A method for preparing and applying a polyglutamic acid and chitosan hydrogel matrix

Abstract

The present invention relates to a preparation method and application of a polyglutamic acid and chitosan hydrogel base material, belonging to the technical field of biopolymer materials. It mainly includes the preparation of polyglutamic acid and chitosan hydrogel base materials and the application after modification. This scheme prepares reaction solution 1 and reaction solution 2, changes the mechanical strength of the hydrogel base material by adjusting the concentration of polyglutamic acid and carboxymethyl chitosan, adds various active substances during the preparation process to meet the needs of different application scenarios, and also modifies the polyglutamic acid and chitosan hydrogel base materials so that they can better absorb and combine other components, thereby further solidifying the hydrogel and improving the mechanical strength of the hydrogel.

Description

Technical Field

[0001] The invention belongs to the technical field of biopolymer materials and relates to a preparation method and application of a polyglutamic acid and chitosan hydrogel base material. Background Art

[0002] Polyglutamic acid is a linear polymer formed by glutamic acid connected by γ-amide bonds. It is highly hydrophilic and biodegradable and is widely used in moisturizing, drug carriers and tissue engineering. However, the current polyglutamic acid hydrogel still has shortcomings in preparation. At present, ethers, aldehydes and other toxic cross-linking agents are commonly used to prepare chemically cross-linked γ-PGA hydrogels. Even if γ-PGA hydrogels can be successfully prepared, the remaining toxic and harmful reagents or toxic by-products make the various advantages of γ-PGA hydrogels dim. In addition, insufficient removal of impurities during the preparation process or unstable structure of the hydrogel can easily lead to differences in the adsorption amount and sustained release performance of active substances. These problems need to be solved. Summary of the invention

[0003] The object of the present invention is to provide a preparation method and application of a polyglutamic acid and chitosan hydrogel base material. In this scheme, reaction liquid 1 and reaction liquid 2 are prepared, the mechanical strength of the hydrogel base material is changed by adjusting the concentrations of polyglutamic acid and carboxymethyl chitosan, and various active substances are added during the preparation process to meet the needs of different application scenarios. In addition, the polyglutamic acid and chitosan hydrogel base materials are modified so that they can better adsorb and combine with other components, thereby further solidifying the hydrogel and improving the mechanical strength of the hydrogel.

[0004] The purpose of the present invention can be achieved through the following technical solutions:

[0005] A method for preparing a polyglutamic acid and chitosan hydrogel base material, the preparation method comprising the following steps:

[0006] (1) Preparation of reaction solution 1: polyglutamic acid is added to a reaction tank, a solvent is introduced, and the pH is adjusted to obtain a polyglutamic acid solution. The motor is started for stirring, and cooling water is introduced for cooling. N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are then added to the reaction tank, and the reaction solution is obtained after stirring for 20-40 minutes. Pure water is added for dilution to obtain a diluted solution, and a liquid pump is started and a valve is opened to allow the diluted solution to enter a hollow fiber ultrafiltration column for ultrafiltration until the volume of the ultrafiltrate in the reaction tank is close to the volume of the reaction solution. The conductivity value is read from a conductivity meter to determine whether it meets the standard. If it does not meet the standard, pure water is continued to be added for dilution and ultrafiltration. If it meets the standard, reaction solution 1 is obtained.

[0007] (2) Preparation of reaction solution 2: Carboxymethyl chitosan is placed in a reaction tank, a solvent is introduced, the pH is adjusted, and a motor is started for stirring to obtain reaction solution 2 after the mixture is fully dissolved;

[0008] (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, start the liquid pump, open the valve, and introduce reaction liquid 1 and reaction liquid 2 respectively, start the motor for stirring, open the valve to release the mixed liquid, and let it stand for 8-12 minutes to obtain polyglutamic acid and chitosan hydrogel base materials.

[0009] Furthermore, the solvent in step (1) is pure water or pure water and an active substance with a polyglutamic acid concentration of 0.01-0.09wt%; the pH adjustment refers to adjusting the pH to 4-6 using 0.8-1.2% hydrochloric acid solution and 0.8-1.2% sodium hydroxide solution; the mass ratio of the polyglutamic acid, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 9-11:1:1.

[0010] Furthermore, when pure water is used for dilution, the amount of pure water used is 3-5 times the volume of the reaction solution.

[0011] Furthermore, the molecular weight cut-off of the hollow fiber ultrafiltration column is 10,000-200,000.

[0012] Furthermore, the conductivity value in step (1) meets the standard value of ≦20μs / cm.

[0013] Furthermore, the final volume of the reaction solution 1 is equal to the volume of the polyglutamic acid solution. If it is not reached, pure water can be added for adjustment. During the preparation of the reaction solution 1, the temperature of the reaction solution 1 is kept at 10-20° C. by controlling the cooling water in the jacket.

[0014] Furthermore, the solvent in step (2) is pure water or pure water and an active substance with a carboxymethyl chitosan concentration of 40-60wt%; the pH adjustment refers to adjusting the pH to 7-8 using 0.8-1.2% sodium hydroxide solution; and the concentration of carboxymethyl chitosan in the reaction solution 2 is 1-10g / L.

[0015] Furthermore, the active substances in step (1) and step (2) are selected from one or more of restorative materials and allergy-relieving substances.

[0016] Furthermore, after the preparation of the polyglutamic acid and chitosan hydrogel base material, the device needs to be cleaned, and the cleaning operation more specifically includes the following steps:

[0017] After the preparation of polyglutamic acid and chitosan hydrogel base materials is completed, washing water is introduced into the water inlet pipe 42, and flushing is carried out after the valve is opened. After flushing, the waste water is collected in the waste liquid pipe 35 and discharged. The hollow fiber ultrafiltration column is cleaned by water entering through valve No. 20 and valve No. 4, and then discharged through valve No. 13.

[0018] Furthermore, the preparation method obtains polyglutamic acid and chitosan hydrogel base materials.

[0019] Furthermore, the polyglutamic acid and chitosan hydrogel base materials are applied to repair and soothing products after being modified.

[0020] Furthermore, the modification method is:

[0021] The polyglutamic acid and chitosan hydrogel base materials are subjected to plasma treatment to obtain a treated base material, the treated base material is immersed in a sodium alginate solution, stirred for 10-20 minutes, hydroxyapatite and catalase are added, and stirred at 36-38°C for 30-40 minutes. The obtained product is immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

[0022] Furthermore, the parameters of the plasma treatment are: treatment power is 50-250W, and treatment time is 3-7min.

[0023] Furthermore, the mass ratio of the polyglutamic acid and chitosan hydrogel base material, the sodium alginate solution, the hydroxyapatite and the catalase is 1:1.2-1.5:0.1-0.2:0.005-0.01.

[0024] Furthermore, the raw materials of the sodium alginate solution include, by percentage, 10-20wt% sodium alginate, 0.2-2wt% vitamin C solution and the balance pure water; the vitamin C solution consists of 1-5wt% vitamin C and the balance pure water; the particle diameter of the hydroxyapatite is 60-100nm.

[0025] Beneficial effects of the present invention:

[0026] (1) This solution prepares reaction solution 1 and reaction solution 2, and changes the mechanical strength of the hydrogel base material by adjusting the concentrations of polyglutamic acid and carboxymethyl chitosan to meet the needs of different application scenarios. In addition, various active substances, such as repair and soothing substances, can be added during the preparation process. The present invention provides a cooling water cooling treatment during the preparation process. For some active substances that are sensitive to temperature, the functionality of these active substances can be protected and maintained, thereby preparing hydrogels with different functional effects.

[0027] (2) The present invention modifies the polyglutamic acid and chitosan hydrogel base materials to enhance their biocompatibility, improve the hydrophilicity and adhesion of the hydrogel, enable it to better adsorb and combine with other components, thereby further solidifying the hydrogel and improving the mechanical strength of the hydrogel. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to facilitate understanding by those skilled in the art, the present invention is further described below with reference to the accompanying drawings.

[0029] Figure 1 A schematic diagram of a device for preparing a polyglutamic acid and chitosan hydrogel base material of the present invention;

[0030] Figure 2 Schematic diagram of the preparation of reaction solution 1 and reaction solution 2 of the present invention;

[0031] Figure 3 This is a schematic diagram of cooling water during the preparation of the reaction solution 1 of the present invention;

[0032] Figure 4 Schematic diagram of mixing reaction solution 1 and reaction solution 2 of the present invention;

[0033] Figure 5 A schematic diagram of cleaning the device for preparing the polyglutamic acid and chitosan hydrogel base material of the present invention;

[0034] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0035] 1. Reactor No. 1; 2. Stirring No. 1; 3. Sight glass No. 1; 4. Conductivity meter; 5. pH meter No. 1; 6. Thermometer; 7. Valve No. 1; 8. Feeding port No. 1; 9. Stirring motor; 10. Valve No. 2; 11. Valve No. 3; 12. Valve No. 4; 13. Hollow fiber ultrafiltration column; 14. Valve No. 5; 15. Valve No. 6; 16. Valve No. 7; 17. Liquid pump No. 1; 18. Valve No. 8; 19. Valve No. 9; 20. Valve No. 10; 21. Valve No. 11; 22. Valve No. 12; 23. Valve No. 13; 2 4. Liquid flow meter No. 1; 25. Motor No. 1; 26. Liquid mixer; 27. Valve No. 14; 28. Valve No. 15; 29. ​​Stirring No. 2; 30. Liquid flow meter No. 2; 31. Valve No. 16; 32. Liquid pump No. 2; 33. Valve No. 17; 34. Valve No. 18; 35. Waste liquid pipe; 36. Sight glass No. 2; 37. pH meter No. 2; 38. Reactor No. 2; 39. Feeding port No. 2; 40. Motor No. 2; 41. Valve No. 19; 42. Water inlet pipe; 43. Jacket; 44. Valve No. 20. DETAILED DESCRIPTION

[0036] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in conjunction with the accompanying drawings and preferred embodiments.

[0037] The polyglutamic acid in all the embodiments and comparative examples of the present invention is purchased directly from the market, and is purchased from Shandong Freda Biotechnology Co., Ltd., with a CAS number of 25513-46-6; N-hydroxysuccinimide is purchased directly from the market, and is purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., with a CAS number of 6066-82-6; 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is purchased directly from the market, and is purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., with a CAS number of 22572-40-3; carboxymethyl chitosan is purchased directly from the market, and is purchased from Shaanxi Baichuan Biotechnology Co., Ltd., CAS No. 83512-85-0; sodium alginate was purchased directly from the market, all purchased from Guangzhou Huayu Biotechnology Co., Ltd.; hydroxyapatite was purchased directly from the market, all purchased from Hebei Qiansheng Biotechnology Co., Ltd.; peroxidase was purchased directly from the market, all purchased from Hebei Qiansheng Biotechnology Co., Ltd., CAS No. 9003-99-0; vitamin C was purchased directly from the market, all purchased from Jiangsu Yuanshengtong Bioengineering Co., Ltd.; human epidermal growth factor was purchased directly from the market, all purchased from Guangzhou Jinyinyuan Sino-US Biotechnology Group Co., Ltd., model OE95 hEGF; dipotassium glycyrrhizinate was purchased directly from the market, all purchased from Guangdong Mingtong Biotechnology Co., Ltd.

[0038] Example 1

[0039] A method for preparing a polyglutamic acid and chitosan hydrogel base material, the preparation method of this embodiment comprises the following steps:

[0040] (1) In reaction tank No. 1, polyglutamic acid is added from feed port No. 1, and pure water is introduced into reaction tank No. 1 through water inlet pipe 42, valve No. 2, and valve No. 3. The pH value is adjusted to 5.6 using 1% hydrochloric acid solution and 1% sodium hydroxide solution to obtain a polyglutamic acid solution. The volume of the polyglutamic acid solution is V1 as read from the scale of sight glass No. 1, and stirring motor 9 is started to drive stirring motor No. 1 to rotate so that the polyglutamic acid is evenly dissolved. At this time, the jacket 43 is supplied with water through valve No. 6. Cooling water was passed into the reaction mixture to cool it down. After cooling down, the cooling water flowed out from valve No. 1. N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride were added into the feeding port No. 1, wherein the mass ratio of polyglutamic acid, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride was 10:1:1. After stirring for 30 minutes, a reaction solution was obtained. Pure water 3 times the volume of the reaction solution was added to dilute the solution to obtain a diluted Release the liquid, then start the No. 1 liquid pump, open valves 8, 12 and 3, so that the diluted liquid enters the hollow fiber ultrafiltration column 13 for ultrafiltration. The molecular weight cutoff of the hollow fiber ultrafiltration column is 100,000. When the volume of the ultrafiltrate in the No. 1 reaction tank is close to the volume of the reaction liquid, read the conductivity value from the conductivity meter 4 to see if it is ≦20μs / cm. If not, continue to add 3 times the volume of the reaction liquid for dilution and ultrafiltration until the conductivity value is ≦20μs / cm. / cm to obtain a reaction solution 1, the volume of the reaction solution 1 is 100L, if it is not reached, pure water can be added for adjustment, and during the preparation of the reaction solution 1, the temperature of the reaction solution 1 is controlled by controlling the cooling water in the jacket so that the temperature is 15°C, and the concentration of polyglutamic acid in the 100L reaction solution 1 is 20g / L, the concentration of N-hydroxysuccinimide is 2g / L, and the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 2g / L;

[0041] (2) Preparation of reaction solution 2: Carboxymethyl chitosan was added to reaction tank No. 1 from feed port No. 2, and pure water was introduced into reaction tank No. 2 through water inlet pipe 42 and valve No. 19. After the pH value was adjusted to 7.4 using 1% sodium hydroxide solution, motor No. 2 was started for stirring. After sufficient dissolution, reaction solution 2 was obtained with a volume of 100 L. At this time, the substance concentration of carboxymethyl chitosan in reaction solution 2 was 2 g / L;

[0042] (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, and allow reaction liquid 1 to flow to liquid mixer 26 through liquid pump No. 1, valve No. 8, valve No. 11, and liquid flow meter No. 1. Allow reaction liquid 2 to flow to liquid mixer 26 through valve No. 17, liquid pump No. 2, valve No. 16, and liquid flow meter No. 2. Start motor No. 1 to stir and rotate motor No. 2. The mixed liquid is discharged from valve No. 15. After standing for 10 minutes, polyglutamic acid and chitosan hydrogel base materials are obtained.

[0043] The polyglutamic acid and chitosan hydrogel base materials prepared by the preparation method of this embodiment.

[0044] The polyglutamic acid and chitosan hydrogel base materials of this embodiment are used in repair and soothing products after modification.

[0045] The modification method of this embodiment is:

[0046] The polyglutamic acid and chitosan hydrogel base materials were plasma treated to obtain a treated base material, the treated base material was immersed in a sodium alginate solution, stirred for 15 minutes, hydroxyapatite and catalase were added, and stirred at 37°C for 35 minutes. The obtained product was immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

[0047] The parameters of the plasma treatment in this embodiment are: treatment power is 150W, and treatment time is 5 minutes.

[0048] The mass ratio of polyglutamic acid and chitosan hydrogel base material, sodium alginate solution, hydroxyapatite and catalase in this embodiment is 1:1.35:0.15:0.007.

[0049] The raw materials of the sodium alginate solution of this embodiment include, by percentage, 15wt% sodium alginate, 1wt% vitamin C solution and the balance pure water; the vitamin C solution consists of 3wt% vitamin C and the balance pure water; the particle diameter of hydroxyapatite is 80nm.

[0050] The tensile properties of the modified polyglutamic acid and chitosan hydrogel base obtained in this example were tested. The testing method was to use the modified polyglutamic acid and chitosan hydrogel base prepared in this example as a sample, immerse the sample in water to saturate it, then take it out, wipe the surface of the sample with filter paper, cut it into rectangular strips with a diameter of about 2 cm and a thickness of about 0.3 mm, and stretch it at a speed of 25 mm / min at room temperature. The tensile strength was measured to be 2.79 MPa.

[0051] Example 2

[0052] A method for preparing a polyglutamic acid and chitosan hydrogel base material, the preparation method of this embodiment comprises the following steps:

[0053] (1) In reaction tank No. 1, polyglutamic acid is added from feed port No. 1, and pure water and an active substance with a polyglutamic acid concentration of 0.05 wt% are introduced into reaction tank No. 1 through water inlet pipe 42, valve No. 2, and valve No. 3. The active substance is a repair material, i.e., human epidermal growth factor. The pH is adjusted to 5.8 using 1% hydrochloric acid solution and 1% sodium hydroxide solution to obtain a polyglutamic acid solution. The volume of the polyglutamic acid solution is V1 as read from the scale of sight glass No. 1, and stirring motor 9 is started to drive stirring motor No. 1 to rotate. , so that the polyglutamic acid is dissolved uniformly, at this time, cooling water is introduced into the jacket 43 through valve No. 6 to cool it down, and the cooling water flows out from valve No. 1 after cooling, and N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are added to the No. 1 feeding port, wherein the mass ratio of polyglutamic acid, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 10:1:1, and the reaction solution is obtained after stirring for 30 minutes, and pure The first reaction tank is diluted with water to obtain a dilution solution, and then the No. 1 liquid pump is started, and the No. 8 valve, the No. 12 valve, and the No. 3 valve are opened to allow the dilution solution to enter the hollow fiber ultrafiltration column 13 for ultrafiltration. The molecular weight cutoff of the hollow fiber ultrafiltration column is 100000. When the volume of the ultrafiltrate in the No. 1 reaction tank is close to the volume of the reaction solution, the conductivity value is read from the conductivity meter 4 to see whether it is ≦20μs / cm. If not, pure water 3 times the volume of the reaction solution is continued to be added for dilution and ultrafiltration until the conductivity value is ≦20μs / cm, and the reaction solution is obtained. The volume of the reaction solution 1 is 100 L. If the volume is not reached, pure water can be added for adjustment. In the process of preparing the reaction solution 1, the temperature of the reaction solution 1 is kept at 15° C. by controlling the cooling water in the jacket. Finally, the concentration of polyglutamic acid in the 100 L reaction solution 1 is 20 g / L, the concentration of N-hydroxysuccinimide is 2 g / L, the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 2 g / L, and the concentration of human epidermal growth factor is 10 mg / L.

[0054] (2) Preparation of reaction solution 2: Carboxymethyl chitosan was added to reaction tank No. 1 from feed port No. 2, and pure water was introduced into reaction tank No. 2 through water inlet pipe 42 and valve No. 19. After the pH value was adjusted to 7.3 using 1% sodium hydroxide solution, motor No. 2 was started for stirring. After sufficient dissolution, reaction solution 2 was obtained with a volume of 100 L. At this time, the substance concentration of carboxymethyl chitosan in reaction solution 2 was 2 g / L;

[0055] (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, and allow reaction liquid 1 to flow to liquid mixer 26 through liquid pump No. 1, valve No. 8, valve No. 11, and liquid flow meter No. 1. Allow reaction liquid 2 to flow to liquid mixer 26 through valve No. 17, liquid pump No. 2, valve No. 16, and liquid flow meter No. 2. Start motor No. 1 to stir and rotate motor No. 2. The mixed liquid is discharged from valve No. 15. After standing for 10 minutes, polyglutamic acid and chitosan hydrogel base materials are obtained.

[0056] The polyglutamic acid and chitosan hydrogel base materials prepared by the preparation method of this embodiment.

[0057] The polyglutamic acid and chitosan hydrogel base materials of this embodiment are used in repair and soothing products after modification.

[0058] The modification method of this embodiment is:

[0059] The polyglutamic acid and chitosan hydrogel base materials were plasma treated to obtain a treated base material, the treated base material was immersed in a sodium alginate solution, stirred for 15 minutes, hydroxyapatite and catalase were added, and stirred at 37°C for 35 minutes. The obtained product was immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

[0060] The parameters of the plasma treatment in this embodiment are: treatment power is 150W, and treatment time is 5 minutes.

[0061] The mass ratio of polyglutamic acid and chitosan hydrogel base material, sodium alginate solution, hydroxyapatite and catalase in this embodiment is 1:1.35:0.15:0.007.

[0062] The raw materials of the sodium alginate solution of this embodiment include, by percentage, 15wt% sodium alginate, 1wt% vitamin C solution and the balance pure water; the vitamin C solution consists of 3wt% vitamin C and the balance pure water; the particle diameter of hydroxyapatite is 80nm.

[0063] Referring to the test method of Example 1, the tensile strength of the hydrogel obtained in this example was tested, and the tensile strength was measured to be 2.80 MPa; at the same time, its repair effect was tested, and the test method was that the modified polyglutamic acid and chitosan hydrogel base prepared in this example were dissolved in pure water to prepare a hydrogel solution with a concentration of 10% as sample B; 10 rats weighing 250±2g were taken, and the hair on the left side of the dorsal spine of each rat was trimmed. Under anesthesia, a scalpel was used to form a wound with a diameter of 1.5cm and a depth of 2mm, and sample B was applied to the wound with a coating amount of 2mL. It was observed that the wound healing rate of the rats reached 38.6% after the 7th day.

[0064] Example 3

[0065] A method for preparing a polyglutamic acid and chitosan hydrogel base material, the preparation method of this embodiment comprises the following steps:

[0066] (1) In reaction tank No. 1, polyglutamic acid is added from feed port No. 1, and pure water is introduced into reaction tank No. 1 through water inlet pipe 42, valve No. 2, and valve No. 3. After the pH is adjusted to 5.2 using 1% hydrochloric acid solution and 1% sodium hydroxide solution, a polyglutamic acid solution is obtained. The volume of the polyglutamic acid solution is 100 L as read from the scale of sight glass No. 1, and stirring motor 9 is started to drive No. 1 to stir and rotate, so that the polyglutamic acid is dissolved uniformly. At this time, cooling water is introduced into jacket 43 through valve No. 6 to cool the temperature. After cooling, the cooling water flows out from valve No. 1, and N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are added into feed port No. 1. After stirring for 30 minutes, a reaction solution is obtained. Pure water 3 times the volume of the reaction solution is added to dilute the solution to obtain a diluted solution. Then, liquid pump No. 1 is started, and valves No. 8, No. 12, and No. 3 are opened. The diluted solution enters the hollow fiber ultrafiltration column 13 for ultrafiltration, and the molecular weight cutoff of the hollow fiber ultrafiltration column is 100000. When the volume of the ultrafiltrate in the No. 1 reaction tank is close to the volume of the reaction solution, the conductivity value is read from the conductivity meter 4 to see whether it is ≦20μs / cm. If not, pure water with a volume of 3 times the volume of the reaction solution is added for dilution and ultrafiltration until the conductivity value is ≦20μs / cm, that is, the reaction solution 1 is obtained. The volume of the reaction solution 1 is 100L. If it is not reached, pure water can be added for adjustment. In the preparation process of the reaction solution 1, the temperature of the reaction solution 1 is 15°C by controlling the cooling water in the jacket. Finally, the concentration of polyglutamic acid in the 100L reaction solution 1 is 20g / L, the concentration of N-hydroxysuccinimide is 2g / L, and the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 2g / L;

[0067] (2) Preparation of reaction solution 2: In reaction tank No. 1, carboxymethyl chitosan is added from feed port No. 2, and at the same time, solvent pure water and active substance with a carboxymethyl chitosan concentration of 50 wt% are introduced into reaction tank No. 2 through water inlet pipe 42 and valve No. 19. The active substance is a soothing material, i.e., dipotassium glycyrrhizinate. After the pH is adjusted to 7.5 using 1% sodium hydroxide solution, motor No. 2 is started for stirring. After sufficient dissolution, reaction solution 2 with a volume of 100 L is obtained. At this time, the substance concentration of carboxymethyl chitosan in reaction solution 2 is 2 g / L, and the concentration of dipotassium glycyrrhizinate is 1 g / L;

[0068] (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, and allow reaction liquid 1 to flow to liquid mixer 26 through liquid pump No. 1, valve No. 8, valve No. 11, and liquid flow meter No. 1. Allow reaction liquid 2 to flow to liquid mixer 26 through valve No. 17, liquid pump No. 2, valve No. 16, and liquid flow meter No. 2. Start motor No. 1 to stir and rotate motor No. 2. The mixed liquid is discharged from valve No. 15. After standing for 10 minutes, polyglutamic acid and chitosan hydrogel base materials are obtained.

[0069] The polyglutamic acid and chitosan hydrogel base materials prepared by the preparation method of this embodiment.

[0070] The polyglutamic acid and chitosan hydrogel base materials of this embodiment are used in repair and soothing products after modification.

[0071] The modification method of this embodiment is:

[0072] The polyglutamic acid and chitosan hydrogel base materials were plasma treated to obtain a treated base material, the treated base material was immersed in a sodium alginate solution, stirred for 15 minutes, hydroxyapatite and catalase were added, and stirred at 37°C for 35 minutes. The obtained product was immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

[0073] The parameters of the plasma treatment in this embodiment are: treatment power is 150W, and treatment time is 5 minutes.

[0074] The mass ratio of polyglutamic acid and chitosan hydrogel base material, sodium alginate solution, hydroxyapatite and catalase in this embodiment is 1:1.35:0.15:0.007.

[0075] The raw materials of the sodium alginate solution of this embodiment include, by percentage, 15wt% sodium alginate, 1wt% vitamin C solution and the balance pure water; the vitamin C solution consists of 3wt% vitamin C and the balance pure water; the particle diameter of hydroxyapatite is 80nm.

[0076] Referring to the test method of Example 1, the tensile strength of the hydrogel obtained in this embodiment was tested, and the tensile strength was measured to be 2.78 MPa; at the same time, the soothing effect was tested, and the test method was to dissolve the modified polyglutamic acid and chitosan hydrogel base prepared in this embodiment in a phosphate buffer solution with a pH of 7.4 to obtain a hydrogel solution with a concentration of 10% as sample A; HaCaT (human keratinocytes) were cultured with RPMI-1640 medium containing 10% bovine serum, 100U / ml penicillin, and 100μg / ml streptomycin at 37°C, 5% CO2 saturated with 98% humidity, and then the cells in the logarithmic growth phase were selected and inoculated into a 96-well culture plate. After culturing for 24 hours, the cells were completely attached to the wall, and the culture medium was replaced with RPMI-1640 medium containing sample A (final concentration of 0.1 mg / mL), and after incubation for 24 hours, the cells were incubated at 20mJ / cm 2 The cells were irradiated with a dose of UVB at a distance of 12 cm. After irradiation, the cell culture supernatant was collected and the concentration of IL-1α in the supernatant was detected by human ELISA kit. The average value was taken after repeating 3 times, and the inhibition rate of IL-1α was measured to be 37.9%.

[0077] Example 4

[0078] A method for preparing a polyglutamic acid and chitosan hydrogel base material, the preparation method of this embodiment comprises the following steps:

[0079] (1) In reaction tank No. 1, polyglutamic acid is added from feed port No. 1, and pure water is introduced into reaction tank No. 1 through water inlet pipe 42, valve No. 2, and valve No. 3. The pH value is adjusted to 5.3 using 1% hydrochloric acid solution and 1% sodium hydroxide solution to obtain a polyglutamic acid solution. The volume of the polyglutamic acid solution is V1 as read from the scale of sight glass No. 1, and stirring motor 9 is started to drive stirring motor No. 1 to rotate so that the polyglutamic acid is evenly dissolved. At this time, the jacket 43 is supplied with water through valve No. 6. Cooling water is introduced into the reaction mixture to cool it down. After cooling down, the cooling water flows out from valve No. 1. N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are added into the feeding port No. 1, wherein the mass ratio of polyglutamic acid, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 10:1:1. After stirring for 30 minutes, a reaction solution is obtained, and pure water 3 times the volume of the reaction solution is added to dilute the solution to obtain a diluted solution. Liquid, then start No. 1 liquid pump, open No. 8 valve, No. 12 valve, No. 3 valve, so that the diluted liquid enters the hollow fiber ultrafiltration column 13 for ultrafiltration, the molecular weight cutoff of the hollow fiber ultrafiltration column is 100000, until the volume of the ultrafiltrate in the No. 1 reaction tank is close to the volume of the reaction liquid, read the conductivity value from the conductivity meter 4 to see if it is ≦20μs / cm, if not, continue to add 3 times the volume of the reaction liquid for dilution and ultrafiltration, until the conductivity value is ≦20μs / cm to obtain a reaction solution 1, the volume of the reaction solution 1 is 100 L, if it is not reached, pure water can be added for adjustment, and in the process of preparing the reaction solution 1, the temperature of the reaction solution 1 is controlled by controlling the cooling water in the jacket so that the temperature is 15° C., and the concentration of polyglutamic acid in the 100 L reaction solution 1 is 50 g / L, the concentration of N-hydroxysuccinimide is 5 g / L, and the concentration of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 5 g / L;

[0080] (2) Preparation of reaction solution 2: In reaction tank No. 1, carboxymethyl chitosan was added from feed port No. 2, and pure water was introduced into reaction tank No. 2 through water inlet pipe 42 and valve No. 19. After the pH value was adjusted to 7.2 using 1% sodium hydroxide solution, motor No. 2 was started for stirring. After sufficient dissolution, reaction solution 2 was obtained with a volume of 100 L. At this time, the substance concentration of carboxymethyl chitosan in reaction solution 2 was 5 g / L;

[0081] (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, and allow reaction liquid 1 to flow to liquid mixer 26 through liquid pump No. 1, valve No. 8, valve No. 11, and liquid flow meter No. 1. Allow reaction liquid 2 to flow to liquid mixer 26 through valve No. 17, liquid pump No. 2, valve No. 16, and liquid flow meter No. 2. Start motor No. 1 to stir and rotate motor No. 2. The mixed liquid is discharged from valve No. 15. After standing for 10 minutes, polyglutamic acid and chitosan hydrogel base materials are obtained.

[0082] The polyglutamic acid and chitosan hydrogel base materials prepared by the preparation method of this embodiment.

[0083] The polyglutamic acid and chitosan hydrogel base materials of this embodiment are used in repair and soothing products after modification.

[0084] The modification method of this embodiment is:

[0085] The polyglutamic acid and chitosan hydrogel base materials were plasma treated to obtain a treated base material, the treated base material was immersed in a sodium alginate solution, stirred for 15 minutes, hydroxyapatite and catalase were added, and stirred at 37°C for 35 minutes. The obtained product was immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

[0086] The parameters of the plasma treatment in this embodiment are: treatment power is 150W, and treatment time is 5 minutes.

[0087] The mass ratio of polyglutamic acid and chitosan hydrogel base material, sodium alginate solution, hydroxyapatite and catalase in this embodiment is 1:1.35:0.15:0.007.

[0088] The raw materials of the sodium alginate solution of this embodiment include, by percentage, 15wt% sodium alginate, 1wt% vitamin C solution and the balance pure water; the vitamin C solution consists of 3wt% vitamin C and the balance pure water; the particle diameter of hydroxyapatite is 80nm.

[0089] Referring to the test method of Example 1, the tensile strength obtained in this example was tested, and the measured tensile strength was 3.68 MPa.

[0090] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A method for preparing a polyglutamic acid and chitosan hydrogel base material, characterized in that: The preparation method comprises the following steps: (1) Preparation of reaction solution 1: polyglutamic acid is added to a reaction tank, a solvent is introduced, and the pH is adjusted to obtain a polyglutamic acid solution. The motor is started for stirring, and cooling water is introduced for cooling. N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride are then added to the reaction tank, and the reaction solution is obtained after stirring for 20-40 minutes. Pure water is added for dilution to obtain a diluted solution, and a liquid pump is started and a valve is opened to allow the diluted solution to enter a hollow fiber ultrafiltration column for ultrafiltration until the volume of the ultrafiltrate in the reaction tank is close to the volume of the reaction solution. The conductivity value is read from a conductivity meter to determine whether it meets the standard. If it does not meet the standard, pure water is continued to be added for dilution and ultrafiltration. If it meets the standard, reaction solution 1 is obtained. (2) Preparation of reaction solution 2: Carboxymethyl chitosan is placed in a reaction tank, a solvent is introduced, the pH is adjusted, and a motor is started for stirring to obtain reaction solution 2 after the carboxymethyl chitosan is fully dissolved; (3) Mixing of reaction liquid 1 and reaction liquid 2: Prepare reaction liquid 1 and reaction liquid 2 in a volume ratio of 1:1, start the liquid pump, open the valve, and introduce reaction liquid 1 and reaction liquid 2 respectively, start the motor for stirring, open the valve to release the mixed liquid, and let it stand for 8-12 minutes to obtain polyglutamic acid and chitosan hydrogel base materials.

2. The method for preparing a polyglutamic acid and chitosan hydrogel base according to claim 1, characterized in that: The solvent in step (1) is pure water or pure water and an active substance with a polyglutamic acid concentration of 0.01-0.09wt%; the pH adjustment refers to adjusting the pH to 4-6 using 0.8-1.2% hydrochloric acid solution and 0.8-1.2% sodium hydroxide solution; the mass ratio of the polyglutamic acid, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is 9-11:1:

1.

3. The method for preparing a polyglutamic acid and chitosan hydrogel base according to claim 1, characterized in that: The conductivity value in step (1) meets the standard value of ≦20μs / cm.

4. The method for preparing a polyglutamic acid and chitosan hydrogel base according to claim 1, characterized in that: The solvent in step (2) is pure water or pure water and an active substance with a carboxymethyl chitosan concentration of 40-60wt%; the pH adjustment refers to adjusting the pH to 7-8 using 0.8-1.2% sodium hydroxide solution; the concentration of carboxymethyl chitosan in the reaction solution 2 is 1-10g / L.

5. An application of polyglutamic acid and chitosan hydrogel base prepared by the preparation method according to claims 1-4, characterized in that: The polyglutamic acid and chitosan hydrogel base materials are applied to repair and soothing products after being modified.

6. The use of a polyglutamic acid and chitosan hydrogel base material according to claim 5, characterized in that: The modification method is: The polyglutamic acid and chitosan hydrogel base materials are subjected to plasma treatment to obtain a treated base material, the treated base material is immersed in a sodium alginate solution, stirred for 10-20 minutes, hydroxyapatite and catalase are added, and stirred at 36-38°C for 30-40 minutes. The obtained product is immersed in pure water for 24 hours, and then freeze-dried at -20°C for 24 hours to obtain the modified polyglutamic acid and chitosan hydrogel base materials.

7. The use of a polyglutamic acid and chitosan hydrogel base material according to claim 6, characterized in that: The parameters of the plasma treatment are: treatment power of 50-250W, and treatment time of 3-7min.

8. The use of a polyglutamic acid and chitosan hydrogel base material according to claim 6, characterized in that: The mass ratio of the polyglutamic acid and chitosan hydrogel base material, the sodium alginate solution, the hydroxyapatite and the catalase is 1:1.2-1.5:0.1-0.2:0.005-0.

01.

9. The use of a polyglutamic acid and chitosan hydrogel base material according to claim 6, characterized in that: The raw materials of the sodium alginate solution include 10-20wt% sodium alginate, 0.2-2wt% vitamin C solution and the balance pure water according to percentage; the vitamin C solution is composed of 1-5wt% vitamin C and the balance pure water; the particle diameter of the hydroxyapatite is 60-100nm.

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