Self-assembled nano-particles, preparation method and application thereof, exosome-coated nano-material and application of exosome-coated nano-material

Abstract

The invention discloses a self-assembled nano-particle, a preparation method and an application, an exosome coated nano-material containing the self-assembled nano-particle and an application, the self-assembled nano-particle comprises schisanhenol and chlorogenic acid, and the schisanhenol and the chlorogenic acid are mixed to form the nano-particle. The nanoparticles are formed through self-assembly of schisanhenol and chlorogenic acid, the self-assembled nanoparticles have excellent oxidation resistance, the self-assembled nanoparticles are wrapped by the exosome to form the nano-material, and the nano-material can treat liver injury and promote formation of specific immune tolerance.

Description

Technical Field

[0001] The present invention belongs to the technical field of self-assembling materials, and specifically relates to self-assembling nanoparticles, a preparation method and applications thereof, and exosome-encapsulated nanomaterials containing self-assembling nanoparticles and applications thereof. Background Art

[0002] In daily work and life, acute liver damage may occur due to excessive drinking or improper diet; acute liver damage can cause loss of appetite, abdominal pain and other phenomena, seriously affecting normal life.

[0003] Existing treatments for acute liver injury rely on Western medications, such as antioxidants and anti-inflammatory drugs, to reduce oxidative stress in the liver and inhibit stem cell inflammation. These medications can have significant side effects, such as elevated liver enzyme levels, monovascular stasis jaundice, drug-induced fatty liver disease, and gastrointestinal bleeding.

[0004] Although Chinese herbal medicine can be used to relieve and regulate the liver with fewer side effects, it cannot be used directly to treat acute liver injury because it takes effect slowly and the symptoms of acute liver injury are urgent and obvious. Summary of the Invention

[0005] The object of the present invention is to avoid the shortcomings of the prior art and provide a self-assembled nanoparticle, a preparation method and application, an exosome-encapsulated nanomaterial containing the self-assembled nanoparticle, and its application. The nanoparticles are formed by self-assembly of schisandra phenol and chlorogenic acid. The self-assembled nanoparticles have excellent antioxidant properties. The self-assembled nanoparticles are encapsulated by exosomes to form nanomaterials. The nanomaterials can treat liver damage and promote the formation of specific immune tolerance.

[0006] To achieve the above object, the present invention provides a self-assembled nanoparticle, comprising schisandra phenol and chlorogenic acid, wherein the schisandra phenol and chlorogenic acid are mixed to form the nanoparticle.

[0007] Furthermore, the mass ratio of schisandra phenol to chlorogenic acid is 1:0.5-1.5.

[0008] Furthermore, the particle size of the self-assembled nanoparticles is 80-90 nm.

[0009] The present application also provides a method for preparing self-assembled nanoparticles, comprising:

[0010] Mixing schisandra phenol and chlorogenic acid, adding them into anhydrous ethanol and ultrasonically dispersing them to obtain a first mixed solution;

[0011] adding the first mixed solution dropwise into pure water for ultrasonic dispersion to obtain a second mixed solution;

[0012] The anhydrous ethanol in the second mixed solution is removed to obtain self-assembled nanoparticles.

[0013] The present application also provides an application of self-assembled nanoparticles, wherein the self-assembled nanoparticles described above are used for in vitro antioxidant treatment.

[0014] The present application also provides an exosome-encapsulated nanomaterial, comprising exosomes and self-assembled nanoparticles encapsulated by the exosomes, wherein the self-assembled nanoparticles are the self-assembled nanoparticles described above.

[0015] Furthermore, the mass ratio of the exosomes to the self-assembled nanoparticles is 1:90-110.

[0016] Furthermore, the particle size of the nanomaterial is 100-120 nm.

[0017] The present application also provides an application of an exosome-encapsulated nanomaterial, wherein the nanomaterial is used to treat liver damage.

[0018] The present application also provides an application of an exosome-encapsulated nanomaterial, wherein the nanomaterial is used to promote the formation of specific immune tolerance.

[0019] The present invention has the following beneficial effects:

[0020] In the present application, schisandra phenol and chlorogenic acid are self-assembled by ultrasonic mixing to form uniformly dispersed nanoparticles. The self-assembled nanoparticles exhibit excellent multiple antioxidant properties against various free radicals in vitro; the self-assembled nanoparticles are encapsulated by exosomes to form nanomaterials with uniform particle size. The exosome-encapsulated nanomaterials can effectively reduce liver tissue damage and necrosis, and have a significant therapeutic effect on liver damage; at the same time, the exosome-encapsulated nanomaterials can effectively regulate the inflammatory response, and by inhibiting the expression of pro-inflammatory factors and promoting the production of anti-inflammatory factors, they have a positive effect on improving the liver inflammation condition and promoting effective specific immune tolerance. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 1 is a transmission electron microscopy image of the self-assembled nanoparticles in Example 1;

[0022] Figure 2 is the Zeta potential diagram of the self-assembled nanoparticles in Example 1;

[0023] Figure 3 Schematic diagram of the free radical scavenging ability of self-assembled nanoparticles at different concentrations in Example 2;

[0024] Figure 4 This is the electron spin resonance spectrum of the nanomaterial in Example 2;

[0025] Figure 5 This is a projection electron microscope image of the nanomaterial in Example 3;

[0026] Figure 6 The results of ALT and AST in the serum of mice corresponding to the blank control group, the induced control group, and the drug-treated control group in Example 4; n=6-8; ***p<0.001, ****p<0.0001;

[0027] Figure 7 Schematic diagram of the histopathological observation results of the blank control group, induced control group and drug-treated control group in Example 4, wherein Figure 7 A is a typical image of H&E staining; Figure 7 B is the statistical result of liver cell necrosis area; Figure 7 C is a typical image of TUNEL staining; Figure 7 D is the statistical result of the number of hepatocyte apoptosis; n = 3-4; **p < 0.01, ****p < 0.0001, scale bar = 100 μm;

[0028] Figure 8 Schematic diagram of the ELISA method for detecting the levels of inflammatory factors in mouse serum in Example 4; **p<0.01, ***p<0.001, ****p<0.0001;

[0029] Figure 9 The immunofluorescence staining images of CD86 (red), CD206 (red), F4 / 80 (green), and DAPI (blue) in Example 5 are shown;

[0030] Figure 10 The levels of inflammatory factors and anti-inflammatory factors in mouse liver detected by qRT-PCR in Example 5; *p<0.05, **p<0.01, ***p<0.001;

[0031] Figure 11 Results and statistics of flow cytometry analysis; *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0032] The preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although preferred embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. Rather, these embodiments are provided to make the present invention more thorough and complete and to fully convey the scope of the present invention to those skilled in the art.

[0033] The present application provides a method for preparing self-assembled nanoparticles, comprising:

[0034] S1: Mix schisandra phenol and chlorogenic acid, add them into anhydrous ethanol and disperse them by ultrasonication to obtain a first mixed solution; specifically, the mass ratio of schisandra phenol to chlorogenic acid is 1:0.5-1.5; the mass ratio of schisandra phenol to anhydrous ethanol is 1:0.5-1.5; and the ultrasonication time is 10-20 minutes.

[0035] S2: adding the first mixed solution dropwise to pure water for ultrasonic dispersion to obtain a second mixed solution; the mass ratio of schisandra phenol to pure water in the first mixed solution is 1:0.5-1.5; and the ultrasonic time is 10-20 minutes.

[0036] S3: removing anhydrous ethanol from the second mixed solution to obtain self-assembled nanoparticles. Specifically, the ethanol from the second mixed solution can be removed by a rotary evaporator.

[0037] The present application also provides self-assembled nanoparticles prepared by the above method. The particle size of the self-assembled nanoparticles is 80-90 nm and is used for in vitro antioxidant effect.

[0038] The present application also provides an exosome-encapsulated nanomaterial, comprising exosomes and self-assembled nanoparticles as described above, encapsulated by the exosomes. The specific exosomes can be exosomes derived from mesenchymal stem cells or other exosomes known in the art. The role of exosomes in the nanomaterial in this application is twofold: first, exosomes facilitate the targeted aggregation of nanomaterials to sites of inflammation. Second, exosomes themselves have anti-inflammatory protective effects, which synergize with the self-assembled nanoparticles to provide a therapeutic effect.

[0039] The mass ratio of exosomes and self-assembled nanoparticles in the nanomaterial of the present application is 1:90-110, and the particle size of the formed nanomaterial is 100-120nm. It is used to treat liver damage and promote the formation of specific immune tolerance.

[0040] Example 1: Self-assembled nanoparticles and preparation methods

[0041] This embodiment provides a method for preparing self-assembled nanoparticles, comprising:

[0042] S1: 5 mg of schisandra chinensis phenol and 5 mg of chlorogenic acid were mixed and added into 5 ml of anhydrous ethanol and ultrasonically dispersed for 15 minutes to obtain a first mixed solution;

[0043] S2: Slowly add the first mixed solution dropwise to 5 ml of pure water and ultrasonically disperse for 15 minutes to obtain a second mixed solution;

[0044] S3: removing anhydrous ethanol from the second mixed solution by a rotary evaporator to obtain self-assembled nanoparticles (SAL-CA).

[0045] like Figure 1, which is a TEM image of SAL-CA prepared in this example, it can be seen that the self-assembled nanoparticles (SAL-CA) form uniformly dispersed nanoparticles with a particle size of about 80-90 nm. Figure 2 Zeta potential analysis of SAL-CA revealed a zeta potential of +10.45 mV for the self-assembled nanoparticles (SAL-CA), indicating good stability in aqueous media. These results demonstrate that the SAL-CA prepared in this example possesses a uniformly dispersed nanostructure and good stability, making it suitable for biomedical applications.

[0046] It should be noted that, by varying the volume of pure water in step S2, the present invention can prepare self-assembled nanoparticle solutions of varying concentrations. Alternatively, the present invention can prepare self-assembled nanoparticles of varying concentrations by dissolving the prepared self-assembled nanoparticles in varying volumes of pure water.

[0047] Example 2: Application of self-assembled nanoparticles

[0048] Self-assembled nanoparticle solutions of 50 μg / mL, 100 μg / mL, 200 μg / mL, 400 μg / mL, 600 μg / mL, and 800 μg / mL were prepared respectively in the manner of Example 1. It should be noted that to prepare self-assembled nanoparticle solutions of different concentrations, it is only necessary to change the volume of pure water added in step S2. The concentration of the self-assembled nanoparticle solution prepared in Example 1 was 2000 μg / mL.

[0049] The elimination rate of ABTS free radicals by the self-assembled nanoparticle solutions of different concentrations prepared above was calculated. The results are as follows: Figure 3 A. The elimination rate of DPPH free radicals by the self-assembled nanoparticle solutions of different concentrations prepared above was calculated. The results are shown in Figure 3 B. Calculate the effect of the self-assembled nanoparticle solutions of different concentrations on superoxide anions (O2 - ) free radical elimination rate, the results are as follows Figure 3 As shown in C.

[0050] Combine Figure 3 It can be seen that the self-assembled nanoparticles (SAL-CA) can effectively scavenge ABTS and DPPH free radicals in a concentration-dependent manner. Specifically, as the concentration of the self-assembled nanoparticles (SAL-CA) increases from 50 μg / mL to 800 μg / mL, the elimination rate of ABTS free radicals rapidly increases from 12% to 86%.

[0051] Similarly, when the concentration of self-assembled nanoparticles (SAL-CA) reached 800 μg / mL, the scavenging rate of DPPH radicals also reached 87%, showing obvious concentration dependence.

[0052] In addition, the self-assembled nanoparticles (SAL-CA) reacted with superoxide anions (O2 - ) free radicals also showed good scavenging effect, with a scavenging rate of up to 45% at a concentration of 800 μg / mL. These results fully demonstrate that the self-assembled nanoparticles (SAL-CA) have excellent and rapid free radical scavenging ability.

[0053] In order to further verify the antioxidant properties of self-assembled nanoparticles (SAL-CA), electron spin resonance (ESR) technology was used in this example to obtain the capture of hydroxyl radicals (·OH), DPPH radicals and superoxide anions (O2 - ) ESR spectra of free radical adducts, such as Figure 4 shown.

[0054] like Figure 4 As shown in Figure A, in the presence of SAL-CA, the ESR signal peak of hydroxyl radical (·OH) is significantly reduced, which intuitively reflects the efficient scavenging ability of SAL-CA for ·OH. Figure 4 As shown in B, in the presence of SAL-CA, the ESR spectrum of O2 - The peak intensity of the adduct also weakened, which indicated that O2 - The adducts are effectively consumed. Figure 4 As shown in Figure C, under the action of SAL-CA, the ESR signal peak of DPPH also showed a significant decrease, further confirming the excellent scavenging ability of SAL-CA for DPPH·.

[0055] In summary, the self-assembled nanoparticles (SAL-CA) in this application have an in vitro - , ABTS free radical, DPPH free radical, OH) can exhibit excellent multiple antioxidant properties and can be used for in vitro antioxidant purposes.

[0056] Example 3: Preparation of exosome-encapsulated nanomaterials

[0057] The present embodiment provides an exosome-encapsulated nanomaterial, which is described by taking exosomes derived from mesenchymal stem cells, i.e., mesenchymal stem cell exosomes, as an example:

[0058] S1: Preparation of Exosomes (EVs) from Mesenchymal Stem Cells:

[0059] S11: Human mesenchymal stem cells (hMSCs) were cultured in an appropriate culture medium to promote exosome production. The culture medium was centrifuged at 500 g for 10 minutes to remove cell debris and larger particles. The supernatant was then further purified by centrifugation at 12,000 g for 20 minutes.

[0060] S12: Exosomes were pelleted by ultracentrifugation at 100,000 g for 70 min. The pellet was carefully removed and resuspended in an appropriate buffer. For further purification, the resuspended exosomes were subjected to another ultracentrifugation at 100,000 g for 70 min, and the supernatant was collected to obtain purified exosomes.

[0061] S2: At 4°C, the purified EVs were mixed with the SAL-CA prepared in Example 1 at a ratio of 1:100 and stirred overnight to form mesenchymal stem cell exosome-encapsulated nanomaterials, which are represented as SAL-CAEVs in the following text and figures.

[0062] like Figure 5 As shown, transmission electron microscopy shows the binding of SAL-CA and EVs in the nanomaterial. SAL-CA appears as a relatively round nanoparticle with exosome structures clearly attached to its surface. The particle size of the nanomaterial is about 100-120nm. Figure 5 The effective binding of SAL-CA to EVs was confirmed, laying the foundation for subsequent studies on the biological functions and potential applications of the SAL-CAEVs complex.

[0063] Example 4: Exosome-encapsulated nanomaterials for the treatment of liver injury

[0064] Blank control group: mice were not induced with APAP and were not administered with SAL-CAEVs. Figure 6 As shown in the first column (expressed as APAP- and SAL-CAEVs- in the accompanying drawings).

[0065] Induction control group: mice were induced with APAP and not administered with SAL-CAEVs, e.g. Figure 6 As shown in the second column (indicated as APAP+ and SAL-CAEVs- in the accompanying drawings).

[0066] Drug control group: mice were induced with APAP and administered with SAL-CAEVs, as Figure 6 As shown in the third column (indicated as APAP+ and SAL-CAEVs+ in the accompanying drawings).

[0067] In order to explore the therapeutic effect of SAL-CAEVs on acute liver injury, this example constructed an acetaminophen (APAP)-induced acute liver injury model in mice. ALT and AST are very important biochemical indicators in liver function testing and are mainly used to evaluate liver health and liver function. During the onset of acute liver injury, the levels of ALT and AST in serum will increase significantly. Therefore, the levels of ALT transaminase and AST transaminase in serum were measured to evaluate the degree of liver damage, such as Figure 6 As shown, compared with the blank control group, serum ALT and AST levels in the induced control group mice were significantly elevated, indicating that APAP successfully induced severe liver damage. Furthermore, compared with the induced control group, ALT and AST transaminase levels in the treated control group mice were significantly reduced. This result indicates that SAL-CAEVs can effectively improve APAP-induced transaminase abnormalities in mice, thereby alleviating acute liver injury. Figure 4 Medium, n=6-8; ***p<0.001, ****p<0.0001.

[0068] To further explore the protective effect of SAL-CAEVs against AILI, this example conducted a histopathological evaluation. The liver tissue structure was observed by H&E staining. Figure 7 As shown in A, the liver tissue structure of the blank control group remained intact, with no obvious damage. In contrast, the induced control group showed obvious cell damage, with loose tissue structure, blurred cell outlines, and obvious tissue necrosis areas, which are marked with dotted lines in the figure. The cell outlines in the drug-treated control group were relatively clear, and the area of ​​liver necrosis was significantly reduced. The corresponding liver necrosis areas of the blank control group, induced control group, and drug-treated control group are shown in Figure 1. Figure 7 As shown in B. This indicates that SAL-CAEVs can effectively alleviate liver tissue damage and necrosis.

[0069] The number of apoptotic cells in the liver was quantified by TUNEL staining. Figure 7 As shown in C, the green mark represents TUNEL positive cells. The results showed that no obvious apoptotic cells were found in the blank control group. In contrast, the number of apoptotic cells in the induced control group increased significantly. Compared with the induced control group, the number of apoptotic cells in the liver of mice in the drug control group decreased. The TUNEL content of the blank control group, induced control group and drug control group is shown in Figure 2. Figure 7 As shown in D. This indicates that SAL-CAEVs have an inhibitory effect on APAP-induced cell apoptosis. Figure 7 Middle, n = 3-4; **p < 0.01, ****p < 0.0001, scale bar = 100 μm.

[0070] Based on the above results, SAL-CAEVs showed a significant protective effect in the APAP-induced acute liver injury model, and could effectively reduce cell apoptosis, thereby exerting a therapeutic effect on liver damage.

[0071] To further evaluate the anti-inflammatory ability of SAL-CAEVs in vivo, this example used ELISA to detect inflammatory factors in the serum and liver of mice in the blank control group, induced control group, and drug-treated control group. Figure 8 As shown in the results, after administration of SAL-CAEVs, the abnormal increase of inflammatory factors such as IL-6, TNF-α, MCP-1 and IL-1β in the serum of mice induced by APAP was significantly inhibited. Figure 8 **p<0.01, ***p<0.001, ****p<0.0001. These results demonstrate that SAL-CAEVs can inhibit the inflammatory response in the liver of mice, thereby exerting a hepatoprotective effect.

[0072] Example 5: Exosome-encapsulated nanomaterials are used to promote the formation of specific immune tolerance

[0073] Blank control group: mice were not induced with APAP and were not administered with SAL-CAEVs. Figure 6 As shown in the first column (expressed as APAP- and SAL-CAEVs- in the accompanying drawings).

[0074] Induction control group: mice were induced with APAP and not administered with SAL-CAEVs, e.g. Figure 6 As shown in the second column (indicated as APAP+ and SAL-CAEVs- in the accompanying drawings).

[0075] Drug control group: mice were induced with APAP and administered with SAL-CAEVs, as Figure 6 As shown in the third column (indicated as APAP+ and SAL-CAEVs+ in the accompanying drawings).

[0076] When AILI occurs, the body is able to recognize pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs). The recognition of these molecular patterns triggers a series of inflammatory responses, leading to the recruitment and activation of macrophages in the liver, promoting the M1-M2 conversion of macrophage phenotypes, and thus inducing an inflammatory response. Therefore, this example marks the phenotype of macrophages, using CD86 (a marker for M1 macrophages) and CD206 (a marker for M2 macrophages) for immunofluorescence staining. Figure 9 As shown, bright fluorescence of CD86 and weak fluorescence of CD206 were observed in the induction control group, indicating that macrophages tended to polarize toward the M1 type. In contrast, enhanced CD206 fluorescence signals and decreased CD86 fluorescence signals were observed in the drug-treated control group, confirming that SAL-CAEVs promoted the transition of macrophages from the M1 to the M2 phenotype. These results indicate that SAL-CAEVs can effectively ameliorate inflammatory pathology and improve liver function.

[0077] It should be noted that Figure 9 DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye that strongly binds to DNA and is commonly used in fluorescence microscopy. It can penetrate intact cell membranes and is used to stain both live and fixed cells. DAPI primarily marks the cell nucleus, helping to determine cell morphology and location. It is represented in blue in the image. F4 / 80 is a highly glycosylated G protein-coupled receptor and a marker for mouse macrophages. It is represented in green in the image.

[0078] like Figure 10 The results showed that at the mRNA level, the induced control group significantly increased the expression of Ccl2 and IL-1β compared with the blank control group, while the drug-treated control group decreased the levels of these pro-inflammatory cytokines in the liver.

[0079] Furthermore, the levels of two anti-inflammatory factors, CD206 and Arg-1, were significantly decreased in the induction control group compared to the blank control group. In the drug-treated control group, the expression levels of these anti-inflammatory factors significantly rebounded. These results suggest that SAL-CAEVs can effectively regulate the inflammatory response, inhibiting the expression of pro-inflammatory factors and promoting the production of anti-inflammatory factors, potentially playing a positive role in ameliorating liver inflammation.

[0080] In order to study the immune effect in vivo, the mice in the blank control group, the induced control group and the drug-administered control group were euthanized, spleen cells were extracted, and immune cell analysis was performed using flow cytometry. Figure 11 A- Figure 11 As shown in Figure D, a shift in macrophage polarization from M1 to M2 was observed in the spleens of mice in the control group. Quantitative analysis of M1 and M2 further confirmed this result.

[0081] Comparison of spleen DC cells in each group revealed that the proportion of mature DCs in the drug-treated control group was lower, e.g. Figure 11 E and Figure 11As shown in Figure F, this indicates that SAL-CAEVs can inhibit the maturation of DC cells, keeping them in an immature state of immune tolerance. These immune-tolerant DC cells are called tolerant DC cells (tDCs), and tolerant DC cells can inhibit the excessive immune activation of APAP mice.

[0082] SAL-CA EVs can also reduce T cell activation and proliferation by reducing CD80 and CD86 expression, thereby decreasing T cell co-stimulatory signals, which helps control excessive immune responses, such as Figure 11 G- Figure 11 As shown in H.

[0083] In addition, SAL-CA EVs can also inhibit the expression of Gr1 and NKP46, further regulating the immune response, such as Figure 11 I- Figure 11 As shown in J.

[0084] In summary, SAL-CA EVs can regulate the inflammatory immune response in APAP-treated mice, promote the formation of specific immune tolerance, and thus promote effective specific immune tolerance.

[0085] In the present application, schisandra phenol and chlorogenic acid are self-assembled by ultrasonic mixing to form uniformly dispersed nanoparticles. The self-assembled nanoparticles exhibit excellent multiple antioxidant properties against various free radicals in vitro; the self-assembled nanoparticles are encapsulated by exosomes to form nanomaterials with uniform particle size. The exosome-encapsulated nanomaterials can effectively reduce liver tissue damage and necrosis, and have a significant therapeutic effect on liver damage; at the same time, the exosome-encapsulated nanomaterials can effectively regulate the inflammatory response, and by inhibiting the expression of pro-inflammatory factors and promoting the production of anti-inflammatory factors, they have a positive effect on improving the liver inflammation condition and promoting effective specific immune tolerance.

[0086] It is understandable that the above embodiments only express the preferred implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the patent scope of the present invention. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present invention, the above technical features can be freely combined, and several deformations and improvements can be made, all of which fall within the scope of protection of the present invention. Therefore, all equivalent changes and modifications made to the scope of the claims of the present invention should fall within the scope of coverage of the claims of the present invention.

Claims

1. A self-assembled nanoparticle, characterized in that The invention comprises schisandra phenol and chlorogenic acid, wherein the schisandra phenol and chlorogenic acid are mixed to form nanoparticles.

2. A self-assembled nanoparticle according to claim 1, characterized in that, The mass ratio of schisandra phenol to chlorogenic acid is 1:0.5-1.

5.

3. The self-assembled nanoparticle according to claim 1, characterized in that: The particle size of the self-assembled nanoparticles is 80-90 nm.

4. A method for preparing self-assembled nanoparticles, characterized in that: include: Mixing schisandra phenol and chlorogenic acid, adding them into anhydrous ethanol and ultrasonically dispersing them to obtain a first mixed solution; adding the first mixed solution dropwise into pure water for ultrasonic dispersion to obtain a second mixed solution; The anhydrous ethanol in the second mixed solution is removed to obtain self-assembled nanoparticles.

5. An application of self-assembled nanoparticles, characterized in that: The self-assembled nanoparticles according to any one of claims 1 to 3 are used for in vitro antioxidant effects.

6. An exosome-encapsulated nanomaterial, characterized in that: The method comprises exosomes and self-assembled nanoparticles wrapped by the exosomes, wherein the self-assembled nanoparticles are the self-assembled nanoparticles according to any one of claims 1 to 3.

7. The exosome-encapsulated nanomaterial according to claim 6, characterized in that: The mass ratio of the exosomes to the self-assembled nanoparticles is 1:90-110.

8. The exosome-encapsulated nanomaterial according to claim 6, characterized in that: The particle size of the nanomaterial is 100-120 nm.

9. An application of exosome-encapsulated nanomaterials, characterized in that: The nanomaterial according to any one of claims 6 to 8 is used to treat liver damage.

10. An application of exosome-encapsulated nanomaterials, characterized in that: The nanomaterial according to any one of claims 6 to 8 is used to promote the formation of specific immune tolerance.

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