阅读说明：本技术 一种硫醇-丙烯酸酯生物材料及其制备方法和应用 (Mercaptan-acrylate biomaterial and preparation method and application thereof ) 是由 张耀明 何文扬 王齐华 王廷梅 周栋 顾浩 周晏仪 于 2021-07-08 设计创作，主要内容包括：本发明提供了一种硫醇-丙烯酸酯生物材料及其制备方法和应用,属于生物材料技术领域。将环糊精和PCL混合进行开环聚合,得到c-PCL-(21)-OH聚合物；将c-PCL-(21)-OH聚合物进行丙烯酰化改性,得到末端带有双键的丙烯酸酯(c-PCL-(21)-C＝C丙烯酸酯聚合物)；将所述末端带有双键的丙烯酸酯、硫醇、光引发剂和有机溶剂混合,进行点击化学反应,得到硫醇-丙烯酸酯生物材料。本发明提供的硫醇-丙烯酸酯生物材料可用于3D打印方式进行智能制造,硫醇末端硫基因其硫基反应性强,具有广泛的生物相容性,在生物医学领域具有广泛的应用前景。(The invention provides a mercaptan-acrylate biomaterial as well as a preparation method and application thereof, belonging to the technical field of biomaterials. Mixing cyclodextrin and PCL for ring-opening polymerization to obtain c-PCL 21 -an OH polymer; c-PCL 21 the-OH polymer is subjected to acryloyl modification to obtain the acrylate (c-PCL) with double bonds at the tail end 21 -C ═ C acrylate polymers); the acrylate, the mercaptan and the photoinitiator with double bonds at the tail ends are addedMixing with organic solvent, and carrying out click chemical reaction to obtain the mercaptan-acrylate biomaterial. The mercaptan-acrylate biomaterial provided by the invention can be used for intelligent manufacturing in a 3D printing mode, and the mercaptan-terminal sulfur gene has strong sulfur-based reactivity and wide biocompatibility, and has wide application prospects in the field of biomedicine.)

1. A preparation method of a thiol-acrylate biomaterial is characterized by comprising the following steps:

mixing cyclodextrin and caprolactone to carry out ring-opening polymerization reaction to obtain a 21-star-arm polymer;

performing acryloyl modification on the 21 star-arm polymer to obtain acrylate with double bonds at the tail end;

and mixing the acrylate with the double bond at the tail end, the mercaptan, the photoinitiator and the organic solvent, and carrying out click chemical reaction to obtain the mercaptan-acrylate biomaterial.

2. The method according to claim 1, wherein the molar ratio of cyclodextrin to caprolactone is 1: 20 to 100.

3. The method according to claim 1, wherein the ring-opening polymerization is carried out at a temperature of 120 to 140 ℃ for 20 to 24 hours.

4. The preparation method of claim 1, wherein the acrylation modification is that the 21 star arm polymer, the acid-binding agent and the modifying agent are mixed for chain extension reaction.

5. The preparation method according to claim 1 or 4, wherein the temperature of the acrylation modification is 0-5 ℃ and the time is 18-24 h.

6. The method according to claim 1, wherein the molar ratio of the thiol to the double bond in the acrylate having a double bond at the terminal is 1:1 to 5.

7. The method according to claim 1 or 6, wherein the molar ratio of the thiol to the double bond in the acrylate having a double bond at the terminal is 1:2 to 3.

8. The method according to claim 1, wherein the click chemistry reaction is carried out for a time of 10 to 20 seconds.

9. The thiol-acrylate biomaterial prepared by the preparation method of any one of claims 1 to 8.

10. Use of the thiol-acrylate biomaterial of claim 9 for the preparation of a vascular tissue engineering biomaterial.

Technical Field

The invention relates to the technical field of biological materials, in particular to a mercaptan-acrylate biological material and a preparation method and application thereof.

Background

Cardiovascular disease remains a major cause of death worldwide, with over 1000 million people suffering from these life-threatening diseases. Vascular disease afflicts up to 20% of elderly patients, rendering them unable to function properly, and diseased vessels often require surgery, treatment with balloons, stents, patches, artificial blood vessels, and the like. The intravascular stent is still in a continuous experimental development stage, still faces a plurality of difficult problems which can not be solved clinically in the aspects of radial supporting force, long-term patency rate, fracture resistance, foreign body reaction, approach damage and the like, and has the problems of easy occurrence of restenosis in the stent, low long-term patency rate and the like. In the research process of the stent, the main factors restricting the development of the stent are the selection of materials and the preparation process. Autologous vascular patches are the best choice for repairing blood vessels because they are non-immunogenic and have the same mechanical properties as the recipient's blood vessels. However, due to the patient's medical complications (e.g., illness and previous surgery), the use of autologous patches may be limited in terms of quality and source: also artificial blood vessels based on materials such as expanded polytetrafluoroethylene and polyethylene terephthalate may cause postoperative failure because they are prone to calcification, inflammatory reactions and thrombosis. The traditional metal and high polymer materials have more or less disadvantages. Therefore, it is important to develop a new bioactive material for cardiovascular tissue engineering.

The photo-induced thiol-acrylate system provides the characteristics of photopolymerization and click reaction, and has the advantages of space and time controllability, high selectivity, insensitivity to oxygen and water, and the like. Compared to conventional photopolymerization, thiol-acrylate photocuring systems overcome the common problems of insufficient curing and high shrinkage stress and can produce highly uniform crosslinked networks. Thiol-acrylate chemistry generally exhibits fast reaction rates, is not sensitive to ambient oxygen, and is simple to perform and high in yield. But the prior thiol-acrylate system has the problem of low biological activity.

Disclosure of Invention

In view of the above, the present invention aims to provide a thiol-acrylate biomaterial, a preparation method thereof and an application thereof. The thiol-acrylate biomaterial prepared by the invention has high bioactivity.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a preparation method of a mercaptan-acrylate biomaterial, which comprises the following steps:

mixing cyclodextrin and caprolactone to carry out ring-opening polymerization reaction to obtain a 21-star-arm polymer;

performing acryloyl modification on the 21 star-arm polymer to obtain acrylate with double bonds at the tail end;

and mixing the acrylate with the double bond at the tail end, the mercaptan, the photoinitiator and the organic solvent, and carrying out click chemical reaction to obtain the mercaptan-acrylate biomaterial.

Preferably, the molar ratio of cyclodextrin to caprolactone is 1: 20 to 100.

Preferably, the temperature of the ring-opening polymerization reaction is 120-140 ℃ and the time is 20-24 h.

Preferably, the acrylation modification is to mix the 21 star arm polymer, the acid-binding agent and the modifying agent to carry out chain extension reaction.

Preferably, the temperature of the acryloyl modification is 0-5 ℃, and the time is 18-24 h.

Preferably, the molar ratio of the mercaptan to the double bond in the acrylate with the double bond at the terminal is 1: 1-5.

Preferably, the molar ratio of the mercaptan to the double bond in the acrylate with the double bond at the terminal is 1: 2-3.

Preferably, the time of the click chemistry reaction is 10-20 s.

The invention also provides the mercaptan-acrylate biomaterial prepared by the preparation method in the technical scheme.

The invention also provides the application of the thiol-acrylate biomaterial in the technical scheme in the preparation of the vascular tissue engineering biomaterial.

The invention provides a preparation method of a mercaptan-acrylate biomaterial, which comprises the following steps: mixing cyclodextrin and Caprolactone (CL) to carry out ring-opening polymerization reaction to obtain 21 star-arm polymer (c-PCL)₂₁-OH polymers); performing acryloyl modification on the 21 star-arm polymer to obtain acrylate (c-PCL) with double bonds at the tail end₂₁-C ═ C acrylate polymers); and mixing the acrylate with the double bond at the tail end, the mercaptan, the photoinitiator and the organic solvent, and carrying out click chemical reaction to obtain the mercaptan-acrylate biomaterial. In the present invention, PCL is a crystalline material composed of a biodegradable polymer, has excellent flexibility and processability, has biocompatibility and biodegradability, and is approved by the FDA for therapeutic use in humans; the cyclodextrin molecule has a slightly tapered hollow cylindrical three-dimensional ring structure, and in the hollow structure, the upper end (larger opening end) of the outer side is composed of secondary hydroxyl groups of C2 and C3, and the lower end (smaller opening end) is composed of primary hydroxyl groups of C6, and has a hydrophilic propertyA hydrophobic region is formed in the cavity under the shielding action of a C-H bond, and the hydrophobic core is matched with the molecular size of most organic drugs, so that an ideal drug loading action site can be provided; the mercaptan is an ester product with excellent stability, the pyrolysis gravimetric analysis result shows that the mercaptan starts to be slowly decomposed until the temperature reaches 400 ℃, the thermal stability is high, the volatility is very small, the oxidation resistance, the crosslinking degree reduction and the antistatic effect are realized, and the sulfur element exists in a human body and participates in the chemical combination reaction of the human body, so the mercaptan has certain biocompatibility. The invention utilizes cyclodextrin and caprolactone ring-opening polymerization reaction to synthesize CD and PCL copolymer with 21 star arms, greatly improves the mechanical property of the material, modifies the end of caprolactone to enable the caprolactone to be solidified, adds mercaptan, utilizes click chemical reaction between mercaptan and acrylate to prepare mercaptan/cyclodextrin-caprolactone acrylate polymer through mercaptan-acrylate photopolymerization, namely mercaptan-acrylate biomaterial, and compared with the traditional acrylate double-bond reaction, the mercaptan-acrylate click chemical reaction can inhibit the aggregation of active oxygen, can reduce the photocuring time and oxygen inhibition, has more excellent bioactivity and mechanical strength, and the prepared mercaptan-acrylate biomaterial has low resistance to oxygen and even no resistance to oxygen, can be quickly solidified in ultraviolet light, can form a biological material with good physical and chemical structure network and excellent biocompatibility, and can be applied to vascular tissue engineering.

The invention also provides the mercaptan-acrylate biomaterial prepared by the preparation method in the technical scheme, the mercaptan-acrylate biomaterial provided by the invention can be used for intelligent manufacturing in a 3D printing mode, and the mercaptan-terminal sulfur gene has strong sulfur-based reactivity and wide biocompatibility and has wide application prospect in the field of biomedicine. The invention evaluates the biological activity of the thiol-acrylate biomaterial by performing in-vitro cell biocompatibility experiments and mouse subcutaneous implantation experiments, and proves the potential of the thiol-acrylate biomaterial as a vascular tissue engineering material.

Drawings

FIG. 1 shows c-PCL₂₁-C ═ C polymer structural formula;

FIG. 2 shows PETMP/c-PCL₂₁-structural formula of DA thiol/acrylate biomaterial;

FIG. 3 is a graph of cell viability values of venous endothelial cells of the prepared thiol/acrylate biomaterial;

FIG. 4 is an under-the-erythrocyte contrast chart of the prepared thiol/acrylate biomaterial;

FIG. 5 is a platelet adhesion profile of the thiol/acrylate biomaterial prepared;

FIG. 6 shows tissue proliferation after subcutaneous implantation of the prepared thiol/acrylate biomaterial in mice;

fig. 7 is a photograph of HE stained tissue sections of the prepared thiol/acrylate biomaterial subcutaneously implanted.

Detailed Description

The invention provides a preparation method of a mercaptan-acrylate biomaterial, which comprises the following steps;

mixing cyclodextrin and caprolactone to carry out ring-opening polymerization reaction to obtain a 21-star-arm polymer;

performing acryloyl modification on the 21 star-arm polymer to obtain acrylate with double bonds at the tail end;

and mixing the acrylate with the double bond at the tail end, the mercaptan, the photoinitiator and the organic solvent, and carrying out click chemical reaction to obtain the mercaptan-acrylate biomaterial.

The invention mixes cyclodextrin and Caprolactone (CL) to carry out ring-opening polymerization reaction to obtain 21 star-arm polymer (c-PCL)₂₁-OH polymer).

In the present invention, the molar ratio of cyclodextrin to caprolactone is preferably 1: 20-100, more preferably 1: 40-100, most preferably 1: 60-80. In the present invention, the cyclodextrin is preferably β -cyclodextrin. In the present invention, the cyclodextrin is preferably dried under vacuum at 60 ℃ for 24 hours before use to remove moisture sufficiently.

In the invention, the temperature of the ring-opening polymerization reaction is preferably 120-140 ℃, and more preferably 12 DEG CThe time is preferably 20-24 hours at 5-135 ℃, and more preferably 21-23 hours. In the ring-opening polymerization reaction process, the caprolactone is subjected to ring opening and is polymerized with cyclodextrin to obtain 21 star-arm polymer c-PCL₂₁-an OH polymer;

in the present invention, the polymerization reaction is preferably carried out under the conditions of an organotin catalyst, no water and a protective atmosphere. The specific type and amount of the organotin catalyst used in the present invention are not particularly limited, and those known to those skilled in the art may be used.

In the present invention, the c-PCL₂₁The number average molecular weight of the-OH polymer is preferably 5000 to 80000, more preferably 30000 to 60000.

After obtaining the 21 star arm polymer, the invention carries out acryloyl modification on the 21 star arm polymer to obtain the acrylate (c-PCL) with double bonds at the tail end₂₁-C ═ C polymer).

In the invention, the acrylation modification is preferably to mix the 21 star arm polymer, the acid-binding agent and the modifying agent for chain extension reaction.

In the present invention, the 21 star arm polymer is preferably added in the form of a solution, i.e., the c-PCL is preferably added in the present invention₂₁After dissolution of the-OH Polymer, the c-PCL obtained₂₁Mixing the-OH polymer solution with an acid binding agent and a modifier, and carrying out an acrylation modification reaction at 0-5 ℃ in a protective atmosphere to obtain the c-PCL₂₁-C ═ C polymer.

In the present invention, the dissolution of c-PCL₂₁The solvent for the-OH polymer preferably comprises one or more of dichloromethane, chloroform, THF, toluene, acetonitrile, dichloroethane, trichloroethane, tetrachloroethane, pentachloroethane and hexachloroethane.

In the present invention, the c-PCL₂₁The mass concentration of the-OH polymer solution is preferably 10-30%, and more preferably 20%.

In the present invention, the modifier is preferably an acryl-based organic substance, and more preferably includes one or more of methacryloyl chloride, acryloyl chloride, methacryloyl bromide, and methacrylic anhydride; the acid-binding agent preferably comprises one or more of triethylamine, pyridine, N-diisopropylethylamine and 4-dimethylaminopyridine.

In the present invention, the c-PCL₂₁The molar ratio of hydroxyl, acid-binding agent and modifying agent in the-OH polymer is preferably 1: 2-6: 2-6, more preferably 1: 3-5: 3 to 5.

In the present invention, the protective atmosphere is preferably nitrogen.

In the present invention, the c-PCL₂₁The method for mixing the-OH polymer solution, the acid-binding agent and the modifying agent is preferably as follows: under a protective atmosphere, adding the c-PCL₂₁Adding an acid binding agent into the-OH polymer solution, and stirring for 20-40 min under the ice bath condition of 0-5 ℃ to obtain a mixed solution; and dropwise adding a modifier into the mixed solution to perform an acrylation modification reaction.

In the invention, the temperature of the acryloyl modification is preferably 0-5 ℃, the time is preferably 18-24 h, and the time of the acryloyl modification reaction is calculated after the modifier is added; in the process of the acryloyl modification reaction, the modifier is reacted with c-PCL₂₁Reaction of-OH polymer to produce modified c-PCL₂₁And (3) reacting the-C ═ C polymer with hydrochloric acid, and reacting the hydrochloric acid with an acid binding agent to form a salt, so as to promote the reaction.

After the reaction of the acylation modification is completed, the invention also preferably carries out post-treatment on the obtained reaction liquid of the acylation modification, and the post-treatment preferably comprises the following steps:

sequentially filtering and washing the acryloyl modified reaction solution, and precipitating in glacial ethyl ether; vacuum drying the obtained precipitate to obtain the c-PCL₂₁-C ═ C polymer, said C-PCL₂₁The structure of the-C ═ C polymer is shown in fig. 1.

In the invention, the washing is preferably deionized water, stirred and washed for 2-3 times to remove salts generated in the reaction.

After obtaining the acrylate with double bonds at the tail end, the invention mixes the acrylate with double bonds at the tail end, the mercaptan, the photoinitiator and the organic solvent to carry out click chemical reaction to obtain the mercaptan-acrylate biomaterial (PETMP/c-PC)L₂₁-DA thiol/acrylate biomaterials). PETMP/c-PCL obtained by the invention₂₁The structural formula of the-DA thiol/acrylate biomaterial is shown in figure 2.

In the invention, the molar ratio of the mercaptan to the double bond in the acrylate with the double bond at the terminal is 1: 1-5, and more preferably 1: 2-3.

In the present invention, the organic solvent preferably includes one or more of dichloromethane, trichloromethane, DMF, DMSO, tetrahydrofuran, acetone, 1, 4-dioxahexane, toluene, trichloroethane, and dichloroethane.

In the invention, the acrylic ester with double bonds at the tail end is preferably mixed with an organic solvent, and then the mercaptan and the photoinitiator are added.

In the invention, the mass percentage of the polymer solution obtained by mixing the acrylate with the double bond at the terminal and the organic solvent is preferably 40-70%.

In the present invention, the mercaptan preferably includes one or more of 3, 6-dioxa-1, 8-octane dithiol, pentaerythritol tetrakis-3-mercaptopropionate, and trimethylolpropane tris-3-mercaptopropionate, and more preferably pentaerythritol tetrakis-3-mercaptopropionate.

In the present invention, the photoinitiator preferably comprises one or more of benzil bismethyl ether (DMPA, 651), 2-hydroxy-2-methyl-1-phenyl-1-propanone (HMPP, 1173), 4-isobutylphenyl-4' -methylphenyliodohexafluorophosphate (250), 2-hydroxy-2-methyl-1- [4- (2-hydroxyethoxy) phenyl ] -1-propanone (2959) and bis (2,4, 6-trimethylbenzoyl) phenylphosphine oxide (819); the mass of the photoinitiator is preferably 0.5-5%, more preferably 1.5-3% of that of the acrylate with double bonds at the terminal.

The invention has no special requirements on the mixing method and can ensure uniform mixing.

In the invention, the time of the click chemistry reaction is preferably 10-20 s, and the temperature is preferably room temperature, i.e. no additional heating or cooling is required.

In the present invention, the c-PCL₂₁Light generation in click chemistry of-C ═ C polymers with thiolsAnd (3) curing, wherein the photocuring is not inhibited by oxygen, the crosslinking degree of the cured product is reduced, and the high toughness and tensile strength are still maintained after long-term storage.

The invention also provides the mercaptan-acrylate biomaterial prepared by the preparation method in the technical scheme.

The invention also provides the application of the thiol-acrylate biomaterial in the technical scheme in the preparation of the vascular tissue engineering biomaterial.

In the invention, the application preferably uses the thiol-acrylate biomaterial to make 3D printing ink, and uses 3D direct-writing printing equipment to print vascular stents and patch materials which can be used for vascular tissue engineering.

In the invention, the blood vessel stent is preferably printed on the receiving rotating shaft when being printed, and the printing preferably adopts a 25-gauge needle or a 27-gauge needle; the number of printed layers is preferably 2-4, and the height of a single layer is preferably 0.1-0.35 mm; the printing speed is preferably 2-5 mm/s.

In the invention, the material of the receiving rotating shaft is preferably copper, silver, alloy, ceramic, silica gel or polytetrafluoroethylene; the diameter of the receiving rotating shaft is preferably less than or equal to 6mm, more preferably 3-6 mm, and the inner diameter of the receiving rotating shaft is controlled by selecting the corresponding receiving shaft.

After printing, the invention preferably performs photocuring on the obtained blood vessel stent blank under the irradiation of ultraviolet light to obtain the blood vessel stent for the blood vessel tissue engineering. In the invention, the wavelength of the ultraviolet light is preferably 254nm or 365nm, and the intensity is preferably 8-20 mW/cm²More preferably 10-15 mW/cm²。

In the present invention, the printing of the biological patch material is preferably performed on a common platform, and the printing is preferably performed using a 27-gauge needle or a 30-gauge needle; the surrounding mode is surrounding, the number of printed layers is preferably 2-4, and the height of a single layer is preferably 0.1-0.25 mm; the printing speed is preferably 1-2 mm/s.

After printing, the blood vessel patch blank is preferably subjected to photocuring under ultraviolet irradiation to obtain the biological patch material for blood vessel tissue engineering. In the invention, the wavelength of the ultraviolet light is preferably 254nm or 365nm, and the intensity is preferably 8-20 mW/cm²More preferably 10-15 mW/cm²。

In the invention, the biological patch has a porous structure, and is beneficial to filling and growing of cell tissues.

In the present invention, when applied, the thiol-acrylate biomaterial is preferably mixed with a drug as a drug-loaded system; the content of the medicine in the medicine carrying system is preferably 100-120 mu g/cm². The invention has no special requirement on the type of the medicine, and can adopt clinically common medicines which are well known to the technical personnel in the field, such as paclitaxel, rapamycin, sirolimus, everolimus, quinoa alcohol, heparin and the like. In the invention, the thiol-acrylate shape memory polymer has a hydrophobic inner cavity brought by a cyclodextrin structure, the size of the inner cavity is suitable for the molecular size of most clinical drugs, and an ideal action site can be provided to include the drugs to form a stable host-guest inclusion compound, so that the drug loading and controlled release performance is met, and the drug loading function of the material is realized.

In the invention, the thiol-acrylate biomaterial has good mechanical property, drug loading property and biocompatibility; the epoxy resin has low resistance to oxygen and even no resistance to oxygen, can be rapidly cured in ultraviolet light, and can form a network with excellent physical and mechanical properties and biocompatibility; the intelligent manufacturing can be carried out by a 3D printing mode, the structures such as a bracket, a biological patch and the like for vascular tissue engineering are printed, the sulfur-based reactivity of the sulfur gene at the tail end of the mercaptan is strong, the biocompatibility is wide, and the application prospect is wide in the field of biomedicine.

To further illustrate the present invention, the thiol-acrylate biomaterials provided by the present invention, their preparation and use, are described in detail below with reference to examples, which should not be construed as limiting the scope of the invention.

Example 1

The ultraviolet light fast photocuring mercaptan/acrylate material is prepared through the following steps:

(1) drying beta-cyclodextrin (beta-CD) at 60 deg.C in vacuum for 24 hr to remove water completely;

(2) weighing 1.134g of dried beta-cyclodextrin (beta-CD) and 167.84g of epsilon-caprolactone (epsilon-CL) and adding the mixture into a three-neck flask, mixing and then carrying out ultrasonic homogenization for 2 h;

(3) the three-neck flask is equipped with a mechanical stirrer, stannous octoate 4 drops are added into the prepolymerization mixture at 130 ℃ and N₂Stirring and polymerizing for 24 hours under protection to obtain a transparent viscous polymer solution;

(4) dissolving the polymer solution obtained in the step (3) in dichloromethane, precipitating in glacial ethyl ether, repeatedly washing twice to remove unreacted monomers, and vacuum drying at 35 ℃ for 24h to obtain c-PCL₂₁-OH polymer powder; the number average molecular weight was determined to be 56263.62.

(5) In a three-neck flask, 11.25g of c-PCL is added₂₁-OH polymer powder and methylene chloride (10% solids) in N₂Adding 1.70g of triethylamine under protection, and magnetically stirring for 30min at 4 ℃ in an ice bath; slowly adding 1.76g of methacryloyl chloride into the mixed solution drop by drop, and continuing to add N₂Carrying out modification reaction for 24 hours under protection to obtain turbid modified polymer solution;

(6) filtering the turbid modified polymer solution, washing to remove impurities, precipitating in glacial ethyl ether, and vacuum drying at 35 ℃ for 24h to obtain white c-PCL₂₁-C ═ C polymer powder;

(7) subjecting the obtained c-PCL₂₁Washing the-C polymer powder with deionized water twice repeatedly to remove salts generated in the reaction, and drying in vacuum at 45 ℃ for 24 hours to obtain an acrylate polymer;

(8) dissolving 2g of acrylate polymer powder in chloroform with a solid content of 50%, adding 80.72ug of tetrapentaerythritol tetra-3-mercaptopropionate, adding 20mg of PPO (1% by mass of acrylate polymer powder) as a photoinitiator, and mixing uniformly at a wavelength of 365mm and a concentration of 10mW/cm²Curing for 10s under the ultraviolet light with light intensity, drying in an oven at 40 ℃ for 2 hours,the thiol-acrylate biomaterial was obtained and subjected to tensile testing with the following results: tensile property data are: elastic modulus (MPa): 96.01 ± 22.41; breaking point (%): 665.98 +/-79.31; tensile strength (MPa): 13.04 plus or minus 2.38; the thiol-acrylate shape memory biomaterial has good tensile resistance.

Example 2

The acrylate polymer was the same as in example 1;

dissolving 2g of acrylate polymer in chloroform, wherein the solid content is 60%, adding 80.72ug of tetrapentaerythritol tetra-3-mercaptopropionate, adding 20mg of PPO (1% by mass of the acrylate polymer) as a photoinitiator, uniformly mixing, filling the obtained printing ink into a charging barrel, and printing the support on a receiving rotating shaft by using a 3D printing technology: a 27-gauge needle head is adopted, the printing speed is 3mm/s, 2 layers are printed, the height of a single layer is 0.35mm, a receiving rotating shaft is made of ceramic, and the diameter of the receiving rotating shaft is 5 mm; then 10mW/cm at 365mm wavelength²And (3) curing for 10s under ultraviolet light with light intensity to obtain the 3D printing mercaptan/acrylate biological blood vessel stent with the inner diameter of 5mm, the length of 20mm and the mass of 60 mg.

The printed vascular stent is tested for biocompatibility according to ISO10993-12 standard: HUVECs at 1X 10⁴The density of individual cell wells-1 was seeded onto a 96-well plate rack and cell viability and proliferation quantified using CCK-8. At specific times (days 1, 3, 5), the cell-seeded discs were incubated in 10% CCK-8 at 37 ℃ in the presence of 5% CO₂Was cultured in the atmosphere of (2.5) for 2.5 hours. Until the cell proliferation rate reaches 80-90%. Then, the medium was removed and the test extract (100 μ L) was added. Mixing the obtained mixture with 5% CO₂Incubate at 37 ℃ for 12 hours in a humid atmosphere. Then, the test extract was removed and tetrazolium salt (MTT) solution (20. mu.L) was added. Mixing the obtained mixture with 5% CO₂Incubate for 4 hours at 37 ℃ in a humid atmosphere. The MTT solution was then removed and dimethyl sulfoxide (150. mu.L) was added. The resulting mixture was shaken for 10 minutes at 37 ℃. The absorbance of each well was measured at 450nm using a microplate reader (MultiskanFC, Thermo Electron corporation, MA, USA). All measurements were run 8 times.

The degree of cytotoxicity can be determined by the following formula from the measured optical density values:

wherein RGR is the relative growth rate of the cell. D_t、D_pcAnd D_ncOD values for test samples, positive controls and negative controls, respectively. The positive control is culture medium containing human endothelial cells, and the negative control is blank culture medium. The results are shown in FIG. 3. As can be seen from FIG. 3, the thiol-acrylate intravascular stent obtained in the example has excellent endothelial cell biocompatibility, and the viability values of the cultured human umbilical vein endothelial cells on days 1, 3 and 5 are all above 95%.

Example 3

The acrylate polymer was the same as in example 1;

dissolving 2g of acrylate polymer in chloroform, wherein the solid content is 60%, adding 80.72ug of tetrapentaerythritol tetra-3-mercaptopropionate, adding 20mg of PPO (1% by mass of the acrylate polymer) as a photoinitiator, uniformly mixing, filling the obtained printing ink into a charging barrel, and printing the vascular patch material on a common platform by using a 3D printing technology.

Example 4

Basically the same as the example 3, except that the everolimus drug is added into the mixed ink, and the content is 100-120 mu g/cm². The printed patch biomaterial was subjected to hemolysis test according to astm f756-17, the structure of which is shown in fig. 4, and the materials of example 3(a) and example 4(b) were normal in red blood cell number and morphology compared to the positive control (c) and the negative control (d), indicating that the patch biomaterial had good blood compatibility, and the hemolysis absorbance values were tested, and the results are shown in table 1, and the hemolysis rate of example 3 was calculated to be 0.9%, and the hemolysis rate of example 4 was calculated to be 1.25%, demonstrating that the material is a non-hemolytic material.

Table 1 absorbance data for examples 3 and 4 (n ═ 3)

Example 3	Example 4	Hemolysis of blood	Has no hemolysis
				0.044	0.047	0.360	0.040
0.040	0.038	0.354	0.038
				0.039	0.042	0.359	0.037

Example 5

Essentially the same as example 3, except that square pieces of material 1cm by 2cm in size and 0.1mm apart, optionally with 50. mu.g/cm everolimus, were printed on a 3D printing platform²。

The adhesion results of different parts of the material are shown in figure 5, the material surface has no obvious platelet adhesion, and the material is proved not to cause thrombosis, and is subjected to subcutaneous implantation and pathological section experiments with different magnification, the results are shown in figures 6 and 7, the figure 6 shows that a large amount of cell tissues grow on the material surface, the figure 7 shows that the HE section is full of normal blood vessel tissues, and the material is proved to have good biocompatibility and potential as blood vessel tissue engineering.

The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.