CN112220968B

CN112220968B - A light-curing 3D printing preparation method of a long-term alternative tissue engineering meniscus scaffold

Info

Publication number: CN112220968B
Application number: CN202011425981.6A
Authority: CN
Inventors: 于然; 李昕盼; 黄伟; 何阳阳; 陈东平; 杨欣; 张瑛; 赵晓娟; 丁国成; 孙牧旸
Original assignee: Institute of Chemistry CAS
Current assignee: Institute of Chemistry CAS
Priority date: 2020-12-09
Filing date: 2020-12-09
Publication date: 2021-03-26
Anticipated expiration: 2040-12-09
Also published as: CN112220968A

Abstract

The invention discloses a photocuring 3D printing preparation method of a tissue engineering meniscus scaffold capable of being replaced for a long time. The photosensitive resin for printing the long-term replacement type meniscus scaffold comprises the following raw material components in parts by weight: 10-80 parts of polyurethane acrylate, 20-80 parts of acrylate monomer, 0.5-5.0 parts of free radical photoinitiator and 0.001-0.5 part of defoaming agent. The polyurethane acrylate has a structure as shown in formula I. In the formula I, R1 can be one of isophorone, hexamethylene and dicyclohexyl methyl; r2 can be one or more of ethyl, propyl and n-butyl; r3 may be specifically one of hydrogen or methyl; n is 10 to 100. The meniscus scaffold provided by the invention has good biocompatibility and in-vivo stability, the mechanical property of the scaffold is similar to that of a natural meniscus, and the three-dimensional structure and the porous structure of the scaffold can be accurately controlled through photocuring 3D printing.

Description

Photocuring 3D printing preparation method of long-term replacement type tissue engineering meniscus scaffold

Technical Field

The invention belongs to the technical field of tissue engineering, and particularly relates to a photocuring 3D printing preparation method of a long-term replacement type tissue engineering meniscus scaffold.

Background

Menisci are white hyaline cartilage covering the articular surfaces, the smooth surfaces of which significantly reduce friction between the articular surfaces during movement. In recent years, the incidence of meniscal damage has become higher, but the self-repair ability of the damaged portion is poor due to lack of blood supply, innervation and lymphatic return. Constructing a substitute for the damaged meniscus, i.e., a tissue engineered meniscus, can achieve good therapeutic results. The meniscus scaffold material should have good biocompatibility, reasonable three-dimensional and porous structure, can realize free diffusion of nutrient substances, and also should have good mechanical properties, the mechanical properties of the scaffold should be similar to those of natural menisci, the compression modulus should be within the range of 75-150 kPa, and the tensile modulus should be within the range of 75-150 MPa [ Biomaterials, 126, 18-30, 2017 ].

Meniscal scaffold materials can be classified into alternative and degradable types. The substituted meniscus implant is mainly made of polyvinyl alcohol hydrogel, polycarbonate polyurethane and other materials with good biocompatibility, in vivo stability and excellent biomechanical performance. The degradable meniscus scaffold can be divided into natural materials such as collagen and silk fibroin and synthetic materials such as polylactic acid and polycaprolactone according to different material sources. For elderly people with slower metabolism, the long-term replacement of meniscal grafts has great advantages.

At present, the synthetic materials for preparing the meniscus scaffold mainly comprise polyurethane, polycaprolactone, polylactic acid and the like with good biocompatibility. Wherein, the toughness and elasticity of the polycaprolactone and the polylactic acid are insufficient, and the difference with the mechanical property of the natural meniscus is increased. The polyurethane material with adjustable molecular structure and controllable material performance is a research hotspot for preparing the meniscus scaffold at present. A degradable polycaprolactone/polyurethane meniscal scaffold manufactured by Orteq corporation, England, has been commercialized (under the trade name Atifit)^®） [Biomaterials, 35, 3527-3540, 2014]. Song et al prepared hydroxyapatite hybrid polyurethane type scaffolds with bioactive surfaces, adjusted their mechanical properties by controlling their porosity and used as meniscal grafts [ Macromolecular Materials and Engineering, 304, 2019]. However, none of the above materials allow for precise control of scaffold pore structure and customization of a meniscus scaffold of a specific three-dimensional shape for a patient, and the recent advent of 3D printing technology has made possible the fabrication of artificial meniscus material and precise control of microstructure. At present, there are some patent reports of meniscal scaffolds prepared by 3D printing (chinese patent CN 105013011a, CN 110478527 a), but the prepared meniscal scaffold has great mechanical property difference with natural menisci and has low printing precision.

Disclosure of Invention

The invention aims to provide a photocuring 3D printing preparation method for a long-term replacement polyurethane tissue engineering meniscus scaffold. The tissue engineering meniscus prepared by the invention has good biocompatibility, in vivo experiments prove that the tissue engineering meniscus has good in vivo stability, and the prepared meniscus bracket has mechanical properties similar to those of natural meniscus. In addition, the method adopts photocuring 3D printing with the highest printing precision, the prepared meniscus support has a smooth surface, and the meniscus support with a specific three-dimensional structure and a porous structure can be customized.

The invention provides a photocuring 3D printing preparation method of a tissue engineering meniscus scaffold capable of being replaced for a long time, which comprises the following steps:

1) preparing photosensitive resin, wherein the photosensitive resin comprises the following raw material components in parts by weight:

10-80 parts of urethane acrylate shown as formula I

20-80 parts of acrylate monomer

0.5-5.0 parts of free radical type photoinitiator

0.001-0.5 part of defoaming agent;

2) 3D printing the photosensitive resin obtained in the step 1) to obtain a meniscus model blank;

3) and cleaning and post-curing the meniscus model blank to obtain the tissue engineering meniscus scaffold capable of being replaced for a long time.

In step 1) of the above method, the urethane acrylate has a structure represented by formula I:

formula I

In the formula I, R₁Specifically, the alkyl group may be any of an isophorone group, a hexamethylene group, and a dicyclohexylmethane group.

R₂Specifically, the compound can be one or more of ethyl, propyl and n-butyl.

R₃In particular hydrogen or methyl.

n is a natural number of 10 to 100.

The urethane acrylate shown in the formula I can be prepared according to the following steps:

s1: in the presence of a catalyst, carrying out polycondensation reaction on diisocyanate and polyether glycol to obtain isocyanate group-terminated polyurethane resin; the structural formula of the diisocyanate is shown as formula II:

formula II wherein R₁Is as defined in formula I, said polyether diol having the formula shown in formula III:

formula III wherein R₂N is defined as formula I; the catalyst can be tertiary amine or organic metal catalyst, such as one or a combination of more of triethylene diamine, trimethyl hydroxyethyl propane diamine, dibutyltin dilaurate, stannous octoate and the like; the reaction temperature can be 20-100 ℃, and the reaction time can be 2-14 h;

s2: reacting the prepared isocyanate group-terminated polyurethane resin with hydroxyethyl methacrylate or hydroxyethyl acrylate, and adding hydroquinone serving as a polymerization inhibitor during the reaction to obtain methacrylate or acrylate-terminated polyurethane resin (formula I); the reaction temperature of the reaction can be 50-100 ℃, and the reaction time can be 2-8 h.

In step 1), the acrylate monomer is one or a combination of hydroxyethyl acrylate, isobornyl acrylate, diethylene glycol diacrylate, dipropylene glycol diacrylate, (ethoxylated) trimethylolpropane triacrylate and pentaerythritol tetraacrylate.

In step 1) of the above method, the radical photoinitiator may be one or a combination of several of cleavage photoinitiators such as benzil derivatives, α -hydroxyalkylphenones, acylphosphorus oxide compounds, and the like.

Wherein the benzil derivative is specifically DMPA (alpha, alpha-dimethoxy-alpha-phenylacetophenone), alpha-hydroxyalkylphenones, specifically 1173 (2-hydroxy-2-methyl-1-phenylpropanone) or 184 (1-hydroxycyclohexylphenylketone), and acylphosphorus oxide compound is specifically TPO (2, 4, 6-trimethylbenzoyldiphenylphosphineoxide).

In the step 1) of the method, the defoaming agent is one or a combination of BYK-053N, BYK-055, BYK-057 and BYK-061.

In the step 1) of the method, the preferable raw material components of the photosensitive resin comprise the following components in parts by weight:

50-60 parts of urethane acrylate shown as formula I

35-50 parts of acrylate monomer

0.5 to 5 parts (preferably 1 to 3 parts) of a radical photoinitiator

0.001-0.5 part (preferably 0.01-0.1 part) of defoaming agent.

According to one embodiment of the invention, the photosensitive resin comprises the following raw material components in parts by weight:

polyurethane acrylate PU-160 parts, hydroxyethyl acrylate 34 parts, trimethylolpropane triacrylate 5 parts,

1 part of 2-hydroxy-2-methyl-1-phenyl acetone and 1 part of BYK-0570.01 part.

According to still another embodiment of the present invention, the photosensitive resin comprises the following raw material components in parts by weight:

urethane acrylate PU-255 parts, hydroxyethyl acrylate 34 parts, trimethylolpropane triacrylate 9 parts,

2 parts of 2-hydroxy-2-methyl-1-phenyl acetone and BYK-0610.05 parts.

polyurethane acrylate PU-350 parts, isobornyl acrylate 20 parts, hydroxyethyl acrylate 18 parts, trihydroxy acrylic triacrylate 10 parts, 2,4, 6-trimethylbenzoyl diphenyl phosphorus oxide 2 parts, and BYK-0550.1 parts.

The preparation method of the photosensitive resin in the step 1) of the method comprises the following steps:

a) weighing polyurethane acrylate, an acrylate monomer, a free radical photoinitiator and a defoaming agent according to a ratio;

b) pouring the urethane acrylate, the acrylate monomer, the free radical photoinitiator and the defoaming agent into a stirrer in sequence, and stirring and uniformly mixing at a low speed to obtain transparent photosensitive resin; wherein the rotating speed of the low-speed stirring is 400 r/min, and the stirring time is 2-4 h.

In the step 3), the cleaning may be performed on the meniscus model blank by using ethanol or isopropanol, and specifically may be: and (3) putting the meniscus model blank into ethanol or isopropanol, and ultrasonically cleaning for 10 min.

In step 3), the post-curing process is as follows: and (3) curing for 30 min in an ultraviolet box by adopting ultraviolet light, then putting the ultraviolet box into a drying oven with the temperature of 60-100 ℃ and curing for 0.5-3 h.

The method may further comprise the step of sterilizing the resulting long-term replaceable tissue engineered meniscal scaffold.

The tissue engineering meniscus scaffold capable of being replaced for a long time prepared by the method also belongs to the protection scope of the invention.

In addition, the photosensitive resin provided above also belongs to the protection scope of the present invention.

Compared with the prior art, the invention has the following advantages:

(1) the meniscus scaffold provided by the invention has good biocompatibility and in-vivo stability;

(2) the meniscus support provided by the invention has mechanical properties similar to those of natural meniscus;

(3) the meniscus support is prepared by adopting a photocuring 3D printing mode, the printing precision is high, and the three-dimensional structure and the porous structure of the meniscus support can be accurately controlled.

Drawings

Fig. 1 shows a meniscus support model fabricated by Digital Light Processing (DLP) 3D printing according to the present invention.

Fig. 2 shows the mechanical tensile curve of the meniscus scaffold printed and fabricated in example 8.

FIG. 3 shows a graph of the tensile modulus of the print-fabricated meniscal scaffolds of example 8 compared to the tensile modulus of the natural meniscus.

FIG. 4 shows a photograph of a confocal laser scanning microscope observing the staining of the cell core matrix;

FIG. 5 shows a 3 month post-operative MRI T2 image of a polyurethane meniscus of the present application implanted in a rabbit knee joint;

FIG. 6 shows a photograph of the dissected rabbit after 3 months of implantation.

FIG. 7 shows photographs of pathological HE staining of synovial tissue of joints around scaffolds.

Detailed Description

The present invention will be described below with reference to specific examples, but the present invention is not limited thereto.

The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, materials and the like used in the following examples are commercially available unless otherwise specified.

Example 1 urethane acrylate (R) of formula I₁= hexamethylene; r₂Ethyl, n-butyl; r₃Synthesis of = hydrogen)

In a 250 mL round-bottomed flask equipped with mechanical stirring, a nitrogen inlet tube, a thermometer and a dropping funnel, 16.8 g of Hexamethylene Diisocyanate (HDI) was added, and then a mixture of 40.0 g of polyethylene glycol diol (molecular weight 2000) and 90.0 g of polytetrahydrofuran diol (molecular weight 3000) was dropwise added to the three-necked flask while maintaining the temperature of the reaction system in the flask at 60 ℃. After the addition was complete, the mixture was allowed to react for 5 h. Adding catalyst stannous octoate (600 ppm of the mass of the reactant) into the system to promote the reaction to fully occur. The extent of reaction was monitored by Fourier infrared and when the characteristic infrared absorption peak of the isocyanate groups did not decrease any more, a mixture of 0.1 g of hydroquinone and 13.9 g of hydroxyethyl acrylate was added dropwise to the system, during which the temperature of the system was maintained at 50 ℃. After the dropwise addition is finished, the reaction is continued until the characteristic absorption peak of the isocyanate group in the infrared spectrogram completely disappears, and the polyurethane acrylate is obtained, wherein the designed number average molecular weight is 2179 and is named as PU-1.

Example 2 urethane acrylate (R) of formula I₁= hexamethylene; r₂= isopropyl, n-butyl; r₃Synthesis of = hydrogen)

In a 250 mL round-bottomed flask equipped with mechanical stirring, a nitrogen inlet tube, a thermometer and a dropping funnel, 16.8 g of Hexamethylene Diisocyanate (HDI) was added, and then a mixture of 30.0 g of polypropylene glycol (molecular weight 1000) and 60.0 g of polytetrahydrofuran diol (molecular weight 2000) was dropwise added to the three-necked flask while maintaining the temperature of the reaction system in the flask at 90 ℃. After the addition was complete, the mixture was allowed to react for 2 h. Adding catalyst stannous octoate (600 ppm of the mass of the reactant) into the system to promote the reaction to fully occur. The extent of reaction was monitored by Fourier infrared and when the characteristic infrared absorption peak of the isocyanate groups did not decrease any more, a mixture of 0.1 g of hydroquinone and 9.2 g of hydroxyethyl acrylate was added dropwise to the system while maintaining the temperature of the system at 60 ℃. After the dropwise addition is finished, the reaction is continued until the characteristic absorption peak of the isocyanate group in the infrared spectrogram completely disappears, and the polyurethane acrylate is obtained, wherein the designed number average molecular weight is 2902 and is named as PU-2.

Example 3 urethane acrylate (R) of formula I₁= isofexol group; r₂Ethyl, n-butyl; r₃Synthesis of = methyl)

In a 250 mL round-bottomed flask equipped with mechanical stirring, a nitrogen inlet, a thermometer and a dropping funnel, 22.2 g of isophorone diisocyanate (IPDI) was charged, and then a mixture of 60.0 g of polyethylene glycol diol (molecular weight 2000) and 60.0 g of polytetrahydrofuran diol (molecular weight 2000) was dropwise added to the three-necked flask while maintaining the temperature of the reaction system in the flask at 100 ℃. After the addition was complete, the mixture was allowed to react for 2 h. Adding catalyst stannous octoate (600 ppm of the mass of the reactant) into the system to promote the reaction to fully occur. The extent of reaction was monitored by Fourier infrared and when the characteristic infrared absorption peak of the isocyanate groups did not decrease any more, a mixture of 0.1 g of hydroquinone and 10.4 g of hydroxyethyl methacrylate was added dropwise to the system while maintaining the temperature of the system at 70 ℃. And after the dropwise addition is finished, continuing the reaction until the characteristic absorption peak of the isocyanate group in the infrared spectrogram completely disappears, thus obtaining the polyurethane acrylate, wherein the designed number average molecular weight is 3815 and the name is PU-3.

Example 4 preparation of photosensitive resin

Firstly, weighing the following raw materials according to the formula proportion: the photosensitive resin comprises the following raw material components in parts by weight:

polyurethane acrylate PU-160 parts

34 parts of hydroxyethyl acrylate

Trimethylolpropane triacrylate 5 parts

1 part of 2-hydroxy-2-methyl-1-phenyl acetone

BYK-0570.01 parts

Then, the components are poured into a stirrer in sequence, and are stirred and mixed uniformly at a low speed to obtain transparent photosensitive resin, wherein the stirring speed is 400 r/min, and the stirring time is 4 hours.

Example 5 preparation of photosensitive resin

polyurethane acrylate PU-255 parts

34 parts of hydroxyethyl acrylate

Trimethylolpropane triacrylate 9 parts

2 parts of 2-hydroxy-2-methyl-1-phenyl acetone

BYK-0610.05 parts

Example 6 preparation of photosensitive resin

polyurethane acrylate PU-350 parts

20 parts of isobornyl acrylate

18 portions of hydroxyethyl acrylate

Trihydroxy acrylic triacrylate 10 parts

2 parts of 2,4, 6-trimethylbenzoyldiphenyl phosphorus oxide

BYK-0550.1 parts

Then, the components are poured into a stirrer in sequence, and are stirred and mixed uniformly at a low speed to obtain transparent photosensitive resin, wherein the stirring speed is 400 r/min, and the stirring time is 3 hours.

Examples 7-9 printing of meniscal scaffolds

Step 1, model printing is carried out on the photosensitive resin prepared in the embodiment 4-6 by using DLP 3D printing equipment, and the surface of the model is smooth and the fineness is high; the printing parameters of the 3D printer are set as required, and in embodiments 7-9 of the application, the printing parameters of the 3D printer are the same.

And 2, removing the support of the sample blank, putting the sample blank into ethanol, performing ultrasonic treatment for 10 min, then putting the sample blank into an ultraviolet box, curing for 15 min, finally putting the sample blank into an oven at 80 ℃, and curing for 1 h to finally obtain the meniscus support.

Mechanical property evaluation of the meniscal scaffold.

The mechanical properties of the printed meniscal scaffolds were evaluated according to the GB/T2567-2008 test standard including tensile strength, elongation at break and tensile modulus.

	Example 7	Example 8	Example 9
				Tensile Strength (MPa)	23.5	26.2	24.2
Elongation at break	274.4%	297.1%	280.0%
				Tensile modulus (MPa)	79.8	96.2	85.8

Fig. 1 is a meniscus scaffold model fabricated by Digital Light Processing (DLP) 3D printing.

Fig. 2 is a mechanical tensile curve of the meniscus scaffold printed and fabricated in example 8.

FIG. 3 is a graph comparing the tensile modulus of a print-fabricated meniscal scaffold of example 8 of the present invention to the tensile modulus of a natural meniscus, showing a mechanical tensile modulus similar to that of a natural meniscus.

Fig. 4 shows that in vitro cells (rabbit adipose-derived mesenchymal stem cells) were planted on the meniscal scaffolds provided in example 8, and the staining detection of the cell core skeleton was performed by confocal laser microscopy, which indicates that the cells were well attached to the surface of the material provided by the present invention and the cell skeleton was fully extended on the surface of the material.

FIG. 5 shows the photographs of nuclear magnetic resonance T2 images of a 3-month post-operative meniscus implanted in a rabbit knee joint, showing (arrows) the structural integrity of the meniscal scaffold in the joint space, no apparent destruction of the articular cartilage, and ingrowth of autologous tissue around the scaffold, in accordance with example 8.

FIG. 6 is a 3-month anatomical photograph after the implantation of the meniscus support in a rabbit body, wherein the medial femoral condyle cartilage and the medial tibial plateau cartilage have no obvious osteoarthritis and are in good condition after the 3-month operation, which shows that the meniscus support plays a good role in protecting the articular cartilage.

FIG. 7 is a photograph of pathological HE staining of synovial tissue of joints around a scaffold, and the result shows that the synovial membrane is in a good state and has no obvious inflammatory reaction.

The tissue engineering meniscus prepared by the method has good biocompatibility and in-vivo stability, the mechanical property of the tissue engineering meniscus is similar to that of a natural meniscus, the printing precision is high, and the meniscus scaffold with a specific three-dimensional structure and a porous structure can be customized.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. a photosensitive resin for preparing a long-term alternative tissue engineering meniscus support, the raw material components of the photosensitive resin are as follows in parts by weight:

50~60 parts of urethane acrylate shown in formula I

Acrylate monomer 35~50 parts

Free radical photoinitiator 0.5~5 parts

Defoamer 0.001~0.5 part

Formula I

In formula I, R ₁ is any one in isophorone group, hexamethylene;

R ₂ is one or more of ethylene, isopropylidene and n-butylene;

R ₃ is hydrogen or methyl;

n is a natural number from 10 to 100;

The acrylate monomer is one or more combinations of hydroxyethyl acrylate, isobornyl acrylate and trimethylolpropane triacrylate;

The free-radical photoinitiator is a combination of one or more of benzil derivatives, α-hydroxyalkyl phenones or acyl phosphorus oxide compounds;

The defoamer adopts one or more compositions of BYK-053N, BYK-055, BYK-057 and BYK-061;

The urethane acrylate shown in the formula I is selected from the urethane acrylate prepared in any of the following 1)-3):

1) In a 250 mL round-bottomed flask equipped with mechanical stirring, nitrogen introduction tube, thermometer and dropping funnel, add 16.8 g of hexamethylene diisocyanate, then 40.0 g of polyethylene glycol glycol with a molecular weight of 2000 and 90.0 g of The mixture with a molecular weight of 3000 polytetrahydrofuran diol was added dropwise to the three-necked flask, and the temperature of the reaction system in the flask was kept at 60 °C; after the addition was completed, the mixture continued to react for 5 h; Promote the reaction to fully occur; the degree of reaction is monitored by Fourier transform infrared spectroscopy. When the infrared characteristic absorption peak of the isocyanate group is no longer reduced, a mixture of 0.1 g of hydroquinone and 13.9 g of hydroxyethyl acrylate is added dropwise to the system. Keep system temperature 50 DEG C; After dripping is completed, continue to react, until the characteristic absorption peak of isocyanate group in the infrared spectrum disappears completely, namely obtains urethane acrylate, and the design number-average molecular weight is 2179, named PU-1;

2) In a 250 mL round-bottomed flask equipped with mechanical stirring, nitrogen introduction tube, thermometer and dropping funnel, add 16.8 g of hexamethylene diisocyanate, then 30.0 g of polypropylene glycol with a molecular weight of 1000 and 60.0 g of a molecular weight of The mixture of 2000 polytetrahydrofuran diol was added dropwise to the three-necked flask, and the temperature of the reaction system in the flask was kept at 90 ° C; Fully occurred; the degree of reaction was monitored by Fourier transform infrared. When the characteristic absorption peak of isocyanate group infrared no longer decreased, then the mixture of 0.1 g hydroquinone and 9.2 g hydroxyethyl acrylate was added dropwise to the system, and the system was maintained during the period. Temperature 60 DEG C; After dripping is completed, continue to react, until the characteristic absorption peak of isocyanate group in the infrared spectrum disappears completely, promptly obtains urethane acrylate, and the design number-average molecular weight is 2902, named PU-2;

3) In a 250 mL round-bottomed flask equipped with mechanical stirring, nitrogen introduction tube, thermometer and dropping funnel, add 22.2 g of isophorone diisocyanate, then 60.0 g of polyethylene glycol glycol with a molecular weight of 2000 and 60.0 g of The molecular weight is that the mixture of 2000 polytetrahydrofuran diols is added dropwise to the there-necked flask, and the temperature of the reaction system in the flask is maintained at 100 ° C now; after the addition is complete, the mixture continues to react for 2 h; Prompt the reaction to fully occur; the degree of reaction is monitored by Fourier transform infrared spectroscopy. When the infrared characteristic absorption peak of isocyanate group is no longer reduced, a mixture of 0.1 g hydroquinone and 10.4 g hydroxyethyl methacrylate is added dropwise to the system. , the system temperature was maintained at 70 °C during the period; after the dropwise addition was completed, the reaction was continued until the characteristic absorption peak of the isocyanate group in the infrared spectrum completely disappeared, i.e., the urethane acrylate was obtained, and the design number-average molecular weight was 3815, named PU-3 .

2. The application of the photosensitive resin of claim 1 in the preparation of a long-term replaceable tissue engineering meniscus scaffold.

3. A light-curing 3D printing preparation method of a long-term alternative tissue engineering meniscus scaffold, comprising the following steps:

1) preparing the photosensitive resin of claim 1;

2) 3D printing the photosensitive resin in step 1) to obtain a meniscus model blank;

3) cleaning and post-curing the meniscus model blank to obtain the long-term replaceable tissue engineering meniscus support;

In the step 1), the preparation method of the photosensitive resin is as follows:

a) Weigh urethane acrylate, acrylate monomer, free-radical photoinitiator and defoamer according to the proportion;

b) Pour the urethane acrylate, acrylate monomer, free-radical photoinitiator and defoamer into the mixer in turn, and stir and mix at low speed to obtain a transparent photosensitive resin; wherein, the rotation speed of low-speed stirring is 400 r /min, stirring time is 2~4 h.

4 . The method according to claim 3 , wherein: in the step 3), the meniscus model blank is cleaned with ethanol or isopropanol; the specific cleaning method is: the meniscus model blank The body was placed in ethanol or isopropanol and ultrasonically cleaned for 10 min.

5. The method according to claim 3, characterized in that: in the step 3), the post-curing process is: post-curing with ultraviolet light for 30 min in an ultraviolet oven, and then put it in a 60-100 ºC oven Post-curing for 0.5-3 h.

6. The long-term replaceable tissue-engineered meniscus scaffold prepared by the method of any one of claims 3-5.