A cell membrane modified nanocomposite, and a preparation method and application thereof

Abstract

The application discloses a cell membrane modified nanocomposite as well as a preparation method and application thereof. The nanocomposite comprises, from outside to inside, a membrane structure outer layer, a piezoelectric material middle layer and a magnetostrictive material core structure, wherein the outer surface of the membrane structure contains protein molecules. Under the stimulation of an external magnetic field, the nanocomposite can regulate Th1 / Th2 balance, promote Th2 cell polarization and enhance T cell proliferation, thereby reducing T cell-mediated autoimmune reactions and maintaining host immune homeostasis, and has a wide application prospect in the treatment of arthritis.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and in particular relates to a cell membrane modified nanocomposite material, its preparation method, and its application. Background Technology

[0002] T helper cells can polarize into Th1 and Th2 subsets under various conditions. Unlike the pathogenic role of the Th1 subset in chronic autoimmune diseases such as diabetes, multiple sclerosis, and rheumatoid arthritis, Th2 cells are protective. Therefore, a successful shift from a Th1-dominant response to a Th2-dominant response may have clinical benefits for patients with autoimmune diseases. However, there are no particularly effective therapeutic strategies to achieve a stable in vivo Th1-Th2 conversion.

[0003] Magnetic fields are fundamental to many physiological processes in living organisms. However, CD4... + How T cells respond to magnetic fields and whether external magnetic fields can regulate T cell-mediated autoimmune diseases remain largely unknown.

[0004] The information in the background section is merely intended to illustrate the general background of the invention and should not be construed as an admission or implication in any way that such information constitutes prior art known to those skilled in the art. Summary of the Invention

[0005] To address at least some of the technical problems in the prior art, this invention constructs a nanocomposite material comprising, from the outside to the inside, an outer membrane structure, a piezoelectric material intermediate layer, and a magnetostrictive material core structure. Simultaneously, the outer surface of this membrane structure contains protein molecules targeting T cells. The nanocomposite material of this invention can regulate the Th1 / Th2 balance and alleviate T cell-mediated autoimmune responses, thereby potentially being used to treat arthritis. Specifically, this invention includes the following:

[0006] In a first aspect, the present invention provides the use of a nanocomposite material in the preparation of a medicament for treating arthritis, wherein the nanocomposite material comprises, from the outside to the inside, an outer layer of a membrane structure, an intermediate layer of a piezoelectric material, and a core structure of a magnetostrictive material, wherein the outer surface of the membrane structure contains protein molecules.

[0007] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the membrane structure is derived from a biological membrane or an artificially synthesized membrane structure.

[0008] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the protein molecule is a binding molecule capable of binding to the surface of T cells.

[0009] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the surface of the T cells contains at least one of the following antigens: CD1, CD2, CD3, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD153, CD154, CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, OX 40. GITR, LAG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6 and CCR7.

[0010] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the protein molecule is capable of targeting CD4. + MHC II complex of T cells.

[0011] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the membrane structure is a cell membrane isolated from immune cells.

[0012] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the cell membrane comprises a cell membrane isolated from dendritic cells, B lymphocytes and / or macrophages.

[0013] In some embodiments, the use of the nanocomposite material according to the present invention in the preparation of a medicament for treating arthritis, wherein the piezoelectric material is selected from at least one of ferric acid, niobic acid, titanic acid, silicate and aluminic acid materials, and / or selected from at least one of polyvinylidene fluoride, polyester, polymethyl methacrylate, nylon, polyvinyl chloride, polylactic acid, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), vinylidene fluoride / trifluoroethylene copolymer, vinylidene fluoride / tetrafluoroethylene copolymer, polyvinylidene fluoride-hexafluoropropylene and polydimethylsiloxane materials; the magnetostrictive material is selected from ferric acid materials.

[0014] A second aspect of the present invention provides a method for preparing a nanocomposite material, comprising:

[0015] (1) A magnetostrictive piezoelectric nanocomposite material is prepared, wherein the magnetostrictive piezoelectric nanocomposite material comprises a piezoelectric material intermediate layer and a magnetostrictive material core structure;

[0016] (2) The magnetostrictive piezoelectric nanocomposite material is coated inside the membrane material, wherein the outer surface of the membrane material contains protein molecules.

[0017] In a third aspect, the present invention provides a nanocomposite material obtained by the preparation method described in the second aspect of the present invention.

[0018] A fourth aspect of the present invention provides a method for regulating Th1 / Th2 cell balance in vitro, comprising the step of contacting the nanocomposite material of the present invention with isolated cells in the presence of a magnetic field of 0.01-300 mT.

[0019] The nanocomposite material of the present invention can regulate the Th1 / Th2 balance under external magnetic field stimulation, promote Th2 cell polarization and enhance T cell proliferation, thereby reducing T cell-mediated autoimmune response and maintaining host immune homeostasis, and can be used to treat arthritis. Attached Figure Description

[0020] Figure 1 The preparation and characterization of DC@CFO / BFO nanocomposites are shown, where a is a representative high-resolution transmission electron microscope image of the DC@CFO / BFO nanocomposites; b and c are the free radicals ·OH and ·O2 generated by CFO / BFO nanoparticles under ultrasound, respectively. - Electron paramagnetic resonance (EPR) images; d and e represent the free radicals ·OH and ·O2 generated by CFO / BFO nanoparticles, respectively. - f represents electron paramagnetic resonance images; f represents flow cytometry analysis of surface markers of bone marrow-derived dendritic cells (BMDCs); g and h represent particle sizes of CFO / BFO nanoparticles and DC@CFO / BFO nanocomposites; i represents the zeta potential of CFO / BFO nanoparticles and DC@CFO / BFO nanocomposites; j represents the glycoprotein levels on the DC membrane, CFO / BFO nanoparticles, and DC@CFO / BFO nanocomposites as measured by ELISA.

[0021] Figure 2 The biocompatibility evaluation of the DC@CFO / BFO nanocomposite is shown, where ac represents the initial CD4 separation. +T cells were incubated with DC@CFO / BFO nanocomposite for 24 hours with or without a magnetic field, and the viability of T cells was measured at an absorbance wavelength of 450 nm using CCK-8 assay. d and e show the intravenous injection of FITC-labeled DC@CFO / BFO nanocomposite (10 mg / kg) into mice, and the in vivo distribution of DC@CFO / BFO was assessed by bioluminescence imaging. fu shows the intravenous injection of 50 mg / kg of DC@CFO / BFO nanocomposite into C57BL / 6 mice daily, followed by magnetic field stimulation (1.5 mT) for 30 minutes daily. Seven days after injection, mouse serum was collected for complete blood count (CBC) tests. RBC represents red blood cells, WBC represents white blood cells, PLT represents platelets, HGB represents hemoglobin, MCV represents mean corpuscular volume, and HCT represents hematocrit.

[0022] Figure 3 This study presents a T-cell transcriptional analysis of DC@CFO / BFO treatment and magnetic field stimulation, where a represents CD4+ in different treatment groups. + PCA analysis of T cell transcripts; b represents CD4 under different treatments. + Heatmap of T cell gene expression; c is a volcano plot analysis of paired comparisons of transcriptional analysis between the DC@CFO / BFO+M group (DC@CFO / BFO and magnetic field treatment) and the DC@CFO / BFO group.

[0023] Figure 4 This study demonstrates how DC@CFO / BFO nanocomposites promote Th2 cell polarization and proliferation under external magnetic field stimulation. In this model, ae represents naïve T cells (CD4+) activated with plate-bound anti-CD3 and anti-CD28 antibodies. + CD25 - CD62L hi CD44 lo Flow cytometry analysis of IFNγ, IL4, IL17A, FOXP3, and CD25 staining (to assess differentiation efficiency) of T cells cultured under various polarization conditions with or without DC@CFO / BFO nanocomposite and magnetic field treatment, followed by restimulation with PMA and ionomycin for 5 hours; f represents activation of CD4 with anti-CD3 / CD28 antibody. + T cells were induced to differentiate into Th2 cells, and then these cells were treated with DC@CFO / BFO nanocomposite materials with or without magnetic field stimulation. Under Th2 differentiation conditions, and with or without magnetic field stimulation, the DC@CFO / BFO nanocomposite materials treated CD4 cells... + GSEA analysis of genes expressed in T cells, where ES represents the enrichment score and NES represents the normalized enrichment score; g represents activation of CD4 with an antibody against CD3 / CD28.+ T cells were induced to differentiate into Th2 cells, and then these cells were treated with DC@CFO / BFO nanocomposites in the presence or absence of magnetic field stimulation. This demonstrated the effect of differentiating CD4 cells in the presence or absence of DC@CFO / BFO nanocomposites and magnetic field stimulation. + A heatmap of genes expressed in T cells related to T helper 2 cell differentiation; h and i represent activation of CD4 cells with antibodies targeting CD3 / CD28, respectively. + T cells were induced to differentiate into Th2 cells, and then these cells were treated with DC@CFO / BFO nanocomposite material with or without magnetic field stimulation to induce CD4+ differentiation. + Flow cytometry analysis of CFSE dilution staining and Ki-67 expression in T cells, MFI represents mean fluorescence intensity.

[0024] Figure 5 This study illustrates how magnetoelectric nanocomposites improve arthritis under external magnetic field stimulation. Specifically, a) is a schematic diagram of DC@CFO / BFO nanocomposite treatment in a CIA mouse model. DBA / 1 mice were intradermally injected with bovine type II collagen (CII) emulsified in Freund's complete or incomplete adjuvant at days 0 and 21. 10 mg / kg of DC@CFO / BFO nanocomposites was then intravenously injected into the mice, and a 1.5 mT magnetic field was applied for 30 minutes at a specified time point after nanocomposite injection. b) shows the arthritis scores of the animals every other day after disease onset, with a maximum clinical score of 16 points. c) shows representative images of the ankle joints from different treatment groups, with a scale bar of 200 μm. d) shows the measurement of paw thickness in arthritis mice from different treatment groups. e) shows micro-CT analysis of bone mineral density (BMD) of the ankle joints from different treatment groups. f) shows the measurement of proteoglycan loss in the ankle joints based on Safranin-O / Fast Green staining images. g and h show flow cytometry analysis of CD4+ in the spleens of mice from different treatment groups. + IFNγ + Cells and CD4 + IL4 + The proportion of cells; i and j represent the proportions of CD4+ cells in mice with drained LN (dLN) from different treatment groups analyzed by flow cytometry. + IFNγ + Cells and CD4 + IL4 + The proportion of cells; k and l are the percentages of CD4+ in the ankle joints of mice from different treatment groups analyzed by flow cytometry. + IFNγ + Cells and CD4 + IL4 +The proportion of cells; m is the mouse serum obtained and the anti-type 2 collagen (anti-CII) specific antibody measured by ELISA; n is the mouse serum obtained and the inflammatory factors assessed.

[0025] Figure 6 CD4 in arthritis mice is shown. + Flow cytometry analysis of T cell subsets, where 'a' represents CD4+ from the spleen of mice from different treatment groups. + Flow cytometry analysis of T cell proportions; b shows CD4 counts in mice with drained lymph nodes (dLNs) from different treatment groups. + Flow cytometry analysis of T cell proportions; c represents CD4+ in the ankle joints of mice from different treatment groups. + Flow cytometry analysis of T cell proportions.

[0026] Figure 7 This paper presents an analysis of T cell subsets and a serum inflammation assessment in arthritic mice, where a and b represent CD4+ in the spleens of mice from different treatment groups, respectively. + IL17A + Cells and CD4 + FOXP3 + Flow cytometry analysis of cell proportions; c and d are flow cytometry analyses of CD4+ in LN mice drained from different treatment groups. + IL17A + Cells and CD4 + FOXP3 + The proportion of cells; e and f are the percentages of CD4+ in the ankle joints of mice from different treatment groups analyzed by flow cytometry. + IL17A + Cells and CD4 + FOXP3 + The proportion of cells; g represents the amount of mouse serum obtained and inflammatory factors assessed.

[0027] Figure 8 The illustration shows the inhibition of arthritis in mice by Th2 cells treated with magnetoelectric nanocomposites. Figure a is a schematic diagram of Th2 cell administration pretreated with DC@CFO / BFO nanocomposites in a CIA mouse model. DBA / 1 mice were intradermally injected with bovine type II collagen (CII) emulsified in either Freund's complete or incomplete adjuvant at days 0 and 21, and CD4 cells were activated with an antibody targeting CD3 / CD28. +T cells were induced to differentiate into Th2 cells, treated with DC@CFO / BFO nanocomposite material (10 μg / mL) and stimulated with a magnetic field (1.5 mT) for 30 minutes. The DC@CFO / BFO nanocomposite-pretreated Th2 cells were intravenously injected into arthritic mice. b shows the arthritis score of the animals every other day after the onset of the disease, with a maximum clinical score of 16 points per animal. c shows representative images of the ankle joint from different treatment groups, with a scale bar of 200 μm. d shows the paw thickness of the arthritic mice from different treatment groups. e shows the micro-CT analysis of bone mineral density (BMD) of the ankle joint from different treatment groups. f shows the proteoglycan loss of the ankle joint. g shows the acquisition of mouse serum and the quantification of anti-type 2 collagen (anti-CII) specific antibodies by ELISA. Detailed Implementation

[0028] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0029] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that the upper and lower limits of the range and each intermediate value between them are specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0030] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0031] use

[0032] In one aspect, the present invention provides the use of a nanocomposite material in the preparation of a medicament for treating arthritis, wherein the nanocomposite material comprises, from the outside to the inside, an outer layer of a membrane structure, an intermediate layer of a piezoelectric material, and a core structure of a magnetostrictive material, wherein the outer surface of the membrane structure contains protein molecules.

[0033] The nanocomposite material of this invention can promote Th2 cell polarization and enhance T cell proliferation under external magnetic field stimulation, thereby alleviating T cell-mediated autoimmune responses and maintaining host immune homeostasis. It should be understood that Th2 cells pretreated with the nanocomposite material or Th2 cells containing nanoparticles (cobalt ferrite-bismuth ferrite) are also within the scope of this invention. Based on this, the present invention further provides the use of Th2 cells pretreated with the nanocomposite material or Th2 cells containing cobalt ferrite-bismuth ferrite in the preparation of medicaments for treating arthritis.

[0034] In this invention, arthritis includes, but is not limited to, degenerative arthritis (e.g., but not limited to osteoarthritis), autoimmune arthritis (e.g., but not limited to rheumatoid arthritis, rheumatic arthritis), and infectious arthritis (e.g., but not limited to Neisseria gonorrhoeae arthritis, non-gonococcal arthritis, etc.). In a preferred embodiment, the arthritis is autoimmune arthritis, particularly T-cell-induced arthritis.

[0035] In this invention, the source of the membrane structure is not particularly limited, as long as its surface contains protein molecules capable of binding to the surface of T cells. Examples of membrane structures include, but are not limited to, naturally occurring biological membranes or artificially synthesized membrane structures. In a preferred embodiment, the biological membrane is a cell membrane isolated from immune cells, including but not limited to cell membranes isolated from dendritic cells, B lymphocytes, or macrophages. In a preferred embodiment, the biological membrane is a dendritic cell cell membrane.

[0036] In this invention, the surface of the T cells contains at least one of the following antigens: CD1, CD2, CD3, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD153, CD154, CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, OX40, GITR, L AG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6 and CCR7.

[0037] Those skilled in the art know how to make the outer surface of the cell membrane contain or express substances that can target and bind CD4. +Protein molecules that are T-cell surface antigens. In a preferred embodiment, the protein molecule is capable of targeting CD4. + MHC II complex of T cells.

[0038] In this invention, the piezoelectric material is selected from at least one of ferric, niobic, titanic, silicatic, and aluminic materials, and / or from at least one of polyvinylidene fluoride, polyester, polymethyl methacrylate, nylon, polyvinyl chloride, polylactic acid lactide, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), vinylidene fluoride / trifluoroethylene copolymer, vinylidene fluoride / tetrafluoroethylene copolymer, polyvinylidene fluoride-hexafluoropropylene, and polydimethylsiloxane. In a preferred embodiment, the intermediate layer of the piezoelectric material is bismuth ferrite.

[0039] In this invention, the magnetostrictive material is selected from ferric acid materials, and the ferric acid material has the following structure: N a Fe 1-a O b N includes at least one of Co, Mn, Zn, and Ni, a is a number from 0.25 to 0.75, and b is 3. In a preferred embodiment, the magnetostrictive material is cobalt ferrite.

[0040] It is understood that the magnetostrictive piezoelectric nanocomposite material of the present invention may or may not contain doped elements. When it contains doped elements, the doped elements include, but are not limited to, at least one of carbon, nitrogen, phosphorus, sulfur, silicon, aluminum, iron, titanium, nickel, manganese, copper, silver and zinc, thereby improving the piezoelectric properties and / or magnetostrictive properties of the magnetostrictive piezoelectric nanocomposite material.

[0041] In this invention, the size of the nanocomposite material is 100-500nm, preferably 100-450nm, even more preferably 100-400nm, further preferably 100-350nm, more preferably 100-300nm, and even more preferably 100-250nm, for example 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, and 250nm.

[0042] In this invention, external magnetic field stimulation refers to treating the nanocomposite particles in the presence of a magnetic field of 0.01-300 mT. The magnetic field strength is preferably 0.1-200 mT, more preferably 0.5-100 mT, even more preferably 0.5-50 mT, and most preferably 0.5-10 mT, for example, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, and 10 mT. In a preferred embodiment, the magnetic field is applied at the above-mentioned magnetic field strength once a day for 15-30 minutes each time, for three consecutive days.

[0043] Preparation method

[0044] One aspect of the present invention provides a method for preparing a nanocomposite material, comprising:

[0045] (1) A magnetostrictive piezoelectric nanocomposite material is prepared, wherein the magnetostrictive piezoelectric nanocomposite material comprises a piezoelectric material intermediate layer and a magnetostrictive material core structure;

[0046] (2) The magnetostrictive piezoelectric nanocomposite material is coated inside the membrane material, wherein the outer surface of the membrane material contains protein molecules.

[0047] Step (1) of the present invention is to prepare the core structure of the magnetostrictive material and the intermediate layer of the piezoelectric material.

[0048] In one specific embodiment, the preparation of the core structure of the magnetostrictive material in step (1) of the present invention includes: dissolving FeCl3, CoCl2, and CTAB in deionized water, wherein the molar ratio of FeCl3, CoCl2, and CTAB is (2-6):1:(3-9), preferably (2-5):1:(3-8), and even more preferably (2-4):1:(3-6), for example 2:1:3, 2:1:4, 2:1:5, 2:1:6, 3:1:3, 3:1:4, 3:1:5, 3:1:6, 4:1:3, 4:1:4, 4:1:5, 4:1:6; then, under stirring and ultrasonic dispersion, adding NaOH to the above solution, wherein the molar ratio of FeCl3 to NaOH is... The solution is prepared by mixing 1:(20-100), preferably 1:(30-90), more preferably 1:(40-80), for example 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, and then the resulting solution is transferred to a sealed hydrothermal reactor and heated at 150-250°C, preferably 160-240°C, more preferably 170-230°C, for example 170°C, 180°C, 190°C, 200°C, 210°C, 220°C, 230°C for 12-36 hours, preferably 14-34 hours, even more preferably 16-32 hours, and more preferably 18-30 hours, for example 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 hours. The obtained black powder is then washed with deionized water and ethanol, and then dried overnight at 80-110°C, preferably 85-105°C, and even more preferably 90-100°C, for example 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C, or 100°C, to obtain the core structure of the magnetostrictive material, namely cobalt ferrite (CFO) nanoparticles.

[0049] In a preferred embodiment, the preparation of the piezoelectric material interlayer in step (1) of the present invention includes: dissolving Bi(NO3)·5H2O and Fe(NO3)·9H2O in ethylene glycol to prepare a bismuth ferrite (BFO) precursor, wherein the molar ratio of Bi(NO3)·5H2O and Fe(NO3)·9H2O is (1-5):1, for example 1:1, 2:1, 3:1, 4:1, 5:1, and then drying CF CFO nanoparticles are dispersed in a BFO precursor solution and sonicated for 1-5 hours (e.g., 1, 2, 3, 4, 5 hours), wherein the molar ratio of CFO nanoparticles to BFO precursor is (1-5):1, for example 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1. The solution is then dried for 6-20 hours, preferably 8-18 hours, and even more preferably 10-16 hours, for example 10, 11 hours. 12, 13, 14, 15, 16 hours later, the dried powder is then heated to 500-700°C, preferably 520-680°C, and more preferably 540-660°C, at a heating rate of 5-15°C / min, preferably 7-13°C / min, for example 7, 8, 9, 10, 11, 12, 13°C / min, such as 540°C, 550°C, 560°C, 570°C, 580°C, 590°C, 600°C, 610°C. Anneal at 620℃, 630℃, 640℃, 650℃, and 660℃ for 1-5 hours, for example, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, and 5 hours. Finally, the annealed powder is ultrasonically treated for 15-45 minutes, preferably 20-40 minutes, and even more preferably 25-35 minutes, for example, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, and 35 minutes, and then filtered through a filter membrane.

[0050] In some embodiments, the membrane material described in step (2) of the present invention is an artificially synthesized membrane structure. The artificially synthesized membrane structure can be prepared by methods known in the art, and is not particularly limited thereto. Examples include, but are not limited to, self-assembly, template method, charge-induced method, etc.

[0051] In some embodiments, the membrane material described in step (2) of the present invention is a cell membrane, which is obtained through cell culture, cell separation, and cell lysis.

[0052] In one specific embodiment, the method for preparing the cell membrane includes: firstly, isolating bone marrow from the femur and tibia of mice, and using 10-30 ng / ml, preferably 12-28 ng / ml, and even more preferably 14-26 ng / ml, such as 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 ng / ml of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and 1-20 ng / ml, preferably 2-18 ng / ml, even more preferably 4-16 ng / ml, and more preferably 6-14 ng / ml, such as 6, 7, 8, 9, 10, 11, 12, 13, 14 ng / ml of recombinant interleukin-4 to induce dendritic cell (DC) differentiation, and then sorting the DCs (CD11c) + MHC II + The cells are lysed, and the lysate is sonicated for 1-10 min, preferably 2-9 min, and even more preferably 3-8 min, for example 3, 4, 5, 6, 7, 8 min, and the cell membrane is collected. The lysis method is not particularly limited, and the sorted dendritic cells can be lysed by freezing and thawing in liquid nitrogen for several cycles (preferably 1-5 cycles, for example 1, 2, 3, 4, 5 cycles).

[0053] In a preferred embodiment, step (2) of the present invention includes continuously passing the membrane material through a porous membrane (e.g., a polycarbonate porous membrane) at 300-500 nm, preferably 320-480 nm, even more preferably 340-460 nm, and more preferably 360-440 nm, such as 360, 370, 380, 390, 400, 410, 420, 430, 440 nm, and then extruding it through a porous membrane at 100-300 nm, preferably 120-280 nm, even more preferably 140-260 nm, and more preferably 160-240 nm, such as 160, 170, 180, 190, 200, 210, 220, 230, 240 nm, into the magnetostrictive piezoelectric nanocomposite material obtained in step (1).

[0054] Methods for regulating Th1 / Th2 cell balance in vitro

[0055] One aspect of the present invention provides a method for in vitro regulation of Th1 / Th2 cell balance, comprising the step of contacting the nanocomposite material described herein with isolated cells in the presence of a magnetic field of 0.01-300 mT. The isolated cells are T cells, particularly CD4 cells. + T cells.

[0056] The present invention further provides a method for promoting the differentiation of T cells into Th2 cell subsets in vitro, which includes the step of contacting the nanocomposite material of the present invention with isolated cells in the presence of a magnetic field of 0.01-300 mT.

[0057] Example 1

[0058] The following exemplarily illustrates the preparation and performance analysis of DC@CFO / BFO nanocomposites.

[0059] I. Experimental Materials

[0060] DBA / 1 mice were purchased from Vital River Laboratory Animal Technology Co., Ltd. All animals were housed and maintained under specific pathogen-free conditions. All animal experiments were conducted according to protocols approved by the Ethics Committee of Peking University School of Medicine.

[0061] II. Preparation Method

[0062] 1. Preparation of CFO / BFO

[0063] (1) Preparation of core-structured CFO nanoparticles: 0.092 M FeCl3, 0.046 M CoCl2, and 0.14 M CTAB were dissolved in deionized water. Then, under thorough mechanical stirring and ultrasonic dispersion, 6 M NaOH was added to the above solution, and the resulting solution was transferred to a sealed polytetrafluoroethylene hydrothermal reactor and heated at 200 °C for 24 hours. The solution was then washed with deionized water and ethanol to obtain a black powder, which was then dried overnight at 95 °C.

[0064] (2) Preparation of core-shell CFO / BFO nanoparticles: 0.011 M Bi(NO3)·5H2O and 0.01 M Fe(NO3)·9H2O were dissolved in ethylene glycol to prepare BFO precursors. Then, 0.15 g of dried CFO nanoparticles were dispersed in 100 mL of the BFO precursor solution and sonicated for 3 hours. The solution was then dried overnight. Subsequently, the dried powder was heated to 600 °C at a heating rate of 10 °C / min and annealed for 2.5 hours. Finally, the annealed powder was sonicated for 30 minutes and filtered through a 0.22 μm filter membrane to obtain magnetoelectric core-shell CFO / BFO nanoparticles.

[0065] 2. Preparation of DC@CFO / BFO nanocomposites

[0066] Preparation of DC membrane-modified CFO / BFO nanocomposites: First, bone marrow from mouse femurs and tibias was isolated, and DC differentiation was induced using 20 ng / ml recombinant GM-CSF and 10 ng / ml recombinant IL4. Then, DCs (CD11c) were sorted by flow cytometry. + MHC II +Cells were lysed by freezing and thawing in liquid nitrogen for three cycles, and the lysate was sonicated for 5 minutes using a bath sonicator. The cell membranes were then collected and continuously extruded through a 400 nm and then a 200 nm polycarbonate porous membrane into the magnetoelectric core-shell CFO / BFO nanoparticles to obtain DC@CFO / BFO nanocomposite material.

[0067] III. Performance Analysis Methods

[0068] 1. Characterization of CFO / BFO nanoparticles and DC@CFO / BFO nanocomposites

[0069] The surface morphology of CFO / BFO nanoparticles was characterized by scanning electron microscopy (SEM). The structure of core-shell CFO / BFO nanoparticles and DC-coated CFO / BFO composites was observed using high-resolution transmission electron microscopy (TEM). The composition of the CFO / BFO nanoparticles was determined using energy-dispersive X-ray spectroscopy (EDS) or X-ray diffraction spectroscopy with TEM. Piezoelectric response force microscopy (PFM) measurements were recorded using a commercial atomic force microscope equipped with a ferroelectric testing system. Measurements were performed using a vibrating sample magnetometer (VSM). The nanocomposites were placed in ddH₂O or DMSO, with or without magnetic field stimulation or sonication, for use with ·OH and ·O₂, respectively. - Detection. 5,5-Dimethyl-1-pyrrolidone-N-oxide (DMPO) was added to a solution, and reactive substances generated by the nanocomposite were detected using EPR technology. The size and surface potential of the nanocomposite were measured by instrumentation in an aqueous environment at room temperature.

[0070] 2. Immunoblot analysis

[0071] Cells were lysed with RIPA lysis buffer supplemented with a protease inhibitor mixture and then subjected to SDS-PAGE. The antibodies used in this example were as follows: anti-CD11b (abcam, ab133357, 1:1000), anti-β-catenin (abcam, ab32572, 1:5000), anti-HDAC1 (Santa Cruz Biotechnology, sc-81598, 1:1000), anti-purinemycin (Merck-millipore, MABE341, 1:1000), and anti-GAPDH (RayAntibody, RM2002, 1:5000).

[0072] 3. Sample preparation and flow cytometry analysis

[0073] Immune cells from the spleen or lymph nodes were isolated by grinding tissue in PBS containing FBS and then filtering it through a cell filter.

[0074] To analyze immune cells in the joints, the ankle was cut from above the heel down to the middle of the toe, minced, and incubated in DMEM medium containing collagenase A, with occasional mixing. The dissociated cells were then washed and filtered through a cell filter.

[0075] To analyze the surface proteins of the DC@CFO / BFO nanocomposite, the DC@CFO / BFO nanocomposite was incubated with a specific antibody at room temperature and analyzed by flow cytometry.

[0076] To detect cellular uptake of the materials, CFO / BFO nanoparticles and DC@CFO / BFO nanocomposites were mixed with equal masses of FITC aqueous solution and stirred. Magnetic separation was performed, and the mixture was washed three times with deionized water to remove excess FITC. The FITC-conjugated materials were then incubated with mouse naïve T cells or spleen cells, and FITC levels were detected by flow cytometry. + cell.

[0077] To detect T cell proliferation, isolated T cells were stained with CFSE at room temperature. Cell activation and proliferation were then induced, and the cells were collected for flow cytometry analysis.

[0078] To analyze cell surface marker expression (CD45, CD4, CD8, CD25, B220, CD11b, Ly6G, CD11c, MHC II, F4 / 80), cells were incubated with specific antibodies at room temperature. For intracellular marker staining (IFNγ, IL4, IL17A), cells were treated with a mixture of protein transport inhibitors, then fixed and permeabilized, followed by staining with specific antibodies. To detect nucleoprotein (Ki-67, FOXP3) expression, cells were fixed and permeabilized, then stained with specific antibodies. The antibodies used include: anti-CD45 (eBioscience, 30-F11), anti-CD4 (BioLegend, GK1.5), anti-CD8 (eBioscience, 53-6.7), anti-CD25 (BioLegend, 3C7), anti-B220 (BioLegend, RA3-6B2), anti-CD11b (BioLegend, M1 / 70), anti-Ly6G (BioLegend, RB6-8C5), anti-CD11c (BioLegend, N418), anti-F4 / 80 (BioLegend, BM8), and anti-MHC. II (BioLegend, IA / IE) (M5 / 114.15.2), anti-IFNγ (BioLegend, XMG1.2), anti-IL4 (eBioscience, 11B11), anti-IL17A (eBioscience, eBio17B7), anti-Ki-67 (BioLegend, 16A8), and anti-FOXP3 (eBioscience, FJK-16s).

[0079] 4. In vivo imaging of DC@CFO / BFO nanocomposites

[0080] To investigate the in vivo distribution of the nanocomposite material, FITC-labeled DC@CFO / BFO nanocomposite material (10 mg / kg) was intravenously injected into C57BL / 6 mice. The mouse heart, liver, kidney, lung, and spleen were isolated at specified time points and then subjected to bioluminescence imaging. The images were analyzed using software.

[0081] 5. Biosafety assessment

[0082] To perform in vitro cytotoxicity assays, mice were initially charged with CD4+, with or without external magnetic field stimulation. + T cells were exposed to a range of concentrations of nanocomposite materials for 24 hours, and T cell viability was assessed using a cell counting kit-8.

[0083] For in vivo biocompatibility assessment, C57BL / 6 mice were administered a high concentration (50 mg / kg) of DC@CFO / BFO nanocomposite material daily and subjected to magnetic field stimulation for 30 minutes. Seven days after injection, blood biochemistry and complete blood count were performed, and histological analysis of major organs was conducted.

[0084] 6. In vitro T cell differentiation

[0085] Initial CD4 counts in mice were sorted from peripheral lymph nodes by flow cytometry. + T cells (CD4) + CD25 - CD62L hi CD44 lo The cells were then activated with 2 μg / mL plate-bound anti-CD3 antibody and 1 μg / mL anti-CD28 antibody. For Th1 polarization, 10 μg / mL anti-IL4 antibody and 10 ng / mL IL-12 were used. For Th2 polarization, 10 μg / mL anti-IFN-γ antibody and 20 ng / mL IL4 were used. For Th17 polarization, 20 ng / mL IL-6, 5 ng / mL TGF-β, 10 μg / mL anti-IFN-γ antibody, and 10 μg / mL anti-IL4 antibody were used. For iTreg cell differentiation, 1 ng / mL TGF-β, 4 ng / mL IL-2, 10 μg / mL anti-IFN-γ antibody, and 10 μg / mL anti-IL4 antibody were used. T cells were incubated with 10 μg / mL DC@CFO / BFO nanocomposite and treated with a 1.5 mT magnetic field for 30 minutes daily for 48 or 72 hours. Cells were then collected for flow cytometry analysis.

[0086] IV. Experimental Results

[0087] 1. Preparation and characterization of cell membrane-coated magnetoelectric nanocomposites

[0088] This embodiment synthesized a DC@CFO / BFO nanocomposite material, comprising a magnetostrictive cobalt(II) ferrite CoFe2O4 (CFO) core and a piezoelectric bismuth(III) ferrite BiFeO3 (BFO) shell (CFO / BFO). The surface morphology and core-shell structure of the film-modified CFO / BFO nanocomposite material were confirmed by high-resolution transmission electron microscopy. Figure 1 (a) Furthermore, under an external magnetic field, EPR spectroscopy was used to detect oxide species (·OH and ·O2) generated by the CFO / BFO nanocomposite in magnetoelectrocatalysis. - Furthermore, due to the piezoelectric properties of BFO, magnetoelectric CFO / BFO nanocomposites and BFO nanoparticles can generate ·OH and ·O2 in the presence of external ultrasound. -The characteristic signal of ROS, but individual nanoparticles cannot trigger the generation of ROS. Figure 1 (be). Therefore, these findings elucidate the role of the magnetic field in inducing magnetostrictive strain in the CFO nucleus and initiating potential polarization in the BFO shell, thus confirming effective magnetoelectric conversion.

[0089] To achieve CD4 + T-cell targeting specificity was studied, using bone marrow-derived dendritic cells as the basis for research. The expression of their major histocompatibility complex (MHC) II was manipulated to interact with CD4+. + T cell receptor (TCR) binding on T cells. Mouse bone marrow cells were first isolated and induced to differentiate into dendritic cells using GM-CSF and IL-4. CD11c... + MHC II + Fluorescence-activated cell sorting (FACS) of cells and collection of cell membranes ( Figure 1 f). Then, a film-modified magnetoelectric nanocomposite material (DC@CFO / BFO) was prepared by repeatedly extruding the freshly extracted BMDC film onto the prepared CFO / BFO nanoparticles. As revealed by TEM, a transparent cell membrane layer of approximately 10 nm was observed on the surface of the film-modified CFO / BFO nanomaterials. Figure 1 (a). Dynamic light scattering experiments showed that the size of the modified DC@CFO / BFO composite material was approximately 170 nm. Figure 1 (g and h). Then, zeta potential measurements showed that the surface charge of the DC@CFO / BFO nanocomposite was smaller than the negative charge of the CFO / BFO nanoparticles (g and h). Figure 1 Furthermore, quantification of glycoproteins revealed that the glycoprotein content on the DC@CFO / BFO nanocomposite was approximately 90% of that in the free BMDC membrane, indicating that the membrane exhibited correct orientation when modified onto the nanoparticles. Figure 1 (j). As can be seen, this embodiment successfully prepared a dendritic cell membrane-modified magnetoelectric nanocomposite material, which exhibits excellent magnetic field responsiveness and magnetostrictive piezoelectric properties.

[0090] 2. Biodistribution characteristics and safety evaluation of DC film-coated magnetoelectric nanocomposites

[0091] First, standard cell counting kit-8 (CCK-8) and mouse initial CD4 were used. + T cell assays were performed to assess the cytotoxicity of the DC@CFO / BFO nanocomposite material with and without magnetic field stimulation, and it was found that the DC@CFO / BFO nanocomposite material exhibited good biocompatibility at concentrations below 100 μg / mL. Figure 2 (ac). In order to maintain CD4 +The high activity of T cells led to the selection of a DC@CFO / BFO nanocomposite material at a concentration of 10 μg / mL for subsequent studies. Next, CD4 counts were measured. + T cell uptake of nanocomposite materials. FITC-labeled DC@CFO / BFO or CFO / BFO nanoparticles were combined with mouse naïve CD4 cells. + T cell incubation was performed, and fluorescence signals were assessed by flow cytometry. The DC membrane accelerated T cell phagocytosis of the nanocomposite, starting at approximately 2 hours and peaking at 8 hours. After 12 hours of co-incubation with mouse spleen cells, most of the DC@CFO / BFO nanocomposite was phagocytosed by CD4+. + T cells took up the nanoparticles, while most of the naked CFO / BFO nanoparticles were taken up by macrophages. Therefore, these data suggest that modifying CFO / BFO nanoparticles with a DC membrane increases their targeting specificity and promotes CD4 activation. + T-cell uptake.

[0092] To investigate the biodistribution characteristics of the nanocomposite material in vivo, FITC-labeled DC@CFO / BFO nanocomposite material was administered intravenously to C57BL / 6 mice. In vivo imaging showed that the nanocomposite material was enriched in the liver, kidneys, lungs, and spleen, and remained in vivo for 4 days post-injection. Figure 2 (d and e). Consistent with low in vitro cytotoxicity, supplementation with high concentrations (50 mg / kg) of DC@CFO / BFO nanocomposite with or without magnetic field stimulation had almost no effect on mouse hematopoietic cells 7 days after injection. Figure 2 Furthermore, the DC@CFO / BFO nanocomposite material had negligible effects on the functional parameters of the liver and kidneys, and no significant abnormalities were observed in the major organs through histological analysis. Therefore, the data from this embodiment validate the excellent in vivo safety of the DC@CFO / BFO nanocomposite material.

[0093] 3. DC@CFO / BFO nanocomposites promote Th2 cell polarization and proliferation under external magnetic field.

[0094] To further investigate the role of DC@CFO / BFO nanocomposites in CD4 + The role of T cell activation and differentiation, and the activation of CD4 cells with anti-CD3 and anti-CD28 antibodies under various helper T cell polarization conditions. + Naïve T cells. Flow cytometry analysis showed that in T cells treated with an external magnetic field (DC@CFO / BFO nanocomposite), IL4 production was enhanced while IFN-γ production was reduced, indicating that the combination of magnetoelectric nanocomposite and magnetic field promoted Th2 differentiation and inhibited Th1 differentiation. Figure 4Furthermore, in the DC@CFO / BFO group and the DC@CFO / BFO+M group, T cells produced less IL17A, and the proportion of induced Treg cells (iTreg) did not differ significantly among all groups. Figure 4 (a, d, and e).

[0095] To more accurately describe the role of magnetoelectric nanocomposites in Th2 polarization, RNA sequencing was performed in this embodiment to analyze the transcriptome of T cells with magnetoelectric nanocomposites and magnetic field treatment under Th2 differentiation conditions.

[0096] Principal component analysis (PCA) and differentially expressed genes (DEGs) analysis showed that, under magnetic field-free conditions, activated T cells exhibited similar gene expression patterns in the presence or absence of DC@CFO / BFO nanocomposites. Figure 3 (a and b). However, T cells in the DC@CFO / BFO+M group were clearly distinguished from T cells without nanocomposite materials or magnetic field stimulation by their transcripts ( Figure 3 (a and b). Subsequent GO enrichment showed that, relative to naive T cells, genes associated with DNA replication, T cell differentiation, and T cell proliferation were significantly upregulated in T cells under Th2 cell polarization conditions (vector group and DC@CFO / BFO group), indicating favorable T cell polarization induction.

[0097] Subsequently, this embodiment analyzed the overall gene expression changes of T cells treated with DC@CFO / BFO in the presence or absence of a magnetic field, and found that after DC@CFO / BFO+M treatment, 898 genes were upregulated and 1703 genes were downregulated in T cells. Figure 3 (c). Consistent with previous data, the upregulated genes were enriched in "type II immune response" and "Th2 cell differentiation" compared to T cells in the DC@CFO / BFO group, reflecting the stimulatory effect of DC@CFO / BFO treatment and accompanying magnetic field stimulation on Th2 cell polarization. Figure 4 (f and g). Furthermore, the DC@CFO / BFO nanocomposite promoted signaling pathways associated with T cell proliferation under magnetic field loading. Subsequent flow cytometry analysis also confirmed that CD4+ was involved under Th2 cell polarization conditions. + T cells in response to DC@CFO / BFO+M treatment underwent more cell division, as detected by CFSE dilution assay and Ki-67 staining. Figure 4 (h and i). In summary, these findings provide evidence that treatment with magnetoelectric nanocomposites combined with magnetic field stimulation promotes Th2 cell polarization induction and enhances T cell proliferation in vitro.

[0098] Example 2

[0099] The following example illustrates the application of DC@CFO / BFO nanocomposites in arthritis.

[0100] I. Experimental Methods

[0101] 1. Collagen-induced arthritis (CIA) model

[0102] 100 μg of bovine type II collagen (CII) emulsified in Freund's complete adjuvant containing 4 mg / ml heat-inactivated mycobacteria was injected intradermally into DBA / 1 mice. On day 21, a reinforcing emulsion prepared from CII and Freund's incomplete adjuvant was administered intradermally near the main injection site. 10 mg / kg of DC@CFO / BFO nanocomposite was injected intravenously into mice at designated time points, and a 1.5 mT magnetic field was applied for 30 minutes daily following nanocomposite injection. Arthritis animals were clinically evaluated every other day after disease onset, with a maximum clinical score of 16 points per animal.

[0103] 2. Enzyme-linked immunosorbent assay (ELISA)

[0104] Perform enzyme-linked immunosorbent assay (ELISA) of glycoproteins on cell membranes or nanoparticles according to the manufacturer's instructions.

[0105] Type 2 collagen-specific antibodies (anti-CII) in serum collected from immunized mice were measured by enzyme-linked immunosorbent assay (ELISA). Serum samples were added to plates pre-coated with 10 μg / mL CII, followed by the addition of goat anti-mouse IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA.

[0106] 3. Serum inflammatory cytokine detection

[0107] Mouse serum samples were collected from CIA mice, and inflammatory cytokines were measured according to the manufacturer's instructions.

[0108] 4. Histological analysis

[0109] For histological evaluation of arthritis in mice, the hind paws and femurs were fixed with paraformaldehyde, decalcified in EDTA solution, dehydrated in graded ethanol, embedded in paraffin, and tissue sections were used for histological evaluation. H&E staining, safranin-O / fast green, and toluidine blue staining were performed to score the histopathology of the inflammatory arthritis. Images were acquired using a microscope.

[0110] 5. RNA sequencing

[0111] To investigate the role of magnetoelectric nanocomposites in T cell-mediated responses, CD4+ was incorporated into the study. +T cells were incubated with magnetoelectric nanocomposites and subjected to magnetic field treatment under Th2 differentiation conditions. Total RNA was then purified using poly-T oligonucleotide-linked magnetic beads. RNA-seq libraries were constructed using a library preparation kit and reads were sequenced on the Illumina platform. Clean data was obtained by removing reads containing adapters and poly-N molecules, as well as low-quality reads, from the raw data, according to the manufacturer's instructions. Clean reads were aligned to the Hisat2 reference genome based on gene model annotation files. Fragments per kilobase per million mapped reads (FPKM) were calculated for each gene based on gene length and the number of reads mapped to that gene. Overlap between gene sets from RNA-seq data and selected datasets in MSigDB was analyzed.

[0112] 6. Quantitative proteomics analysis

[0113] Separate CD4 + T cells were then incubated with magnetoelectric nanocomposites under Th2 differentiation conditions and magnetic field treatment. LC-MS / MS analysis was performed on 100 μg protein samples from each subject.

[0114] 7. Non-targeted metabolomics analysis

[0115] Separate CD4 + T cells were then incubated with a magnetoelectric nanocomposite material under Th2 differentiation conditions and magnetic field treatment. To extract T cell metabolites, the cell pellet was resuspended in methanol (pre-cooled to -80°C) and frozen in liquid nitrogen. These quenched cells were then thawed and vortexed. After centrifugation, the supernatant was transferred to a new tube and lyophilized under vacuum. The dried sample was then reconstituted in methanol and incubated. The sample was then centrifuged, and the supernatant was used for LC-MS / MS analysis.

[0116] 8. Statistical Analysis

[0117] Statistical analysis was performed using software. A two-tailed Student's t-test was used to analyze differences between the two groups. P < 0.05 was considered significant.

[0118] II. Experimental Results

[0119] Considering CD4 + T cells play a crucial role in regulating the host's autoimmune response. This embodiment further evaluates the anti-inflammatory effect of DC@CFO / BFO nanocomposites under repeated magnetic field exposure. This embodiment explores the therapeutic application of magnetoelectric nanocomposites in a collagen-induced arthritis (CIA) model and monitors the dynamic pathological process. Figure 5 (a). After disease onset, mice treated with DC@CFO / BFO+M showed a significant reduction in arthritis scores ( Figure 5 (b). Data shows that under external magnetic field stimulation, treatment with DC@CFO / BFO nanocomposite materials reduced tissue swelling caused by local inflammation. Figure 5 (c and d). Micro-CT showed that the joint structures in the DC@CFO / BFO+M group were well preserved, and the bone mineral density around the ankle joint was restored ( Figure 5 (c and e). Subsequent histological analysis of ankle injuries showed that treatment of arthritis in mice with DC@CFO / BFO nanocomposite material under magnetic field stimulation significantly reduced the degree of synovitis and the overall extent of cartilage and bone damage (c and e). Figure 5 (c and f).

[0120] It is noteworthy that in the spleen of mice in the DC@CFO / BFO+M group ( Figure 5 g and h), draining lymph nodes (dLN) ( Figure 5 (i and j) and ankle joint ( Figure 5 In both k and l), a higher frequency of IL4 production was consistently observed, along with IFNγ. + The cell proportion was lower. As shown by flow cytometry analysis, DC@CFO / BFO treatment also slightly reduced IL17A in CIA mice. + T cells and increased FOXP3 + T cells. Furthermore, the detection of humoral anti-type 2 collagen (anti-CII) specific antibodies and serum cytokine profiles showed that DC@CFO / BFO+M inhibited the production of pathogenic anti-CII IgG and pro-inflammatory cytokines. Figure 5 (m and n).

[0121] To further verify the finding that the anti-inflammatory effect of the magnetoelectric nanocomposite material is mainly reflected in its stimulation of Th2 cell polarization, this embodiment uses the magnetoelectric nanocomposite material to treat mouse nascent CD4 cells. + T cells were stimulated with a magnetic field under Th2 differentiation conditions. Subsequently, these cells were transferred to mice after an arthritis attack. Figure 8 a). Through arthritis scores ( Figure 8 b) Gross observation and histological analysis Figure 8 Evaluation using the cf) method showed that T cells pretreated with DC@CFO / BFO nanocomposites and a magnetic field significantly reduced the incidence and severity of arthritis in CIA mice. Lower levels of anti-CII antibodies were detected in the DC@CFO / BFO+M group, indicating the inhibitory effect of the magnetoelectric nanocomposites on arthritis development. Figure 8 (g). In summary, these data thus demonstrate the anti-arthritis efficacy of magnetoelectric nanocomposites under the influence of an external magnetic field.

[0122] III. Conclusion

[0123] The DC membrane-modified magnetoelectric nanocomposite material of this invention can regulate the Th1 / Th2 balance and promote Th2 cell proliferation, thereby improving T cell-mediated autoimmune responses and maintaining host immune homeostasis. This invention, through DC membrane modification, enhances the T cell targeting specificity of the magnetoelectric particles, making it applicable to T cell-induced arthritis.

[0124] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of magnetically stimulated nanocomposite materials in the preparation of drugs that promote Th2 cell polarization and proliferation for the treatment of arthritis, characterized in that, The nanocomposite material comprises, from the outside in, an outer membrane structure, a piezoelectric intermediate layer, and a magnetostrictive core structure. The piezoelectric intermediate layer and the magnetostrictive core achieve magnetoelectric conversion under magnetic field stimulation. The membrane structure is a cell membrane isolated from dendritic cells, and the outer surface of the cell membrane contains components capable of targeting CD4. + The MHC II complex of T cells, wherein the piezoelectric material is bismuth ferrite and the magnetostrictive material is cobalt ferrite.

2. The use according to claim 1, characterized in that, The strength of the magnetic field is 0.01-300 mT.

3. The use according to claim 1, characterized in that, The nanocomposite material was prepared by the following method: (1) Prepare a magnetostrictive piezoelectric nanocomposite material, wherein the magnetostrictive piezoelectric nanocomposite material comprises a piezoelectric material intermediate layer and a magnetostrictive material core structure; (2) The magnetostrictive piezoelectric nanocomposite material is coated within a membrane material, wherein the outer surface of the membrane material contains protein molecules.

4. The use of Th2 cells pretreated with magnetically stimulated nanocomposite materials in the preparation of drugs for treating arthritis, characterized in that, The nanocomposite material comprises, from the outside in, an outer membrane structure, a piezoelectric intermediate layer, and a magnetostrictive core structure. The piezoelectric intermediate layer and the magnetostrictive core achieve magnetoelectric conversion under magnetic field stimulation. The membrane structure is a cell membrane isolated from dendritic cells, and the outer surface of the cell membrane contains components capable of targeting CD4. + The MHC II complex of T cells, wherein the piezoelectric material is bismuth ferrite and the magnetostrictive material is cobalt ferrite.

5. The use according to claim 4, characterized in that, The strength of the magnetic field is 0.01-300 mT.

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