CN111956868A - Small-caliber composite multilayer artificial blood vessel and preparation method thereof - Google Patents
Small-caliber composite multilayer artificial blood vessel and preparation method thereof Download PDFInfo
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- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
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Abstract
The invention discloses a small-caliber composite multilayer artificial blood vessel and a preparation method thereof. The chitosan/gelatin composite gel coating is prepared by preparing a chitosan/gelatin mixed solution as an electrolyte and depositing chitosan/gelatin composite gel on the surface of a needle electrode by utilizing an electrophoretic deposition technology; then, the composite gel coating is used as a substrate, and a sodium alginate/chitosan outer layer is constructed on the surface of the composite gel coating by utilizing the layer-by-layer self-assembly function to form chitosan/gelatin/sodium alginate composite multilayer gel; after the composite multilayer gel is taken down, the chitosan/gelatin/sodium alginate composite multilayer artificial blood vessel which can simulate the special multilayer structure of the natural blood vessel can be obtained by freeze drying. By the mode, the artificial blood vessel with small caliber and good mechanical property can be quickly and conveniently prepared; and the whole preparation process is easy to regulate and control, and the prepared composite multilayer artificial material has good biocompatibility and high use safety, and can meet the requirements of practical application.
Description
Technical Field
The invention relates to the technical field of artificial blood vessels, in particular to a small-caliber composite multilayer artificial blood vessel and a preparation method thereof.
Background
Vascular prostheses are a replacement for many severely stenotic or occluded blood vessels and play an important role in the repair and treatment of cardiovascular diseases. At present, large-caliber and medium-caliber artificial blood vessels made of synthetic materials such as nylon, terylene, polytetrafluoroethylene and the like are successfully applied to clinic and obtain satisfactory effects, but because small-caliber blood vessels have low blood flow speed and low blood pressure, the synthetic materials cannot support the adhesion and growth of endothelial cells, do not have good compliance, easily cause platelet enrichment, cause the problems of acute thrombus, intimal hyperplasia at anastomotic sites, infection and the like, and are difficult to meet the requirements of practical application. Meanwhile, in the preparation process of the small-caliber artificial blood vessel, the preparation process and the preparation difficulty are multiplied as the dimension is smaller, and the large-scale application is difficult. Therefore, there is still a need for materials and methods suitable for small-bore (diameter < 6mm) vascular prostheses.
In recent years, rapidly developed 3D printing technology and electrospinning technology are gradually applied to the preparation of small-caliber artificial blood vessels. For example, patent publication No. CN106806945A provides a 3D printing preparation method of a small-caliber bioartificial blood vessel and an artificial blood vessel, which successfully prepares a small-caliber bioartificial blood vessel by printing a first stent and a second stent in advance, then printing endothelial cells, assembling the stents, and planting the cells in the stents. Patent publication No. CN111000661A provides a composite artificial blood vessel and a method for preparing the same, in which a first nanofiber membrane and a second nanofiber membrane are prepared by electrospinning, and are combined by winding, casting and crosslinking, so as to form a small-caliber artificial blood vessel having a double-layer tubular structure.
Although the method can successfully prepare the small-caliber composite multilayer artificial blood vessel in a mode of compounding multilayer materials and accurately regulate and control the micro-nano scale structure of the small-caliber composite multilayer artificial blood vessel, the preparation process still has the problems of high equipment requirement, limited material types, long time consumption and the like, and the large-scale market demand is difficult to meet.
In view of the above, there is still a need for a small-caliber composite multilayer artificial blood vessel and a method for preparing the same to solve the above problems.
The preparation method of the small-aperture blood vessel material is developed, can accurately regulate and control the aperture and the internal structure of the blood vessel material, has the advantages of being green, rapid, convenient and fast, and capable of being produced in mass, and has important value and significance for repairing cardiovascular diseases.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention provides a small-caliber composite multilayer vascular prosthesis and a method for preparing the same. The chitosan/gelatin composite gel is deposited on the surface of the needle electrode in an electrophoretic deposition mode, and the sodium alginate/chitosan outer layer is constructed by taking the composite gel coating on the surface of the needle electrode as a substrate and adopting the layer-by-layer self-assembly effect, so that the chitosan/gelatin/sodium alginate composite multilayer artificial blood vessel which can simulate the special multilayer structure of a natural blood vessel and has a small caliber is formed in a rapid and convenient method, and the requirements of practical application are met.
In order to achieve the aim, the invention provides a preparation method of a small-caliber composite multilayer artificial blood vessel, which comprises the following steps:
s1, dissolving chitosan and gelatin in an acid solution, heating and stirring to fully dissolve the chitosan and gelatin to obtain a chitosan/gelatin mixed solution;
s2, taking the chitosan/gelatin mixed solution obtained in the step S1 as electrolyte, constructing a two-electrode system, and performing electrophoretic deposition to deposit the chitosan/gelatin composite gel on the surface of the needle electrode to form a chitosan/gelatin composite gel coating;
s3, cleaning the chitosan/gelatin composite gel coating obtained in the step S2, cooling to room temperature, alternately soaking the chitosan/gelatin composite gel coating in a sodium alginate solution and a chitosan solution, and forming a sodium alginate/chitosan gel layer on the outer surface of the chitosan/gelatin composite gel coating by utilizing a layer-by-layer self-assembly technology to obtain chitosan/gelatin/sodium alginate composite multilayer gel;
and S4, taking the composite multilayer gel obtained in the step S3 from the surface of the needle electrode, removing residual solution in the composite multilayer gel cavity, and performing freeze drying treatment to obtain the small-caliber composite multilayer artificial blood vessel.
Further, in step S1, the chitosan concentration in the chitosan/gelatin mixed solution is 0.5 w/v% -2 w/v%, and the gelatin concentration is 0.1 w/v% -4 w/v%; dissolving the chitosan and the gelatin by hydrochloric acid and adjusting the pH value to 3.0-5.5; the chitosan/gelatin mixed solution does not contain salt ions.
Further, in step S2, the chitosan/gelatin composite gel coat is one or more layers; the number of layers of the chitosan/gelatin composite gel coating is regulated and controlled in a mode of continuous deposition or interruption of electrical signals.
Further, in step S2, the needle electrode is a cathode in the two-electrode system, and is made of stainless steel with a radius of 0.2-3 mm; the anode in the two-electrode system is a platinum wire electrode.
Further, in step S2, the distance between the two electrodes is 1-60 cm, and the immersion depth of the two electrodes in the electrolyte is 1-100 cm; the deposition voltage in the electrophoretic deposition process is 1-10V, the deposition temperature is 37-60 ℃, and the deposition time is 1-30 min.
Further, in step S3, the concentration of the chitosan solution is 0.05 w/v% -4 w/v%, and the concentration of the sodium alginate solution is 0.05 w/v% -4 w/v%.
Further, in step S3, the number of the alternate soaking is 1-100, and the time for each soaking is 0.5-30 min.
Further, in step S1, the temperature in the heating process is 80 to 100 ℃, and the temperature in the stirring process is 37 to 60 ℃.
In order to realize the aim, the invention also provides a small-caliber composite multilayer artificial blood vessel which is prepared according to any one of the technical schemes and comprises a chitosan/gelatin composite gel coating and a sodium alginate/chitosan gel layer coated on the outer surface of the chitosan/gelatin composite gel coating; the sodium alginate/chitosan gel layer is an alternating sodium alginate gel layer and a chitosan gel layer from inside to outside.
Further, the caliber of the small-caliber composite multilayer artificial blood vessel is 0.4-6 mm.
Compared with the prior art, the invention has the beneficial effects that:
1. the chitosan/gelatin composite gel coating is prepared by preparing a chitosan/gelatin mixed solution as an electrolyte and depositing chitosan/gelatin composite gel on the surface of a needle electrode by utilizing an electrophoretic deposition technology; then, the composite gel coating is used as a substrate, and a sodium alginate/chitosan outer layer is constructed on the surface of the composite gel coating by utilizing the layer-by-layer self-assembly function to form chitosan/gelatin/sodium alginate composite multilayer gel; after the composite multilayer gel is taken down, the chitosan/gelatin/sodium alginate composite multilayer artificial blood vessel which can simulate the special multilayer structure of the natural blood vessel can be obtained by freeze drying. The whole preparation process is green, rapid, convenient and fast, and can be produced in mass, and the requirements of practical application can be met.
2. The invention can rapidly construct the tubular gel material with small caliber by carrying out electrophoretic deposition on the surface of the needle electrode, the hollow structure of the tubular gel material can be accurately controlled by the size of the needle electrode, the composite layer number and the thickness of each layer of gel can also be accurately controlled by regulating and controlling relevant parameters in the electrophoretic deposition process, and the whole process is convenient and controllable and is convenient for practical application. Meanwhile, the chitosan/gelatin mixed solution without salt ions is prepared, so that the chitosan gel formed in an electrophoresis state and the gelatin gel formed after cooling are interpenetrated and inserted to form an interpenetrating network structure by utilizing the electronic responsiveness of the chitosan and the temperature responsiveness of the gelatin, and the influence of the salt ions on the chitosan hydrogel structure can be avoided, so that the prepared chitosan/gelatin composite gel coating has higher crystallinity and stronger mechanical property, is easier to completely take down from the surface of an electrode, and solves the problem that the coating material is difficult to take down or is easy to damage when taking down due to poor mechanical property. In addition, the composite gel coating formed by electrophoretic deposition has a porous structure which can provide a three-dimensional porous environment similar to an extracellular matrix, and is beneficial to the attachment and proliferation of cells.
3. According to the invention, the composite gel coating obtained by electrophoretic deposition is alternately soaked in the sodium alginate solution and the chitosan solution, and can be combined through electrostatic interaction by utilizing opposite charges carried by the sodium alginate and the chitosan, so that the layers of self-assembly process not only can form mutually-alternating chitosan/sodium alginate composite layers, but also can wrap the composite gel coating surface obtained by electrophoretic deposition, thereby further improving the mechanical property of the finally prepared artificial blood vessel. Meanwhile, the number of layers of the formed chitosan/sodium alginate composite layer can be regulated and controlled by controlling the alternate soaking times and soaking time in the layer-by-layer self-assembly process, so that the compliance of the prepared artificial blood vessel is regulated, and the problem of poor compliance of the artificial blood vessel in the prior art is solved.
4. The chitosan, the gelatin and the sodium alginate used in the invention are all natural medical polymer materials, and no toxic cross-linking agent is used, so that the prepared artificial blood vessel has good biocompatibility; the gelatin also has the functions of promoting cell adhesion and growth and preventing blood coagulation, so that the prepared artificial blood vessel can induce intima formation after implantation and prevent blood coagulation, thrombus is not easy to generate, and the application safety is high.
5. The invention combines the electrophoretic deposition technology and the layer-by-layer self-assembly technology, can rapidly prepare the small-caliber composite multilayer artificial blood vessel, and has low energy consumption, safe material and no pollution in the whole process; the minimum caliber of the prepared composite multilayer artificial blood vessel can reach 0.4 mm.
Drawings
FIG. 1 is a schematic cross-sectional view of a small-caliber composite multilayer vascular prosthesis provided in accordance with the present invention;
FIG. 2 is a schematic diagram of the preparation of a small-caliber composite multilayer artificial blood vessel by combining an electrophoretic deposition technology and a layer-by-layer self-assembly technology in the invention;
FIG. 3 is a scanning electron microscope image of a small-caliber composite multilayer artificial blood vessel prepared in example 1 of the present invention;
FIG. 4 is a scanning electron micrograph and an optical photograph of a single-layered chitosan/gelatin composite gel tube prepared in example 2 of the present invention;
FIG. 5 is an optical photograph of a multi-layered chitosan/gelatin composite gel tube prepared in example 3 of the present invention;
the parts in the drawings are numbered as follows: 1. chitosan/gelatin composite gel coating; 2. a sodium alginate gel layer; 3. a chitosan gel layer.
Detailed Description
The following detailed description of the preferred embodiments of the present invention, taken in conjunction with the accompanying drawings, will make the advantages and features of the invention easier to understand by those skilled in the art, and thus will clearly and clearly define the scope of the invention. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without any inventive step, are within the scope of the present invention.
The invention provides a preparation method of a small-caliber composite multilayer artificial blood vessel, which comprises the following steps:
s1, dissolving chitosan and gelatin in an acid solution, heating and stirring to fully dissolve the chitosan and gelatin to obtain a chitosan/gelatin mixed solution;
s2, taking the chitosan/gelatin mixed solution obtained in the step S1 as electrolyte, constructing a two-electrode system, and performing electrophoretic deposition to deposit the chitosan/gelatin composite gel on the surface of the needle electrode to form a chitosan/gelatin composite gel coating;
s3, cleaning the chitosan/gelatin composite gel coating obtained in the step S2, cooling to room temperature, alternately soaking the chitosan/gelatin composite gel coating in a sodium alginate solution and a chitosan solution, and forming a sodium alginate/chitosan gel layer on the outer surface of the chitosan/gelatin composite gel coating by utilizing a layer-by-layer self-assembly technology to obtain chitosan/gelatin/sodium alginate composite multilayer gel;
and S4, taking the composite multilayer gel obtained in the step S3 from the surface of the needle electrode, removing residual solution in the composite multilayer gel cavity, and performing freeze drying treatment to obtain the small-caliber composite multilayer artificial blood vessel.
In step S1, the chitosan concentration in the chitosan/gelatin mixed solution is 0.5 w/v% -2 w/v%, and the gelatin concentration is 0.1 w/v% -4 w/v%; dissolving the chitosan and the gelatin by hydrochloric acid and adjusting the pH value to 3.0-5.5; the chitosan/gelatin mixed solution does not contain salt ions.
In step S2, the chitosan/gelatin composite gel coat is one or more layers; the number of layers of the chitosan/gelatin composite gel coating is regulated and controlled in a mode of continuous deposition or interruption of electrical signals.
In step S2, the needle electrode is a cathode in the two-electrode system, and is made of stainless steel with a radius of 0.2-3 mm; the anode in the two-electrode system is a platinum wire electrode.
In step S2, the distance between the two electrodes is 1-60 cm, and the immersion depth of the two electrodes in the electrolyte is 1-100 cm; the deposition voltage in the electrophoretic deposition process is 1-10V, the deposition temperature is 37-60 ℃, and the deposition time is 1-30 min.
In step S3, the concentration of the chitosan solution is 0.05 w/v% -4 w/v%, and the concentration of the sodium alginate solution is 0.05 w/v% -4 w/v%.
In step S3, the number of the alternate soaking is 1-100, and the time for each soaking is 0.5-30 min.
In step S1, the temperature in the heating process is 80 to 100 ℃, and the temperature in the stirring process is 37 to 60 ℃.
The invention also provides a small-caliber composite multilayer artificial blood vessel which is prepared by the preparation method and comprises a chitosan/gelatin composite gel coating and a sodium alginate/chitosan gel layer coated on the outer surface of the chitosan/gelatin composite gel coating; the sodium alginate/chitosan gel layer is composed of an alternating sodium alginate gel layer and a chitosan gel layer from inside to outside in sequence, and the schematic structural diagram is shown in figure 1.
The caliber of the small-caliber composite multilayer artificial blood vessel is 0.4-6 mm.
The small-caliber composite multilayer artificial blood vessel and the preparation method thereof provided by the invention are described below by referring to the examples and the accompanying drawings.
Example 1
The embodiment provides a preparation method of a small-caliber composite multilayer artificial blood vessel, which comprises the following steps:
s1, dissolving chitosan and gelatin in hot water (85 ℃), adjusting the pH value to about 5 by hydrochloric acid, stirring at 37 ℃ to fully dissolve the chitosan and the gelatin, and preparing a chitosan/gelatin mixed solution containing 1 w/v% of chitosan and 0.5 w/v% of gelatin, wherein NaCl or other salt ions are not contained in the chitosan/gelatin mixed solution;
s2, taking the chitosan/gelatin mixed solution obtained in the step S1 as an electrolyte, taking a platinum electrode as an anode and a stainless steel needle-shaped electrode with the radius of 0.2mm as a cathode, and carrying out electrophoretic deposition, wherein the immersion depth of the two electrodes is 1cm, and the distance between the two electrodes is 2 cm; setting the deposition voltage of electrophoretic deposition to be 3V, the deposition temperature to be 37 ℃, the single deposition time to be 1min, continuously depositing for 4 times, and forming a chitosan/gelatin composite gel coating on the surface of the needle electrode;
s3, washing the chitosan/gelatin composite gel coating obtained in the step S2 with deionized water, cooling to room temperature, alternately soaking the chitosan/gelatin composite gel coating in a 0.5 w/v% sodium alginate solution and a 0.5 w/v% chitosan solution for 2min each time for 6 times, and forming a sodium alginate/chitosan gel layer on the outer surface of the chitosan/gelatin composite gel coating by utilizing a layer-by-layer self-assembly technology to obtain chitosan/gelatin/sodium alginate composite multilayer gel;
s4, taking the composite multilayer gel obtained in the step S3 from the surface of the needle electrode, washing the composite multilayer gel with deionized water to remove residual solution in the gel lumen, and then placing the gel lumen at-40 ℃ for vacuum freeze drying for 12 hours to obtain the small-caliber composite multilayer artificial blood vessel.
A schematic diagram of a combined process of electrophoretic deposition and layer-by-layer self-assembly in steps S2-S3 is shown in FIG. 2.
A scanning electron microscope image of the small-caliber composite multilayer artificial blood vessel prepared in step S4 is shown in fig. 3. As can be seen from FIG. 3, the small-caliber composite multilayer artificial blood vessel is a hollow tubular structure, the caliber of the small-caliber composite multilayer artificial blood vessel is about 0.4mm, the thickness of the tube wall is about 160 μm, and the tube wall is of a multilayer structure; meanwhile, the artificial blood vessel still keeps a good circular cavity shape after freeze drying, which shows that the artificial blood vessel has strong mechanical property.
Therefore, the small-caliber composite multilayer artificial blood vessel with the caliber of only 0.4mm is successfully prepared by the embodiment, can simulate the special multilayer structure of a natural blood vessel, has strong mechanical property, and can meet the requirements of practical application.
Examples 2 to 3
In order to further analyze the performance of the chitosan/gelatin composite gel coating prepared in the electrophoretic deposition process, embodiments 2 to 3 respectively provide a preparation method of a chitosan/gelatin composite gel tube, wherein the embodiment 2 prepares a single-layer chitosan/gelatin composite gel tube, and the embodiment 3 prepares a multi-layer chitosan/gelatin composite gel tube.
Specifically, the preparation method of the single-layer chitosan/gelatin composite gel tube provided in example 2 is as follows:
a chitosan/gelatin mixed solution containing 1 w/v% of chitosan and 1 w/v% of gelatin was prepared as an electrolyte in the same manner as in example 1; then taking a platinum wire electrode as an anode and a stainless steel needle-shaped electrode with the radius of 0.2mm as a cathode, and carrying out electrophoretic deposition, wherein the immersion depth of the two electrodes is 1cm, and the distance between the two electrodes is 2 cm; the deposition voltage of electrophoretic deposition is set to be 5V, the deposition temperature is set to be 45 ℃, and the deposition time is set to be 3 min. After the deposition was completed, the surface of the needle electrode was washed with deionized water and cooled to room temperature, and it was found that a chitosan/gelatin composite gel coating was generated on the surface of the needle electrode, as shown in fig. 4 (a). And taking the chitosan/gelatin composite gel coating off the electrode by using forceps to obtain the single-layer chitosan/gelatin composite gel tube.
After the single-layer chitosan/gelatin composite gel tube prepared in example 2 was placed at-40 ℃ for vacuum freeze-drying for 12 hours, the microscopic morphology of the single-layer chitosan/gelatin composite gel tube prepared in example 2 was observed by using a scanning electron microscope and an optical microscope, respectively, to obtain a scanning electron microscope image and an optical photograph thereof, which are respectively shown in (b) and (c) of fig. 4.
It can be observed from (b) in fig. 4 that the gel tube has an interfacial hollow morphology, and the diameter of the gel tube is about 0.4mm, and the thickness of the tube wall is about 100 μm; compared with the shape of the circular cavity still maintained after freeze drying in the embodiment 1, the shape of the cavity of the single-layer chitosan/gelatin composite gel tube prepared in the embodiment 2 is deformed after freeze drying, which shows that the mechanical property of the single-layer chitosan/gelatin composite gel tube prepared only by electrophoretic deposition is insufficient, but the mechanical property of the single-layer chitosan/gelatin composite gel tube can be effectively improved after the surface of the gel tube is coated with the sodium alginate/chitosan gel layer by utilizing the layer-by-layer self-assembly technology, so that the prepared artificial blood vessel has stronger mechanical property, and the requirement of practical application is met.
As can be seen from (c) of fig. 4, the dextran solution with fluorescent label can flow in the single-layered chitosan/gelatin composite gel tube, indicating that it has a continuous cavity inside.
Compared with example 2, the preparation method of the multilayer chitosan/gelatin composite gel tube provided in example 3 is different in that the deposition voltage set during electrophoretic deposition is 3V, the deposition time is 1min each time, the deposition is performed for 5 times continuously, and the rest steps are the same as those in example 2 and are not repeated herein.
The multilayer chitosan/gelatin composite gel tube prepared in example 3 was observed using an optical microscope, and the result is shown in fig. 5. As can be seen from FIG. 5, the multilayer chitosan/gelatin composite gel tube has an interface hollow appearance, the caliber of the tube is about 0.4mm, the thickness of the tube wall is about 120 μm, and the multilayer chitosan/gelatin composite gel tube has a distinct layered structure (about 5 layers); the internal cavity structure of the multilayer chitosan/gelatin composite gel tube can be demonstrated by trapping air bubbles.
Therefore, in the preparation method of the small-caliber composite multilayer artificial blood vessel, the chitosan/gelatin composite gel coating prepared in the electrophoretic deposition process has a good internal cavity structure and can meet the requirement of blood circulation; and the number of the tube wall layers of the composite gel tube can be regulated and controlled by adjusting the continuous deposition times in the electrophoretic deposition process or by interrupting the electric signal so as to meet different application requirements. Meanwhile, the composite gel tube prepared by electrophoretic deposition has insufficient mechanical properties, and the mechanical properties of the composite gel tube can be obviously improved by combining the composite gel tube with a layer-by-layer self-assembly technology, so that the finally prepared artificial blood vessel has stronger mechanical properties to meet the requirements of practical application.
It should be noted that, as will be understood by those skilled in the art, the pH of the acidic solution in step S1 may be adjusted to be 3.0 to 5.5, the gelatin dissolution temperature may be adjusted to be 80 to 100 ℃, and the heating and mixing temperature may be adjusted to be 37 to 60 ℃, so that the chitosan and the gelatin are fully dissolved to form a mixed solution. Meanwhile, in order to regulate and control the performance of the chitosan/gelatin gel and the sodium alginate/chitosan gel, the concentration of the chitosan in the chitosan/gelatin mixed solution prepared in the step S1 can be regulated between 0.5 w/v% and 2 w/v%, and the concentration of the gelatin can be regulated between 0.1 w/v% and 4 w/v%; the concentration of the sodium alginate solution in the step S3 can be adjusted between 0.05 w/v% and 4 w/v%, and the concentration of the chitosan solution can be adjusted between 0.05 w/v% and 4 w/v%.
Similarly, in order to regulate and control the number of layers and the thickness of a chitosan/gelatin composite gel coating formed in the electrophoretic deposition process, relevant parameters of the electrophoretic deposition process can be regulated, wherein the distance between two electrodes can be 1-60 cm, the immersion depth of the electrodes in electrolyte can be 1-100 cm, the deposition voltage can be 1-10V, the deposition temperature can be 37-60 ℃, and the deposition time can be 1-30 min. In order to regulate and control the number of layers and the thickness of each layer of the sodium alginate/chitosan gel layer formed by layer-by-layer self-assembly, the number of alternate soaking times can be regulated within 1-100 times, and the time for each soaking can be 0.5-30 min.
In addition, the caliber of the small-caliber composite multilayer artificial blood vessel prepared by the method can be accurately controlled through the size of the needle electrode, so that the size of the needle electrode can be selected from 0.2-3 mm in order to meet the requirements of artificial blood vessels with different calibers, and the method belongs to the protection range of the method.
In conclusion, the chitosan/gelatin composite gel coating is prepared by preparing a chitosan/gelatin mixed solution as an electrolyte and depositing the chitosan/gelatin composite gel on the surface of the needle electrode by utilizing an electrophoretic deposition technology; then, the composite gel coating is used as a substrate, and a sodium alginate/chitosan outer layer is constructed on the surface of the composite gel coating by utilizing the layer-by-layer self-assembly function to form chitosan/gelatin/sodium alginate composite multilayer gel; after the composite multilayer gel is taken down, the chitosan/gelatin/sodium alginate composite multilayer artificial blood vessel which can simulate the special multilayer structure of the natural blood vessel can be obtained by freeze drying. By the mode, the artificial blood vessel with small caliber and good mechanical property can be quickly and conveniently prepared; and the whole preparation process is easy to regulate and control, and the prepared composite multilayer artificial material has good biocompatibility and high use safety, and can meet the requirements of practical application.
The above description is only for the purpose of illustrating the technical solutions of the present invention and is not intended to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; all the equivalent structures or equivalent processes performed by using the contents of the specification and the drawings of the invention, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. A preparation method of a small-caliber composite multilayer artificial blood vessel is characterized by comprising the following steps:
s1, dissolving chitosan and gelatin in an acid solution, heating and stirring to fully dissolve the chitosan and gelatin to obtain a chitosan/gelatin mixed solution;
s2, taking the chitosan/gelatin mixed solution obtained in the step S1 as electrolyte, constructing a two-electrode system, and performing electrophoretic deposition to deposit the chitosan/gelatin composite gel on the surface of the needle electrode to form a chitosan/gelatin composite gel coating;
s3, cleaning the chitosan/gelatin composite gel coating obtained in the step S2, cooling to room temperature, alternately soaking the chitosan/gelatin composite gel coating in a sodium alginate solution and a chitosan solution, and forming a sodium alginate/chitosan gel layer on the outer surface of the chitosan/gelatin composite gel coating by utilizing a layer-by-layer self-assembly technology to obtain chitosan/gelatin/sodium alginate composite multilayer gel;
and S4, taking the composite multilayer gel obtained in the step S3 from the surface of the needle electrode, removing residual solution in the composite multilayer gel cavity, and performing freeze drying treatment to obtain the small-caliber composite multilayer artificial blood vessel.
2. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1, wherein the method comprises the following steps: in step S1, the chitosan concentration in the chitosan/gelatin mixed solution is 0.5 w/v% -2 w/v%, and the gelatin concentration is 0.1 w/v% -4 w/v%; dissolving the chitosan and the gelatin by hydrochloric acid and adjusting the pH value to 3.0-5.5; the chitosan/gelatin mixed solution does not contain salt ions.
3. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1, wherein the method comprises the following steps: in step S2, the chitosan/gelatin composite gel coat is one or more layers; the number of layers of the chitosan/gelatin composite gel coating is regulated and controlled in a mode of continuous deposition or interruption of electrical signals.
4. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1 or 3, wherein the method comprises the following steps: in step S2, the needle electrode is a cathode in the two-electrode system, and is made of stainless steel with a radius of 0.2-3 mm; the anode in the two-electrode system is a platinum wire electrode.
5. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1 or 4, wherein the method comprises the following steps: in step S2, the distance between the two electrodes is 1-60 cm, and the immersion depth of the two electrodes in the electrolyte is 1-100 cm; the deposition voltage in the electrophoretic deposition process is 1-10V, the deposition temperature is 37-60 ℃, and the deposition time is 1-30 min.
6. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1, wherein the method comprises the following steps: in step S3, the concentration of the sodium alginate solution is 0.05 w/v% -4 w/v%, and the concentration of the chitosan solution is 0.05 w/v% -4 w/v%.
7. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1, wherein the method comprises the following steps: in step S3, the number of the alternate soaking is 1-100, and the time for each soaking is 0.5-30 min.
8. The method for preparing a small-caliber composite multilayer artificial blood vessel according to claim 1, wherein the method comprises the following steps: in step S1, the temperature in the heating process is 80 to 100 ℃, and the temperature in the stirring process is 37 to 60 ℃.
9. A small-caliber composite multilayer artificial blood vessel is characterized in that: the small-caliber composite multilayer artificial blood vessel is prepared by the preparation method of any one of claims 1 to 8 and comprises a chitosan/gelatin composite gel coating and a sodium alginate/chitosan gel layer coated on the outer surface of the chitosan/gelatin composite gel coating; the sodium alginate/chitosan gel layer is an alternating sodium alginate gel layer and a chitosan gel layer from inside to outside.
10. The small-caliber composite multilayer prosthesis according to claim 9, wherein: the caliber of the small-caliber composite multilayer artificial blood vessel is 0.4-6 mm.
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