CN113694253A - Preparation method of small-caliber artificial blood vessel - Google Patents
Preparation method of small-caliber artificial blood vessel Download PDFInfo
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- CN113694253A CN113694253A CN202111111410.XA CN202111111410A CN113694253A CN 113694253 A CN113694253 A CN 113694253A CN 202111111410 A CN202111111410 A CN 202111111410A CN 113694253 A CN113694253 A CN 113694253A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3826—Muscle cells, e.g. smooth muscle cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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
- A61L27/58—Materials at least partially resorbable by the body
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/06—Wet spinning methods
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/08—Melt spinning methods
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The invention provides a preparation method of a small-caliber artificial blood vessel. The preparation method of the small-caliber artificial blood vessel comprises the following steps: s10: preparing a tubular stent; s20: seeding smooth muscle cells in the tubular scaffold; s30: placing the tubular stent inoculated with the smooth muscle cells into a near-physiological pulsating flow environment for culturing to obtain a vascular stent; s40: and carrying out decellularization treatment on the vascular stent to obtain the small-caliber artificial blood vessel. The tubular stent inoculated with the smooth muscle cells is placed in a near-physiological pulsating flow environment for culture to obtain the vascular stent, so that the real physiological environment of the vascular stent can be simulated, on one hand, stimulation can promote the smooth muscle cells to express more collagen and elastin, and on the other hand, the vascular stent can also be endowed with good mechanical properties matched with the real physiological environment. Then the small-caliber artificial blood vessel is obtained by cell removal treatment, and the immune response after transplantation can be reduced.
Description
Technical Field
The invention belongs to the technical field of medical instruments, and particularly relates to a preparation method of a small-caliber artificial blood vessel with a acellular matrix.
Background
In modern society, cardiovascular diseases seriously endanger human health, vascular remodeling plays a very important role in clinical surgery, and approximately more than 60 million people worldwide need to undergo various vascular surgeries each year, most of which require appropriate vascular grafts. At present, the clinically applied vascular grafts mainly include autologous blood vessels, allogeneic blood vessels and artificially synthesized blood vessels, and the artificial vascular graft is valued due to the limited source of the autologous blood vessels and the rejection reaction of the allogeneic blood vessels. At present, large-caliber artificial blood vessels with the diameter of more than 6 millimeters are commercialized, and the clinical application of small-caliber artificial blood vessels with the diameter of less than 6 millimeters is very disappointing.
In more than 10 years, the research on small-caliber artificial blood vessels has been greatly advanced. In recent years, acellular matrix vascular stents have received the attention of researchers. The tissue engineering blood vessel composed of extracellular matrix is prepared successfully by using in vitro cell culture technology. However, the vascular stents without the cell matrixes have the problems of poor mechanical properties and incomplete matching with the mechanical properties of autologous vessels.
Disclosure of Invention
The invention mainly aims to provide a preparation method of a small-caliber artificial blood vessel, which aims to solve the problem that the mechanical property of a blood vessel stent in the prior art is not matched with the practical application environment.
In order to achieve the above object, the present invention provides a method for preparing a small-caliber artificial blood vessel, comprising: s10: preparing a tubular stent; s20: seeding smooth muscle cells in the tubular scaffold; s30: placing the tubular stent inoculated with the smooth muscle cells into a bionic environment for culturing to obtain a vascular stent; s40: and (3) carrying out decellularization treatment on the vascular stent to obtain the small-caliber artificial blood vessel.
In one embodiment, in S30, the biomimetic environment simulates a mechanical environment corresponding to a vascular stent.
In one embodiment, the mechanical environment includes physiological pulsatile flow to the vascular stent and axial mechanical tension to the vascular stent.
In one embodiment, in S10, the tubular scaffold is prepared by one or more of electrospinning, wet spinning, melt spinning, or 3D printing techniques using biodegradable polymer as a raw material.
In one embodiment, the tubular scaffold is ordered or disordered.
In one embodiment, the tubular scaffold is nano-sized or micro-sized.
In one embodiment, in S40, the vascular stent is treated multiple times in a decellularization solution containing 1.8mM SDS, 25mM EDTA, 1M NaCl, and 0.12M NaOH.
In one embodiment, the vascular stent is placed in a decellularization solution for at least 5 treatments.
In one embodiment, the vascular stent is treated with the decellularization solution for at least 2 hours at a time.
By applying the technical scheme of the invention, the tubular stent inoculated with the smooth muscle cells is placed in a bionic environment to be cultured to obtain the vascular stent, the environment in a real application scene of the vascular stent can be simulated, on one hand, stimulation can promote the smooth muscle cells to express more collagen and elastin, and on the other hand, the vascular stent can also be endowed with good mechanical properties matched with the real application scene. Then the small-caliber artificial blood vessel is obtained by cell removal treatment, and the immune response after transplantation can be reduced.
In addition to the objects, features and advantages described above, other objects, features and advantages of the present invention are also provided. The present invention will be described in further detail below with reference to the drawings.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:
fig. 1 shows a flow chart of a method for preparing a small-caliber artificial blood vessel according to the present invention.
Detailed Description
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances for describing embodiments of the invention herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
In order to solve the defects in the prior art, researches show that periodic mechanical tension can improve the contraction function of smooth muscle cells inoculated on a vascular stent and the expression of elastin, and radial pulsating stress can improve the structural strength of a tissue engineering blood vessel. Therefore, in the technical scheme of the invention, the artificial blood vessel which better meets the application requirement can be obtained by performing the bionic environment simulation on the application scene, and therefore, the technical scheme of the invention is provided. Fig. 1 shows a method for preparing a small-caliber artificial blood vessel according to the present invention, the method comprising: s10: preparing a tubular stent; s20: seeding smooth muscle cells in the tubular scaffold; s30: placing the tubular stent inoculated with the smooth muscle cells into a bionic environment for culturing to obtain a vascular stent; s40: and carrying out decellularization treatment on the vascular stent to obtain the small-caliber artificial blood vessel.
By applying the technical scheme of the invention, the tubular stent inoculated with the smooth muscle cells is placed in a bionic environment to be cultured to obtain the vascular stent, the environment in a real application scene of the vascular stent can be simulated, on one hand, stimulation can promote the smooth muscle cells to express more collagen and elastin, and on the other hand, the vascular stent can also be endowed with good mechanical properties matched with the real application scene. Then the small-caliber artificial blood vessel is obtained by cell removal treatment, and the immune response after transplantation can be reduced.
The small-caliber artificial blood vessel can be used for repairing the small-caliber blood vessel and establishing a hemodialysis access.
Optionally, in S30, the biomimetic environment simulates a mechanical environment corresponding to the vascular stent. Under the mechanical stimulation of the mechanical environment, smooth muscle cells can be promoted to express more collagen and elastin, so that good mechanical properties are endowed to blood vessels. Preferably, the mechanical environment comprises physiological pulsatile flow to the vascular stent and axial mechanical tension to the vascular stent. The physiological pulsating flow can provide radial tension for smooth muscle cells in the vascular stent, and the axial mechanical tension can provide axial tension for the smooth muscle cells in the vascular stent, so that the finally obtained small-caliber artificial blood vessel is matched with a real application scene.
It should be noted that in S30, physiological pulsating flow and axial mechanical tension of different arterial segments of different animals can be simulated to prepare corresponding vascular stents.
Optionally, in S10, the biodegradable polymer is used as a raw material, and the tubular scaffold is prepared by one or more of electrospinning, wet spinning, melt spinning, or 3D printing techniques.
As an alternative embodiment, the tubular stents are ordered. As another alternative, the tubular stent may also be unordered. In addition, the tubular stent may be fabricated in nano-or micro-scale as desired.
Specifically, in the technical scheme of the embodiment, in S40, the vascular stent is placed in a cell-removing solution containing 1.8mM SDS, 25mM EDTA, 1M NaCl and 0.12M NaOH for treatment for multiple times. To ensure that the decellularization is sufficiently complete, the vascular stent is treated with a decellularization solution at least 5 times. As another condition to ensure sufficient and thorough decellularization, the vascular stent is treated with a decellularization solution for at least 2 hours each time.
Based on the above technical solution, the present invention provides the following embodiments:
example 1
a) Preparing an electrostatic spinning tubular stent:
dissolving silk fibroin sponge in hexafluoroisopropanol to prepare a mixed solution with the concentration of 8%. The solution was placed in a syringe at an applied voltage of 20kV, and the fiber was received by a rotating metal rod at a receiving distance of 15cm, the diameter of the receiving rod was selected to be 3mm, the length was 300mm, the rotation speed was 500rpm, and the syringe advance speed was 0.05 mm/s. Placing the silk fibroin blood vessel received on the metal bar in 99% methanol, inducing the silk fibroin to convert to a beta sheet structure, and vacuum drying for 24 hours to obtain a silk fibroin tubular scaffold with the tube wall thickness of 0.2mm and the tube diameter of 3 mm;
b) seeding of smooth muscle cells:
separating smooth muscle cells from the neck of the beagle dog, inoculating the smooth muscle cells on the silk fibroin tubular bracket, and performing rotary culture for 24 hours;
c) culturing in a bionic environment:
placing the above tubular stent inoculated with smooth muscle cells in a bionic environment, simulating beagle carotid artery pulsation frequency, pressure, flow waveform and axial tension, and culturing for 2 months under the above conditions;
d) and (3) cell removal treatment:
the vascular stent obtained by the above culture was subjected to decellularization treatment, and the cultured vascular stent was treated in an aqueous solution containing 1.8mM SDS, 25mM EDTA, 1M NaCl and 0.12M NaOH for 2 hours, and repeated 5 times. And finally, preserving the small-caliber artificial blood vessel obtained after the cell removal treatment in physiological saline for 4 degrees. The prepared small-caliber artificial blood vessel can be used for replacing carotid artery blood vessels of beagle dogs.
Example 2
a) Preparing an electrostatic spinning tubular stent:
dissolving silk fibroin sponge in hexafluoroisopropanol to prepare a mixed solution with the concentration of 8%. The solution was placed in a syringe at an applied voltage of 20kV, and the fiber was received by a rotating metal rod at a receiving distance of 15cm, the diameter of the receiving rod was selected to be 3mm, the length was 300mm, the rotation speed was 500rpm, and the syringe advance speed was 0.05 mm/s. Placing the silk fibroin blood vessel received on the metal bar in 99% methanol, inducing the silk fibroin to convert to a beta sheet structure, and vacuum drying for 24 hours to obtain a silk fibroin tubular scaffold with the tube wall thickness of 0.2mm and the tube diameter of 3 mm;
b) seeding of smooth muscle cells:
smooth muscle cells isolated from the neck of the pig were seeded on the silk fibroin tubular scaffolds. Rotating and culturing for 24 hours;
c) culturing in a bionic environment:
placing the above tubular scaffold inoculated with smooth muscle cells in a bionic environment, simulating carotid artery pulsation frequency, pressure, flow waveform and axial tension of pigs, and maintaining the above conditions for culturing for 2 months;
d) and (3) cell removal treatment:
the vascular stent obtained by the above culture was subjected to decellularization treatment, and the cultured vascular stent was treated in an aqueous solution containing 1.8mM SDS, 25mM EDTA, 1M NaCl and 0.12M NaOH for 2 hours, and repeated 5 times. And finally, preserving the small-caliber artificial blood vessel obtained after the cell removal treatment in physiological saline for 4 degrees. The prepared small-caliber artificial blood vessel can be used for replacing the carotid artery blood vessel of a pig.
Example 3
a) Preparing an electrostatic spinning tubular stent:
dissolving silk fibroin sponge in hexafluoroisopropanol to prepare a mixed solution with the concentration of 8%. The solution was placed in a syringe at an applied voltage of 20kV, and the fiber was received by a rotating metal rod at a receiving distance of 15cm, the diameter of the receiving rod was selected to be 3mm, the length was 300mm, the rotation speed was 500rpm, and the syringe advance speed was 0.05 mm/s. Placing the silk fibroin blood vessel received on the metal bar in 99% methanol, inducing the silk fibroin to convert to a beta sheet structure, and vacuum drying for 24 hours to obtain a silk fibroin tubular scaffold with the tube wall thickness of 0.2mm and the tube diameter of 3 mm;
b) seeding of smooth muscle cells:
separating out smooth muscle cells from abdominal aorta of a rat, inoculating the smooth muscle cells on the silk fibroin tubular bracket, and performing rotary culture for 24 hours;
c) culturing in a bionic environment:
placing the above tubular scaffold inoculated with smooth muscle cells in a bionic environment, simulating abdominal aorta pulsation frequency, pressure, flow waveform and axial tension of a rat, and culturing for 2 months under the above conditions;
d) and (3) cell removal treatment:
the vascular stent obtained by the above culture was subjected to decellularization treatment, and the cultured vascular stent was treated in an aqueous solution containing 1.8mM SDS, 25mM EDTA, 1M NaCl and 0.12M NaOH for 2 hours, and repeated 5 times. And finally, preserving the small-caliber artificial blood vessel obtained after the cell removal treatment in physiological saline for 4 degrees. The prepared small-caliber artificial blood vessel can be used for abdominal aorta blood vessel replacement of rats.
From the above, the advantages of the present invention are:
the tubular stent inoculated with the smooth muscle cells is placed into a bionic environment to be cultured to obtain the vascular stent, the physiological pulsating flow and the axial mechanical force can provide radial and axial tension for the smooth muscle cells in the vascular stent, and the mechanical stimulation can promote the smooth muscle cells to express more collagen and elastin, so that the vascular stent has good mechanical property; the vascular tissue engineering reactor can construct small-caliber artificial blood vessels meeting the mechanical property requirements of different parts in vitro; the small-caliber artificial blood vessel prepared by the invention can meet different blood vessel transplantation conditions by preparing blood vessel stents with different calibers and lengths; the small-caliber artificial blood vessel prepared by the invention can be used for repairing small-caliber blood vessels, tissue engineering and establishing hemodialysis access.
The relative arrangement of the components and steps, the numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present invention unless specifically stated otherwise. Meanwhile, it should be understood that the sizes of the respective portions shown in the drawings are not drawn in an actual proportional relationship for the convenience of description. Techniques, methods, and apparatus known to those of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate. In all examples shown and discussed herein, any particular value should be construed as merely illustrative, and not limiting. Thus, other examples of the exemplary embodiments may have different values. It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, further discussion thereof is not required in subsequent figures.
Spatially relative terms, such as "above … …," "above … …," "above … …," "above," and the like, may be used herein for ease of description to describe one device or feature's spatial relationship to another device or feature as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is turned over, devices described as "above" or "on" other devices or configurations would then be oriented "below" or "under" the other devices or configurations. Thus, the exemplary term "above … …" can include both an orientation of "above … …" and "below … …". The device may be otherwise variously oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.
In the description of the present invention, it is to be understood that the orientation or positional relationship indicated by the orientation words such as "front, rear, upper, lower, left, right", "lateral, vertical, horizontal" and "top, bottom", etc. are usually based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplicity of description, and in the case of not making a reverse description, these orientation words do not indicate and imply that the device or element being referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore, should not be considered as limiting the scope of the present invention; the terms "inner and outer" refer to the inner and outer relative to the profile of the respective component itself.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. A method for preparing a small-caliber artificial blood vessel is characterized by comprising the following steps:
s10: preparing a tubular stent;
s20: seeding smooth muscle cells in the tubular scaffold;
s30: placing the tubular stent inoculated with the smooth muscle cells into a near-physiological pulsating flow environment for culturing to obtain a vascular stent;
s40: and carrying out decellularization treatment on the vascular stent to obtain the small-caliber artificial blood vessel.
2. The method for preparing a small-caliber artificial blood vessel according to claim 1, wherein in S30, the near-physiological pulsating flow environment simulates a biomechanical environment corresponding to the vascular stent.
3. The method for preparing a small-caliber artificial blood vessel according to claim 2, wherein the biomechanical environment comprises a near-physiologic pulsatile flow to the vascular stent and an axial tension to the vascular stent.
4. The method for preparing the small-caliber artificial blood vessel according to claim 1, wherein in S10, the biodegradable polymer is used as a raw material, and the tubular scaffold is prepared by one or more of electrospinning, wet spinning, melt spinning or 3D printing techniques.
5. The method for preparing a small-caliber artificial blood vessel according to claim 4, wherein the tubular stent is ordered or unordered.
6. The method for preparing a small-caliber artificial blood vessel according to claim 4, wherein the tubular stent is nano-sized or micro-sized.
7. The method for preparing a small-caliber artificial blood vessel according to claim 1, wherein the vascular stent is treated a plurality of times in S40 in a decellularizing solution containing 1-2mM SDS, 20-50mM EDTA, 1-2M NaCl, and 0.1-0.2M NaOH.
8. The method for preparing a small-caliber artificial blood vessel according to claim 7, wherein the blood vessel stent is put into a decellularization solution for 3 to 5 times.
9. The method for preparing a small-caliber artificial blood vessel according to claim 8, wherein the blood vessel stent is treated with the decellularizing solution for 2 to 3 hours at a time.
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Cited By (1)
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| WO2024021869A1 (en) * | 2022-07-29 | 2024-02-01 | 海迈医疗科技(苏州)有限公司 | In-vivo rapid recellularization tissue engineered blood vessel and preparation method therefor |
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