CN113278120A

CN113278120A - Anti-fatigue amphiphilic organic hydrogel and preparation method and application thereof

Info

Publication number: CN113278120A
Application number: CN202110573865.7A
Authority: CN
Inventors: 巫林平; 侯金飞; 孙家明
Original assignee: Guangzhou Institute of Biomedicine and Health of CAS
Current assignee: Guangzhou Institute of Biomedicine and Health of CAS
Priority date: 2021-05-25
Filing date: 2021-05-25
Publication date: 2021-08-20
Anticipated expiration: 2041-05-25
Also published as: CN113278120B

Abstract

The invention provides an anti-fatigue amphiphilic organic hydrogel and a preparation method and application thereof. The preparation method includes the following steps: (1) adding bis-mercapto polyethylene glycol to a solution of double-bond functionalized polyhydroxy fatty acid ester , after dissolving, adding polyethylene glycol diacrylate, dissolving to obtain a polymer solution; (2) adding graphdiyne to the polymer solution obtained in step (1), then adding sodium dodecyl sulfate, dispersing to obtain dispersion liquid; (3) adding 2,2-dimethylolpropionic acid and a photoinitiator to the dispersion liquid in step (2), and curing with ultraviolet light to obtain the anti-fatigue amphiphilic organic hydrogel. The anti-fatigue amphiphilic hydrogel prepared by the preparation method of the present invention can solve the problems of water absorption swelling and poor mechanical properties of the current hydrogel.

Description

Anti-fatigue amphiphilic organic hydrogel and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biological materials, and relates to an anti-fatigue amphiphilic organic hydrogel and a preparation method and application thereof.

Background

The hydrogel is a very hydrophilic three-dimensional network structure material, has good biocompatibility and biodegradability, and has a structure similar to extracellular matrix. The method is widely used in the field of biological materials, can be used as a tissue engineering scaffold, and is used for repairing tissue and organ regeneration.

Although hydrogel is a mature material, the conventional hydrogel still has the following disadvantages: firstly, under the condition of body fluid, hydrogel is easy to swell after absorbing water, so that the stability is reduced and the hydrogel is easy to break; secondly, the mechanical property of the hydrogel is poor, and particularly, the hydrogel is easy to fatigue, so that the hydrogel is easy to break in the repeated stretching process. Therefore, how to prepare a hydrogel which has fatigue resistance, amphiphilic property, no swelling in water phase and stability is one of the key problems to be solved.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide an anti-fatigue amphiphilic organic hydrogel and a preparation method and application thereof. The anti-fatigue amphiphilic hydrogel prepared by the preparation method can solve the problems of water absorption swelling, poor mechanical property and the like of the existing hydrogel.

In order to achieve the purpose, the invention adopts the following technical scheme:

in one aspect, the invention provides a preparation method of an anti-fatigue amphiphilic organic hydrogel, which comprises the following steps:

(1) adding dimercaptopolyethylene glycol into a double-bond functionalized Polyhydroxyalkanoate (PHA) solution, dissolving, adding polyethylene glycol diacrylate, and dissolving to obtain a polymer solution;

(2) adding graphite alkyne into the polymer solution obtained in the step (1), then adding a water-phase dispersing agent, and dispersing to obtain a dispersion liquid;

(3) and (3) adding 2, 2-dimethylolpropionic acid and a photoinitiator into the dispersion liquid obtained in the step (2), and carrying out ultraviolet curing to obtain the anti-fatigue amphiphilic organic hydrogel.

The invention takes PEGDA (polyethylene glycol diacrylate) hydrogel as a main material, and reacts with double-bond polyhydroxyalkanoate organic gel in a click chemistry mode to improve the swelling characteristic of the PEGDA hydrogel, and graphite alkyne nano-particles are mixed to enhance the mechanical property of the PEGDA hydrogel. The anti-fatigue amphiphilic hydrogel prepared by the method can solve the problems of water absorption swelling, poor mechanical property and the like of the existing hydrogel.

Preferably, the solvent in the polyhydroxyalkanoate solution in step (1) is any one or a combination of at least two of tetrahydrofuran, toluene, dimethyl sulfoxide, dichloromethane or chloroform.

Preferably, the double bond functionalized polyhydroxyfatty acids of step (1) have a double bond content of 15% to 45%, such as 15%, 18%, 20%, 23%, 25%, 28%, 30%, 33%, 35%, 40%, 43% or 45%.

Preferably, the concentration of the double bond functionalized polyhydroxyalkanoate solution of step (1) is 3% to 10%, such as 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%.

Preferably, the double bond functionalized polyhydroxyalkanoate of step (1) has a number average molecular weight of 10000 to 300000Da, such as 10000Da, 12000Da, 15000Da, 20000Da, 50000Da, 80000Da, 100000Da, 130000Da, 200000Da, 250000Da, 280000Da or 300000 Da.

Preferably, the mass ratio of the dimercaptopolyethylene glycol to the double-bond functionalized polyhydroxyalkanoate in the step (1) is 1:0.5 to 3, for example 1:0.5, 1:0.8, 1:1, 1:1.5, 1:1.8, 1:2, 1:2.5, 1:2.8 or 1: 3.

Preferably, the dimercaptopolyethylene glycol has a number average molecular weight of 1000 to 20000Da, such as 2000Da, 5000Da, 8000Da, 10000Da, 13000Da, 15000Da, 18000Da or 20000 Da.

Preferably, the mass ratio of the polyethylene glycol diacrylate to the double-bond functionalized polyhydroxyalkanoate in the step (1) is 1: 0.5-3, such as 1:0.5, 1:0.8, 1:1, 1:1.5, 1:1.8, 1:2, 1:2.5, 1:2.8 or 1: 3.

Preferably, the number average molecular weight of the polyethylene glycol diacrylate in the step (1) is 5000-30000 Da, 5000Da, 8000Da, 10000Da, 13000Da, 15000Da, 18000Da, 20000Da, 23000Da, 25000Da, 28000Da or 30000 Da.

Preferably, the amount of the graphdiyne added in step (2) is 0.3 to 0.45mg, such as 0.3mg, 0.35mg, 0.4mg, 0.45mg, etc., relative to 3 to 5mL of the polymer solution.

Preferably, the aqueous phase dispersant added in step (2) is any one or a combination of at least two of sodium dodecyl sulfate, tween 20 or benzalkonium bromide.

Preferably, the concentration of the aqueous phase dispersant in step (2) is 1% to 5%, such as 1%, 1.5%, 1.8%, 2%, 2.5%, 2.8%, 3%, 3.5%, 4%, 4.5%, or 5%, relative to 3 to 5mL of the polymer solution.

Preferably, the dispersion of step (2) is ultrasonic dispersion.

Preferably, the dispersion in step (2) is performed at a temperature of 0-10 ℃ (e.g., 0 ℃,2 ℃, 5 ℃, 8 ℃ or 10 ℃) and a power of 500-1000W (e.g., 500W, 700W, 800W or 1000W) for 1-1.5 h (e.g., 1h, 1.2h, 1.4h or 1.5 h). In the invention, the graphite alkyne is uniformly dispersed into nano particles in the solution through dispersion.

Preferably, step (3) is carried out under exclusion of light.

Preferably, the 2, 2-dimethylolpropionic acid is added in an amount of 20 to 30mg, for example 20mg, 22mg, 25mg, 28mg or 30mg, relative to 3 to 5mL of the dispersion in step (3).

Preferably, the photoinitiator in the step (3) is Irgacure 2959.

Preferably, the photoinitiator is added in step (3) in an amount of 30 to 40mg, for example 30mg, 32mg, 35mg, 38mg or 40mg, relative to 3 to 5mL of the dispersion.

Preferably, the UV curing of step (3) is irradiating with a UV lamp at a power of 100-500 mW (e.g., 100mW, 130mW, 150mW, 200mW, 250mW, 280mW, 300mW, 400mW or 500mW) for 6-10min (e.g., 6min, 7min, 8min, 9min or 10 min).

In the invention, before the ultraviolet curing in the step (3), a solution obtained by adding 2, 2-dimethylolpropionic acid and a photoinitiator into the dispersion liquid in the step (2) is poured into a transparent glass mold under the condition of keeping out of the sun.

Preferably, after the ultraviolet curing in the step (3), the obtained anti-fatigue amphiphilic organic hydrogel is placed in chloroform for soaking for 1-2 hours, and is repeatedly soaked and washed in 75% alcohol and deionized water for 3-5 times respectively until the organic solvent is completely removed, and the organic hydrogel is placed in deionized water for storage.

In another aspect, the present invention provides the anti-fatigue amphiphilic organic hydrogel prepared by the preparation method as described above.

In another aspect, the present invention provides a tissue engineering scaffold comprising the fatigue-resistant amphiphilic organic hydrogel as described above.

Compared with the prior art, the invention has the following beneficial effects:

the anti-fatigue amphiphilic hydrogel prepared by the preparation method can solve the problems of water absorption swelling, poor mechanical property and the like of the existing hydrogel, so that the hydrogel has good fatigue resistance.

Drawings

Fig. 1 is a schematic external view and a microscopic structure diagram of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1, wherein a is a schematic external view, B is a microscopic frozen electron microscope image, and C is a microscopic scanning electron microscope image after freeze-drying.

FIG. 2 is a graph showing swelling results obtained in the test of example 4 on the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 and a general hydrogel.

FIG. 3 is a graph showing the results of the phase contact angle measurement of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 with a conventional hydrogel;

FIG. 4A is a graph showing the results of the elastic modulus test of the fatigue resistant amphiphilic organic hydrogel prepared in example 1 and a conventional hydrogel gel;

FIG. 4B is a graph showing the maximum tolerable stress test results of the fatigue resistant amphiphilic organic hydrogel prepared in example 1 and a conventional hydrogel gel;

FIG. 4C is a graph showing the results of a fracture strain test of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 and a conventional hydrogel gel;

FIG. 5 is a graph showing the results of the loss modulus (G ') and storage modulus (G') tests of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 and a conventional hydrogel gel;

FIG. 6A is a graph showing the results of an elastic modulus test on a conventional hydrogel gel;

FIG. 6B is a graph showing the results of an elastic modulus test of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1;

FIG. 7 is an immunofluorescence staining pattern of the general hydrogel and the anti-fatigue amphiphilic organic hydrogel prepared in example 1 for vascular stents prepared by implanting vascular endothelial cells in vitro, perfusing for 14 days, slicing and then performing endothelial cell marker protein CD31 and nuclear DAPI;

FIG. 8 is a graph showing the general appearance of a common hydrogel and the HE staining after slicing after the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 was implanted into rats and the materials were taken at different time points.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

Example 1

(1) Adding 100mg PHA family polymer P3HB10U (poly 3-hydroxybutyrate-co-3-hydroxyundecylenic acid) (Beijing blue crystal Microbiol. Co., Ltd., China) into 3mL tetrahydrofuran solution, placing into a round flask, and stirring in a magnetic stirrer until completely dissolving;

(2) adding 160mg of HS-PEG-SH (dimercaptopolyethylene glycol) with the molecular weight of 2000Da (Shanghai Aladdin Biotechnology Co., Ltd., China) into the solution P3HB10U prepared in the step (1), and stirring at normal temperature until the solution is completely dissolved;

(3) 260mg of 10000Da molecular weight PEGDA powder (polyethylene glycol (diol) diacrylate) (Sigma-Aldrich, USA) is added into the solution obtained in the step (2) and further mixed until completely dissolved;

(4) preparing a graphite alkyne nanoparticle dispersion liquid, adding 0.3mg of graphite alkyne powder (Nanjing Xian Feng nanometer material science and technology Co., Ltd., China) into the solution obtained in the step (3), then adding 5mg of SDS (sodium dodecyl sulfate) (Shanghai Aladdin Biotechnology science and technology Co., Ltd., China) powder, placing the powder into an ultrasonic dispersion instrument, and dispersing for 1.5 hours at a low temperature (0 ℃) until the graphite alkyne powder is uniformly dispersed into nanoparticles in the solution;

(5) placing the solution obtained in the step (4) in a dark condition, respectively adding 20mg of DMPA (2, 2-dimethylolpropionic acid) and 40mg of photoinitiator 2959(Irgacure2959), uniformly mixing, and immediately carrying out the next operation;

(6) pouring the solution obtained in the step (5) into a transparent glass mold under the condition of keeping out of the sun, and irradiating for 6min by using an ultraviolet curing lamp until the gel is completely changed into a solid;

(7) and (4) soaking the anti-fatigue amphiphilic organic hydrogel obtained in the step (6) in chloroform for 2h, repeatedly soaking and washing in 75% alcohol and deionized water respectively until the organic solvent is completely removed, and storing in deionized water.

The general appearance and microstructure of the fatigue-resistant amphiphilic organic hydrogel prepared in example 1 were studied, wherein a in fig. 1 is a schematic diagram of the general appearance of the fatigue-resistant amphiphilic organic hydrogel of example 1; in FIG. 1, B and C are a cryo-electron micrograph (S-4800, Hitachi, Japan) and a scanning electron micrograph (SU-70, Hitachi, Japan) of the fatigue-resistant amphiphilic organic hydrogel in example 1, respectively. As is apparent from the B and C diagrams in FIG. 1, the surface of the gel has a porous structure with a pore size of about 20um, which is suitable for cell penetration and capillary growth, and helps the gel regenerate in vivo tissue to form blood vessels.

Example 2

(1) Adding 150mg of PHA polymer P3HB10U into 4.5mL of tetrahydrofuran solution, placing the mixture into a round flask, and stirring the mixture in a magnetic stirrer until the mixture is completely dissolved;

(2) adding 240mg of HS-PEG-SH (dimercaptopolyethylene glycol) with the molecular weight of 2000Da into the P3HB10U solution prepared in the step (1), and stirring at normal temperature until the solution is completely dissolved;

(3) adding 390mg of PEGDA powder with 10000Da molecular weight into the solution obtained in the step (2), and further uniformly mixing until the PEGDA powder is completely dissolved;

(4) preparing a graphite alkyne nanoparticle dispersion liquid, adding 0.45mg of graphite alkyne powder into the solution obtained in the step (3), then adding 10mg of SDS (sodium dodecyl sulfate) powder, placing the mixture into an ultrasonic dispersion instrument, and dispersing for 1.2 hours at a low temperature (5 ℃) until the graphite alkyne powder is uniformly dispersed into nanoparticles in the solution;

(5) placing the solution obtained in the step (4) in a dark condition, respectively adding 30mg of 2, 2-dimethylolpropionic acid (DMPA) and 30mg of photoinitiator 2959(Irgacure2959), uniformly mixing, and immediately carrying out the next operation;

(6) pouring the solution obtained in the step (5) into a transparent glass mold under the condition of keeping out of the sun, and irradiating for 10min by using an ultraviolet curing lamp until the gel is completely changed into a solid;

(7) and (4) soaking the gel intravascular stent obtained in the step (6) in chloroform for 2h, repeatedly soaking and washing in 75% alcohol and deionized water respectively until the toxic organic solvent is completely removed, and storing in the deionized water.

Example 3

(1) Adding 120mg of PHA polymer P3HB10U into 4mL of tetrahydrofuran solution, placing the mixture into a round flask, and stirring the mixture in a magnetic stirrer until the mixture is completely dissolved;

(2) adding 200mg of HS-PEG-SH (dimercaptopolyethylene glycol) with the molecular weight of 2000Da into the P3HB10U solution prepared in the step (1), and stirring at normal temperature until the solution is completely dissolved;

(3) adding 300mg of PEGDA powder with 10000Da molecular weight into the solution obtained in the step (2), and further uniformly mixing until the PEGDA powder is completely dissolved;

(4) preparing a graphite alkyne nanoparticle dispersion liquid, adding 0.4mg of graphite alkyne powder into the solution obtained in the step (3), then adding 8mg of SDS (sodium dodecyl sulfate) powder, placing the mixture into an ultrasonic dispersion instrument, and dispersing for 1 hour at a low temperature (10 ℃) until the graphite alkyne powder is uniformly dispersed into nanoparticles in the solution;

(5) placing the solution obtained in the step (4) in a dark condition, respectively adding 30mg of 2, 2-dimethylolpropionic acid (DMPA) and 40mg of photoinitiator 2959(Irgacure2959), uniformly mixing, and immediately carrying out the next operation;

(6) pouring the solution obtained in the step (5) into a transparent glass mold under the condition of keeping out of the sun, and irradiating for 8min by using an ultraviolet curing lamp until the gel is completely changed into a solid;

Example 4

The fatigue-resistant amphiphilic organic hydrogel prepared in example 1 and a common hydrogel (PEGDA hydrogel) were cut into disc-shaped gel pieces having a diameter of about 5mm and a height of 2mm, weighed and recorded mi immediately after drying the surface liquid with a filter paper, and the diameter di thereof was measured, and then placed in an aqueous solution (deionized water) and an organic phase solution (chloroform) respectively for 24h and 2h, weighed and recorded ma immediately after drying the surface liquid with a filter paper, and the diameter da thereof was measured after completion of the soaking and calculated according to the following formula:

swelling degree (%) (ma-mi)/mi

Rate of change (%) of diameter (da-di)/di

Calculations and statistical analyses were performed according to the above formula, and a graph was plotted showing that fig. 2 (n-4, P <0.001, n.s. has no statistical difference).

As can be seen from a comparison of the results in fig. 2, the anti-fatigue amphiphilic organic hydrogel can be maintained stable in the aqueous phase, exhibits significant resistance to swelling, and helps maintain stability under body fluid conditions after implantation into the body.

Example 5

The contact angle of the anti-fatigue amphiphilic organic hydrogel prepared in example 1 was studied. Cutting the anti-fatigue amphiphilic organic hydrogel prepared in the example 1 and a common hydrogel (PEGDA hydrogel) into a flat plate with the thickness of 2-4mm, and accurately controlling the volume of the aqueous phase and the organic phase drops to be 2 mu L; the photograph of the contact angle thus obtained is shown in FIG. 3.

As can be seen by comparison in fig. 3, the fatigue resistant amphiphilic organic hydrogel exhibits significant amphiphilicity.

Example 6

Cutting the anti-fatigue amphiphilic organic hydrogel prepared in the example 1 and a common hydrogel (PEGDA hydrogel) into a rectangular gel block with the length of 30mm, the width of 10mm and the thickness of 2mm, wrapping two ends of a long shaft by using filter paper to increase friction force, fixing the long shaft on polystyrene clamps at two sides, and stretching at a constant speed of 2mm/min until a sample is broken; the tensile properties of the samples include modulus of elasticity (the slope of the tangent to the stress-strain curve), ultimate stress (stress at failure), maximum strain or expandability (strain level at failure), and the data are collated and plotted in FIG. 4A (modulus of elasticity for each group of gels), FIG. 4B (maximum sustainable stress for each group of gels), and FIG. 4C (strain at break for each group of gels). (n-3, P < 0.001).

As can be seen from the comparison of the fatigue-resistant amphiphilic organic hydrogel with the ordinary hydrogel in FIG. 4A, FIG. 4B and FIG. 4C, the elastic modulus of the fatigue-resistant amphiphilic organic hydrogel is significantly increased, the maximum bearable stress is increased, the breaking strain is increased, and the fatigue resistance is enhanced.

Example 7

The hydrogel sample obtained in example 1 and a normal hydrogel (PEGDA hydrogel) sample were cut into a disk-shaped structure having a diameter of 15mm and a thickness of 4mm, placed on a 15mm rheometer vibration test plate, subjected to small amplitude vibration at a frequency of 0.001Hz to 1000Hz, and recorded for loss modulus (G ') and storage modulus (G') values, to plot FIG. 5.

As can be seen from the comparison between the hydrogel sample obtained in example 1 in FIG. 5 and the conventional hydrogel, the amphiphilic organic hydrogel has significantly increased storage modulus, enhanced shear resistance, and better mechanical properties as a vascular stent.

Example 8

The gel sample obtained in example 1 and a normal hydrogel (PEGDA hydrogel) sample were cut into a strip shape having a length of 30mm, a width of 10mm and a thickness of 3mm, and repeated fatigue tests were performed at a rate of 10mm/min in a 50% tensile range, and FIG. 6A and FIG. 6B were plotted based on the experimental data.

As can be seen by comparing FIG. 6A and FIG. 6B, the fatigue resistance of the anti-fatigue amphiphilic organic hydrogel is significantly better, and the elastic modulus can still be kept after 2000 cycles of stretching.

Example 9

The gel sample in example 1 and a common hydrogel (PEGDA hydrogel) sample are prepared into a tubular structure, the tubular structure is planted with vascular endothelial cells and then placed in a bioreactor for further amplification culture, the section is sliced after 14 days of culture, CD31 protein immunofluorescence staining is carried out to identify the proliferation condition of the cells, and figure 7 is arranged according to an immunofluorescence photograph.

As can be seen from comparison between the gel sample obtained in example 1 in FIG. 7 and a common hydrogel, the anti-fatigue amphiphilic organic hydrogel has better cell proliferation and higher expression level of the marker protein, and is more suitable for being used as a material of a scaffold for vascular tissue engineering.

Example 10

After the hydrogel obtained in example 1 and a sample of a general hydrogel (PEGDA hydrogel) were cut into small disks of 10mm in diameter and 3mm in thickness and implanted into SD rats, the sections were removed for 14 days and HE-stained to observe biocompatibility in vivo, and fig. 8 was prepared based on the photomicrograph.

As can be seen by comparing the gel sample of example 1 with a conventional hydrogel in FIG. 8, the gel sample of example 1 has significantly more cell infiltration, and collagen tissue regeneration, collagen remodeling and small vessel regeneration around the gel are observed, which are all sufficient to demonstrate that the fatigue-resistant amphiphilic organic hydrogel is more biocompatible in vivo than a conventional hydrogel.

The applicant states that the present invention is illustrated by the above examples to the preparation method and application of the anti-fatigue amphiphilic organic hydrogel of the present invention, but the present invention is not limited to the above examples, i.e. it does not mean that the present invention must be implemented by the above examples. It will be apparent to those skilled in the art that any modification of the present invention, equivalent substitutions of selected materials and additions of auxiliary components, selection of specific modes and the like, which are within the scope and disclosure of the present invention, are contemplated by the present invention.

Claims

1. A preparation method of an anti-fatigue amphiphilic organic hydrogel is characterized by comprising the following steps:

(1) adding dimercapto polyethylene glycol into the double-bond functionalized polyhydroxy fatty acid ester solution, dissolving, adding polyethylene glycol diacrylate, and dissolving to obtain a polymer solution;

2. The method according to claim 1, wherein the solvent in the polyhydroxyalkanoate solution in step (1) is any one or a combination of at least two of tetrahydrofuran, toluene, dimethyl sulfoxide, dichloromethane or chloroform;

preferably, the double bond content of the double bond functionalized polyhydroxy fatty acid in the step (1) is 15-45%;

preferably, the concentration of the double bond functionalized polyhydroxyalkanoate solution in the step (1) is 3% -10%;

preferably, the number average molecular weight of the double-bond functionalized polyhydroxyalkanoate in the step (1) is 10000-300000 Da;

preferably, the mass ratio of the dimercaptopolyethylene glycol to the double-bond functionalized polyhydroxyalkanoate in the step (1) is 1:0.5 to 3;

preferably, the number average molecular weight of the dimercaptopolyethylene glycol in the step (1) is 1000-20000 Da.

3. The preparation method according to claim 1 or 2, wherein the mass ratio of the polyethylene glycol diacrylate to the double-bond functionalized polyhydroxyalkanoate in step (1) is 1: 0.5-3.

Preferably, the number average molecular weight of the polyethylene glycol diacrylate in the step (1) is 5000-30000 Da.

4. The method according to any one of claims 1 to 3, wherein the amount of the graphdiyne added in step (2) is 0.3 to 0.45mg per 3 to 5mL of the polymer solution;

preferably, the water-phase dispersing agent added in the step (2) is sodium dodecyl sulfate, tween 20 or benzalkonium bromide;

preferably, in the step (2), the concentration of the aqueous phase dispersant is 1-5% relative to 3-5 mL of the polymer solution.

5. The production method according to any one of claims 1 to 4, wherein the dispersion of step (2) is ultrasonic dispersion;

preferably, the dispersion in the step (2) is performed at a temperature of 0-10 ℃ and a power of 500-1000W for 1-1.5 h.

6. The production method according to any one of claims 1 to 5, wherein the step (3) is carried out under light-shielding conditions;

preferably, in the step (3), the addition amount of the 2, 2-dimethylolpropionic acid is 20-30 mg relative to 3-5 mL of the dispersion liquid.

7. The production method according to any one of claims 1 to 6, wherein the photoinitiator in step (3) is preferably Irgacure 2959;

preferably, in the step (3), the addition amount of the photoinitiator is 30-40 mg relative to 3-5 mL of the dispersion liquid;

preferably, the ultraviolet curing in the step (3) is performed by irradiating for 6-10min with an ultraviolet lamp at a power of 100-500 mW.

8. The preparation method according to any one of claims 1 to 7, wherein after the ultraviolet curing in the step (3), the obtained anti-fatigue amphiphilic organic hydrogel is soaked in chloroform for 1-2 hours, and is repeatedly soaked and washed in 75% alcohol and deionized water for 3-5 times until the organic solvent is completely removed, and is stored in deionized water.

9. The anti-fatigue amphiphilic organic hydrogel prepared by the preparation method according to any one of claims 1 to 8.

10. A tissue engineering scaffold, comprising the fatigue resistant amphiphilic organic hydrogel of claim 9.