NL2025687B1 - Superparamagnetic iron oxide nanoclusters and preparation method and application thereof - Google Patents

Superparamagnetic iron oxide nanoclusters and preparation method and application thereof Download PDF

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NL2025687B1
NL2025687B1 NL2025687A NL2025687A NL2025687B1 NL 2025687 B1 NL2025687 B1 NL 2025687B1 NL 2025687 A NL2025687 A NL 2025687A NL 2025687 A NL2025687 A NL 2025687A NL 2025687 B1 NL2025687 B1 NL 2025687B1
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iron oxide
drying oven
superparamagnetic iron
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Sun Xilin
Wu Lina
Li Yingbo
Wang Kai
Yang Lili
Yang Jie
Liu Shuang
Li Xiaona
Jiang Meng
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Univ Harbin Medical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
    • A61K49/10—Organic compounds
    • A61K49/12—Macromolecular compounds
    • A61K49/126—Linear polymers, e.g. dextran, inulin, PEG
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1887—Agglomerates, clusters, i.e. more than one (super)(para)magnetic microparticle or nanoparticle are aggregated or entrapped in the same maxtrix
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
    • C01G49/00—Compounds of iron
    • C01G49/02—Oxides; Hydroxides
    • C01G49/08—Ferroso-ferric oxide [Fe3O4]
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2004/00—Particle morphology
    • C01P2004/01—Particle morphology depicted by an image
    • C01P2004/03—Particle morphology depicted by an image obtained by SEM

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  • Compounds Of Iron (AREA)

Abstract

The present invention discloses a superparamagnetic iron oxide nanoclusters and a preparation method and an application thereof. The superparamagnetic iron oxide nanoclusters are synthesized by an improved solvothermal method, ferric chloride hexahydrate (FeC1306H20) is used as a raw material, a cluster-shaped product Fe304 is synthesized by a hydrothermal method, and a size of the cluster-shaped product Fe304 is adjustable within a range of 60-200 nm by changing a proportion of a mixed solvent. Because of characteristics of good biocompatibility, unique magnetic characteristic, pH response and the like, the superparamagnetic iron oxide nanoclusters prepared by the present invention not only can be used as a Magnetic Resonance Imaging (MRI)-T2 weighted nanoprobe for magnetic resonance molecular imaging, but also has the characteristic of pH response.

Description

SUPERPARAMAGNETIC IRON OXIDE NANOCLUSTERS AND PREPARATION METHOD AND APPLICATION THEREOF
FIELD OF TECHNOLOGY The present invention belongs to the technical field of nano magnetic materials, and relates to a superparamagnetic iron oxide nanoclusters and a preparation method and an application thereof.
BACKGROUND ART In the prior art, along with rapid development of manufacturing, assembly and modification technologies, the application of nanomaterials shows a bright future in the high and new technical fields, such as high-sensitivity and high-selectivity separators and sensors, etc. The research on unique physical properties and mechanisms of the nanomaterials and the utilization on characteristics of the nanomaterials are constituted into core contents of nanoscience and nanotechnology. In recent years, owing to the development of the nanomaterials and particularly novel nanoparticles for cancer diagnosis and treatment, the nanobiotechnology has made an unprecedented progress in aspects of magnetic separation between proteins and cells, replacement of fluorescems by quantum dot, and research, development and medical diagnosis of magnetic- resonance molecular imaging probes. Superparamagnetic Iron Oxide Nanoparticles (SPIONs) are nanomaterials that are most functional and researched in biomedical application. With a high biocompatibility, a negligible toxicity to healthy tissues and excellent surface chemical properties, the SPIONs have been widely applied to Magnetic Resonance Imaging (MRI), tissue repair, immunoassay, thermal therapy, drug delivery and cell separation. As a magnetic nanoparticle, nano Fe;O, has a single magnetic domain structure and a very high coercivity; and when prepared into a magnetic recording material, it may improve a signal-to-noise ratio and improve the image quality. Because of such characteristics as a high saturated magnetization intensity, the Fe;O4 nanoparticles are often used for magnetic- resonance T2 weighed imaging, Till now, many processes for preparing the nano Fe;O, have been researched by people, including a coprecipitation method. a sol-gel method, a micro-emulsion method, a microwave synthesis method, a polyol method, a mechanical grinding method, a pyrolysis method. a hydrothermal method, etc. The SPIONSs are widely applied to the field of biomedicine, for example, as MRI-T2 weighed imaging nanomaterial, they are used in imaging of livers and spleens, lymph nodes, tumors and other tissues, and magnetic-heat treatment, magnetic separation, and tumor diagnosis and treatment. Different from normal tissues, the Tumor Microenvironment (TME) has the characteristics of non-uniform nutritional distribution, insufficient oxygenation (oxygen deprivation), acidic pH (acidosis) and high redox state, thus leads to weakly acidic environment. In case of synthesis of the Fe;0, nanoparticles that have the MRI-T2 weighed imaging characteristic and the TME response characteristic, and can be dissociated into undersized nanoparticles in a weakly acidic condition to penetrate into a deep tumor part to release iron ions, and thus accelerates apoptosis of tumor cells in combination with other therapies such as a chemotherapy, it will be of a great significance to integrated research of tumor diagnosis and treatment. However, the Fe;Oy researched and synthesized at present fails to have the TME response characteristic.
SUMMARY One technical problem to be solved by the present invention is to research and develop a superparamagnetic iron oxide nanoclusters and a preparation method and an application thereof for the above shortages of the prior art. Fe;Qy synthesized by the method has a controllable size, and the nanoclusters may respond to a weakly acidic TME. To achieve the above objective, the present invention researches and develops the following technical solutions: A preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps: (1) weighing FeCl;+6H,0 in a container, adding an Ethylene Glycol (EG) solution and a Diethylene Glycol (DEG) solution, and magnetically stirring to dissolve the FeCl;*6H,0; (2) adding Polyacrylic Acid (PPA) to a solution, and stirring to dissolve the PPA; (3) adding sodium acetate (CH:COONa) and sodium hydroxide (NaOH), stirring, and heating to dissolve the CH;COONa and the NaOH to form a precursor; (4) transferring the above precursor to a hydrothermal reactor, sealing the hydrothermal reactor, then putting into a blast drying oven, heating to 180-280°C, and after 4-16 h of reaction, closing the blast drying oven and cooling naturally; and (5) taking out a colloidal product, adding anhydrous ethanol and water, ultrasonically washing, separating with a magnet, and washing and separating once again with a same method; and placing a separated solid into a vacuum drying oven and drying for 12-36 h at a room temperature, and sealing obtained Fe; 0, dry powder for storage. In the above solutions, a total volume of the EG and DEG solutions 1s 30 mL, and a ratio of the EG solution to the DEG solution is 30/0-10/20 by volume (v/v). Preferably, a preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps:
(1) weighing FeCl;*6H,0 in a beaker, adding an EG solution and a DEG solution, and magnetically stirring for 10 min to dissolve the FeCl;*6H,0; (2) adding PPA to a solution, and stirring for 30 min to dissolve the PPA; (3) adding CH;COONa and NaOH, stirring for 40 min, and heating to dissolve the CH3COONa and the NaOH to form a precursor; (4) transferring the above precursor to a hydrothermal reactor, sealing the hydrothermal reactor, then putting into a blast drying oven, heating to 210°C, and after 10 h of reaction, closing the blast drying oven and cooling naturally: and (5) taking out a colloidal product, adding anhydrous ethanol and water, ultrasonically washing, separating with a magnet, and washing and separating once again with a same method; and placing a separated solid into a vacuum drying oven and drying for 24 h at a room temperature, and sealing obtained Fe:O, dry powder for storage.
The present invention has the following advantages:
1. The present invention is simple and easy in operation, and low in raw material cost: and synthesis reaction is carried out in an airtight environment, thereby being environment-friendly. 2. The synthesized nanoclusters has strong size controllability, and good monodispersity and biocompatibility. 3. The synthesized nanoclusters has a pH response characteristic, may be decomposed into iron oxide nanoparticles of different particle sizes in different pH conditions, and shows different T2 signal intensities and T2 relaxation rates on MR. 4. The synthesized nanoclusters have a high T2 relaxation rate, which provides new method and technology for MR diagnosis or treatment.
BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic diagram of a preparation route according to the present invention.
Fig. 2 is a Scanning Electron Microscope (SEM) image of SPIONSs of different particle sizes.
Fig. 3 is a phantom in-vitro MR image of SPIONSs in different pH conditions.
Fig. 4 is a value of a T2 relaxation rate measured in different pH conditions.
DETAILED DESCRIPTION OF THE EMBODIMENTS Embodiment | A preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps: (1) 0.2162 g of FeCl;+6H,0 was weighed in a beaker, an EG solution and a DEG solution were added, and magnetic stirring was carried out for 10 min to dissolve the FeCl;*6H,0, a ratio of the
EG to the DEG being 30 mL to 0 mL by volume. (2) 0.5794 g of PPA was added to a solution, and dissolved with stirring for 30 min. (3) 2.5874 g of CH3COONa and 0.1402 g of NaOH were added and dissolved with stirring for 40 min and appropriate heating to form a precursor.
(4) The above precursor was transferred to a hydrothermal reactor; the hydrothermal reactor was sealed, put into a blast drying oven, heated to 210°C; and after 10 h of reaction, the blast drying oven was closed and cooled naturally.
(5) A colloidal product was taken out, 100 mL of anhydrous ethanol and water were added, ultrasonic washing was carried out for 5 min, a magnet was used for separating, and a same method was used for washing and separating once again; and a separated solid was placed into a vacuum drying oven and dried at a room temperature, and obtained Fe;O, dry powder was sealed for storage. Thereafter, further detection on particle size, shape and MR performance was carried out, with a specific result as shown in Fig. 2 (200 nm).
During synthesis, the PPA is used as a stabilizer, and a synthesized Fe30, particle is self- assembled under a cross-linking action of the PPA to form the nanoclusters; and as the PPA has pH response characteristic, the svnthesized nanoclusters may be decomposed into iron oxide nanoparticles of different particle sizes in different pH conditions.
By controlling different volume ratios of the EG to the DEG, the synthesized Fe30: has a controllable size.
In an acidic condition, the PPA cross-linked on a surface of the nanoclusters is dissociated, and the covered Fe;0, nanoparticles are separated into nanoparticles of a smaller size from the clusters. Therefore. the nanoclusters may respond to a weakly acidic TME.
Embodiment 2 A preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps: (1) 0.2162 g of FeC:1::6H;0 was weighed in a beaker, an EG solution and a DEG solution were added, and magnetic stirring was carried out for 10 min to dissolve the FeCl;+6H,0, a ratio of the EG to the DEG being 20 mL to 10 mL by volume.
(2) 0.5794 g of PPA was added to a solution, and dissolved with stirring for 30 min.
(3) 2.5874 g of CH:COONa and 0.1402 g of NaOH were added and dissolved with stirring for 40 min and appropriate heating to form a precursor.
(4) The above precursor was transferred to a hydrothermal reactor; the hydrothermal reactor was sealed, put into a blast drying oven, heated to 210°C; and after 10 h of reaction, the blast drving oven was closed and cooled naturally.
(5) A colloidal product was taken out, 100 mL of anhydrous ethanol and water were added. ultrasonic washing was carried out for 5 min, a magnet was used for separating, and a same method was used for washing and separating once again; and a separated solid was placed into a vacuum drying oven and dried for 24 h at a room temperature, and obtained Fe:O, dry powder was sealed 5 for storage. Thereafter, further detection on particle size, shape and MR performance was carried out, Thereafter, further detection on particle size, shape and MR performance was carried out, with a specific result as shown in Fig. 2 (140 nm).
Embodiment 3 A preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps: (1) 0.2162 g of FeC;:6H;0 was weighed in a beaker, an EG solution and a DEG solution were added. and magnetic stirring was carried out for 10 min to dissolve the FeCl;6H;0. a ratio of the EG to the DEG being 15 mL to 15 mL by volume.
(2) 0.5794 g of PPA was added to a solution, and dissolved with stirring for 30 min.
(3) 2.5874 g of CH;COONa and 0. 1402 g of NaOH were added and dissolved with stirring for 40 min and appropriate heating to form a precursor.
(4) The above precursor was transferred to a hydrothermal reactor; the hydrothermal reactor was sealed, put into a blast drying oven, heated to 210°C; and after 10 h of reaction, the blast drying oven was closed and cooled naturally.
(5) A colloidal product was taken out, 100 mL of anhydrous ethanol and water were added, ultrasonic washing was carried out for 5 min, a magnet was used for separating, and a same method was used for washing and separating once again; and a separated solid was placed into a vacuum drying oven and dried for 24 h at a room temperature, and obtained FeO, dry powder was sealed for storage. Thereafter, further detection on particle size, shape and MR performance was carried out. with a specific result as shown in Fig. 2 (80 nm).
Embodiment 4 A preparation method of a superparamagnetic iron oxide nanoclusters includes the following steps: (1) 0.2162 g of FeCl;+6H,0 was weighed in a beaker, an EG solution and a DEG solution were added, and magnetic stirring was carried out for 10 min to dissolve the FeCl;*6H,0, a ratio of the EG to the DEG being 10 mL to 20 mL by volume.
(2) 0.5794 g of PPA was added to a solution, and dissolved with stirring for 30 min.
(3) 2.5874 g of CH;COONa and 0. 1402 g of NaOH were added and dissolved with stirring for 40 min and appropriate heating to form a precursor.
(4) The above precursor was transferred to a hydrothermal reactor; the hydrothermal reactor was sealed, put into a blast drying oven, heated to 210°C; and after 10 h of reaction, the blast drying oven was closed and cooled naturally. (5) A colloidal product was taken out, 100 mL of anhydrous ethanol and water were added, ultrasonic washing was carried out for 5 min, a magnet was used for separating, and a same method was used for washing and separating once again; and a separated solid was placed into a vacuum drying oven and dried for 24 h at a room temperature, and obtained Fe;O, dry powder was sealed for storage. Thereafter, further detection on particle size, shape and MR performance was carried out, with a specific result as shown in Fig. 2 (60 nm).
Experimental Example | In-vitro MRI detection of superparamagnetic iron oxide nanoclusters: a superparamagnetic iron oxide nanoclusters having a particle size of 60 nm was used, and respectively prepared, in a PBS or diluted hydrochloric acid solution having a pH value of 7.4/6.5/5.5, into 200 pL of superparamagnetic iron oxide nanoclusters aqueous solution having a concentration of 50 pg Fe/mL, 25 ug Fe/mL, 12.5 pg Fe/mL, 6.25 ug Fe/mL and 3.125 pg Fe/mL; and the solution was loaded into an EP tube having a capacity of 250 uL, and a T2 image and a T2 mapping were detected by a 9.4 T MR scanner. The 9.4 T MR scanner uses the following scanning parameters: TurboRARE-T2 has repetition time=2000 ms, echo time=8 ms, FOV=60*32 mm, thickness=1 mm, matrix=256*256 and the Number of Excitation (NEX)=1; and the RARE-T2 mapping has repetition time= 2000 ms, echo time=8-64 ms, FOV=60*32 mm, thickness=1 mm, matrix=256*256 and NEX=I1. Upon the completion of test, image data was processed, and determination of a relaxation rate and contrast in T2 signal enhancement were carried out in different pH conditions. Thereafter, further detection on particle size, shape and MR performance was carried out, with a specific result as shown in Fig. 3 and Fig. 4.
The present invention is simple and easy in operation, and low in raw material cost. The synthesized nanoclusters have good monodispersity and biocompatibility. The synthesized nanoclusters may be decomposed into iron oxide nanoparticles of different particle sizes in different pH conditions, and shows different T2 signal intensities and T2 relaxation rates on MR.
The above descriptions are merely specific embodiments of the present invention, and various illustrations are not intended to limit a substantial content of the present invention.

Claims (3)

ConclusiesConclusions 1. Een bereidingswerkwijze voor superparamagnetische ijzeroxide nanoclusters, omvattende de volgende stappen: (1) wegen van FeCl;s6H;0 in een reservoir, toevoegen van een ethyleenglycol (EG)-oplossing en een di-ethvleenglycol (DE G)-oplossing en magnetisch roeren om het FeCl;*6H,O op te lossen; (2) toevoegen van polyaerylzuur (PPA) aan een oplossing en roeren om de PPA op te lossen; (3) toevoegen van natriumacetaat (CH;COONa) en natriumhydroxide (NaOH); roeren en verwarmen om het CH;COONa en het NaOH op te lossen om een voorloper te vormen; (4) overbrengen van de bovenstaande voorloper naar een hydrothermische reactor; afdichten van de hydrothermische reactor, en vervolgens plaatsen in een sneldroogoven, verhitten tot 180- 280°C; en na 4-16 uur van reageren, sluiten van de sneldroogoven en op natuurlijke wijze laten afkoelen; en (5) uitnemen van een colloïdaal product, toevoegen van watervrije ethanol en water: ultrasoon wassen; scheiden met een magneet, en opnieuw wassen en scheiden met een zelfde methode; en plaatsen van een afgescheiden vaste stof in een vacuümdroogoven en drogen gedurende 12-36 uur bij kamertemperatuur: en afdichten van verkregen Fe30, droog poeder voor opslag.A preparation method for superparamagnetic iron oxide nanoclusters, comprising the following steps: (1) weighing FeCl 3 6 H 2 O in a reservoir, adding an ethylene glycol (EG) solution and a diethylene glycol (DE G) solution and magnetically stirring to dissolve the FeCl 2 *6H 2 O; (2) adding polyaerylic acid (PPA) to a solution and stirring to dissolve the PPA; (3) adding sodium acetate (CH;COONa) and sodium hydroxide (NaOH); stirring and heating to dissolve the CH2COONa and the NaOH to form a precursor; (4) transferring the above precursor to a hydrothermal reactor; sealing the hydrothermal reactor, then placing in a quick drying oven, heating to 180-280°C; and after 4-16 hours of reacting, closing the quick drying oven and allowing to cool naturally; and (5) taking out a colloidal product, adding anhydrous ethanol and water: ultrasonic washing; separating with a magnet, and again washing and separating with the same method; and placing a separated solid in a vacuum drying oven and drying for 12-36 hours at room temperature: and sealing obtained Fe 3 O, dry powder for storage. 2. De bereidingswerkwijze voor de superparamagnetische ijzeroxide nanoclusters volgens conclusie 1, waarbij een totaal volume van de EG- en DEG-oplossingen 30 ml is, en een verhouding van de EG-oplossing tot de DEG-oplossing 30/0-10/20 naar volume is.The preparation method for the superparamagnetic iron oxide nanoclusters according to claim 1, wherein a total volume of the EG and DEG solutions is 30 ml, and a ratio of the EG solution to the DEG solution is 30/0-10/20 to volume. 3. De bereidingswerkwijze voor de superparamagnetische ijzeroxide nanoclusters volgens conclusie 1, omvattende: (1) wegen van FeCl;*6H,0 in een beker, toevoegen van een EG-oplossing en een DEG- oplossing, en magnetisch roeren gedurende 10 min om het FeCl;«6H,0 op te lossen; (2) toevoegen van PPA aan een oplossing en 30 min roeren om de PPA op te lossen; (3) toevoegen van CH;COONa en NaOH; 40 min roeren en verwarmen om het CH;COONa en het NaOH op te lossen om een voorloper te vormen;The preparation method for the superparamagnetic iron oxide nanoclusters according to claim 1, comprising: (1) weighing FeCl 2 *6H 2 O in a beaker, adding an EG solution and a DEG solution, and magnetically stirring for 10 min to FeCl;«6H0.0 to be dissolved; (2) adding PPA to a solution and stirring for 30 min to dissolve the PPA; (3) addition of CH;COONa and NaOH; Stir and heat for 40 min to dissolve the CH2COONa and the NaOH to form a precursor; (4) overbrengen van de bovenstaande voorloper naar een hydrothermische reactor; afdichten van de hydrothermische reactor; vervolgens plaatsen in een sneldroogoven, verhitten tot 210°C, en na 10 uur van reageren, sluiten van de sneldroogoven en op natuurlijke wijze laten afkoelen; en(4) transferring the above precursor to a hydrothermal reactor; sealing the hydrothermal reactor; then placing in a quick-drying oven, heating to 210°C, and after reacting for 10 hours, closing the quick-drying oven and allowing to cool naturally; and (5) uitnemen van een colloïdaal product; toevoegen van watervrije ethanol en water; ultrasoon wassen, scheiden met een magneet: en opnieuw wassen en scheiden met een zelfde methode; en plaatsen van cen afgescheiden vaste stof in een vacuümdroogoven en drogen gedurende 24 uur bij kamertemperatuur; en afdichten van verkregen Fe;O, droog poeder voor opslag.(5) taking out a colloidal product; adding anhydrous ethanol and water; ultrasonic washing, separating with a magnet: and again washing and separating with the same method; and placing a separated solid in a vacuum drying oven and drying at room temperature for 24 hours; and sealing obtained Fe;O, dry powder for storage.
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