Preparation method and application of Fe3O4 nanoparticles modified by amyloid-like albumin coating

Abstract

The invention discloses a preparation method and application of Fe3O4 nanoparticles modified by an amyloid-like albumin coating, and relates to the technical field of nano-medicines. The preparation method comprises the following steps: S1, performing amyloid-like transformation on human serum albumin to prepare an amyloid-like albumin solution; and S2, modifying the surface of a Fe3O4 nanoparticle with the amyloid-like albumin solution prepared in the step S1 to prepare the nanoparticle modified by the amyloid-like albumin coating. According to the preparation method and the application of the Fe3O4 nano-particles modified by the amyloid-like albumin coating, the Fe3O4 nano-particles modified by the amyloid-like albumin coating, which have macrophage phagocytosis and inflammation bidirectional regulation and control effects, are innovatively constructed through an amyloid-like protein self-assembly technology.

Description

Technical Field

[0001] The present invention relates to the field of nanomedicine technology, and in particular to a preparation method and application of Fe3O4 nanoparticles modified with an amyloid albumin coating. Background Art

[0002] In recent years, nanoparticle-based targeted cancer therapy has become a research hotspot both domestically and internationally. Because nanoparticles can overcome the tumor-stromal barrier and, by combining with antibodies and drugs, can target cancer tissue, they have broad applications in targeted cancer therapy. Among them, ferroferric oxide nanoparticles (Fe3O4 nanoparticles) are the most promising magnetic nanoparticles. They can serve not only as drug carriers for targeted tumor therapy but also as contrast agents for magnetic resonance imaging (MRI) and ultrasound examinations. They also possess magnetic therapy and thermal therapy capabilities, enabling both early diagnosis and targeted therapy. They have broad applications in the field of targeted tumor therapy.

[0003] Precisely balancing macrophage phagocytosis and inflammatory responses is a key challenge in advancing the clinical translation of therapeutic nanoparticles. As key effectors of innate immunity, macrophages eliminate pathogens and regulate inflammatory homeostasis through phagocytosis. However, excessive uptake of nanoparticles often triggers a storm of proinflammatory cytokines (such as IL-1β and TNF-α), leading to tissue damage and treatment resistance.

[0004] Among the many surface modification molecules, human serum albumin (HSA) has shown great application potential in the field of drug delivery due to its low immunogenicity and long circulation characteristics. (albumin-bound paclitaxel) improves the therapeutic effect of breast cancer through an active targeting mechanism. HSA can also achieve active targeted drug delivery by binding to specific ligands. However, traditional HSA coatings also face three difficulties in practical applications:

[0005] (1) Physical instability: Physically adsorbed albumin has a low affinity with nanomaterials and is easily desorbed in the serum environment and replaced by other serum proteins, causing it to lose its preset immune regulatory function.

[0006] (2) Conformation-dependent immunogenicity: Differences in the surface physicochemical properties of nanoparticles can induce albumin to present different conformations.

[0007] (3) Chemical cross-linking induces conformational changes: Although chemical cross-linking improves the stability of adsorption, chemical reagents will cause changes in the structure of albumin.

[0008] Amyloid-like protein self-assembly technology provides a new idea to overcome the above limitations. Summary of the Invention

[0009] To solve the above problems, the present invention provides a preparation method and application of Fe3O4 nanoparticles modified with an amyloid albumin coating. Through the amyloid albumin self-assembly technology, an amyloid albumin coating with dual synergistic regulatory effects on macrophage phagocytosis and inflammation is innovatively constructed.

[0010] To achieve the above object, the present invention provides a method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating, comprising the following steps:

[0011] S1. performing amyloid-like conversion on human serum albumin to prepare an amyloid-like albumin solution;

[0012] S2. Modify the amyloid albumin solution prepared in S1 on the surface of Fe3O4 nanoparticles to prepare nanoparticles A modified with an amyloid albumin coating.

[0013] Preferably, the specific steps of S1 include:

[0014] S1-1. Prepare a HEPES dilution solution at a volume ratio of HEPES to deionized water of 1:20; prepare a solution at a ratio of TCFP to HEPES dilution solution of 0.1 mmol to 10 mL and adjust the pH to acidic to obtain solution A.

[0015] S1-2. Prepare solution B according to the ratio of human serum albumin: HEPES dilution solution = 20 mg: 1 mL; mix equal volumes of solution A and solution B, and stir evenly to prepare an amyloid-like albumin solution.

[0016] Preferably, in S1-2, the stirring is to magnetically stir the mixed solution of solution A and solution B at 300 rpm for 1 hour.

[0017] Preferably, in S2, the modification of the amyloid albumin solution on the surface of the Fe3O4 nanoparticles specifically includes:

[0018] The Fe3O4 nanoparticles were evenly mixed with 0.01-1 mg / mL amyloid albumin solution, shaken in a shaker at 100 rpm for 30 minutes, and then magnetically separated and the supernatant was discarded. The precipitate was washed with deionized water to obtain Fe3O4 nanoparticles modified with an amyloid albumin coating.

[0019] Preferably, the amyloid-like albumin solution prepared in S1 is diluted with deionized water to a concentration of 0.01-1 mg / mL.

[0020] Preferably, the mass volume ratio of Fe3O4 nanoparticles to 0.01-1 mg / mL amyloid albumin solution is 1 g:1 mL.

[0021] The preparation method and application of Fe3O4 nanoparticles modified with an amyloid albumin coating of the present invention have the following beneficial effects:

[0022] (1) The present invention converts human serum albumin into amyloid-like proteins. Amyloid-like proteins can rapidly self-assemble on the surface of Fe3O4 nanoparticles, achieving a stable bond with the Fe3O4 nanoparticles. Compared with conventional albumin modification, amyloid-like proteins can effectively increase the adhesion between the proteins and the nanoparticles, reduce displacement in the serum environment and desorption during subsequent processing of the nanoparticles, and maintain the stability of the properties of the composite nanoparticles.

[0023] (2) The Fe3O4 nanoparticles modified with an amyloid albumin coating prepared by the present invention have a bidirectional regulatory effect on macrophage phagocytosis and inflammatory response, which can promote the uptake of nanoparticles by macrophages while inhibiting excessive inflammatory response;

[0024] (3) The preparation method of the present invention is fast to operate, has a wide range of applications, does not use organic solvents in the preparation process, is green and environmentally friendly, and is suitable for large-scale commercial production.

[0025] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 To characterize the amyloid transformation of HSA and ALH modification at different concentrations;

[0027] Figure 2 Circular dichroism spectra of composite nanoparticles prepared at different HSA and ALH concentrations;

[0028] Figure 3 The particle size distribution and micromorphology of composite nanoparticles prepared at different HSA and ALH concentrations, where A-1 is the particle size distribution of HSA-Fe3O4 nanoparticles, A-2 is the particle size distribution of ALH-Fe3O4 nanoparticles, B-1 is the micromorphology of pure Fe3O4 nanoparticles, B-2 is the micromorphology of HSA-Fe3O4 nanoparticles, and B-3 is the micromorphology of ALH-Fe3O4 nanoparticles;

[0029] Figure 4 Characterization of the amount of protein desorption on the surface of the Fe3O4 nanoparticles, ALH-Fe3O4 nanoparticles prepared in Example 1 and the HSA-Fe3O4 nanoparticles prepared in Comparative Example 1 under SDS and ultrasonic treatment;

[0030] Figure 5 The fluorescence intensity changes of ALH-Fe3O4 nanoparticles and HSA-Fe3O4 nanoparticles in serum environment;

[0031] Figure 6 The survival rate of macrophages was evaluated after treatment of macrophages with pure Fe3O4 nanoparticles, HSA-Fe3O4 nanoparticles with different protein concentrations, and ALH-Fe3O4 nanoparticles. The control group was cell culture without nanoparticles.

[0032] Figure 7 The effect of ALH-Fe3O4 nanoparticles and HSA-Fe3O4 nanoparticles on macrophage uptake in the presence or absence of serum;

[0033] Figure 8 : Effects of ALH coating and HSA coating on macrophage inflammation, where A is the mRNA expression of IL-1β, TNF-α, MyD88, and Socs3, B is the concentration of IL-1β, and C is the concentration of TNF-α. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solutions and advantages disclosed in the embodiments of the present invention more clear, the embodiments of the present invention are further described in detail below with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are only used to explain the embodiments of the present invention and are not intended to limit the embodiments of the present invention. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0035] Example 1

[0036] A method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating comprises the following steps:

[0037] (1) Preparation of Fe3O4 nanoparticles:

[0038] Add 30 mL of deionized water to a three-necked flask connected to N2, then add 1.85 mmol of FeCl2·4H2O and 3.7 mmol of FeCl3·6H2O in sequence. Place in a 30°C water bath, add 10 mL of ammonia water dropwise into the flask, magnetically stir at 300 rpm for 15 min, and magnetically separate. Wash the product with deionized water several times until the pH value reaches 7 and store in a N2 environment.

[0039] (2) Preparation of amyloid-like albumin ALH: 0.1 mmol of disulfide bond reducing agent Tris (2-carboxyethyl) phosphine (TCEP) was dissolved in 10 mL of N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) diluted solution (HEPES: deionized water = 1:20 dilution), and the pH was adjusted to acidic.

[0040] Dissolve 100 mg of human serum albumin (HSA) in 5 mL of HEPES (HEPES: deionized water = 1:20 dilution) solution and mix well.

[0041] The above TCEP solution and HSA solution were mixed in equal volumes, and magnetically stirred at 300 rpm for 1 h to obtain a 10 mg / mL ALH solution, which was then diluted with deionized water to a 1 mg / mL ALH solution.

[0042] (3) 1 mg of the above Fe3O4 nanoparticles was mixed evenly with 1 mL of the 1 mg / mL ALH solution obtained in (2). The mixture was shaken at 100 rpm for 30 min, and then magnetically separated and the supernatant was discarded. After multiple washings, Fe3O4 nanoparticles modified with an amyloid-like albumin coating (ALH-Fe3O4) were obtained.

[0043] Example 2

[0044] A method for preparing Fe3O4 nanoparticles modified with an amyloid albumin-like coating is the same as that of Example 1, except that the ALH solution is diluted to 0.1 mg / mL, 1 mg of Fe3O4 nanoparticles is uniformly mixed with 1 mL of the 0.1 mg / mL ALH solution, and the mixture is shaken at 100 rpm on a shaker. The supernatant is then magnetically separated and discarded. The mixture is washed multiple times to obtain ALH-Fe3O4 nanoparticles.

[0045] Example 3

[0046] A method for preparing Fe3O4 nanoparticles modified with an amyloid albumin-like coating is similar to that of Example 1, except that the ALH solution is diluted to 0.01 mg / mL, 1 mg of Fe3O4 nanoparticles is uniformly mixed with 1 mL of the 0.01 mg / mL ALH solution, and the mixture is shaken at 100 rpm on a shaker. The supernatant is then magnetically separated and discarded. The mixture is washed multiple times to obtain ALH-Fe3O4 nanoparticles.

[0047] Comparative Example 1

[0048] A method for preparing albumin-coated nanoparticles is the same as that of Example 1, except that amyloid-like albumin is replaced by conventional albumin HSA, and the solubility of the HSA solution is 1 mg / mL to prepare HSA-Fe3O4 nanoparticles.

[0049] Comparative Example 2

[0050] A method for preparing albumin-coated nanoparticles is the same as that of Example 1, except that amyloid-like albumin is replaced by conventional albumin HSA, and the solubility of the HSA solution is 0.1 mg / mL to prepare HSA-Fe3O4 nanoparticles.

[0051] Comparative Example 3

[0052] A method for preparing albumin-coated nanoparticles is the same as that of Example 1, except that amyloid-like albumin is replaced by conventional albumin HSA, and the solubility of the HSA solution is 0.01 mg / mL to prepare HSA-Fe3O4 nanoparticles.

[0053] Experimental testing

[0054] (I) ThT staining was used to evaluate the successful surface modification of ALH on the Fe3O4 nanoparticles. ThT fluorescent dye can be used to investigate the formation of amyloid structures within proteins attached to the nanoparticle surface, reflecting the successful preparation of ALH on the nanoparticle surface.

[0055] In the experiment, three different concentrations of ALH and HSA (1 mg / mL, 0.1 mg / mL, and 0.01 mg / mL) were used to modify Fe3O4 nanoparticles. The results showed that for these three concentrations, the binding amount of HSA and ALH modified nanoparticles to ThT was significantly different, indicating that different concentrations of ALH were successfully modified on the surface of Fe3O4 nanoparticles. Especially when the ALH protein concentration was 1 mg / mL, the binding difference of ThT was most significant compared with HSA, as shown in Figure 2. Figure 1 As shown in .

[0056] At the same time, for free protein, the fluorescence intensity of ALH solution was significantly higher than that of HSA solution, indicating that ALH solution contained rich β-sheet amyloid structures.

[0057] (II) The secondary structures of HSA and ALH adsorbed at different concentrations were analyzed using circular dichroism (CD) spectroscopy. Figure 2 shown.

[0058] First, the secondary structures of pure HSA and ALH proteins were examined as a control. In pure protein solutions, HSA exhibited a positive peak at 192 nm and negative peaks at 208 nm and 222 nm, indicating a typical α-helix structure. ALH exhibited a positive peak at 190 nm and a negative peak at 210 nm, indicating a typical β-sheet structure. When these two proteins were adsorbed on the Fe3O4 NPs surface, CD data revealed that the positions of the relevant peaks remained largely unchanged, with only a partial red shift, indicating that the HSA and ALH proteins did not undergo structural changes before and after adsorption.

[0059] (III) In order to explore the effect of ALH and HSA adhesion on the particle size and microstructure of composite nanoparticles, dynamic light scattering (DLS) and transmission electron microscopy (TEM) were used for characterization analysis. The experimental results showed that the concentration and type of protein had a certain effect on the particle size of composite nanoparticles, such as Figure 3 As shown in part A. Figure 3 A-1 is the particle size distribution of HSA-Fe3O4 nanoparticles at different HSA and ALH concentrations; A-2 is the particle size distribution of ALH-Fe3O4 nanoparticles at different HSA and ALH concentrations.

[0060] For HSA, the particle size of the composite nanoparticles gradually decreased with decreasing protein concentration, reaching a peak of approximately 100 nm for 0.01 mg / mL HSA-Fe₃O₄ composite nanoparticles. For ALH, the overall trend was similar to that of HSA, but at 0.1 mg / mL ALH, the composite nanoparticles exhibited significant aggregation around 1000 nm. At protein concentrations of 1 mg / mL and 0.01 mg / mL, the particle sizes of the HSA-Fe₃O₄ and ALH-Fe₃O₄ composite nanoparticles were similar.

[0061] TEM observation of the microscopic morphology of the composite nanoparticles showed that the modification of HSA and ALH hardly changed the microstructure of the nanoparticles. The single particles of pure Fe3O4, HSA-Fe3O4 and ALH-Fe3O4 were all spherical structures of about 20nm. In aqueous solution, the three particles all agglomerated to a certain extent, and the diameter of the particles after agglomeration was between 100 and 200nm. Figure 3 As shown in part B, the results are basically consistent with the DLS results.

[0062] Figure 3 B-1 is the micromorphology characterization of pure Fe3O4 nanoparticles; Figure 3 B-2 is the micromorphology characterization of HSA-Fe3O4 nanoparticles; Figure 3 Figure B-3 is the micromorphology characterization of ALH-Fe3O4 nanoparticles.

[0063] (IV) The Fe3O4 nanoparticles, ALH-Fe3O4 nanoparticles and HSA-Fe3O4 nanoparticles prepared in Example 1 and Comparative Example 1 were placed in a 5% SDS solution and superimposed with ultrasonic treatment. After 30 minutes of treatment, the supernatant was collected after centrifugation and the concentration of desorbed protein in the supernatant was detected using a BCA kit. The desorption rate of proteins on the surface of different nanoparticles was calculated based on the protein concentration adsorbed on the surface of the nanoparticles (protein solution concentration before modification - protein concentration in the centrifuged supernatant after modification). The results are shown in Figure 4. Figure 4 shown.

[0064] SDS is a common emulsifier and a commonly used reagent in the preparation and modification of nanomaterials. It is also a protein denaturant that changes the secondary and tertiary structures of proteins, thereby separating the proteins from the surface to which they adhere.

[0065] The experimental results showed that conventional HSA-modified nanoparticles were easily eluted in the presence of SDS protein denaturant, further confirming that their adhesion strength was insufficient and easily affected by subsequent operations.

[0066] However, the modified ALH protein showed significantly lower protein desorption from the nanoparticle surface compared to HSA. For nanoparticles with a modified protein concentration of 1 mg / mL, the protein desorption rate decreased from 7.02% for HSA to 0.50% for ALH; for nanoparticles with a modified protein concentration of 0.1 mg / mL, the protein desorption rate decreased from 26.14% for HSA to 4.08% for ALH; and for nanoparticles with a modified protein concentration of 0.01 mg / mL, the protein desorption rate decreased from 88.90% for HSA to 11.26% for ALH. All three concentrations of protein-nanoparticle complexes demonstrated that amyloid-like albumin could be firmly adsorbed to the nanoparticle surface, forming a stable coating.

[0067] (V) HSA and ALH were fluorescently labeled to further verify the stability of the ALH coating in the serum environment. Figure 5 shown.

[0068] The results showed that nearly 50% of the albumin in the HSA coating was replaced by serum proteins, while the fluorescence intensity of the ALH coating did not change significantly after treatment in a serum environment. These experiments demonstrate that the ALH coating can achieve stable self-assembly on the nanoparticle surface through strong anchoring mediated by β-sheet.

[0069] (VI) 25,000 macrophages (RAW 264.7) were seeded into 96-well plates and allowed to adhere overnight at 37°C, 5.0% CO2 in whole blood culture medium.

[0070] Prepare serum-free culture medium containing different nanoparticles. Add 300 μL of 0.5 mg / mL ALH-Fe₃O₄ nanoparticles, HSA-Fe₃O₄ nanoparticles, and pure Fe₃O₄ nanoparticles to 2 mL of serum-free DMEM, vortex to mix, and sterilize under UV light for half an hour.

[0071] The cell supernatant in the 96-well plate was aspirated and serum-free culture medium containing different nanoparticles was added. After incubation for 24 hours, cell viability was measured using a CCK8 assay.

[0072] The results are as follows Figure 6 As shown, the control group was a cell culture without nanoparticles. The results indicate that surface modification of pure Fe3O4 nanoparticles with either HSA or ALH does not increase cytotoxicity. The effects of HSA and ALH on cytotoxicity are similar, and increasing the concentration of the modified proteins helps reduce the cytotoxicity of Fe3O4 nanoparticles. Modification of Fe3O4 nanoparticles with ALH at a concentration of 1 mg / mL significantly improves cell survival, demonstrating the good cytocompatibility of ALH-Fe3O4 nanoparticles.

[0073] (VII) Evaluate the effects of nanoparticles prepared by different modification methods on macrophage uptake. FITC-labeled HSA protein was used for nanoparticle preparation, and HSA and ALH coatings were prepared. Macrophages were co-cultured with pure Fe3O4 nanoparticles, HSA-coated Fe3O4 nanoparticles, and ALH-coated Fe3O4 nanoparticles in the presence and absence of serum for 3 hours. Flow cytometry was then used to measure the fluorescence intensity in the macrophages to reflect the nanoparticle uptake rate.

[0074] The results are as follows Figure 7 As shown, the uptake rate of ALH-coated Fe3O4 nanoparticles by macrophages was significantly higher than that of HSA-coated Fe3O4 nanoparticles, regardless of the presence or absence of serum.

[0075] Notably, in a serum-free environment, the uptake rate of ALH-coated Fe3O4 nanoparticles by macrophages was approximately 45 times higher than that of HSA-coated Fe3O4 nanoparticles. In a serum-containing environment, the uptake rate of ALH-coated Fe3O4 nanoparticles by macrophages was approximately 70 times higher than that of HSA-coated Fe3O4 nanoparticles.

[0076] For HSA-modified nanoparticles, there was almost no difference in macrophage phagocytosis in the presence or absence of serum; while for ALH-modified nanoparticles, the uptake rate in the serum environment increased significantly, nearly twice the uptake rate in the serum-free environment.

[0077] In order to exclude the influence of the fluorescence intensity of the nanoparticles themselves, the fluorescence difference of the nanoparticles themselves was analyzed by flow cytometry ( Figure 5 The results showed that the fluorescence of the ALH-coated particles was nearly 6 times that of the HSA group.

[0078] Therefore, the actual uptake rate of nanoparticles by macrophages is: in a serum-free environment, the uptake rate of Fe3O4 nanoparticles modified with ALH coating by macrophages is about 7 times that of those modified with HSA coating, while in a serum environment, the uptake rate of Fe3O4 nanoparticles modified with ALH coating by macrophages is about 11 times that of those modified with HSA coating.

[0079] (8) The effect of ALH coating on macrophage inflammation was further analyzed by analyzing the transcription and expression of macrophage inflammatory factors.

[0080] Real-time fluorescence PCR data such as Figure 8 As shown, the ALH coating significantly inhibited the transcription of pro-inflammatory factors IL-1β and TNF-α, among which the inhibitory effect on IL-1β transcription was more obvious. The IL-1β gene expression in the ALH group was only 16.3% of that in the HSA group.

[0081] At the same time, ELISA data further confirmed that the secretion of pro-inflammatory factors in the ALH group was significantly lower than that in the HSA group. The secretion of TNF-α was 30% of the HSA group, and IL-1β was 25%, which was consistent with the PCR results, confirming that the ALH coating effectively inhibited the inflammatory storm at the gene transcription and protein secretion levels.

[0082] To further identify anti-inflammatory targets, the gene transcription levels of MyD88 (myeloid differentiation primary response protein 88) and Socs3 (a negative inflammatory signaling molecule) were measured. MyD88, a core adaptor protein in the TLR signaling pathway, was also lower in the ALH group. This inhibitory effect suggests that the ALH coating may alleviate the inflammatory response by blocking the TLR / MyD88 pathway. Socs3 mRNA expression in the ALH group was significantly lower than in the HSA group, suggesting that the early anti-inflammatory mechanism is independent of Socs3 negative feedback.

[0083] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the same. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solutions of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating, characterized in that: The steps include: S1. performing amyloid-like conversion on human serum albumin to prepare an amyloid-like albumin solution; S2. Modify the surface of Fe3O4 nanoparticles with the amyloid-like albumin solution prepared in step S1 to prepare nanoparticles modified with an amyloid-like albumin coating.

2. The method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating according to claim 1, characterized in that: S1 specifically includes: S1-1. Prepare a HEPES dilution solution at a volume ratio of HEPES to deionized water of 1:20; prepare a solution at a ratio of TCFP to HEPES dilution solution of 0.1 mmol to 10 mL and adjust the pH to acidic to obtain solution A. S1-2. Prepare solution B according to the ratio of human serum albumin: HEPES dilution solution = 20 mg: 1 mL; mix equal volumes of solution A and solution B, and stir evenly to prepare an amyloid-like albumin solution.

3. The method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating according to claim 2, characterized in that: In S1-2, the stirring was performed by magnetically stirring the mixed solution of solution A and solution B at 300 rpm for 1 h.

4. The method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating according to claim 1, wherein: In S2, the amyloid albumin solution is modified on the surface of Fe3O4 nanoparticles, specifically including: The Fe3O4 nanoparticles were evenly mixed with a 0.01-1 mg / mL amyloid albumin solution, shaken in a shaker at 100 rpm, and then magnetically separated and the supernatant was discarded. The precipitate was washed with deionized water to obtain Fe3O4 nanoparticles modified with an amyloid albumin coating.

5. The method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating according to claim 4, characterized in that: The amyloid-like albumin solution was diluted with deionized water to a concentration of 0.01-1 mg / mL.

6. The method for preparing Fe3O4 nanoparticles modified with an amyloid albumin coating according to claim 4, characterized in that: The mass-to-volume ratio of Fe3O4 nanoparticles to 0.01-1 mg / mL amyloid albumin solution is 1 g:1 mL.

7. Nanoparticles modified with an amyloid-like albumin coating prepared by the method for preparing Fe3O4 nanoparticles modified with an amyloid-like albumin coating according to any one of claims 1 to 6.

8. Use of the Fe3O4 nanoparticles modified with an amyloid albumin coating as claimed in claim 7 in coordinating the balance between macrophage phagocytosis and inflammatory response.

9. Use of the Fe3O4 nanoparticles modified with an amyloid albumin coating as claimed in claim 7 in the preparation of nanocarriers for regulating inflammatory response.

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