CN114053479B - Preparation method of bionic artificial blood vessel based on self-healing hydrogel - Google Patents
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Abstract
The invention relates to the technical field of tissue engineering, in particular to a preparation method of a bionic artificial blood vessel based on self-healing hydrogel. The method comprises the following steps: forming dynamic covalent bonds in the hydrogel to prepare a self-healing hydrogel material; preparing a self-healing hydrogel material into a bionic artificial blood vessel preform; and curing the bionic artificial blood vessel preform to obtain the bionic artificial blood vessel. In order to solve the problems of unclosed structure, time-consuming and high-cost preparation process, poor reprocessing capability, inconformity of the artificial blood vessel structure and the autologous blood vessel structure and the like in the conventional preparation of the bionic artificial blood vessel, the invention provides a preparation method of the bionic artificial blood vessel based on self-healing hydrogel, and a simple, convenient and efficient method for preparing and reprocessing the closed artificial blood vessel based on the self-healing hydrogel.
Description
Technical Field
The invention relates to the technical field of tissue engineering, in particular to a preparation method of a bionic artificial blood vessel based on self-healing hydrogel.
Background
The health of blood vessels plays an important role in maintaining normal physiological activities of the human body. According to the latest statistics of the world health organization, cardiovascular diseases are seriously threatening human health as one of the diseases with the highest morbidity and mortality worldwide. Compared with medical treatment, the blood vessel transplantation is more effective in treating blood vessel failure caused by cardiovascular diseases. The primary blood vessel transplantation operation uses autologous blood vessels, but the source and quality of the autologous blood vessel transplantation are difficult to guarantee in some cases, and the requirements of the operation cannot be completely met. In recent years, biomimetic artificial blood vessels have become the first alternative treatment for many cardiovascular diseases.
At present, researchers have developed various methods for preparing biomimetic artificial blood vessels. For example, a bacterial cellulose membrane is curled for three times by a mould curling method, and after freeze drying and mould removal, a hollow tubular structure with three layers of pipe walls is obtained, and different types of cells can be loaded in each layer of pipe walls (adv. healthcare mater, 2017,1601343). The construction of artificial blood vessels can also be achieved by spontaneous crimping using scaffold materials with deformability (adv. funct. mater.,2018,1801027). It can be summarized that the bionic artificial blood vessel obtained by artificial curling or spontaneous curling is not closed, and the inner structure and the outer structure of the artificial blood vessel wall obtained based on the curling method are consistent, and have a larger difference with the real structure of the autologous blood vessel with compact outside and loose and porous inside. The above disadvantages severely limit the practical application value of these artificial blood vessels. On the other hand, the electrospinning technology can easily convert the solution of polymer or natural raw material into the closed tubular structure material, but the method takes a long time to prepare the artificial blood vessel, and the obtained artificial blood vessel has poor mechanical properties (Acta biometer, 2014,10, 2739-2749). Moreover, the preparation of multilayered vascular prostheses using electrospinning techniques requires multiple spinning processes or further combination with other preparation techniques (such as phase separation, 3D printing, etc.), which are more complicated (Acta Biomat, 2010,6, 110-. In addition, Wanghong (patent number: 201611189284.9) of Qingdao Sandi Biotechnology Ltd utilizes a new 3D printing technology to prepare the three-layer bionic artificial blood vessel, but the technology has high requirements on printing equipment and operation process, has high preparation cost and is not beneficial to wide application. Therefore, it is urgently needed to develop a simple and low-cost preparation and processing method of the closed artificial blood vessel, and simultaneously make the bionic artificial blood vessel have the structure and performance which are comparable to those of the autologous blood vessel.
Disclosure of Invention
In order to solve the problems of unclosed structure, time-consuming and high-cost preparation process, poor reprocessing capability, inconformity of the artificial blood vessel structure and the autologous blood vessel structure and the like in the conventional preparation of the bionic artificial blood vessel, the invention provides a preparation method of the bionic artificial blood vessel based on self-healing hydrogel, and a simple, convenient and efficient method for preparing and reprocessing the closed artificial blood vessel based on the self-healing hydrogel.
One of the technical schemes of the invention is a preparation method of a bionic artificial blood vessel based on self-healing hydrogel, which comprises the following steps:
(1) forming dynamic covalent bonds in the hydrogel to prepare a self-healing hydrogel material;
(2) preparing a self-healing hydrogel material into a bionic artificial blood vessel preform;
(3) and curing the bionic artificial blood vessel preform to obtain the bionic artificial blood vessel.
Further, in the step (1), the matrix of the hydrogel is any one of sodium alginate, chitosan, gelatin, cellulose, bacterial cellulose and lignin, and the dynamic covalent bond is any one of borate bond, disulfide bond, imine bond, acylhydrazone bond, Diels-Alder reversible covalent bond and the like;
further, the self-healing hydrogel material is prepared into a bionic artificial blood vessel preform by adopting a mould curling method in the step (2);
further, in the step (3), the curing treatment is specifically a metal cation aqueous solution soaking crosslinking curing treatment.
In the technical scheme of the invention, the dynamic covalent bond has the following functions: the hydrogel and the self-healing function are formed, and in the curing treatment process, metal cations and hydroxyl groups in the hydrogel matrix have chelation to form a new network structure, so that the curing treatment is realized. Therefore, in the invention, 1. the hydrogel matrix is not limited, as long as a dynamic covalent bond can be introduced, and a large number of hydroxyl groups can be provided (sodium alginate, chitosan, gelatin, cellulose, bacterial cellulose and lignin are all satisfied); 2. the variety of dynamic covalent bonds is not limited, and the preparation of the closed artificial blood vessel can be realized by various dynamic covalent bonds; 3. curing the treated solutionWithout limitation, metal cations such as Ca 2+ 、Mg 2+ 、Zn 2+ Divalent metal ion and Fe 3+ 、Al 3+ 、Cr 3+ The trivalent metal ions can realize the curing treatment. Moreover, the dynamic covalent bond is not influenced in the curing process, and only a new network structure is formed.
The process of dynamic covalent bond formation is reversible, involving the breaking and formation of covalent bonds. The phenylboronic acid diol ester bond in the hydrogel is reversible, so that the obtained hydrogel has strong self-healing capability. The dynamic covalent bond in the curing process can be considered to have no influence, but after the curing process, the self-healing capability is influenced due to the newly formed network structure: specifically, the longer the curing time, the lower the self-healing ability. Revascularization, which may be performed prior to curing, followed by curing; or the port is not solidified when being solidified, and the port is solidified after a new blood vessel is formed by healing processing.
Furthermore, the shape of the bionic artificial blood vessel is any one of a linear type, a Y type, a well shape and a tree shape, and the bionic artificial blood vessel is formed by two or more bionic artificial blood vessel prefabricated bodies through contact and self-healing of ports. For example, a port of two bionic artificial blood vessel prefabricated bodies with the same diameter is subjected to contact healing, so that an artificial blood vessel with increased length can be obtained.
Further, the method specifically comprises the following steps:
placing a hydrogel matrix, a coupling agent and dopamine hydrochloride into water, mixing, and stirring in a nitrogen atmosphere to obtain a precursor solution A; placing the hydrogel matrix, the coupling agent and the phenylboronic acid into water, mixing, and stirring in a nitrogen atmosphere to obtain a precursor solution B;
dialyzing the precursor solution A and the precursor solution B, and freeze-drying to obtain a precursor A and a precursor B;
dissolving the precursor A with water, adding the precursor B, uniformly stirring, dropwise adding an alkaline aqueous solution, and stirring to obtain a self-healing hydrogel material;
fourthly, the self-healing hydrogel material is curled on a mould to obtain a bionic artificial blood vessel prefabricated body;
soaking the bionic artificial blood vessel preform in a metal cation aqueous solution, and then demolding to obtain the bionic artificial blood vessel based on the self-healing hydrogel.
The technical scheme of the invention modifies dopamine and 3-aminophenylboronic acid into a sodium alginate structure, forms a dynamic covalent bond by complexing the two precursors under an alkaline condition, prepares a sodium alginate-based self-healing hydrogel material, and quickly, simply and conveniently prepares the self-healing hydrogel into the artificial blood vessel by adopting a mould curling method. Further, the self-healing capability of the artificial blood vessel port and CaCl are combined 2 The surface treatment of the artificial blood vessel by the aqueous solution can realize the reprocessing of the artificial blood vessel.
Further, in the step I, the mass ratio of the hydrogel matrix, the coupling agent and the dopamine hydrochloride in the precursor solution A is 1:0.96:0.44, and the mass ratio of the hydrogel matrix, the coupling agent and the phenylboronic acid in the precursor solution B is 1:0.96: 0.39; the hydrogel matrix is sodium alginate, and the coupling agent is 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride. Before the synthesis of the precursor solution A and the precursor solution B, vacuumizing and introducing nitrogen for protection, and shading treatment is needed when the precursor solution A is synthesized.
Further, in the step (II), dialysis is preferably carried out by using a dialysis bag with the molecular weight cut-off of 3500Da and the size of 15cm multiplied by 3cm, wherein the dialysis time is 5 days, the freeze-drying temperature is-18 ℃, and the freeze-drying time is 4 days.
Further, in the third step, the mass ratio of the hydrogel matrix in the precursor A to the hydrogel matrix in the precursor B is 1:2, and the alkaline aqueous solution is 0.1mol/L sodium hydroxide solution. The NaOH aqueous solution can complex the two precursors, and a phenylboronic acid-catechol dynamic covalent bond is induced to be formed, so that the prepared hydrogel has self-healing property.
Further, in the fourth step, the diameter of the die is 1-20 mm;
furthermore, in the fifth step, the metal cation aqueous solution is a calcium chloride aqueous solution with the concentration of 50mM, and the soaking time is 0-100 s. By controlling the soaking time, the artificial blood vessel with different inner and outer wall structures can be obtained. Specifically, calcium chloride and hydroxyl groups and other groups in the hydrogel matrix are chelated to form a new network structure. The longer the soaking time is, the more compact the outer wall pore structure of the artificial blood vessel is, and the inner wall pore structure hardly changes because the inner wall pore structure is not contacted with the calcium chloride aqueous solution, so that the real blood vessel-like structure with compact outer structure and loose and porous inner part can be obtained.
According to the second technical scheme, the bionic artificial blood vessel is prepared by the preparation method of the bionic artificial blood vessel based on the self-healing hydrogel.
Compared with the prior art, the invention has the beneficial effects that:
the self-healing hydrogel material is prepared by forming dynamic covalent bonds in the hydrogel; then preparing the self-healing hydrogel material into a bionic artificial blood vessel preform; the prepared hydrogel shows good self-healing capability, so that the closed artificial blood vessel can be prepared simply and conveniently by a curling method.
The diameter and the length of the artificial blood vessel prepared by crimping the die can be adjusted by the size of the die.
The closed artificial blood vessel prepared by the invention still has self-healing capability at the port, and can be reprocessed to prepare the artificial blood vessel with the required length.
The artificial blood vessel prepared by the invention can be mixed with CaCl 2 And (3) crosslinking the aqueous solution to ensure that the structure of the outer surface of the blood vessel is compact, and the inner surface of the blood vessel is a loose porous structure to obtain a structure inosculated with the three-layer structure of the autologous blood vessel. The dense structure of the outer surface is beneficial to increasing the mechanical strength of the blood vessel, while the loose porous structure of the inner surface can promote the adhesion, differentiation, material exchange with blood and the like of endothelial cells.
The invention takes natural high molecular material as the hydrogel matrix, which is beneficial to constructing the artificial blood vessel with better biocompatibility, thereby inhibiting possible immunological rejection after transplantation.
The bionic artificial blood vessel based on the self-healing hydrogel can be prepared into artificial blood vessels with standardized length and then assembled to obtain artificial blood vessels with different shapes, so that the standardization and the industrial production of the artificial blood vessels are realized.
Drawings
FIG. 1 shows the preparation of Alg-PBA and Alg-DA as the precursors of sodium alginate-based self-healing hydrogel in example 1 of the present invention 1 H NMR characterization spectrum, wherein a is of Alg-PBA precursor 1 H NMR characterization spectrum, b is of Alg-DA precursor 1 H NMR characterization spectrogram;
FIG. 2 is a photograph of a self-healing hydrogel synthesized from two precursors, Alg-PBA and Alg-DA, in example 1 of the present invention;
FIG. 3 is a demonstration of the self-healing potential of a sodium alginate-based hydrogel of example 1 of the present invention;
FIG. 4 is a photograph showing a bionic artificial blood vessel prepared by crimping a sodium alginate-based hydrogel according to example 1 of the present invention;
FIG. 5 is a photograph showing different diameters and lengths of the bionic artificial blood vessels prepared from the sodium alginate-based hydrogel in example 1 of the present invention, wherein a is the bionic artificial blood vessels with different diameters and b is the bionic artificial blood vessels with different lengths;
fig. 6 is a photo of two bionic artificial blood vessels of the same diameter self-healing at the through-port to obtain a bionic artificial blood vessel in example 1 of the present invention;
FIG. 7 is a CaCl-treated sodium alginate-based hydrogel of example 1 of the present invention 2 SEM photographs after soaking for different times;
FIG. 8 shows a CaCl-crossing of a biomimetic artificial blood vessel in example 1 of the present invention 2 SEM photographs of the outer wall, the inner wall and the section after soaking for different times, wherein a is the SEM photograph of the outer wall, b is the SEM photograph of the inner wall, and c is the SEM photograph of the section;
FIG. 9 is a graph showing the results of the cytotoxicity test of the sodium alginate-based hydrogel in example 1 of the present invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. It is intended that the specification and examples be considered as exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
(1) Synthesis of self-healing hydrogel precursor: weighing 1g of sodium alginate, 0.39g of 3-aminophenylboronic acid and 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride under the conditions of nitrogen atmosphere and dark, placing the weighed materials into 100mL of deionized water, and stirring the materials at room temperature for 24 hours to synthesize Alg-PBA precursor solution; under the conditions of nitrogen atmosphere and dark place, 1g of sodium alginate, 0.44g of dopamine hydrochloride and 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride are weighed, placed in 100mL of deionized water, and stirred at room temperature for 24 hours to synthesize Alg-DA precursor solution.
(2) Respectively placing the Alg-DA precursor solution and the Alg-PBA precursor solution in a dialysis bag (the molecular weight cutoff is 3500Da, the size is 15cm multiplied by 3cm, boiling in 500mL of 2% (w/v) sodium bicarbonate and 1mmol of EDTA (ethylene diamine tetraacetic acid) (about 80 ℃) for 30min, and washing with distilled water for 3 times) for dialysis for 5d, wherein water is required to be changed frequently during dialysis. Transferring into a culture dish after dialysis, freeze drying in a-18 deg.C refrigerator, freeze drying in a freeze drier for 4 days to obtain two precursors of self-healing hydrogel Alg-PBA and Alg-DA (sodium alginate-based self-healing hydrogel precursors Alg-PBA and Alg-DA) 1 The characterization spectrum of H NMR is shown in figure 1, wherein a is of Alg-PBA precursor 1 H NMR characterization spectrum, b is of Alg-DA precursor 1 H NMR characterization spectrum).
(3) Firstly, putting 50mg of Alg-PBA precursor sample obtained in the step (2) into a 5ml beaker, adding 2ml of deionized water, adding 40mg of Alg-DA precursor sample after the Alg-PBA precursor sample is completely dissolved, stirring for two hours at room temperature, then adding 300 mu L of 0.1mol/L NaOH aqueous solution into the beaker, continuously stirring, and forming Alg hydrogel within 30s (the picture of the process of synthesizing the self-healing hydrogel by two precursors of Alg-PBA and Alg-DA is shown in figure 2).
(4) The prepared sodium alginate-based hydrogel is respectively prepared into 3 regular small balls, two small balls are respectively dyed by methylene blue and rhodamine B, the other small ball is not treated, and then the three small balls can automatically heal within 2min without applying external force (the picture for demonstrating the self-healing capacity of the sodium alginate-based hydrogel is shown in figure 3).
(5) And (3) uniformly coating the Alg hydrogel prepared in the step (3) into a hydrogel film with uniform thickness (the thickness is 2mm), wrapping the hydrogel film on polytetrafluoroethylene rods with different diameters (4mm, 6mm, 8mm and 10mm), enabling two ends of the wrapped hydrogel film to be in contact healing, and demolding to obtain the bionic artificial blood vessel preform. The bionic artificial blood vessel preforms with different lengths can be obtained by adjusting the size of the hydrogel film, and the bionic artificial blood vessel preforms with different diameters can be obtained by adjusting the diameter of the polytetrafluoroethylene rod.
(6) Reprocessing of the bionic artificial blood vessel: preparing two bionic artificial blood vessel preforms with the same diameter according to the method in the step (5), and performing contact healing on one port openings of the two bionic artificial blood vessel preforms to obtain a bionic artificial blood vessel preform with increased length.
(7) Inner and outer wall treatment of the bionic artificial blood vessel: soaking the artificial blood vessel wrapped on the polytetrafluoroethylene rod in the steps (5) and (6) in 50mM CaCl 2 Taking out the artificial blood vessels from the water solution (soaking time is 20s, 40s and 60s), demolding to obtain the bionic artificial blood vessels with different inner and outer wall structures, and taking a process photograph as shown in figure 4 and a prepared bionic artificial blood vessel photograph with different diameters and lengths as shown in figure 5, wherein a is the bionic artificial blood vessels with different diameters, and b is the bionic artificial blood vessels with different lengths, soaking the bionic artificial blood vessels in 50mM CaCl after contact healing in the step (6) 2 The photo of the bionic artificial blood vessel obtained in the aqueous solution is shown in FIG. 6. SEM analysis of the bionic artificial blood vessels obtained at different soaking times showed that the results are shown in FIGS. 7-8. In FIG. 8, a is an SEM photograph of the outer wall, b is an SEM photograph of the inner wall, and c is a sectional SEM photograph. As can be seen from FIG. 8, the outer wall and the inner wall of the artificial blood vessel are in CaCl 2 The water solution has loose and porous structure before soaking, and Ca after soaking 2+ And the gel and hydroxyl and other groups in the molecular structure of the sodium alginate in the gel are chelated to form a new network structure, so that the outer wall of the artificial blood vessel becomes more compact. The inner wall of the artificial blood vessel is not mixed with CaCl 2 In direct contact without significant structural change. The variation trend of the structure can be further confirmed by SEM pictures of the section of the artificial blood vessel. Thus by simple CaCl 2 The outer wall structure is compact, the inner wall is loose and porous, and the structure is matched with the structure of the real artificial blood vessel.
(8) Testing cytotoxicity of the bionic artificial blood vessel: freeze-drying the Alg hydrogel prepared in step (3), cutting into small pieces with the size of 2 x 1mm, soaking in water for 2 days, and replacing every 6 hours to removeAnd (5) sub-water. After sterilization, the cells were placed in a 96-well plate at 3X 10 3 Cell seeding was performed at a density of individual cells/well using mouse fibroblasts (NIH/3T3), NIH/3T3 cells in DMEM medium containing 10% fetal bovine serum FBS + 1% penicillin-streptomycin, and control group with 100. mu.L DMEM medium. At 37 deg.C, 5% CO 2 After 0 and 24 hours of incubation in an incubator saturated with humidity, proliferation of NIH/3T3 cells was measured by MTT (Sigma) colorimetry. The results are shown in FIG. 9. Fig. 9 can conclude that the number of cells corresponding to the Alg hydrogel sample increases significantly when the incubation time is from 0h to 24h, and is slightly higher than the control group, indicating that the hydrogel has no significant cytotoxicity.
Example 2
The difference from the embodiment 1 is that: sodium carboxymethyl cellulose (CMC) is used in the synthesis process of the self-healing hydrogel precursor. And (3) synthesizing a CMC-PBA precursor: 1g of sodium carboxymethylcellulose, 0.39g of 3-aminophenylboronic acid and 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride are weighed and placed in a 250ml round-bottom flask, 100ml of deionized water is added, and stirring is carried out at room temperature for 24 hours under the protection of nitrogen. Synthesizing a CMC-DA precursor: 1g of sodium carboxymethylcellulose, 0.44g of dopamine hydrochloride and 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride are weighed and placed in a 250ml round-bottom flask, 100ml of deionized water is added, and stirring is carried out at room temperature for 24 hours under the protection of nitrogen. And the reaction needs to be carried out under protection from light. After the reaction is finished, the obtained sample needs to be dialyzed. Directly pouring the reaction solution into a dialysis bag, dialyzing in a beaker for 5 d. And after dialysis is finished, pouring the sample into a culture dish, putting the culture dish into a refrigerator at the temperature of 18 ℃ below zero for freezing and shaping, and then putting the sample into a freeze drier for freeze drying for 4 days to obtain two precursors of the self-healing hydrogel, namely a CMC-PBA precursor and a CMC-DA precursor.
The CMC-PBA precursor and the CMC-DA precursor samples are prepared into the bionic artificial blood vessel by adopting the same steps as the example 1.
The bionic artificial blood vessel prepared in the embodiment 1 is subjected to the same scanning electron microscope analysis and self-healing detection, and the result shows that the bionic artificial blood vessel has similar properties to the artificial blood vessel prepared in the embodiment 1.
Example 3
The difference from the embodiment 1 is that: sodium alginate and polyvinyl alcohol (PVA) are used in the synthesis process of the self-healing hydrogel precursor. Synthesizing an Alg-PBA precursor: weighing 0.39g of sodium alginate, 0.96g of 3-aminophenylboronic acid and 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride under the conditions of nitrogen atmosphere and light shielding, placing the mixture in 100mL of deionized water, and stirring the mixture at room temperature for 24 hours to synthesize an Alg-PBA precursor solution; after the reaction is finished, the obtained sample needs to be dialyzed. Directly pouring the reaction solution into a dialysis bag, dialyzing in a beaker for 5 d. And after the dialysis is finished, pouring the sample into a culture dish, putting the culture dish into a refrigerator at the temperature of-18 ℃ for freezing and shaping, and then putting the sample into a freeze dryer for freeze drying for 4 days to obtain the self-healing hydrogel precursor Alg-PBA.
Putting 50mg of the Alg-PBA precursor sample obtained in the step (1) into a 5mL beaker, adding 2mL of deionized water, adding 1mL of PVA aqueous solution with the mass fraction of 10% after the sample is completely dissolved, stirring for two hours at room temperature, adding 300 mu L of 0.1mol/L NaOH aqueous solution into the beaker, and continuously stirring for 30 seconds to form the Alg-PVA hydrogel.
The PVA contains a large amount of hydroxyl groups in the molecule, has similarity with Alg-DA synthesized in example 1, and therefore, the PVA can still be used for preparing self-healing hydrogel, and the gel forming principle is the same and is based on borate bond formation.
The same procedure as in example 1 was used to prepare a biomimetic artificial blood vessel from the Alg-PVA hydrogel.
The bionic artificial blood vessel prepared in the embodiment 1 is subjected to the same scanning electron microscope analysis and self-healing detection, and the result shows that the bionic artificial blood vessel has similar properties to the artificial blood vessel prepared in the embodiment 1.
In further experimental verification, it is found that in the present invention, 1, the hydrogel matrix is not limited, as long as a dynamic covalent bond can be introduced, and a large number of hydroxyl groups can be provided (sodium alginate, chitosan, gelatin, cellulose, bacterial cellulose and lignin are all satisfied); 2. the variety of dynamic covalent bonds is not limited, and the preparation of the closed artificial blood vessel can be realized by various dynamic covalent bonds(ii) a 3. Solution for solidification treatment is not limited, Ca 2+ 、Mg 2+ 、Zn 2+ Divalent metal ion and Fe 3+ 、Al 3+ 、Cr 3+ The trivalent metal ions can realize the curing treatment.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A preparation method of a bionic artificial blood vessel based on self-healing hydrogel is characterized by comprising the following steps:
(1) forming dynamic covalent bonds in the hydrogel to prepare a self-healing hydrogel material;
(2) preparing a self-healing hydrogel material into a bionic artificial blood vessel preform;
(3) curing the bionic artificial blood vessel preform to obtain the bionic artificial blood vessel;
in the step (1), the matrix of the hydrogel is any one of sodium alginate, chitosan, gelatin, cellulose, bacterial cellulose and lignin, and the dynamic covalent bond is any one of borate bond, disulfide bond, imine bond, acylhydrazone bond and Diels-Alder reversible covalent bond; grafting the dynamic covalent bonds onto the matrix of the hydrogel by means of a coupling agent which is 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride;
preparing the self-healing hydrogel material into a bionic artificial blood vessel preform by adopting a mould curling method in the step (2);
in the step (3), the curing treatment is specifically a metal cation aqueous solution soaking crosslinking curing treatment;
the metal cation aqueous solution is a calcium chloride aqueous solution with the concentration of 50mM, and the soaking time is 20-100 s.
2. A method for preparing a bionic artificial blood vessel based on a self-healing hydrogel according to claim 1, wherein the shape of the bionic artificial blood vessel is any one of linear, Y-shaped, well-shaped and dendritic, and the bionic artificial blood vessel is formed by contacting and self-healing more than two bionic artificial blood vessel preforms through ports.
3. The preparation method of the bionic artificial blood vessel based on the self-healing hydrogel according to claim 1, which comprises the following steps:
placing a hydrogel matrix, a coupling agent and dopamine hydrochloride into water, mixing, and stirring in a nitrogen atmosphere to obtain a precursor solution A; placing the hydrogel matrix, the coupling agent and the phenylboronic acid into water, mixing, and stirring in a nitrogen atmosphere to obtain a precursor solution B;
dialyzing the precursor solution A and the precursor solution B, and freeze-drying to obtain a precursor A and a precursor B;
dissolving the precursor A with water, adding the precursor B, uniformly stirring, dropwise adding an alkaline aqueous solution, and stirring to obtain a self-healing hydrogel material;
fourthly, the self-healing hydrogel material is curled on a mould to obtain a bionic artificial blood vessel prefabricated body;
soaking the bionic artificial blood vessel preform in a metal cation aqueous solution, and then demolding to obtain the bionic artificial blood vessel based on the self-healing hydrogel.
4. The method for preparing a biomimetic artificial blood vessel based on a self-healing hydrogel according to claim 3, wherein in the step (r), the mass ratio of the hydrogel matrix, the coupling agent and dopamine hydrochloride in the precursor solution A is 1:0.96:0.44, and the mass ratio of the hydrogel matrix, the coupling agent and phenylboronic acid in the precursor solution B is 1:0.96: 0.39; the hydrogel matrix is sodium alginate, and the coupling agent is 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride.
5. A method for preparing a bionic artificial blood vessel based on a self-healing hydrogel according to claim 3, wherein in the second step, the dialysis time is 5 days, the freeze-drying temperature is-18 ℃, and the freeze-drying time is 4 days.
6. A method for preparing a biomimetic artificial blood vessel based on self-healing hydrogel according to claim 3, wherein in the step (iii), the alkaline aqueous solution is 0.1mol/L sodium hydroxide solution.
7. A method for preparing a biomimetic artificial blood vessel based on self-healing hydrogel according to claim 3, wherein in the step (iv), the diameter of the mold is 1-20 mm.
8. A biomimetic artificial blood vessel prepared by the preparation method of the biomimetic artificial blood vessel based on self-healing hydrogel according to any one of claims 1-7.
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