Preparation method of magnetic targeting ultra-small Fe3O4 nanoprobe SPIONPs

Abstract

The invention discloses a preparation method of a magnetic targeting ultra-small Fe3O4 nanoprobe SPIONPs, relates to the field of immune targeting biomedical diagnosis and treatment, and adopts S1 to prepare a NaOH solution and S2 to prepare a triton solvent. And S3, triton drops by drops. Heating is performed; s5, weighing FeCl3. 6H2O and FeCl2. 4H2O according to a molar ratio to prepare a solution; S6, slowly dropwise adding the mixed solution into the NaOH solution; according to the preparation method, magnetic nanoparticles with the particle size of 2-30 nm are prepared, an ultra-small Fe3O4 nanoprobe SPIONPs realizes loading functionalization, and the obtained magnetic targeting nanoparticles and probes subjected to surface modification functionalization modification treatment have the characteristics of long acting, stability, high efficiency, simplicity in preparation and high yield. Particularly, the invention has important application in the fields of immune targeted biomedical diagnosis and treatment, magnetic imaging, nuclear magnetic resonance enhanced imaging, gene editing, gene protection, gene differentiation, nuclear magnetic contrast fluorescence microscopy and targeted immune medical treatment.

Description

Technical Field

[0001] The present invention relates to the field of immune-targeted biomedical diagnosis and treatment, and in particular to a method for preparing magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs. Background Art

[0002] Chip-encapsulated magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs have superparamagnetic properties. Superparamagnetic ferrimagnetic oxide Fe3O4 exhibits excellent half-metallic properties. [1] The research on magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs is of great significance in magnetic targeting, immunology, and biomedical diagnosis and treatment.

[0003] Magnetic targeting ultra-small Fe3O4 nanoprobe SPIONPs is an important type of nanomaterial. In addition to its small size effect, large specific surface effect and nano-quantum effect, it also has specific superparamagnetic properties. Its unique biomedical, targeted diagnosis and treatment, immunology, and physical and chemical functional properties make magnetic targeting ultra-small Fe3O4 nanoprobe SPIONPs widely used in many fields, including biomedicine. [1] , magnetic fluid [2] , information storage [3] , magnetic resonance imaging, etc. At present, an inevitable problem with magnetic nanoparticles is that they cannot remain stable for a long time, cannot have long-term effects, cannot pass through the blood-brain barrier, and cannot avoid redox reaction degradation. Moreover, larger magnetic nanoparticles and nanoparticles with high specific surface area are more prone to agglomeration. In addition, exposed nanoparticles have high chemical activity and are easily oxidized in the air, resulting in a decrease in magnetism and dispersibility. The magnetic targeted ultrasmall Fe3O4 nanoprobe SPIONPs of this project has the following advantages: 1. The preparation method is simple and efficient, and the prepared Fe3O4 has a unique ultrasmall size structure and excellent biomedical, physical and chemical properties. 2. Nano-Fe3O with special morphology 4】 Methods; 3. With the advantages of magnetically targeted Fe3O4 surface modification and loading modification, it has excellent biomedical, pharmaceutical, diagnostic and immunological, targeted nano and magnetocaloric effects. Magnetic targeted ultrasmall Fe3O4 nanoprobes SPIONPs have important applications and impacts in the fields of biomedicine, targeted diagnostics and immunology, and physical and chemical functional properties. Chip-packaged magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs have a spinel structure. Ultrasmall Fe3O4 has superparamagnetic semimetallic properties and superparamagnetism. It is one of the most influential and potential magnetic superparamagnetic semimetallic materials. It is simple to prepare, has a high Curie temperature, a large spin polarization rate at room temperature, is ultrasmall and long-lasting, has stable magnetoelectric properties, and has long-lasting, stable and efficient excellent characteristics. It has attracted great attention from the medical, pharmaceutical, academic and industrial communities.

[0004] Research Significance and Application Value: Chip-encapsulated magnetically targeted ultrasmall Fe3O4 nanoprobes (SPIONPs) were used to prepare uniform Fe3O4 magnetic nanoparticles. Further surface chemical modification to form core-shell Fe3O4 magnetic nanocomposite particles stabilized the Fe3O4 nanoparticles against oxidation, improving their dispersibility and allowing for further functionalization. These magnetically targeted ultrasmall Fe3O4 nanoprobes (SPIONPs) not only retain the unique superparamagnetic properties of magnetic nanoparticles, making them suitable for targeted drug delivery, but also enhance several excellent properties inherent to the coated nanoparticles, offering promising applications in catalysis, biolabeling, and magnetic separation. Furthermore, while the magnetoelectric properties of semimetallic Fe3O4 nanoprobes have been extensively studied, detailed calculations of these materials' magnetoelectric properties, such as the band structure and density of states, in their stable phases are crucial for discovering more highly spin-polarized materials and predicting their potential applications. However, the iron oxide provided by magnetically targeted ultrasmall Fe₃O₄ nanoprobes (SPIONPs) has widespread applications in many fields. They are a promising candidate for microscopy and nuclear magnetic resonance imaging, significantly impacting not only genome sequencing, bioproteins, pharmaceuticals, industrial catalysts, and resource exploration, but also immunotherapy, targeted drug delivery, RNA and DNA editing, genome sequencing, and bioprotein editing. Research on magnetically targeted ultrasmall Fe₃O₄ nanoprobes (SPIONPs) not only has implications for targeted drug delivery, sustained release, targeted therapy, RNA and DNA editing, genome sequencing, and bioprotein editing, but also could represent a landmark scientific revolution in immunotherapy, targeted biomedical treatment, targeted drug diagnosis and therapy, tumor diagnosis, and the development of new drugs. They also have profound implications for biomedicine, targeted diagnosis and therapy, immunology, physicochemical functions, biopharmaceuticals, magnetic fluids, information storage, nuclear magnetic resonance imaging, semiconductor integration, micro-nano-NMR technology, nano-optics, and laser plasma micromechanical MEMS technology. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for preparing chip-encapsulated magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs, and in particular to a method for preparing detectors for magnetic imaging and targeted medical methods under manufacturing technology in the field of immune-targeted biomedical diagnosis and treatment.

[0006] The present invention provides a method for preparing a magnetic targeting ultrasmall Fe3O4 nanoprobe SPIONPs composite material.

[0007] The present invention is implemented by the following technical solution: comprising the following steps:

[0008] S1 prepares NaOH solution;

[0009] S2 prepares 1-1.5 mL of 0.1-0.98 g / mL Triton solvent.

[0010] S3: Add the aforementioned Triton dropwise into the prepared NaOH solution.

[0011] S4 Prepare a mixed solution of Triton and NaOH and heat it to 50-80°C;

[0012] S5 Prepare a solution of 10-50 g each of FeCl3·6H2O and FeCl2·4H2O;

[0013] S6 slowly adding the mixed solution dropwise to the above NaOH solution;

[0014] S7: Heating the mixed solution;

[0015] S8 washes the black product;

[0016] S9 vacuum drying of nano-Fe3O4.

[0017] S10 magnetically targeted ultrasmall Fe3O4 nanoprobe SPIONPs system editing technology delivery carrier;

[0018] S11 Surface modification of Fe3O4 nanoprobe SPIONPs.

[0019] By using the method of the present invention to prepare magnetic nanoparticles, ultra-small Fe3O4 nanoprobes SPIONPs achieve functionalization of the load, and obtain surface-modified magnetic targeting nanoparticles and probes. DETAILED DESCRIPTION

[0020] A method for preparing a magnetically targeted ultrasmall Fe3O4 nanoprobe SPIONPs composite material. Prepare 250 mL of a 1 mol / L NaOH solution. Prepare 1.5 mL of a 0.98 g / mL Triton solvent. Add 1.5 mL of the aforementioned solution containing 0.98 g / mL Triton dropwise to the prepared 250 mL of a 1 mol / L NaOH solution. NaOH, Triton solvent. Add 1.5 mL of the aforementioned solution containing 0.98 g / mL Triton dropwise to the prepared 250 mL of a 1 mol / L NaOH solution. Prepare a mixed solution of Triton and NaOH and heat it to 80°C. Weigh 50g each of FeCl3·6H2O and FeCl2·4H2O in a 1:1 molar ratio and slowly add the solution dropwise to the NaOH solution. Heat the mixture and stir at 80°C for 0.5 hours. Wash the black product twice with deionized water and once with anhydrous ethanol. Dry the nano-Fe3O4 under vacuum at 40°C. The magnetically targeted ultrasmall Fe3O4 nanoprobes, SPIONPs, are delivered via system editing technology.

[0021] The implementation steps are broken down as follows:

[0022] Surface-coated PZS-modified Fe₃O₄: 0.1 g of the resulting Fe₃O₄@PZS nanoparticles was placed in 50 mL of deionized water. Then, 50 mL of a 0.01 mol / L HAuCl₄ solution was added and sonicated at room temperature for 30 minutes. 50 mL of a 0.01 mol / L Na₃Ct solution and 50 mL of a freshly prepared 0.1 mol / L NaBH₄ solution were quickly added and stirred for 3 minutes, immediately producing a black flocculent substance accompanied by numerous bubbles. The black flocculent Fe₃O₄@PZS@Au seeds were collected using a magnet, washed three times with water, and sonicated for another 3 minutes to obtain a black suspension, which was then dried under vacuum. This completed the Fe₃O₄@PZS modification.

[0023] Fe3O4@PZS@Au modification: Weigh 0.02 g of Fe3O4@PZS@Au seeds, add 120 mL of deionized water, and stir for 10 minutes; then add 50 mL of 0.01 mol / L HAuCl4 solution, heat to 100°C in a water bath and reflux for 10 minutes; then add 12 mL of Na3Ct, the solution immediately turns purple, and stir for 5 minutes to obtain the product Fe3O4@PZS@Au composite nanoparticles.

[0024] Targeted release of Fe3O4 nanoprobe SPIONPs delivery carrier: guided by an external magnetic field, 5 times the Fe3O4 iron oxide nanoparticles are directionally enriched to specific tissues or cells (such as tissue sites) through cell membranes, cells and lipid cell membranes to reduce off-target effects. Fe3O4 nanoprobe SPIONPs encapsulation: avoids degradation by serum nucleases and prolongs the in vivo circulation time by 10-60x hours. Fe3O4 nanoprobe SPIONPs has a protective layer. The surface modification of Fe3O4 nanoprobe SPIONPs designed as a carrier: PEI (polyethyleneimine) is positively charged and adsorbs negatively charged nucleic acids (sgRNA) through electrostatic interaction. SPIONPs lipid coating: enhances biocompatibility and promotes cell membrane fusion. SPIONPs targeting ligands: folic acid, RGD peptide, further enhance tumor targeting.

[0025] Preparation of cell membrane loaded magnetic Fe3O4 nanoprobe SPIONPs: First, extract the cells and immerse them in 75% ethanol for 5 minutes. Take them out and blow dry, wash them twice with PBS, and blow dry. Put them in pre-cooled 0.1 mol glucose in PBS, pass the mixed liquid through a 200nm membrane filter by syringe, and extrude it mechanically, and repeat 10 times. Finally, a culture medium with cell membrane loaded magnetic Fe3O4 nanoprobe SPIONPs is obtained. It includes the implementation method of surface modification and modification of magnetic targeted ultra-small Fe3O4 nanoprobe SPIONPs system, surface functionalization, delivery carrier Fe3O4 targeting, Fe3O4 SPIONPs protective layer, and Fe3O4 nanoprobe SPIONPs multifunctional integrated steps.

Claims

1. A method for preparing magnetically targeted ultrasmall Fe3O4 nanoprobes SPIONPs, characterized in that: The following steps are involved: S1 prepares NaOH solution; S2 prepares 1-1.5 mL of 0.1-0.98 g / mL Triton solvent; S3: Add the aforementioned Triton dropwise into the prepared NaOH solution. S4 Prepare a mixed solution of Triton and NaOH and heat it to 50-80°C; S5 Prepare a solution of 10-50 g each of FeCl3·6H2O and FeCl2·4H2O; S6 slowly adding the mixed solution dropwise to the above NaOH solution; S7: Heating the mixed solution; S8 washes the black product; S9 vacuum drying of nano-Fe3O4; S10 magnetically targeted ultrasmall Fe3O4 nanoprobe SPIONPs system editing technology delivery carrier; S11 Surface modification of Fe3O4 nanoprobe SPIONPs.

2. The preparation method according to claim 1, wherein: (S1) Prepare 50-250 mL of 0.1-1 mol / L NaOH solution.

3. The preparation method according to claim 1, wherein: Step (S5) Weigh 50 g of each of FeCl 3 ·6H 2 O and FeCl 2 ·4H 2 O in a molar ratio of 2:1-1:1 and prepare a solution.

4. The preparation method according to claim 1, characterized in that (S7) The mixed solution is heated and stirred at 50-80° C. for half an hour.

5. The preparation method according to claim 1, characterized in that Step (S8) The black product is washed twice with deionized water and once with anhydrous ethanol.

6. The preparation method according to claim 1, characterized in that Step (S9) vacuum drying the nano-Fe3O4 at 10-40°C.

7. The preparation method according to claim 1, characterized in that Step (S11) is specifically: First, the surface was modified by coupling PZS. The resulting Fe3O4@PZS nanoparticles were placed in deionized water, and then HAuCl4 solution was added. Ultrasonication was performed at room temperature for 30 minutes. 10-50 mL of 0.01 mol / L Na3Ct was added, followed by 10-50 mL of the prepared 0.1 mol / L NaBH4 solution. The mixture was stirred for 3 minutes, and the black flocculent Fe3O4@PZS@Au seeds were collected by magnet. Then, Fe3O4@PZS@Au seeds were weighed, 10-120 mL of deionized water was added, and the mixture was stirred for 1-10 minutes. 10-50 mL of 0.01 mol / L HAuCl4 solution was added, and the mixture was heated in a water bath to 20-100°C and refluxed for 10 minutes. 6-12 mL of Na3Ct was added, and the solution immediately turned purple. The mixture was stirred for 1-5 minutes to obtain the product, Fe3O4@PZS@Au composite nanoparticles. Secondly, Fe3O4 nanoprobes are wrapped in SPIONPs to prolong the circulation time in the body. Afterwards, Fe3O4 iron oxide nanoparticles are enriched in specific tissues or cells to reduce off-target effects. Magnetic nanoparticles carry complex MOFs and are enriched in tumors under the guidance of magnetic fields. Thirdly, Fe3O4 nanoprobes are fixed to the surface of nanoparticles through covalent coupling and chemical bonds. Finally, a culture medium with cell membrane-loaded magnetic Fe3O4 nanoprobes was obtained.

8. The preparation method according to claim 7, characterized in that Weigh 0.05-0.1 g of the resulting Fe3O4@PZS nanoparticles and place them in 10-50 mL of deionized water.

9. The preparation method according to claim 7, characterized in that Add 10-50 mL of 0.005-0.01 mol / L HAuCl4 solution.

10. The preparation method according to claim 7, characterized in that Weigh 0.02 g of Fe3O4@PZS@Au seeds.