Anti-inflammatory lipid phoyunbene C nanomaterial, preparation method and application

Abstract

This invention discloses an anti-inflammatory lipid Phoyunbene C nanomaterial, its preparation method, and its applications. The nanomaterial mainly consists of Phoyunbene C nanodot cores, DSPE-PEG2000, soybean lecithin, and cholesterol. In the preparation process, Phoyunbene C is dissolved in phosphate buffer solution, heated, and stirred to obtain an aqueous phase. DSPE-PEG2000, soybean lecithin, and cholesterol are dissolved in ethanol, heated, stirred, and ultrasonically dispersed to obtain an organic phase. The organic and aqueous phase solutions are loaded separately into syringes, and a lipid Phoyunbene C nanodot solution is obtained through a biphase microfluidic mixing process. The preparation method of the lipid Phoyunbene C nanomaterial provided by this invention is simple and the preparation conditions are mild, achieving stronger anti-inflammatory effects both in vitro and in vivo, effectively improving osteoarthritis symptoms.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological materials, and particularly relates to an anti-inflammatory lipid Phoyunbene C nanomaterial, a preparation method and application. BACKGROUND

[0002] Osteoarthritis (OA) is a common chronic degenerative joint disease, which is more common in the elderly population and the incidence significantly increases with age. The disease is characterized by gradual wear and tear of articular cartilage, accompanied by structural changes in surrounding bone tissue (such as osteophyte formation), synovial membrane (such as synovial hyperplasia), and increased inflammation levels (such as the release of pro-inflammatory factors IL-1β and TNF-α). OA can affect multiple joints, with weight-bearing joints such as the knee and hip being the most common, but it can also affect small joints in the hands and joints in the spine. As the disease progresses, patients often experience joint pain (especially after activity), stiffness (morning stiffness usually lasts no more than 30 minutes), limited mobility, and swelling, which severely impairs quality of life.

[0003] Although the exact cause of OA is not fully understood, a variety of factors have been shown to be associated with its risk of onset. These factors include age (decreased cartilage repair capacity), obesity (increased joint mechanical load and inflammation levels), joint injury history (such as ligament tears or fractures), genetic predisposition (such as COL2A1 gene mutations), and long-term overuse of joints (such as occupational or sports-related injuries). In addition, metabolic abnormalities (such as diabetes) and gender (women are more susceptible) are also considered potential risk factors.

[0004] Currently, the treatment of OA is still mainly focused on relieving symptoms, and there is a lack of radical treatment. Conventional treatments include non-drug therapy (such as exercise therapy, weight management) and drug therapy (such as non-steroidal anti-inflammatory drugs, intra-articular injection of hyaluronic acid or glucocorticoids). In recent years, the development of regenerative medicine and biomaterials has provided new ideas for the treatment of OA, such as stem cell therapy, tissue engineered cartilage, and nanomaterial delivery systems. However, these methods still face many challenges, such as the long-term stability of biomaterials, immune rejection, and the controllability of treatment effects. Therefore, the development of an efficient and safe biomaterial to promote cartilage repair, inhibit inflammatory response, and improve joint function has important scientific significance and clinical value for the treatment of OA. SUMMARY

[0005] In order to solve the problems in the background art, in view of the difficulty of continuous anti-inflammation in the joint in the prior art, the purpose of the present application is to provide an anti-inflammatory lipid Phoyunbene C nanodot and a preparation method and application thereof.

[0006] The application adopts the following technical solutions:

[0007] One kind of lipid Phoyunbene C nanomaterial:

[0008] It comprises a lipid spheroid shell and a Phoyunbene C nanoparticle core enclosed inside the lipid spheroid shell.

[0009] The lipid spheroid shell comprises 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000 (DSPE-PEG 2000-NH2), soybean lecithin and cholesterol.

[0010] Soybean lecithin is the main framework of the lipid shell, cholesterol stabilizes the lipid structure, and the introduction of DSPE-PEG 2000-NH2 increases the hydrophilicity.

[0011] The mass ratio relationship of the Phoyunbene C nanoparticle core, DSPE-PEG 2000-NH2, soybean lecithin and cholesterol is 8-12:1-3:30-40:5-10.

[0012] The lipid Phoyunbene C nanomaterial is in the form of a particle solution and can be a preparation, as a kind of solution, injection.

[0013] Two, a preparation method of a lipid Phoyunbene C nanomaterial:

[0014] (1) Preparation of aqueous phase: disperse the original Phoyunbene C monomer in phosphate buffer, vortex and ultrasonic until completely dissolved to obtain a Phoyunbene C solution, then heat the Phoyunbene C solution to 60-70℃, keep warm for 5-8h and stir at the same time to obtain an aqueous solution;

[0015] Here, the monomer refers to the original Phoyunbene C molecule without treatment, and the Phoyunbene C nanoparticle core refers to the original Phoyunbene C molecule that is polymerized and self-assembled into a nanometer-sized nanoparticle when heated.

[0016] (2) Preparation of organic phase: according to the mass ratio relationship, sequentially dissolve the required mass of DSPE-PEG 2000-NH2, soybean lecithin and cholesterol balanced to room temperature in anhydrous ethanol, heat to 40-60℃ and stir for a period of time, then ultrasonic dispersion to obtain an organic phase solution;

[0017] (3) preparing liposome: the water phase solution in step (1) and the organic phase solution in step (2) are loaded into two syringes respectively, then the outlets of the two syringes are communicated with two inlets of a microfluidic chip through pipes, under the control of a microfluidic pressure pump, the organic phase solution and the water phase solution are mixed in the microfluidic chip at a preset flow rate ratio, and the mixed solution is collected at the outlet of the microfluidic chip, so that an original liposome solution is obtained;

[0018] (4) dialysis filtration: the original liposome solution in step (3) is added into a dialysis bag, the dialysis bag is immersed in a dialysate, ethanol is removed, and after sterilization filtration through a filter membrane, a final lipid Phoyunbene C nanomaterial is obtained.

[0019] In the step (1), the concentration of the Phoyunbene C solution is 5-30 mg / ml, the pH of the phosphate buffer is 7.3-7.5, the vortexing time is 0.1-1 min, the ultrasonic power is greater than 20W, the ultrasonic time is 1-10 min, and the stirring rate is greater than 1000 revolutions / min.

[0020] In the step (2), the total concentration of the organic phase solution is 2-20 mg / ml, the stirring rate is greater than 300 revolutions / min, the stirring time is 1-10 min, the ultrasonic power is greater than 20W, and the ultrasonic time is 1-10 min.

[0021] In the step (3), the preset flow rate ratio is the flow rate ratio of the organic phase solution to the water phase solution = 1:1-1:20, the total flow rate of the mixed solution is 5-20 ml / min, and the microfluidic chip is fishbone-shaped or Y-shaped.

[0022] In the step (4), the molecular weight cut-off of the dialysis bag is 1000-10000 Da, the dialysate is a phosphate buffer with a pH of 7.3-7.5, the dialysis time is 48-72 h, and the pore diameter of the filter membrane is mainly 0.22 mu m.

[0023] III. Application of the lipid Phoyunbene C nanomaterial in preparing a drug for treating osteoarthritis. The lipid Phoyunbene C nanodot has an anti-inflammatory effect. The lipid Phoyunbene C nanomaterial is one of a solution and an injection.

[0024] The lipid Phoyunbene C nanomaterial is prepared into a solution or an injection for treating osteoarthritis, and then is injected into the joint cavity.

[0025] Compared with the prior art, the present application has the following advantages:

[0026] 1. The application innovatively designs and uses pi-pi stacking and thermal oxidative polymerization to prepare Phoyunbene C nanodots, which have improved anti-inflammatory efficacy compared to monomeric Phoyunbene C, and form a core-shell structure with the outer lipid.

[0027] 2. The lipid Phoyunbene C nanodots prepared by the application have injectability, cartilage permeability, biocompatibility, biodegradability, and anti-inflammatory effects, which complement each other. The lipid Phoyunbene C nanodot preparation can be directly injected into the joint cavity, and the lipid shell protects the internal core Phoyunbene C nanodots from direct contact with enzymes to avoid uncontrolled degradation, increases the residence time in the joint cavity, suppresses the expression of cartilage cell inflammatory factors, enhances the anti-inflammatory efficacy, reduces cartilage damage, and effectively improves the symptoms of osteoarthritis.

[0028] 3. The lipid Phoyunbene C nanodots are prepared under mild conditions using microfluidic, self-assembly, self-polymerization, and nanocomposite combination processes. The process route is developed according to the actual preparation requirements of the lipid Phoyunbene C nanodots, and the size of the lipid Phoyunbene C nanodots is controllable and has originality.

[0029] 4. The self-designed Phoyunbene C nanodots are combined with multifunctional lipids to precisely, stably, and efficiently treat cartilage in vivo and in vitro, reduce cartilage damage, and inhibit inflammation. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 TEM image of the lipid Phoyunbene C nanodots obtained in Example 1 of the application.

[0031] Figure 2 G1-G7 inflammatory factor protein content chart in Example and Comparative Example of the application.

[0032] Figure 3 TEM image of the lipid Phoyunbene C nanodots obtained in Example 2 of the application.

[0033] Figure 4 TEM image of the lipid Phoyunbene C nanodots obtained in Example 3 of the application.

[0034] Figure 5 TEM image of the lipid Phoyunbene C nanodots obtained in Example 4 of the application.

[0035] Figure 6This is a TEM image of the blank lipid spherical shell without Phoyunbene C nanodots obtained in Comparative Example 1 of this invention. Detailed Implementation

[0036] The present invention will be further described below with reference to the accompanying drawings and embodiments. The specific embodiments described below are used to explain and illustrate the present invention, but should not be construed as limiting the present invention. Any modifications and changes made to the present invention within the spirit and scope of the claims fall within the protection scope of the present invention. All embodiments use reagents of analytical purity or higher.

[0037] The embodiments of the present invention are as follows: Example 1:

[0038] Step S1: Preparation of aqueous phase: Disperse Phoyunbene C monomer in PBS at a concentration of 5 mg / ml, vortex for 1 min, and sonicate at 20 W for 1 min. Heat the solution at 60℃ and stir at 1000 rpm for 5 h to obtain an aqueous phase solution for later use.

[0039] Step S2, Preparation of organic phase: After equilibration to room temperature, DSPE-PEG2000-NH2, soybean lecithin, and cholesterol were dissolved in anhydrous ethanol, heated to 50°C, stirred at 350 r / min for 1 min, and ultrasonically dispersed at 30 W for 1 min to obtain an organic phase solution for later use; wherein, the molar ratio of DSPE-PEG2000-NH2, soybean lecithin, and cholesterol was 1:35:5, and the total lipid concentration was 5 mg / ml;

[0040] Step S3: Load the aqueous solution obtained in step S1 and the organic solution obtained in step S2 into two syringes respectively. Connect the outlets of the two syringes to the two inlets of the microfluidic chip through pipes. At the same time, connect the syringes to the external microfluidic pressure pump. Under the control of the microfluidic pressure pump, mix the organic solution and the aqueous solution in the fishbone-shaped microfluidic chip with an organic phase: aqueous phase flow rate ratio of 1:5 and a total flow rate of 10 ml / min. Collect the original liposome solution at the outlet of the microfluidic chip.

[0041] Step S4: The solution is dialyzed for 48 hours to remove ethanol, the molecular weight cutoff is 1000 Da, and the solution is filtered through a 0.22 μm filter membrane for sterilization to obtain the final lipid Phoyunbene C nanodots.

[0042] The lipid Phoyunbene C nanodots prepared in Example 1 were spherical nanoscale vesicles (e.g., Figure 1As shown), the CCK8 assay showed a 1.05-fold increase in chondrocyte proliferation and a 1.07-fold increase in type II collagen mRNA expression in chondrocytes. In a rat knee osteoarthritis model, 28 days after injection, compared to injections of PBS (G1) and monomeric Phoyunbene C solution (G2), the expression of inflammatory factors in Example 1 (G3) decreased (…). Figure 2 ).

[0043] Implementation: 2:

[0044] Step S1: Preparation of aqueous phase: Disperse Phoyunbene C monomer in PBS (Phoyunbene C concentration 5 mg / ml), vortex for 1 min, and sonicate at 20 W for 1 min. Heat the solution at 60℃ and stir at 1000 rpm for 8 h to obtain an aqueous phase solution for later use.

[0045] Step S2, Preparation of organic phase: After equilibration to room temperature, DSPE-PEG2000-NH2, soybean lecithin, and cholesterol were dissolved in anhydrous ethanol, heated to 50°C, stirred at 350 r / min for 1 min, and ultrasonically dispersed at 30 W for 1 min to obtain an organic phase solution for later use; wherein, the molar ratio of DSPE-PEG2000-NH2, soybean lecithin, and cholesterol was 1:35:5, and the total lipid concentration was 5 mg / ml;

[0046] Step S3: Load the aqueous solution obtained in step S1 and the organic solution obtained in step S2 into two syringes respectively. Connect the outlets of the two syringes to the two inlets of the microfluidic chip through pipes. At the same time, connect the syringes to the external microfluidic pressure pump. Under the control of the microfluidic pressure pump, mix the organic solution and the aqueous solution in the fishbone-shaped microfluidic chip with an organic phase: aqueous phase flow rate ratio of 1:5 and a total flow rate of 10 ml / min. Collect the original liposome solution at the outlet of the microfluidic chip.

[0047] Step S4: The solution is dialyzed for 48 hours to remove ethanol, the molecular weight cutoff is 1000 Da, and the solution is filtered through a 0.22 μm filter membrane for sterilization to obtain the final lipid Phoyunbene C nanodots.

[0048] Compared to Example 1, extending the heating and stirring time of Phoyunbene C resulted in spherical nanoscale vesicles (G4) of the lipid Phoyunbene C nanodots. Figure 3 The CCK8 assay showed a 1.1-fold increase in chondrocyte proliferation and a 1.96-fold increase in type II collagen mRNA expression in chondrocytes. In a rat knee osteoarthritis model, inflammatory factor expression decreased 28 days after injection. Figure 2 ).

[0049] Implementation: 3:

[0050] Step S1: Preparation of aqueous phase: Disperse Phoyunbene C monomer in PBS (Phoyunbene C concentration 10 mg / ml), vortex for 1 min, and sonicate at 20 W for 1 min. Heat the solution at 60℃ and stir at 1000 rpm for 5 h to obtain an aqueous phase solution for later use.

[0051] Step S2, Preparation of organic phase: After equilibration to room temperature, DSPE-PEG2000-NH2, soybean lecithin, and cholesterol were dissolved in anhydrous ethanol, heated to 50°C, stirred at 350 r / min for 1 min, and ultrasonically dispersed at 30 W for 1 min to obtain an organic phase solution for later use; wherein, the molar ratio of DSPE-PEG2000-NH2, soybean lecithin, and cholesterol was 1:35:5, and the total lipid concentration was 5 mg / ml;

[0052] Step S3: Load the aqueous solution obtained in step S1 and the organic solution obtained in step S2 into two syringes respectively. Connect the outlets of the two syringes to the two inlets of the microfluidic chip through pipes. At the same time, connect the syringes to the external microfluidic pressure pump. Under the control of the microfluidic pressure pump, mix the organic solution and the aqueous solution in the fishbone-shaped microfluidic chip with an organic phase: aqueous phase flow rate ratio of 1:5 and a total flow rate of 10 ml / min. Collect the original liposome solution at the outlet of the microfluidic chip.

[0053] Step S4: The solution is dialyzed for 48 hours to remove ethanol, the molecular weight cutoff is 1000 Da, and the solution is filtered through a 0.22 μm filter membrane for sterilization to obtain the final lipid Phoyunbene C nanodots.

[0054] Compared to Example 1, the concentration of Phoyunbene C was increased, and the liposome (G5) was a spherical nanoscale vesicle. Figure 4 The CCK8 assay showed a 1.05-fold increase in chondrocyte proliferation and a 1.08-fold increase in type II collagen mRNA expression in chondrocytes. In a rat knee osteoarthritis model, inflammatory factor expression decreased 28 days after injection. Figure 2 ).

[0055] Implementation 4:

[0056] Step S1: Preparation of aqueous phase: Disperse Phoyunbene C monomer in PBS at a concentration of 5 mg / ml, vortex for 1 min, and sonicate at 20 W for 1 min. Heat the solution at 60℃ and stir at 1000 rpm for 5 h to obtain an aqueous phase solution for later use.

[0057] Step S2, Preparation of organic phase: After equilibration to room temperature, DSPE-PEG2000-NH2, soybean lecithin, and cholesterol were dissolved in anhydrous ethanol, heated to 50°C, stirred at 350 r / min for 1 min, and ultrasonically dispersed at 30 W for 1 min to obtain an organic phase solution for later use; wherein, the molar ratio of DSPE-PEG2000-NH2, soybean lecithin, and cholesterol was 1:35:5, and the total lipid concentration was 5 mg / ml;

[0058] Step S3: Load the aqueous solution obtained in step S1 and the organic solution obtained in step S2 into two syringes respectively. Connect the outlets of the two syringes to the two inlets of the microfluidic chip through pipes. At the same time, connect the syringes to an external microfluidic pressure pump. Under the control of the microfluidic pressure pump, mix the organic solution and the aqueous solution in the fishbone-shaped microfluidic chip at an organic phase: aqueous phase flow rate ratio of 1:5 and a total flow rate of 5 ml / min. Collect the original liposome solution at the outlet of the microfluidic chip.

[0059] Step S4: The solution is dialyzed for 48 hours to remove ethanol, the molecular weight cutoff is 1000 Da, and the solution is filtered through a 0.22 μm filter membrane for sterilization to obtain the final lipid Phoyunbene C nanodots.

[0060] Compared to Example 1, the total flow rate of lipids during preparation was reduced, and the liposomes (G6) were spherical nanoscale vesicles. Figure 5 The CCK8 assay showed a 1.06-fold increase in chondrocyte proliferation and a 1.22-fold increase in type II collagen mRNA expression in chondrocytes. In a rat knee osteoarthritis model, inflammatory factor expression decreased 28 days after injection. Figure 2 ).

[0061] Comparative Example 1: Only lipid globular shell

[0062] Step S1: Prepare the aqueous phase: Take PBS as the aqueous phase solution for later use;

[0063] Step S2, Preparation of organic phase: After equilibration to room temperature, DSPE-PEG2000-NH2, soybean lecithin, and cholesterol were dissolved in anhydrous ethanol, heated to 50°C, stirred at 350 r / min for 1 min, and ultrasonically dispersed at 30 W for 1 min to obtain an organic phase solution for later use; wherein, the molar ratio of DSPE-PEG2000-NH2, soybean lecithin, and cholesterol was 1:35:5, and the total lipid concentration was 5 mg / ml;

[0064] Step S3: Load the aqueous solution from step S1 and the organic solution from step S2 into two syringes respectively. Connect the outlets of the two syringes to the two inlets of the microfluidic chip through pipes. Simultaneously, connect the syringes to an external microfluidic pressure pump. Under the control of the microfluidic pressure pump, mix the organic and aqueous solutions in the fishbone-shaped microfluidic chip at an organic phase:water flow rate ratio of 1:5 and a total flow rate of 10 ml / min. Collect the original liposome solution at the outlet of the microfluidic chip.

[0065] Step S4: The solution is dialyzed for 48 hours to remove ethanol, with a molecular weight cutoff of 1000 Da. After sterilization by filtration through a 0.22 μm filter membrane, blank lipid spherical shells without Phoyunbene C nanodots are obtained.

[0066] The blank lipid spherical shells (G7) without Phoyunbene C nanodots prepared in Comparative Example 1 were spherical nanoscale vesicles. Figure 6 After 28 days of injection into the rat knee osteoarthritis model, there was no significant difference in the expression of inflammatory factors between Comparative Example 1 and Example 1. Figure 2 ).

[0067] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A lipid Phoyunbene C nanomaterial, characterized in that: The lipid globular shell and the Phoyunbene C nanoparticle core enclosed in the lipid globular shell; The lipid globular shell comprises 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000 (DSPE-PEG2000-NH2), soybean lecithin and cholesterol; The mass ratio of the Phoyunbene C nanoparticle core, DSPE-PEG2000-NH2, soybean lecithin and cholesterol is 8-12:1-3:30-40:5-10; The lipid Phoyunbene C nanomaterial is prepared by the following method: (1) Preparation of an aqueous phase: the original Phoyunbene C is dispersed in a phosphate buffer, vortexed and ultrasonicated until dissolved to obtain a Phoyunbene C solution, and then the Phoyunbene C solution is heated to 60-70°C, incubated for 5-8 hours and stirred at the same time to obtain an aqueous phase solution; (2) Preparation of an organic phase: according to the mass ratio, the required mass of DSPE-PEG2000-NH2, soybean lecithin and cholesterol is sequentially dissolved in anhydrous ethanol, heated to 40-60°C and stirred for a period of time, and then ultrasonically dispersed to obtain an organic phase solution; (3) Preparation of liposomes: the aqueous phase solution in step (1) and the organic phase solution in step (2) are loaded into two syringes respectively, and then the outlets of the two syringes are connected to two inlets of a microfluidic chip through pipelines, under the control of a microfluidic pressure pump, the organic phase solution and the aqueous phase solution are mixed in the microfluidic chip at a preset flow rate ratio, and the mixed solution is collected at the outlet of the microfluidic chip to obtain a raw liposome solution; (4) Dialysis filtration: the raw liposome solution in step (3) is added to a dialysis bag, the dialysis bag is immersed in a dialysis solution, and after sterilization by membrane filtration, a lipid Phoyunbene C nanomaterial is obtained.

2. The preparation method of the lipid Phoyunbene C nanomaterial of claim 1, characterized by: The preparation steps are as follows: (1) Preparation of an aqueous phase: the original Phoyunbene C is dispersed in a phosphate buffer, vortexed and ultrasonicated until dissolved to obtain a Phoyunbene C solution, and then the Phoyunbene C solution is heated to 60-70°C, incubated for 5-8 hours and stirred at the same time to obtain an aqueous phase solution; (2) Preparation of an organic phase: according to the mass ratio, the required mass of DSPE-PEG2000-NH2, soybean lecithin and cholesterol is sequentially dissolved in anhydrous ethanol, heated to 40-60°C and stirred for a period of time, and then ultrasonically dispersed to obtain an organic phase solution; (3) Liposome preparation: the aqueous phase solution in step (1) and the organic phase solution in step (2) are loaded into two syringes respectively, then the outlets of the two syringes are connected to two inlets of a microfluidic chip through pipes, under the control of a microfluidic pressure pump, the organic phase solution and the aqueous phase solution are mixed in the microfluidic chip at a preset flow rate ratio, and the mixed solution is collected at the outlet of the microfluidic chip to obtain a raw liposome solution; (4) Dialysis filtration: the raw liposome solution in step (3) is added to a dialysis bag, the dialysis bag is immersed in a dialysate, and after sterilization by filtration membrane, a lipid Phoyunbene C nanomaterial is obtained.

3. The method for preparing the lipid Phoyunbene C nanomaterial according to claim 2, characterized in that: In step (1), the concentration of the Phoyunbene C solution is 5-30 mg / ml, the pH of the phosphate buffer is 7.3-7.5, the vortexing time is 0.1-1 min, the ultrasonic power is greater than 20W, the ultrasonic time is 1-10 min, and the stirring rate is greater than 1000 rpm.

4. The method for preparing the lipid Phoyunbene C nanomaterial according to claim 2, characterized in that: In step (2), the total concentration of the organic phase solution is 2-20 mg / ml, the stirring rate is greater than 300 rpm, the stirring time is 1-10 min, the ultrasonic power is greater than 20W, and the ultrasonic time is 1-10 min.

5. The method for preparing lipid Phoyunbene C nanomaterials according to claim 2, characterized in that: In step (3), the preset flow rate ratio is organic phase solution flow rate ratio:aqueous phase solution flow rate ratio = 1:1-1:20, the total flow rate of the mixed solution is 5-20 ml / min, and the microfluidic chip is fishbone-shaped or Y-shaped.

6. The method for preparing lipid Phoyunbene C nanomaterials according to claim 2, characterized in that: In step (4), the molecular weight cut-off of the dialysis bag is 1000-10000 Da, the dialysate is a phosphate buffer with a pH of 7.3-7.5, the dialysis time is 48-72 h, and the pore diameter of the filter membrane is mainly 0.22 μm.

7. Use of the lipid Phoyunbene C nanomaterial of claim 1 or the lipid Phoyunbene C nanomaterial prepared by the method of any one of claims 2-6 in the preparation of a drug for treating osteoarthritis.

8. Use according to claim 7, characterized in that: The lipid Phoyunbene C nanomaterial is prepared into a solution for treating osteoarthritis.

9. Use according to claim 7, characterized in that: The lipid Phoyunbene C nanomaterial is prepared into an injection for treating osteoarthritis.

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