A method for preparing a cartilage repair material, and products and uses thereof

Abstract

The application relates to a preparation method of a cartilage repair material, a product and application thereof, in particular to a preparation method of a hydrogel for repairing a cartilage microenvironment. The application has the advantages that calcium phosphate particles are induced to deposit to form an organic-inorganic composite structure through a three-dimensional structure of an organic hydrogel, and the rigidity of the hydrogel is improved. MgH2@ZIF-8 NPs contained in the hydrogel can release hydrogen under an acidic pH condition, the inflammatory microenvironment of cartilage damage is adjusted, and the effect of immunorepair is achieved.

Description

Technical Field

[0001] This invention relates to a method for preparing a cartilage repair material, its products, and applications, specifically to a method for preparing a hydrogel for cartilage microenvironment repair. Background Technology

[0002] The quality of bone / cartilage injuries is a major challenge in orthopedics, often affecting articular cartilage and subchondral bone. Articular cartilage has a unique structure, making it difficult to repair after injury, leading to severe joint dysfunction. Hydrogels possess a three-dimensional network structure similar to the extracellular matrix of bone / cartilage cells. They can carry bioactive molecules to regulate the immune response in cartilage injuries, creating a microenvironment conducive to tissue regeneration and repairing the damaged area. However, the relatively weak mechanical strength of hydrogels severely limits their application in the biomedical field. Summary of the Invention

[0003] The purpose of this invention is to provide a method for preparing a cartilage repair material, a cartilage repair material with high mechanical strength and immune microenvironment repair, and the method for preparing the same.

[0004] Another object of the present invention is to provide a cartilage repair material product prepared by the above method.

[0005] Another object of the present invention is to provide an application of the above-mentioned product.

[0006] The objective of this invention is achieved through the following scheme: a method for preparing a cartilage repair material, comprising preparing metal-organic framework-coated magnesium hydride nanoparticles (MgH2@ZIF-8 NPs), followed by using potassium persulfate (PPS) to initiate the polymerization of acrylic acid (AA), N,N,-methylenebisacryloyl (MBA), α-tricalcium phosphate (α-TCP) and MgH2@ZIF-8, depositing calcium phosphate on a hydrogel network to form a brick-cement-like structure, thereby improving the mechanical properties of the hydrogel, including the following steps:

[0007] (1) 20 mL of polyvinylpyrrolidone (PVP) modified MgH2NPs were thoroughly mixed with 40 mL of 0.1~0.2% (m / v) 2-MM methanol solution, and then 40 mL of 0.2~0.3% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1~3 hours, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100 ℃ for 1 hour to obtain MgH2@ZIF-8 NPs;

[0008] (2) Under nitrogen (N2) conditions, 20-50% (w / v) AA is thoroughly mixed with sodium hydroxide solution (NaOH); a small amount of citric acid is thoroughly mixed with 5-20% (w / v) sodium carboxymethyl cellulose, and then MgH2@ZIF-8 NPs and α-TCP are thoroughly mixed.

[0009] (3) After the two solutions are mixed evenly, add 0.3~0.6% (w / v) MBA and PPS dropwise. After mixing thoroughly, remove all air bubbles under vacuum and react at 60~70℃ for 2~4 hours. Demold the polymerized gel, remove unreacted material, and dry.

[0010] The amount of MgH2@ZIF-8 NPs used in this invention is 1~5% (w / v).

[0011] The amount of α-TCP used in this invention is 20~50 (w / v).

[0012] This invention provides a cartilage repair material, prepared according to any of the methods described above.

[0013] This invention provides the application of a cartilage repair material in the preparation of materials required for cartilage repair materials.

[0014] The advantages of this invention are:

[0015] (1) The present invention improves the stiffness of hydrogel by inducing the deposition of calcium phosphate particles in the three-dimensional structure of organic hydrogel to form an organic-inorganic composite structure.

[0016] (2) The MgH2@ZIF-8 NPs contained in this invention can release hydrogen gas under acidic pH conditions, regulate the inflammatory microenvironment of cartilage damage, and achieve the effect of immune repair. Attached Figure Description

[0017] Appendix Figure 1 This is a TEM image of the MgH2@ZIF-8 NPs prepared in Example 1;

[0018] Appendix Figure 2 These are cell viability diagrams of the hydrogels prepared in Examples 1-3.

[0019] Depend on Figure 2 It can be seen that, compared with the control group, the hydrogels prepared in Examples 1-3 of this application have good biocompatibility. Detailed Implementation

[0020] The technical solution of the present invention will be further described below through specific embodiments. These embodiments are further illustrations of the present invention and do not limit the scope of the invention. Example 1

[0021] A cartilage repair material was prepared by fabricating magnesium hydride nanoparticles (MgH2@ZIF-8NPs) coated with a metal-organic framework, followed by polymerization of acrylic acid (AA), N,N,-methylenebisacryloyl (MBA), α-tricalcium phosphate (α-TCP), and MgH2@ZIF-8 initiated by potassium persulfate (PPS); calcium phosphate was then deposited on the hydrogel network to form a brick-cement-like structure. The preparation was carried out according to the following steps:

[0022] (1) 20 mL of polyvinylpyrrolidone (PVP) modified MgH2NPs were thoroughly mixed with 40 mL of 0.1~0.2% (m / v) 2-MM methanol solution, and then 40 mL of 0.2~0.3% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hour, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100℃ for 1 hour to obtain MgH2@ZIF-8 NPs; Figure 1 This is a TEM image of the prepared MgH2@ZIF-8 NPs. The image shows that the size of the MgH2@ZIF-8 NPs is 500 nm.

[0023] (2) Under N2 conditions, 30% (w / v) acrylic acid AA was thoroughly mixed with NaOH solution; a small amount of citric acid was thoroughly mixed with 10% (w / v) sodium carboxymethyl cellulose, and then 2% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP were added to the solution and thoroughly mixed.

[0024] (3) After the two solutions are mixed evenly, 0.3~0.6% (w / v) MBA and PPS are added drop by drop. After mixing thoroughly, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain cartilage repair material. Example 2

[0025] A cartilage repair material, prepared according to steps similar to those in Example 1, is as follows:

[0026] (1) 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs were thoroughly mixed with 40 mL of 0.15% (m / v) 2-MM methanol solution, and then 40 mL of 0.2% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hour, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100 °C for 1 hour to obtain MgH2@ZIF-8 NPs;

[0027] (2) Under N2 conditions, 25% (w / v) AA and Na OH solution were thoroughly mixed; a small amount of citric acid and 8% (w / v) sodium carboxymethyl cellulose were thoroughly mixed with 3% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP and thoroughly mixed.

[0028] (3) After the two solutions are mixed evenly, 0.3% (w / v) MBA and PPS are added dropwise. After thorough mixing, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain the cartilage repair material. Example 3

[0029] A cartilage repair material, prepared according to steps similar to those in Example 1, is as follows:

[0030] (1) 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs were thoroughly mixed with 40 mL of 0.1% (m / v) 2-MM methanol solution, and then 40 mL of 0.2% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hr, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100 ℃ for 1 hr to obtain MgH2@ZIF-8 NPs;

[0031] (2) Under N2 conditions, 30% (w / v) AA and Na OH solution were thoroughly mixed; a small amount of citric acid and 8% (w / v) sodium carboxymethyl cellulose were thoroughly mixed with 5% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP and thoroughly mixed.

[0032] (3) After the two solutions are mixed evenly, 0.4% (w / v) MBA and PPS are added dropwise. After thorough mixing, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain the cartilage repair material.

[0033] Performance testing

[0034] The hydrogels prepared in Examples 1-3 were made into small discs with a diameter of 0.3 mm and laid flat on the bottom of a 96-well plate.

[0035] Subsequently, 100 µL of culture medium (10% fetal bovine serum) containing 3000 rat articular chondrocytes was inoculated into each well of a 96-well plate and cultured for 24 hours in a cell culture incubator (37°C, 5% carbon dioxide (CO2) concentration, 95% relative humidity). The culture medium was discarded, and residual culture medium was thoroughly washed away with PBS. 100 µL of culture medium containing the hydrogel extract prepared in Examples 1-3 was added to each well and cultured for 24 hours. The culture medium was discarded, and residual culture medium was thoroughly washed away with PBS. 10% CCK-8 serum-free DMEM medium was added to each well and cultured at room temperature for 2 hours. The absorbance of each well at 450 nm was measured using a microplate reader. Figure 2 The images show the cell viability of the hydrogels prepared in Examples 1-3. Figure 2 It can be seen that, compared with the control group, the hydrogel cartilage repair materials prepared in Examples 1-3 of the present invention have good biocompatibility.

Claims

1. A method for preparing a cartilage repair material, characterized in that, Metal-organic framework-coated magnesium hydride nanoparticles MgH2@ZIF-8 NPs were prepared, followed by polymerization of AA acrylic acid, N,N,-methylenebisacryloyl MBA, α-TCP, and MgH2@ZIF-8 initiated by potassium persulfate PPS; calcium phosphate was deposited on the hydrogel network to form a brick-cement-like structure, including the following steps: (1) After thoroughly mixing 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs with 40 mL of 0.1~0.2% (m / v) 2-MM methanol solution, add 40 mL of 0.2~0.3% (m / v) zinc acetate methanol solution, stir mechanically for 1~3 hours, centrifuge and wash three times with methanol, and dry in an oven at 100℃ for 1 hour to obtain MgH2@ZIF-8 NPs; (2) Under nitrogen (N2) conditions, 20-50% (w / v) acrylic acid (AA) is thoroughly mixed with NaOH solution; a small amount of citric acid is thoroughly mixed with 5-20% (w / v) sodium carboxymethyl cellulose, and then MgH2@ZIF-8 NPs and α-TCP are added and thoroughly mixed. (3) After the two solutions are mixed evenly, 0.3~0.6% (w / v) MBA and PPS are added dropwise. After thorough mixing, all air bubbles are removed by vacuum. The mixture is reacted at 60~70℃ for 2~4 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain the cartilage repair material; wherein, The amount of α-TCP used is 20~50 (w / v).

2. The method for preparing the cartilage repair material according to claim 1, characterized in that... The amount of MgH2@ZIF-8 NPs used is 1~5% (w / v).

3. The method for preparing the cartilage repair material according to any one of claims 1 to 2, characterized in that, Prepare according to the following steps: (1) After thoroughly mixing 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs with 40 mL of 0.1~0.2% (m / v) 2-MM methanol solution, 40 mL of 0.2~0.3% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hour, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100℃ for 1 hour to obtain MgH2@ZIF-8 NPs with a size of 500 nm. (2) Under N2 conditions, 30% (w / v) acrylic acid AA was thoroughly mixed with NaOH solution; a small amount of citric acid was thoroughly mixed with 10% (w / v) sodium carboxymethyl cellulose, and then 2% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP were added to the solution and thoroughly mixed. (3) After the two solutions are mixed evenly, 0.3~0.6% (w / v) MBA and PPS are added drop by drop. After mixing thoroughly, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain cartilage repair material.

4. The method for preparing the cartilage repair material according to any one of claims 1 to 2, characterized in that, Prepare according to the following steps: (1) After 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs were thoroughly mixed with 40 mL of 0.15% (m / v) 2-MM methanol solution, 40 mL of 0.2% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hr, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100 ℃ for 1 hr to obtain MgH2@ZIF-8 NPs. (2) Under N2 conditions, 25% (w / v) AA and Na OH solution were thoroughly mixed; a small amount of citric acid and 8% (w / v) sodium carboxymethyl cellulose were thoroughly mixed with 3% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP and thoroughly mixed. (3) After the two solutions are mixed evenly, 0.3% (w / v) MBA and PPS are added dropwise. After mixing thoroughly, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and the gel is dried to obtain cartilage repair material.

5. The method for preparing the cartilage repair material according to any one of claims 1 to 2, characterized in that, Prepare according to the following steps: (1) 20 mL of polyvinylpyrrolidone (PVP) modified MgH2 NPs were thoroughly mixed with 40 mL of 0.1% (m / v) 2-MM methanol solution, and then 40 mL of 0.2% (m / v) zinc acetate methanol solution was added. After mechanical stirring for 1 hr, the mixture was washed three times by methanol centrifugation and then dried in an oven at 100 ℃ for 1 hr to obtain MgH2@ZIF-8 NPs; (2) Under N2 conditions, 30% (w / v) AA and Na OH solution were thoroughly mixed; a small amount of citric acid and 8% (w / v) sodium carboxymethyl cellulose were thoroughly mixed with 5% (w / v) MgH2@ZIF-8 NPs and 20 (w / v) α-TCP and thoroughly mixed. (3) After the two solutions are mixed evenly, 0.4% (w / v) MBA and PPS are added drop by drop. After mixing thoroughly, all air bubbles are removed by vacuum and the mixture is reacted at 65°C for 3 hours. The polymerized gel is demolded, unreacted material is removed, and dried to obtain cartilage repair material.

6. A cartilage repair material, characterized in that... Prepared according to any one of claims 1-5.

7. The application of the cartilage repair material according to claim 6 in the materials required for preparing the cartilage repair material.

Images