Preparation method of artificial blood vessel and artificial blood vessel

Abstract

The invention provides a preparation method of an artificial blood vessel and the artificial blood vessel, and relates to the technical field of biomedical engineering.The preparation method of the artificial blood vessel comprises the steps that a patterned microstructure pit with a preset pattern is prepared on a culture plate; inoculating cells on the culture plate, and culturing the cells to obtain a cell sheet with patterned micro-structure bulges; preparing a porous support sheet; preparing an adhesive layer on one side of the cell sheet, which is provided with the bulge, and covering the porous support sheet to obtain a unit sheet layer; culturing a unit sheet layer; the unit sheet layers are curled and bonded into the artificial blood vessel. The patterned microstructure can guide cell growth and arrangement, simulates the endothelial structure of a natural blood vessel, and is beneficial to improving the biocompatibility and functionality of the artificial blood vessel. The porous supporting sheet provides mechanical support, the bonding layers connect all the layers into a whole, a tubular structure is constructed through curling forming, and finally the artificial blood vessel is formed.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical engineering, and in particular to a method for preparing an artificial blood vessel and the artificial blood vessel. Background Art

[0002] At present, cardiovascular diseases such as arteriosclerosis, vascular stenosis, and embolism are still one of the diseases with the highest mortality rates worldwide, and vascular transplantation is an important means of treating cardiovascular diseases caused by vascular stenosis or blockage. Although using the patient's own blood vessels for transplantation is the first choice, not all patients have enough healthy blood vessels available. In addition, autologous transplantation may bring additional surgical risks and recovery time. Small-caliber artificial blood vessels (usually refers to blood vessels with an inner diameter of less than 6 mm) have important application value in the medical field, especially in cases where vascular transplantation or vascular bypass surgery is required. Due to vascular disease, trauma, congenital defects or the need for surgical operations, substitutes for small-caliber blood vessels are essential for the treatment of various vascular diseases.

[0003] Natural blood vessels have a complex multilayer structure, including an endothelial cell layer, an elastic layer, a smooth muscle layer, and an adventitia. These layered structures give blood vessels unique mechanical properties and biological functions. Existing methods for preparing small-caliber artificial blood vessels, such as 3D bioprinting and electrospinning technology, are difficult to fully replicate the complex structure and function of natural blood vessels. 3D bioprinting technology has problems such as low printing accuracy, poor mechanical properties, and easy induction of intimal hyperplasia and thrombosis; blood vessels prepared by electrospinning technology have defects such as insufficient mechanical properties, a single structure, and slow endothelialization.

[0004] Therefore, it is necessary to improve the existing artificial blood vessel technology to overcome the defects of the existing technology. Summary of the invention

[0005] In order to overcome the problems existing in the related art, one of the purposes of the present invention is to provide a method for preparing an artificial blood vessel, wherein the method for preparing an artificial blood vessel obtains a cell membrane sheet having a corresponding protruding structure by culturing cells on a culture plate having a patterned microstructure, so as to overcome the problem existing in the prior art that it is difficult to completely replicate the natural blood vessel structure.

[0006] A method for preparing an artificial blood vessel, comprising:

[0007] Step S100, preparing a patterned microstructure depression having a predetermined pattern on a culture plate;

[0008] Step S200, inoculating cells on the culture plate and culturing the cells to obtain a cell membrane sheet having patterned microstructure protrusions;

[0009] Step S300, preparing a porous support sheet;

[0010] Step S400, preparing an adhesive layer on the side of the cell membrane sheet having the protrusions, and covering the porous support sheet to obtain a unit sheet layer;

[0011] Step S500, culturing the unit sheet;

[0012] Step S600, curling and bonding the unit sheets into an artificial blood vessel.

[0013] By culturing cells on a culture plate with a patterned microstructure, a cell membrane sheet with a corresponding protruding structure is obtained. It is then assembled, cultured and rolled into shape with an adhesive layer and a porous support sheet. The patterned microstructure can guide cell growth and arrangement, simulate the endothelial structure of natural blood vessels, and help improve the biocompatibility and functionality of artificial blood vessels. The porous support sheet provides mechanical support, the adhesive layer connects the layers into a whole, and the rolled-up structure constructs a tubular structure, ultimately forming an artificial blood vessel.

[0014] Furthermore, in step S100, the patterned microstructure recesses may be produced by photolithography, electron beam lithography, laser patterning, or the like.

[0015] Furthermore, the step of preparing a patterned microstructure having a predetermined pattern on a culture plate comprises:

[0016] A laser is used to etch a predetermined pattern on the culture plate to form the patterned microstructure.

[0017] Laser etching technology has high precision and can produce a variety of complex patterns and control the size and shape of microstructures, thereby better controlling the growth and arrangement of cells, and ultimately improving the performance and customization of artificial blood vessels. Compared with other micromachining methods, laser etching is more efficient and convenient.

[0018] Furthermore, depending on the material of the culture plate, the laser involved may be a millisecond pulse laser, a microsecond pulse laser, a nanosecond pulse laser, a picosecond pulse laser, a femtosecond pulse laser, etc.; the laser may include an infrared laser, a visible light laser, an ultraviolet laser, etc.

[0019] Furthermore, the material of the culture plate is selected from polystyrene, polytetrafluoroethylene, polypropylene, polycarbonate, polymethyl methacrylate, glass, siliconized glass, polydimethylsiloxane and biodegradable materials.

[0020] These materials have good biocompatibility, are suitable for cell culture, and are easy to modify on the surface, making it convenient to prepare patterned microstructures. The properties of different materials can be selected according to specific needs.

[0021] Furthermore, after rolling up and bonding the unit sheets to form an artificial blood vessel, the method further comprises:

[0022] The artificial blood vessel is subjected to a decellularization treatment.

[0023] Decellularization can remove cellular components and reduce the risk of immune rejection, while retaining the extracellular matrix, maintaining the structure and mechanical properties of artificial blood vessels, and improving their long-term stability and safety in the body.

[0024] Furthermore, the artificial blood vessels after decellularization can be modified, and the modified molecules can include growth factors and cell adhesion molecules to promote cell migration, proliferation and tissue regeneration.

[0025] Furthermore, the step of curling and bonding the unit sheets into an artificial blood vessel specifically includes:

[0026] According to the predetermined size of the artificial blood vessel, a rod of corresponding size is selected;

[0027] According to a predetermined size of the artificial blood vessel, curling a corresponding number of unit sheets onto the rod;

[0028] Bonding the rolled unit sheets into an artificial blood vessel;

[0029] removing the rod;

[0030] The surface of the rod has a hydrophobic film, and the diameter of the rod is 1 mm to 15 mm.

[0031] The use of rods of specific sizes can control the diameter of the artificial blood vessel to meet different clinical needs. The hydrophobic film on the surface of the rod can facilitate the demoulding of the artificial blood vessel and avoid damaging the vascular structure. Controlling the diameter and hydrophobicity of the rod ensures the controllability of the artificial blood vessel preparation process and the consistency of the finished product.

[0032] Furthermore, the inner diameter of the blood vessel can be adjusted by the diameter of the rod, and the diameter range can be 1mm-15mm. The length of the blood vessel can be adjusted by the size of the cell membrane, and the length range can be 1mm-60mm. The thickness of the blood vessel wall can be adjusted by the number of unit sheets, and the thickness of the cell membrane, the adhesive layer and the porous support layer in the unit sheet, wherein the number of unit sheets can be 1-20 layers.

[0033] Further, the width of the patterned microstructure depression ranges from 1 micron to 1000 microns;

[0034] The depth of the recessed patterned microstructure is in a range of 1 micrometer to 1000 micrometers.

[0035] The size of the patterned microstructure depressions affects the morphology, arrangement and function of cells. The width and depth of this range can effectively guide cell growth, simulate the microenvironment of natural vascular endothelial cells, promote cell adhesion and proliferation, and ultimately affect the biocompatibility and function of artificial blood vessels.

[0036] Furthermore, the predetermined pattern of the patterned microstructure is selected from at least one of a diamond shape, a cross shape, a cross grid shape, a stripe shape, a dot shape and a bead shape.

[0037] Different patterns can guide cells to form different arrangements and simulate different types of vascular structures. Choosing appropriate patterns can better simulate the structure and function of natural blood vessels.

[0038] Furthermore, the predetermined pattern of the patterned microstructure is a diamond pattern.

[0039] Furthermore, the adhesive layer is prepared by a method selected from photocuring molding, temperature-sensitive molding, dipping process, chemical crosslinking and ionic covalent crosslinking; the material of the adhesive layer is selected from polylactic acid-glycolic acid copolymer, polydopamine, polymer hydrogel bioadhesive, tannic acid, catechol, polyacrylic acid, arachidonic acid / chitin nanofiber composite material, polyglycerol sebacate acrylate and polymer phase change material.

[0040] These preparation methods can effectively bond the cell membrane sheet and the porous support sheet together to form a stable unit sheet. The listed materials have good biocompatibility and adhesion properties and can meet the preparation requirements of artificial blood vessels.

[0041] Furthermore, the active layer is rich in nutrients such as growth factors, cytokines, small molecule compounds, specific signaling pathway activators, metabolic regulation, plant extracts, special compounds, extracellular vesicles, etc., which help cells to rapidly proliferate, differentiate and migrate to the porous support sheet.

[0042] Furthermore, the porous support sheet is made of a biodegradable polymer.

[0043] Biodegradable materials can be gradually degraded and absorbed in the body, avoiding long-term foreign body residues and promoting the generation of new tissues, ultimately integrating artificial blood vessels with host tissues and improving long-term efficacy.

[0044] Furthermore, the biodegradable polymers may specifically include: polylactic acid (PLA), polyhydroxyalkanoates (PHA), polycaprolactone (PCL), polyurethane (PU), silicone, expanded polytetrafluoroethylene, nylon elastomer materials, elastin, collagen, silk fibroin, hyaluronic acid (HA), fibrin, etc.

[0045] Furthermore, before performing step S200, the culture plate with the patterned microstructure depressions of the predetermined pattern may be sterilized, and the sterilization methods involved include: autoclaving, chemical disinfectants, dry heat sterilization, ultraviolet (UV) irradiation, γ-ray irradiation, ethylene oxide (EO) sterilization, filtration sterilization, flame sterilization, pre-sterilized culture plates, etc. Before sterilization, it is necessary to select a suitable sterilization method according to the material of the culture plate and the experimental requirements.

[0046] Furthermore, before performing step S200, the polystyrene culture plate with the patterned microstructure depressions having the predetermined pattern is sterilized using an ultrasonic cleaning machine, and 75% ethanol solution is used to properly store the culture plate with the patterned culture dish after disinfection to avoid re-contamination. During the cell culture process, the operator should also comply with the aseptic operation procedures to ensure a sterile environment throughout the culture process.

[0047] Furthermore, in step S200, the inoculated cells may include: endothelial cells (ECs), smooth muscle cells (SMCs), pericytes, stem cells, endothelial progenitor cells (EPCs), etc.

[0048] The second object of the present invention is to provide an artificial blood vessel, which is prepared based on the above-mentioned method for preparing an artificial blood vessel, and comprises at least one layer of curled unit sheets, wherein the unit sheets comprise a cell membrane sheet, an adhesive layer, and a porous support sheet;

[0049] The cell membrane sheet layer has patterned microstructure protrusions, and the cell membrane sheet has a side with the protrusions facing inward.

[0050] The patterned microstructure protrudes toward the lumen of the blood vessel, which can simulate the surface morphology of natural vascular endothelial cells, promote blood flow, reduce thrombosis, and improve the patency and long-term use of artificial blood vessels. The multi-layer structure design also improves the strength and stability of artificial blood vessels.

[0051] The beneficial effects of the present invention are:

[0052] The present invention provides a method for preparing an artificial blood vessel, which includes culturing cells on a culture plate with a patterned microstructure to obtain a cell membrane sheet with a corresponding protrusion structure. Subsequently, the cell membrane sheet is assembled, cultured and rolled with an adhesive layer and a porous support sheet. The patterned microstructure can guide cell growth and arrangement, simulate the endothelial structure of natural blood vessels, and is conducive to improving the biocompatibility and functionality of the artificial blood vessel. The porous support sheet provides mechanical support, the adhesive layer connects the layers into a whole, and the rolling molding constructs a tubular structure, ultimately forming an artificial blood vessel. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 is a schematic cross-sectional view of the artificial blood vessel provided in Example 1 of the present application;

[0054] Figure 2 is a schematic cross-sectional view of a unit sheet layer provided in Example 1 of the present application;

[0055] Figure 3 is a schematic diagram of four different predetermined patterns provided in Example 1 of the present application;

[0056] Figure 4 is a schematic plan view of a cell membrane sheet with patterned microstructure protrusions provided in Example 1 of the present application;

[0057] Figure 5 is a schematic cross-sectional view of a cell membrane sheet with patterned microstructure protrusions provided in Example 1 of the present application;

[0058] Figure 6 It is a flow chart of the method for preparing an artificial blood vessel provided in the present application.

[0059] Reference numerals:

[0060] 11. Cell membrane sheet; 12. Adhesion layer; 13. Porous support sheet. DETAILED DESCRIPTION

[0061] The preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the preferred embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0062] Cardiovascular disease is one of the leading causes of death worldwide, and vascular transplantation is a key means of treating vascular stenosis or obstruction. Although autologous vascular transplantation is the preferred option, its source is limited and there is a risk of complications. There is a huge clinical demand for small-caliber artificial blood vessels (inner diameter less than 6 mm), but there is still a lack of ideal substitutes.

[0063] Natural blood vessels have a complex multilayer structure, including an endothelial cell layer, an elastic layer, a smooth muscle layer, and an adventitia. These layered structures give blood vessels unique mechanical properties and biological functions. Existing methods for preparing small-caliber artificial blood vessels, such as 3D bioprinting and electrospinning technology, are difficult to fully replicate the complex structure and function of natural blood vessels. 3D bioprinting technology has problems such as low printing accuracy, poor mechanical properties, and easy induction of intimal hyperplasia and thrombosis; blood vessels prepared by electrospinning technology have defects such as insufficient mechanical properties, a single structure, and slow endothelialization.

[0064] The patterned cell membranes constructed in vitro can mimic the structure and function of natural cell membranes and effectively regulate gene expression and protein secretion. By combining different types of cell membranes, multilayer structures with specific cell arrangements and functions can be constructed, and the overall mechanical properties can be enhanced.

[0065] Example

[0066] like Figures 1 to 6 As shown, this embodiment provides a method for preparing an artificial blood vessel, and the method for preparing an artificial blood vessel includes:

[0067] Step S100, preparing a patterned microstructure depression having a predetermined pattern on a culture plate;

[0068] Step S200, inoculating cells on the culture plate and culturing the cells to obtain a cell membrane sheet 11 having patterned microstructure protrusions;

[0069] Step S300, preparing a porous support sheet 13;

[0070] Step S400, preparing an adhesive layer 12 on the side of the cell membrane sheet 11 having the protrusion, and covering the porous support sheet 13 to obtain a unit sheet; the cross-sectional view of the obtained unit sheet is as follows: Figure 1 shown.

[0071] Step S500, culturing the unit sheet;

[0072] Step S600, the unit sheets are rolled and bonded to form an artificial blood vessel. The cross-sectional view of the obtained artificial blood vessel is as follows: Figure 2 shown.

[0073] Specifically, in step S100, the patterned microstructure recesses can be manufactured by photolithography, electron beam lithography, laser patterning, and the like.

[0074] Furthermore, the step of preparing a patterned microstructure having a predetermined pattern on a culture plate comprises:

[0075] A laser is used to etch a predetermined pattern on the culture plate to form the patterned microstructure.

[0076] Specifically, the material of the culture plate is selected from polystyrene, polytetrafluoroethylene, polypropylene, polycarbonate, polymethyl methacrylate, glass, siliconized glass, polydimethylsiloxane and biodegradable materials.

[0077] More specifically, depending on the material of the culture plate, the laser involved can be a millisecond pulse laser, a microsecond pulse laser, a nanosecond pulse laser, a picosecond pulse laser, a femtosecond pulse laser, etc.; the laser can include an infrared laser, a visible light laser, an ultraviolet laser, etc.

[0078] Furthermore, the width of the patterned microstructure depression ranges from 1 micron to 1000 microns; the depth of the patterned microstructure depression ranges from 1 micron to 1000 microns. By adjusting the size of the patterned microstructure, such as width, depth, pattern spacing, etc., it can be used to adjust the wall thickness of the blood vessel.

[0079] Furthermore, the predetermined pattern of the patterned microstructure is selected from at least one of a diamond shape, a cross shape, a cross grid shape, a stripe shape, a dot shape, and a bead shape. Figure 3 As shown, Figure 3 From left to right, they are cross, cross grid, dot, and stripe. The predetermined pattern can be designed based on the arrangement of cells in the endothelium, smooth muscle layer, and adventitia that mimic natural blood vessels. By adjusting the arrangement of cells through patterned microstructures and creating specific microstructure protrusions, the contact area between blood vessels and surrounding tissues can be increased, and the secretion of specific proteins can be regulated.

[0080] In this embodiment, step S100 specifically includes using a picosecond laser to prepare a patterned microstructure with a predetermined pattern of 55 mm in length and 22 mm in width on a culture dish of polystyrene material, wherein the predetermined pattern is a diamond structure, the width of the patterned microstructure depression is about 50 um, and the depth of the patterned microstructure depression is 1.28 um. The laser parameters used are a pulse overlap rate of 99.6%, a single pulse energy of 44 uJ, a pulse overlap rate of 500 KHz, a scanning speed of 100 mm / s, and an average power of 22 W.

[0081] Before performing step S200, the culture plate with the patterned microstructure depression having a predetermined pattern can be sterilized, and the sterilization methods involved include but are not limited to: high-pressure steam sterilization (Autoclaving), chemical disinfectants, dry heat sterilization, ultraviolet (UV) irradiation, γ-ray irradiation, ethylene oxide (EO) sterilization, filtration sterilization, flame sterilization, and pre-sterilized culture plates; before sterilization, it is necessary to select a suitable sterilization method according to the material of the culture plate and the experimental requirements.

[0082] In this embodiment, an ultrasonic cleaning machine is used to sterilize the polystyrene culture plate with a predetermined pattern of patterned microstructure depressions, and a 75% ethanol solution is used to properly store the culture plate after disinfection to avoid re-contamination. During the cell culture process, the operator should also comply with the aseptic operation procedures to ensure a sterile environment throughout the culture process.

[0083] More specifically, in step S200, the inoculated cells may include: endothelial cells (ECs), smooth muscle cells (SMCs), pericytes, stem cells, endothelial progenitor cells (EPCs), etc.

[0084] In this embodiment, step S200 specifically includes inoculating endothelial cells on a sterilized culture plate, using a carbon dioxide cell culture incubator to culture cells, inoculating 600,000 endothelial cells in the pattern area, using a low-sugar culture medium to prepare a cell suspension, and after the cells have completed attachment, replacing them with HFV culture medium for 7 days of culture, with the culture medium being replaced once a day. After the cell membrane 11 is formed, the following is obtained: Figure 4 Figure 5 The cell membrane sheet 11 with patterned microstructure protrusions is shown to have a thickness of about 150 um.

[0085] Further, in step S300, the preparation method of the porous support sheet 13 includes 3D printing, electrospinning, injection molding, template manufacturing, sacrificial template method, foaming method, etc. The porous support sheet 13 is made of a biodegradable polymer that can be gradually absorbed by the human body. The biodegradable polymer can specifically include: polylactic acid (PLA), polyhydroxyalkanoate (PHA), polycaprolactone (PCL), polyurethane (PU), silica gel, expanded polytetrafluoroethylene, nylon elastomer material, elastin, collagen, silk protein, hyaluronic acid (HA), fibrin (Fibrin), etc. The porous support sheet 13 can be modified with growth factors, cell adhesion molecules and other bioactive molecules to promote cell migration, proliferation and tissue regeneration. In this embodiment, the porous support sheet 13 is made of polyurethane (PU) with a thickness of 50um and is prepared in the form of electrospinning.

[0086] Furthermore, in step S400, the adhesive layer 12 is prepared by a method selected from photocuring, thermosensitive molding, dip coating, chemical crosslinking and ionic covalent crosslinking; the material of the adhesive layer 12 is selected from polylactic acid-glycolic acid copolymer, polydopamine, polymer hydrogel bioadhesive, tannic acid, catechol, polyacrylic acid, oleic acid / chitin nanofiber composite material, polyglycerol sebacate acrylate and polymer phase change material. The adhesive layer 12 has good adhesive properties, biocompatibility, and appropriate mechanical properties, and is used to bond the cell membrane 11 and the porous support sheet 13 in the early stage of cell culture. At the same time, the adhesive layer 12 can be rich in certain nutrients that help cells to rapidly proliferate, differentiate and migrate to the porous support sheet 13.

[0087] In this embodiment, step S400 specifically includes, with the side of the cell membrane sheet 11 having the diamond-patterned microstructure protrusion facing upward, covering it with a layer of adhesive layer 12 and a porous support sheet 13 in sequence; the adhesive layer 12 is made of polylactic acid-glycolic acid copolymer (PLGA), which is rich in growth factors and cytokine nutrients and has a thickness of 50um; the porous support layer is made of polyurethane (PU), which has a thickness of 50um and is prepared by electrospinning.

[0088] In this embodiment, the culture time of step S500 is 2-3 days, during which 50% of the adhesive layer 12 is absorbed in the thickness direction, so that the cell membrane sheet 11 and the porous support sheet 13 are more closely connected, and the thickness of the finally formed unit sheet is 220-230um;

[0089] Furthermore, in step S600, the step of curling and bonding the unit sheets into an artificial blood vessel specifically includes:

[0090] According to the predetermined size of the artificial blood vessel, a rod of corresponding size is selected;

[0091] According to a predetermined size of the artificial blood vessel, curling a corresponding number of unit sheets onto the rod;

[0092] Bonding the rolled unit sheets into an artificial blood vessel;

[0093] removing the rod;

[0094] The surface of the rod has a hydrophobic film, and the diameter of the rod is 1 mm to 15 mm.

[0095] Specifically, the inner diameter of the blood vessel can be adjusted by the diameter of the rod, and the diameter range can be 1-15 mm. The length of the blood vessel can be adjusted by the size of the cell membrane sheet 11, and the length range can be 1-60 mm. The thickness of the blood vessel wall can be adjusted by the number of unit sheets, and the thickness of the cell membrane sheet 11, the adhesive layer 12 and the porous support layer in the unit sheet, wherein the number of unit sheets can be 1-20 layers.

[0096] More specifically, in this embodiment, a rod with a diameter of 5 mm is used to roll up unit sheets with different cells into an artificial blood vessel with a diameter of 5 mm. Since the length of the prepared unit sheet is 55 mm, three turns can be formed during the rolling process, so that the length of the artificial blood vessel is about 50 mm. The excess unit sheet can be removed. Finally, the rolled artificial blood vessel contains three layers of unit sheets, and each unit sheet contains the same patterned microstructure, adhesive layer 12 material and porous support layer material and structure, and finally the thickness of the artificial blood vessel wall is 660-690 um.

[0097] Furthermore, after rolling up and bonding the unit sheets to form an artificial blood vessel, the method further comprises:

[0098] The artificial blood vessel is subjected to a decellularization treatment.

[0099] Furthermore, the artificial blood vessel after decellularization can be modified, and the modified molecules can include growth factors and cell adhesion molecules to promote cell migration, proliferation and tissue regeneration. In this embodiment, dexamethasone is modified on the outer wall of the artificial blood vessel to achieve local release after the artificial blood vessel is implanted to prevent complications after transplantation.

[0100] This embodiment provides a method for preparing a multilayer small-caliber artificial blood vessel, by using picosecond laser to etch a diamond-patterned microstructure on a polystyrene culture plate, culturing endothelial cells to form a cell membrane sheet 11 with a corresponding protruding structure, and then combining the cell membrane sheet 11 with an adhesive layer 12 and a PU porous support sheet 13 prepared by electrospinning to form a unit sheet, after short-term culture, the adhesive layer 12 is partially degraded and the combination of the cell membrane sheet 11 and the porous support sheet 13 is enhanced, and finally multiple unit sheets are wound on a 5mm diameter rod coated with a hydrophobic film to form a tubular structure and bonded, and after removing the rod, an artificial blood vessel with an inner diameter of 5mm, a length of about 50mm, a wall thickness of 660-690um and a diamond-patterned inner surface is obtained. This method accurately controls the microstructure and cell arrangement on the cell membrane sheet 11, realizes the design of a multilayer structure, and combines electrospinning technology to finally prepare a small-caliber artificial blood vessel with a specific size, excellent mechanical properties and good biocompatibility. In addition, by decellularizing the artificial blood vessels and modifying the outer wall with dexamethasone, the immune rejection reaction can be further reduced and post-transplantation complications can be prevented, thereby improving the clinical application potential of artificial blood vessels.

[0101] Unless otherwise specifically stated, the relative arrangement, numerical expressions and numerical values ​​of the parts and steps set forth in these embodiments do not limit the scope of the application. In all examples shown and discussed here, any specific value should be interpreted as being merely exemplary, rather than as a restriction. Therefore, other examples of exemplary embodiments can have different values. It should be noted that similar reference numerals and letters represent similar items in the following drawings, and therefore, once a certain item is defined in an accompanying drawing, it does not need to be further discussed in the subsequent accompanying drawings.

[0102] In addition, it should be noted that the use of terms such as "first" and "second" to limit is only for the convenience of distinction. Unless otherwise stated, the above terms have no special meaning and therefore cannot be understood as limiting the scope of protection of this application.

[0103] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing an artificial blood vessel, characterized in that: include: A patterned microstructure depression having a predetermined pattern is prepared on a culture plate; Inoculating cells on the culture plate and performing cell culture to obtain a cell membrane sheet having patterned microstructure protrusions; preparing a porous support sheet; Preparing an adhesive layer on the side of the cell membrane sheet with the protrusion and covering the porous support sheet to obtain a unit sheet layer; culturing the unit sheet; The unit sheets are rolled and bonded to form an artificial blood vessel.

2. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The method of preparing a patterned microstructure having a predetermined pattern on a culture plate comprises: A laser is used to etch a predetermined pattern on the culture plate to form the patterned microstructure.

3. The method for preparing an artificial blood vessel according to claim 1, characterized in that: After rolling up and bonding the unit sheets into an artificial blood vessel, the method further comprises: The artificial blood vessel is subjected to a decellularization treatment.

4. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The step of curling and bonding the unit sheets into an artificial blood vessel specifically comprises: According to the predetermined size of the artificial blood vessel, a rod of corresponding size is selected; According to a predetermined size of the artificial blood vessel, curling a corresponding number of unit sheets onto the rod; Bonding the rolled unit sheets into an artificial blood vessel; removing the rod; The surface of the rod has a hydrophobic film, and the diameter of the rod is 1 mm to 15 mm.

5. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The width of the patterned microstructure depressions ranges from 1 micron to 1000 microns; The depth of the recessed patterned microstructure is in a range of 1 micrometer to 1000 micrometers.

6. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The predetermined pattern of the patterned microstructure is selected from at least one of a diamond shape, a cross shape, a cross grid shape, a stripe shape, a dot shape, and a connected bead shape.

7. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The material of the culture plate is selected from the group consisting of polystyrene, polytetrafluoroethylene, polypropylene, polycarbonate, polymethyl methacrylate, glass, siliconized glass, polydimethylsiloxane and biodegradable materials.

8. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The porous support sheet is made of a biodegradable polymer.

9. The method for preparing an artificial blood vessel according to claim 1, characterized in that: The adhesive layer is prepared by a method selected from photocuring molding, temperature-sensitive molding, dipping process, chemical crosslinking and ionic covalent crosslinking; the material of the adhesive layer is selected from polylactic acid-glycolic acid copolymer, polydopamine, polymer hydrogel bioadhesive, tannic acid, catechol, polyacrylic acid, arachidonic acid / chitin nanofiber composite material, polyglycerol sebacate acrylate and polymer phase change material.

10. An artificial blood vessel, characterized in that: The artificial blood vessel is prepared according to the preparation method of any one of claims 1 to 9, comprising at least one curled unit sheet layer, wherein the unit sheet layer comprises a cell membrane sheet layer, an adhesive layer, and a porous support sheet; The cell membrane sheet layer has patterned microstructure protrusions, and the cell membrane sheet has a side with the protrusions facing inward.

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