnetic field.

13. The method of claim 9, wherein at least one organic acid is selected from the group consisting of oleic acid, lauric acid, stearic acid, myristic acid, and hexadecanoic acid.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

(2) FIG. 1 is a diagram illustrating a method of preparing M.sub.x-Fe.sub.2O.sub.3 (M=Li, Na, K, Mg, and Ca) nanoparticles according to an embodiment of the present invention.

(3) FIG. 2 shows transmission electron microscopy (TEM) images of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles prepared according to an embodiment of the present invention.

(4) FIG. 3 are diagrams illustrating atomic structure models describing the spin configuration of -Fe.sub.2O.sub.3 (maghemite) and Fe.sub.3O.sub.4 (magnetite) magnetic nanoparticles according to an embodiment of the present invention.

(5) FIG. 4 are diagrams illustrating crystal structure of Mg.sub.0.13-Fe.sub.2O.sub.3 (maghemite) magnetic nanoparticles according to an embodiment of the present invention. An enlarged image indicates that face-centered cubic lattices of oxygen (white sphere). The Fe.sup.3+ (blue sphere) spins at T.sub.d sites aligns in antiparallel with Fe.sup.3+ spins at O.sub.h sites under external magnetic field. The Mg.sup.2+ (red sphere) ions predominantly occupy the Fe vacancy sites existing in the O.sub.h sites of -Fe.sub.2O.sub.3.

(6) FIG. 5 is a graph depicting AC magnetically-induced heating characteristics of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and Fe.sub.3O.sub.4 measured at a f.sub.appl=110 KHz and H.sub.appl=140 Oe according to an embodiment of the present invention.

(7) FIG. 6 shows transmission electron microscopy (TEM) images of MFe.sub.2O.sub.4 (M=Mn, Co, Ni, Fe) nanoparticles prepared by a conventional method and a graph depicting AC magnetically-induced heating characteristics thereof in a low AC magnetic field.

(8) FIG. 7 shows graphs depicting X-ray absorption spectroscopy (left: X-ray absorption near edge structure, right: extended X-ray absorption fine structure) measurement results of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and bulk Fe.sub.3O.sub.4.

(9) FIG. 8 is a diagram showing composition determination of Mg.sub.0.13-Fe.sub.2O.sub.3 by energy dispersive X-ray spectroscopy (EDS).

(10) FIG. 9 shows DC minor hysteresis loops measured at a sweeping field of H.sub.appl=140 Oe (=11.14 KAm.sup.1) of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and Fe.sub.3O.sub.4, and FIG. 10 is a graph depicting a temperature dependent magnetization of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles measured at an excitation magnetic field of 100 Oe.

(11) FIG. 11 shows AC hysteresis loops measured at a f.sub.appl=110 KHz and H.sub.appl=140 Oe of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and Fe.sub.3O.sub.4.

(12) FIG. 12 shows graphs depicting the dependence of M.sub.s, P.sub.total, P.sub.relaxation loss, and on the Mg.sup.2+ cation doping concentration (level) (x) of Mg.sub.x-Fe.sub.2O.sub.3 according to an embodiment of the present invention.

(13) FIG. 13 shows graphs depicting the dependence of anisotropy energy and calculated Neel relaxation time on Mg.sup.2+ cation doping concentration (x) in Mg.sub.x-Fe.sub.2O.sub.3 according to an embodiment of the present invention.

(14) FIG. 14 shows characteristics of AC magnetically-induced heating temperature rise of Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids dispersed in toluene, ethanol, and D.I water measured at a f.sub.appl=110 KHz and H.sub.appl=140 Oe with a concentration of 3 mg/mL according to an embodiment of the present invention are moved to an aqueous solution layer.

(15) FIG. 15 is a graph for comparison of ILP values between previously reported superparamagnetic nanoparticles and Mg.sub.0.13-Fe.sub.2O.sub.3 superparamagnetic nanoparticles according to an embodiment of the present invention and existing representative materials known in the art.

(16) FIG. 16 shows graphs depicting characteristics of AC magnetically-induced heating temperature rise of Li.sub.0.15-Fe.sub.2O.sub.3, Na.sub.0.20-Fe.sub.2O.sub.3, K.sub.0.18-Fe.sub.2O.sub.3, and Ca.sub.0.18-Fe.sub.2O.sub.3 nanoparticles according to an embodiment of the present invention with Li.sup.+, Na.sup.+, K.sup.+, and Ca.sup.2+, respectively, in a low AC magnetic field.

(17) FIGS. 17 to 19 show results obtained by an in-vitro hyperthermia test using Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles according to an embodiment of the present invention.

(18) FIG. 20 shows results obtained by an in-vivo hyperthermia test using Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles according to an embodiment of the present invention.

(19) FIG. 21 shows graphs depicting results obtained by a toxicity test of Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles according to an embodiment of the present invention on U87MG cells and Hep3B cells.

(20) FIG. 22 shows graphs depicting cell survival rate of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles and reported superparamagnetic nanoparticles determined using U87MG cell lines.

DETAILED DESCRIPTION

(21) Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. It should be understood that the present invention is not limited to the following embodiments and may be embodied in different ways, and that the embodiments are provided for complete disclosure and thorough understanding of the invention by those skilled in the art.

(22) According to the present invention, iron oxide nanoparticles are prepared by doping an Fe vacancy site of -Fe.sub.2O.sub.3 with an alkali metal ion or alkali earth metal ion, and generate gigantic heat even in a biocompatible low AC magnetic field. In hyperthermia, a biocompatible AC magnetic field generally has f.sub.appl.Math.H.sub.appl of 5.010.sup.9 Am.sup.1s.sup.1 or less, preferably 3.010.sup.9 Am.sup.1s.sup.1 or less. The iron oxide nanoparticles according to the present invention can generate gigantic heat even in such a biocompatible low AC magnetic field.

(23) As used herein, the expression doped with . . . metal ion means that a metal atom is doped and ion-bonded to surrounding atoms, and thus all of expressions doped with . . . metal ion, doped with . . . metal atom, and doped with . . . metal should be interpreted as having the same meaning.

(24) The iron oxide nanoparticles according to the present invention generate heat in AC magnetic fields, and are preferably used in a biocompatible low AC magnetic field.

(25) Hereinafter, as an example of the iron oxide nanoparticles according to the present invention, Mg.sub.x-Fe.sub.2O.sub.3 will be described.

(26) Mg.sub.x-Fe.sub.2O.sub.3 is prepared by high-temperature thermal decomposition of an Fe.sup.3+ precursor and an Mg.sup.2+ precursor in an oxygen atmosphere, has a crystal structure in which an Fe vacancy site of -Fe.sub.2O.sub.3 is doped with Mg.sup.2+, and generates gigantic heat even in a biocompatible low AC magnetic field.

(27) A process of preparing Mg.sub.x-Fe.sub.2O.sub.3 (x=0.13) will be described in more detail (see FIG. 1).

(28) To prepare Mg.sub.0.13-Fe.sub.2O.sub.3, 0.13 mmol of magnesium (Mg) acetate tetrahydrate, 2.0 mmol of iron (Fe) acetylacetonate, 1.2 mmol of oleic acid, and 20 mL of benzyl ether are mixed in a 50 mL round bottom flask and are magnetically stirred. The mixed solution is heated to 200 C. for 30 minutes (8 C./min, first ramping up rate) in a mixed atmosphere of oxygen and argon (flow rate of 100 mL/min) and is then maintained for 50 minutes (nucleation step). Next, the mixed solution is heated again to 296 C. (boiling point of benzyl ether) for 20 minutes (5 C./min, second ramping rate) and is then maintained for 60 minutes (growth step).

(29) Next, a heating source is removed and the mixed solution is cooled to room temperature.

(30) A polar solvent such as ethanol is added to the mixed solution, followed by centrifugation, thereby precipitating and separating black powder. Separated products (nanoparticles) are dispersed in a nonpolar solvent such as toluene.

(31) To control the Mg.sup.2+ doping concentration (x) of Mg.sub.x-Fe.sub.2O.sub.3, the different amount of Mg.sup.2+/Fe.sup.3+ metal precursor are used under identical experimental conditions. For example, to synthesize the Mg.sub.0.10-Fe.sub.2O.sub.3 nanoparticles, 0.10 mmol of Mg acetate tetrahydrate and 2.0 mmol of Fe acetylacetonate are used under the identical experimental conditions.

(32) FIG. 2 shows transmission electron microscope images of 7 nm Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles prepared through the above processes. Referring to the right-side image, it can be seen that a crystal growth orientation (400) is observed and the corresponding lattice distance is 2.09 , and that a crystal growth orientation (200) is observed and the corresponding lattice distance is 2.95 . These values are the same as those of a reference bulk material of -Fe.sub.2O.sub.3. Therefore, it can be proven that the crystal structure of nanoparticles prepared through the above processes has a typical spinel structure of -Fe.sub.2O.sub.3.

(33) In a conventionally synthesized nanoparticles, MgFe.sub.2O.sub.4 nanoparticles (Mg.sup.2+ doped Fe.sub.3O.sub.4 structure) are prepared.

(34) To prepare MgFe.sub.2O.sub.4 nanoparticles, 1.0 mmol of MgCl.sub.2 and 2.0 mmol of Fe(acac).sub.3 are placed in a 100 mL round bottom flask containing dibenzyl ether and a surfactant (oleic acid and oleylamine). 10.0 mmol of 1,2-hexadecandiol is used as a reductant.

(35) The mixed solution is heated to 200 C. for 25 minutes in an argon atmosphere and maintained for 60 minutes (nucleation step).

(36) Next, the mixed solution is heated again to 296 C. (boiling point of benzyl ether) for 30 minutes and maintained for 60 minutes. A heat source is removed and a reaction mixture is cooled to room temperature.

(37) Ethanol is added to the reaction product, followed by centrifugation, thereby obtaining precipitated black powder. The obtained MgFe.sub.2O.sub.4 nanoparticles are dispersed in a nonpolar solvent such as toluene.

(38) Upon preparation of existing nanoparticles, two chemical reagents, that is, oleic acid and oleylamine are used as size control factors, and oleylamine can be used as reducing agent. The crystal structure may be change -Fe.sub.2O.sub.3 into Fe.sub.3O.sub.4 during synthesis process thereof.

(39) Oleylamine is mainly used in preparation of iron oxide nanoparticles. In order to investigate the role of surfactant, a control experiments were carried out with oleylamine instead of oleic acid. The nanoparticles synthesized with oleylamine showed a similar AC self-heating behavior to MFe.sub.2O.sub.4 (M=Fe.sup.3+, Co.sup.2+, Ni.sup.2+, Mg.sup.2+) nanoparticles due to the reduction of Fe.sup.3+ into Fe.sup.2+ that leads to the Fe.sub.3O.sub.4 lattice in the presence of oleylamine. This result confirms that Mg.sup.2+ ion doped Fe.sub.3O.sub.4 lattice has no contribution to the AC heating properties.

(40) Mg.sub.x-Fe.sub.2O.sub.3, which corresponds to the iron oxide nanoparticles according to the present invention, is obtained by doping Fe vacancy sites of -Fe.sub.2O.sub.3 with Mg.sup.2+.

(41) Unlike Fe.sub.3O.sub.4, -Fe.sub.2O.sub.3 has spaces (vacancy sites), which occupy about 12% of the total volume thereof (see FIG. 3). Since Fe.sub.3O.sub.4 is gradually changed into -Fe.sub.2O.sub.3 upon preparation in an oxygen atmosphere, Fe.sup.2+ ions present in Fe.sub.3O.sub.4 are oxidized to Fe.sup.3+ ions, diffused out, and vacancy sites was formed in -Fe.sub.2O.sub.3. Since oxidation is required to prepare the iron oxide nanoparticles according to the present invention, preparation may be performed in an oxygen atmosphere, or an oxidant may be used. Actual preparation is preferably performed in a mixed atmosphere of oxygen and argon for stability of reaction.

(42) When such vacancy sites of -Fe.sub.2O.sub.3 is doped with an alkali metal or alkali earth metal, the doped -Fe.sub.2O.sub.3 demonstrate change in DC/AC magnetic softness and magnetic properties, specifically magnetic susceptibility, and thus responds to a low AC magnetic field, thereby generating heat (see FIG. 4).

(43) On the other hand, unlike the above case, in the case of a transition metal (Zn, Fe, Mn, Co, Ni, and the like), since it is energetically favorable in terms of thermodynamics that the transition metal is predominantly substituted with Fe.sup.3+ ions at an octahedral site (O.sub.h) and a tetrahedral site (T.sub.h), not in vacancy site, doped -Fe.sub.2O.sub.3 exhibits reduced net magnetic properties and respond negligibly to a low AC magnetic field (see FIGS. 3 and 4).

(44) The iron oxide nanoparticles according to the present invention are nanoparticles in which Fe vacancy sites of -Fe.sub.2O.sub.3 are doped with an alkali metal or alkali earth metal. According to the present invention, a doping metal includes any alkali metal or alkali earth metal without limitation. Preferably, the alkali metal is lithium (Li), sodium (Na), or potassium (K), and the alkali earth metal is magnesium (Mg) or calcium (Ca).

(45) In addition, the iron oxide nanoparticles according to the present invention are doped with at least one alkali metal or alkali earth metal, preferably at least one metal ion selected from the group consisting of Li.sup.+, Na.sup.+, K.sup.+, Mg.sup.2+, and Ca.sup.2+.

(46) The iron oxide nanoparticles according to the present invention may emit gigantic heat even in a biocompatible low AC magnetic field of f.sub.appl.Math.H.sub.appl of 3.010.sup.9 Am.sup.1s.sup.1 or less, and have an intrinsic loss power (ILP) of 13.5 nHm.sup.2/Kg to 14.5 nHm.sup.2/Kg in an AC magnetic field of f.sub.appl.Math.H.sub.appl<1.810.sup.9 Am.sup.1s.sup.1 (f.sub.appl<120 KHz, H.sub.appl<15.12 KA/m).

(47) Further, the iron oxide nanoparticles according to the present invention may be represented by M.sub.x-Fe.sub.2O.sub.3 (M=Li, Na, K, Mg, and Ca), and x may vary with a doping concentration of a metal. x satisfies 0.00<x0.30, preferably 0.10x0.25, more preferably 0.10x0.20.

(48) The iron oxide nanoparticles according to the present invention have an average particle diameter of about 7 nm to about 13 nm, without being limited thereto, and may have various nanometer scale sizes.

(49) FIG. 5 is a graph depicting AC magnetically-induced heating characteristics of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and Fe.sub.3O.sub.4 measured at a low AC magnetic field (f.sub.appl=110 kHz, H.sub.appl=140 Oe) according to an embodiment of the present invention. As shown in FIG. 5, Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles exhibits an exceptionally high T.sub.ac,max of 180 C. While, conventionally prepared MgFe.sub.2O.sub.4 (Mg.sup.2+ ion doped Fe.sub.3O.sub.4) nanoparticles exhibits a low T.sub.ac,max of 22 C. under same AC magnetic field. According to an conventional method have almost no effect of heat emission at a low AC magnetic field.

(50) FIG. 6 shows a graph depicting transmission electron microscopy images and AC magnetically-induced heating characteristics of MFe.sub.2O.sub.4 (M=Co.sup.2+, Fe.sup.2+, Mn.sup.2+, and Ni.sup.2+) nanoparticles, which are prepared by a conventional method under the same AC magnetic conditions (f.sub.appl=110 kHz, H.sub.appl=140 Oe). It can be seen that conventionally synthesized nanoparticles (CoFe.sub.2O.sub.4, Fe.sub.3O.sub.4, MnFe.sub.2O.sub.4, and NiFe.sub.2O.sub.4) have almost no effect of heat emission. CoFe.sub.2O.sub.4, Fe.sub.3O.sub.4, MnFe.sub.2O.sub.4, and NiFe.sub.2O.sub.4 have a structure in which Co.sup.2+, Fe.sup.2+, Mn.sup.2+, and Ni.sup.2+ ions are doped into Fe.sub.3O.sub.4 instead of -Fe.sub.2O.sub.3, respectively.

(51) FIG. 7 shows X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) measurement results of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles. For comparison, conventional MgFe.sub.2O.sub.4 nanoparticles and bulk Fe.sub.3O.sub.4 was prepared. The Fe K-edge XANES spectra (left-side graph in FIG. 7) showed that Mg.sub.0.13-Fe.sub.2O.sub.3, and MgFe.sub.2O.sub.4 nanoparticles have average iron oxidation states of +3, and +2.75, respectively. Comparing to bulk Fe.sub.3O.sub.4, MgFe.sub.2O.sub.4 nanoparticles have a typical local Fe coordination of Fe.sub.3O.sub.4 while, according to the EXAFS analyzed results (right-side graph in FIG. 7), it was confirmed that Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles has a typical -Fe.sub.2O.sub.3 (maghemite). In contrast, conventionally synthesized MgFe.sub.2O.sub.4 nanoparticles are obtained by doping Mg.sup.2+ into a magnetite (Fe.sub.3O.sub.4) structure, and that the Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles according to the present invention are obtained by doping Mg.sup.2+ into a maghemite (-Fe.sub.2O.sub.3) structure.

(52) FIG. 8 shows composition determination results of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles using an energy dispersive X-ray spectroscopy (EDS), and it can be seen that Mg.sup.2+ ions is well presented in -Fe.sub.2O.sub.3 nanoparticles.

(53) FIG. 9 shows DC minor hysteresis loops and FIG. 10 shows the temperature dependent magnetization of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles. Referring to FIGS. 9 and 10, it can be proven that Mg.sub.0.13-Fe.sub.2O.sub.3 according to the present invention exhibits superparamagnetism.

(54) FIG. 11 shows the area of AC hysteresis loops measured at a f.sub.appl=110 KHz and H.sub.appl=140 Oe of Mg.sub.0.13-Fe.sub.2O.sub.3, MgFe.sub.2O.sub.4, and Fe.sub.3O.sub.4. Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles had the much larger area than those of MgFe.sub.2O.sub.4 and Fe.sub.3O.sub.4. The larger AC hysteresis loss area of Mg.sub.0.13-Fe.sub.2O.sub.3 indicates that it has a higher AC magnetic softness (or faster AC magnetic response). Thus, it can be proven that the Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles according to the present invention exhibit gigantic (or exceptionally high) heat emission.

(55) FIG. 12 shows graphs depicting the dependence of M.sub.s, P.sub.total, P.sub.relaxation loss, and .sub.m on the Mg.sup.2+ cation doping concentration (x) of Mg.sub.x-Fe.sub.2O.sub.3 nanoparticles. The .sub.m (and P.sub.relaxation-loss) was dramatically increased from 2.5110.sup.3 emu.Math.g.sup.1Oe.sup.1 (6.8410.sup.6 W/m.sup.3) to 7.57410.sup.3 emu.Math.g.sup.1Oe.sup.1 (3.4210.sup.6 W/m.sup.7) by increasing the doping concentration of Mg.sup.2+ ion up to 0.13 and P.sub.total was correspondingly increased from 7.8210.sup.6 W/m.sup.3 to 410.sup.7 W/m.sup.3. However, the further increase the doping concentration of Mg.sup.2+ ion up to 0.15 led to severe reduction of .sub.m (P.sub.Nel-relaxation-loss), M.sub.s, and P.sub.total. The increase of Mg.sup.2+ ion doping concentration from 0 to 0.13 during the synthesis leads to the acceleration of occupation of Fe vacancy sites by Mg.sup.2+ ions in -Fe.sub.2O.sub.3 lattice so that results in the increase of Mg.sup.2+ doping concentration that would be mainly responsible for the significant enhancement of M.sub.s (.sub.m). On the contrary, the sudden decrease of .sub.m, P.sub.total, and M.sub.s at x=0.15 can be supposed to be due to the reduction of Mg.sup.2+ doping concentration in Fe vacancy sites in -Fe.sub.2O.sub.3 resulted from the substitution of Fe.sup.3+ in the O.sub.h site by Mg.sup.2+ cations. Referring to FIG. 12, it can be seen that the Mg.sub.x-Fe.sub.2O.sub.3 nanoparticles emit a large amount of heat when x satisfies 0.05x0.15. The Mg.sub.x-Fe.sub.2O.sub.3 nanoparticles theoretically emit the largest amount of heat in the case of x=0.13, and it can be seen that this coincides with the experimental results.

(56) FIG. 13 shows graphs depicting the dependence of anisotropy energy and calculated Neel relaxation time on Mg.sup.2+ cation doping concentration (x) in Mg.sub.x-Fe.sub.2O.sub.3 nanoparticles. The physical reason for the obvious increase of .sub.m (P.sub.Nel-relaxation-loss) depending on the Mg.sup.2+ cation doping concentration is thought to be primarily due to the enhanced .sub.N (faster .sub.N) that is resulted from the change of AC magnetic softness or magnetic anisotropy caused by the modification of Mg.sup.2+ doping concentration in Fe vacancy sites of -Fe.sub.2O.sub.3.

(57) FIG. 14 shows characteristics of AC magnetically-induced heating temperature rise of Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids dispersed in toluene, ethanol, and D.I water measured at a f.sub.appl=110 KHz and H.sub.appl=140 Oe with a concentration of 3 mg/mL Referring to FIG. 14, it can be confirmed that the Mg.sub.0.13-Fe.sub.2O.sub.3 iron oxide nanoparticles according to the present invention have an intrinsic loss power (ILP) value of about 13.9 nHm.sup.2 kg.sup.1 (in toluene), a 14.5 nHm.sup.2 kg.sup.1 (in ethanol), and a 14.0 nHm.sup.2 kg.sup.1 (in water), respectively.

(58) FIG. 15 is a graph for comparison of ILP values between iron oxide nanoparticles according to the present invention and previously reported superparamagnetic nanoparticles known in the art. Referring to FIG. 15, it can be confirmed that the iron oxide nanoparticles according to the present invention have an ILP value that is about 100 times higher than that of commercial Fe.sub.3O.sub.4 (Feridex) nanoparticles.

(59) FIG. 16 shows graphs depicting characteristics of AC magnetically-induced heating temperature rise of Li.sub.0.15-Fe.sub.2O.sub.3, Na.sub.0.20-Fe.sub.2O.sub.3, K.sub.0.18-Fe.sub.2O.sub.3, and Ca.sub.0.18-Fe.sub.2O.sub.3 nanoparticles, respectively, in a low AC magnetic field (f.sub.appl=110 kHz, H.sub.appl=140 Oe). It can be confirmed that all the nanoparticles (Li.sub.0.15-Fe.sub.2O.sub.3, Na.sub.0.20-Fe.sub.2O.sub.3, K.sub.0.18-Fe.sub.2O.sub.3, and Ca.sub.0.18-Fe.sub.2O.sub.3) exhibited an exceptionally high T.sub.AC,max above 100 C. Therefore, it can be seen that all of the M.sub.x-Fe.sub.2O.sub.3 (M=Li, Mg, K, Na, and Ca) nanoparticles according to the present invention, which are obtained by doping -Fe.sub.2O.sub.3 with Mg or by doping Fe.sub.2O.sub.3 with a different alkali metal or alkali earth metal from Mg, exhibit high AC self-heating in a low AC magnetic field.

(60) FIG. 17 show results obtained by an in-vitro hyperthermia test using U87MG cells after treating Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles and Resovist.

(61) The U87MG cells were incubated with 700 g/mL of Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids and Resovist, as a control group, for cellular uptake. The cells were placed in the center of an AC magnetic coil, and a magnetic field of f.sub.appl=99 kHz and H.sub.appl=155 Oe (H.sub.appl.Math.f.sub.appl=1.2210.sup.9 Am.sup.1s.sup.1) was applied to the cells for 1500 seconds. Referring to the right-side graph of FIG. 17, it can be confirmed that the cells treated with Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids showed the much higher T.sub.AC,max (63.5 C.) than Resovist (37.5 C.).

(62) Referring to FIG. 18 showing the optical microscope images of U87MG cells before and after magnetic nanofluid hyperthermia with Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids, it can be confirmed that the cell necrosis of U87MG resulted from severe deformation and shrinkage of the cell morphology caused by the applied thermal energy was clearly observed after magnetic nanofluid hyperthermia, all cancer cells were killed by heat. In more detail, it was confirmed that 75% of the cancer cells were killed at 48 C. and all of the cancer cells were completely necrotized at 63.5 C. Thus, bioavailability of the nanoparticles according to the present invention was proven.

(63) On the other hand, FIG. 19 is an image showing the optical microscope image of U87MG cells after magnetic nanofluid hyperthermia with Resovist (control group), and cells suffering from deformation or shrinkage were not observed. Thus, it can be seen that cell viability was strongly dependent on AC heating temperature.

(64) FIG. 20 shows results obtained by an in-vivo hyperthermia test using Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles.

(65) Hep3B cells transfected with luciferase (for bioluminescence imaging, BLI) grew subcutaneously in mice (cancer-xenograft model)

(66) A 100 L of Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids (100 L, 11.5 g/L) was intratumorally injected into cancer cells of the mice (1000 mm.sup.3) through soft tissue using a bent needle and optical thermometers were mounted in the cancer cells and rectum area to monitor the temperature.

(67) For comparison, Resovist (100 L, 11.5 g/L) and normal saline (100 L, 11.5 g/L) were also intratumorally injected into the mice, respectively.

(68) The mice were placed in the center of an AC magnetic coil and exposed to an AC magnetic field (f.sub.appl=99 kHz, H.sub.appl=155 Oe, H.sub.appl.Math.f.sub.appl=1.2210.sup.9 Am.sup.1s.sup.1) for 900 seconds.

(69) The temperature of the rectum and Hep3B injected with Resovist were slightly increased from 34 C. to 36.37 C. and 37.14 C., respectively. However, the temperature of the Hep3B cells injected with the Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids was rapidly increased up to 50.2 C. (thermoablation temperature).

(70) The activity of the Hep3B was analyzed by employing a bioluminescence imaging (BLI) technique. The Hep3B treated with Mg.sub.0.13-Fe.sub.2O.sub.3 nanofluids did not exhibit any BL-intensity from day 2 after magnetic nanofluid hyperthermia, while the control groups still exhibited strong BL-intensity after magnetic nanofluid hyperthermia. No BL-intensity means that the cancer cells was completely necrotized by magnetic nanofluid hyperthermia.

(71) FIG. 21 shows graphs depicting results obtained by a toxicity test of Mg.sub.0.13-Fe.sub.2O.sub.3 magnetic nanoparticles according to the present invention on U87MG cells and Hep3B cells. From the this result, it was confirmed that Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles showed a high biocompatibility (non-toxicity) even at a higher concentration (300 g/mL).

(72) FIG. 22 shows graphs depicting cell survival rate of Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles and reported superparamagnetic nanoparticles determined using U87MG cell lines. Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles had a higher biocompatibility (non-toxicity) with U87Mg cell lines compared to all other reported superparamagnetic nanoparticles even at a higher concentration (300 g/mL). The reported nanoparticles (Fe.sub.3O.sub.4, CoFe.sub.2O.sub.4, CoFe.sub.2O.sub.4@MnFe.sub.2O.sub.4) have a Fe.sub.3O.sub.4 crystal structure. In the case of Fe.sub.3O.sub.4, the Fenton reaction is likely to occur and produce a toxic effect to the cells during cellular internalization due to the Fe.sup.2+ ions in Fe.sub.3O.sub.4 lattice. However, Mg.sub.0.13-Fe.sub.2O.sub.3 nanoparticles, which is fully oxidized forms from Fe.sub.3O.sub.4, has only Fe.sup.3+ ions in -Fe.sub.2O.sub.3 lattice crystal. Hence the possibility to occur Fenton reaction is readily expected to be an extremely low during cellular internalization.

(73) As described above, it was proven through both of the in-vitro and in-vivo tests that cancer cells could be completely killed using magnetic nanoparticles according to the present invention. Therefore, the iron oxide nanoparticles according to the present invention can be clinically used.

(74) Heretofore, the present invention has been described with reference to some embodiments in conjunction with the accompanying drawings. Although specific terms are used herein, it should be understood that the terms are only for the purpose of describing the embodiments of the present invention and are not intended to limit the present invention. In addition, it should be understood that various modifications, changes, alterations, and equivalent embodiments can be made by those skilled in the art without departing from the spirit and scope of the present invention. Therefore, the scope of the present invention should be defined only by the accompanying claims and equivalents thereof.