CN109009561B - A kind of artificial blood vessel and preparation method thereof - Google Patents
A kind of artificial blood vessel and preparation method thereof Download PDFInfo
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- CN109009561B CN109009561B CN201810914099.4A CN201810914099A CN109009561B CN 109009561 B CN109009561 B CN 109009561B CN 201810914099 A CN201810914099 A CN 201810914099A CN 109009561 B CN109009561 B CN 109009561B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- 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
- A61F2/02—Prostheses implantable into the body
- A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
- A61F2/06—Blood vessels
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y80/00—Products made by additive manufacturing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2240/00—Manufacturing or designing of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2240/001—Designing or manufacturing processes
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- Heart & Thoracic Surgery (AREA)
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- Materials For Medical Uses (AREA)
- Prostheses (AREA)
Abstract
The present invention provides a kind of artificial blood vessel and preparation method thereof, which solve the unmatched technical problems of growing multiplication speed of existing artificial blood vessel degradation speed and inoculating cell, it is made of intravascular stent and inoculating cell, the intravascular stent is successively made of close-connected inner stent, middle layer bracket and outer stent, and the inner stent is one layer of porous fibre round tube shape structure being made of polyethylene glycol succinate and anti-coagulants；The middle layer bracket is one layer of porous fibre round tube shape structure being made of Hydrogels material, bioceramic material, growth factor, and the inoculating cell is filled in adherency in the hole of the middle layer bracket；The outer stent is one layer of porous fibre round tube shape structure being made of poly butylene succinate；The preparation method of the disclosed artificial blood vessel simultaneously, can be widely applied to Implantable Medical Device field.
Description
Technical field
The present invention relates to Implantable Medical Device field, in particular to a kind of artificial blood vessel and preparation method thereof.
Background technique
Currently, cardiovascular disease and peripheral arterial disease are the main reason for causing human death in the whole world.It is annual big
About need to implement 600000 vascular transplants, but close to the patient of one third because of blood vessel aging, vascular lesion,
Damage or before operation the reasons such as use and are not available autologous vein.Currently used timbering material mainly includes non-degradable
Vascular stent material and degradable blood vessel bracket.Non-degradable intravascular stent mainly has the materials such as stainless steel, Nitinol, these
Metal material non-degradable in human body needs to increase the risk for causing restenosis in permanent retention body.In such case
Under, become good selection using degradable artificial blood vessel.In the 1950s, since it is observed that cell is immersing
After the phenomenon that growing on the silk thread of blood, the manufacture history of Weave type artificial blood vessel has been begun to.Even to this day, Weave type is artificial
Blood vessel has been widely used in clinic, replaces in human body the blood vessel of corrupted, and clinically achieve preferable medical effect.
Currently, the research contents of artificial blood vessel is mainly to select the natural and synthetic high polymer with good biological performance,
It is built into composite fibre blood vessel by different methods, it is external to guide seed cell proliferation, extension and differentiation, finally move into human body
The reconstruction and regeneration of autologous vein are realized after interior.But the inoculating cell of existing artificial blood vessel is in artificial blood vessel inner stent
Wall proliferation film forming, with inner stent, the continuous degradation of middle layer bracket, outer stent, cell is grown into and is proliferated in formation
Layer and outer layer, in the process, the problem of being primarily present is the degradation speed of intravascular stent and the growing multiplication speed of inoculating cell
It mismatches, i.e. artificial blood vessel inner stent inoculating cell slow growth, and the excessively slow block cell growth of intravascular stent degradation speed
Or degradation speed is too fast causes the problem of being unable to satisfy angiogenic growth demand.
Summary of the invention
The purpose of the present invention is to solve the deficiency of above-mentioned technology, a kind of artificial blood vessel and preparation method thereof is provided.
For this purpose, the present invention provides a kind of artificial blood vessel, be made of intravascular stent and inoculating cell, intravascular stent successively by
Close-connected inner stent, middle layer bracket and outer stent are constituted, which is characterized in that inner stent is by poly-succinic second
One layer of porous fibre round tube shape structure that diol ester and anti-coagulants are constituted；Middle layer bracket is by Hydrogels material, biology pottery
One layer of porous fibre round tube shape structure that ceramic material, growth factor are constituted, middle layer bracket is interior to be equipped with reticulated cell along three-dimensional
Gap, the interior adherency filling inoculating cell of hole；Middle layer branch is provided with rodlike layer and circular layer, and rodlike layer is with the axis of middle layer bracket
Line is that axle center annular array is equipped with more rod-like fibre silks being spaced apart from each other, and rod-like fibre silk and the axis of middle layer bracket are flat
Row, circular layer are equipped at intervals with multiple circular fiber silks in the axial direction of middle layer bracket, and circular fiber silk and middle layer bracket are same
Axis；Along middle layer bracket, successively alternating layers are superimposed connection setting, rodlike layer and ring-type to rodlike layer in the radial direction with circular layer
The hole being distributed along three-dimensional is formed between layer；Outer stent is one layer of porous fibre being made of poly butylene succinate
Round tube shape structure；The porosity of inner stent and the porosity of outer stent are respectively less than the porosity of middle layer bracket.
Preferably, the filament diameter of inner stent is 8-20 μm, porosity 10-70%, with a thickness of 50-300 μm；In
The filament diameter of interbed bracket is 50-300 μm, porosity 40-80%, with a thickness of 400-800 μm；The fiber of outer stent
Silk diameter is 10-80 μm, porosity 10-70%, with a thickness of 400-600 μm.
Preferably, its production raw material of artificial blood vessel is mainly made of the component of following parts by weight: 10-20 parts of anti-coagulants,
20-50 parts of polyethylene glycol succinate, 20-40 parts of Hydrogels material, bioceramic material 10-20, -10 parts of growth factors 5,
20-30 parts of poly butylene succinate.
Hydrogels material is one of collagen, gelatin, polyvinyl alcohol；
Bioceramic material is one of tricalcium phosphate, hydroxyapatite, bio-vitric or more than one mixing；
Anti-coagulants is one of heparin, sodium oxalate, sodium citrate or more than one mixing；
Inoculating cell is one of fat stem cell, multipotential stem cell, mescenchymal stem cell and vascular smooth muscle cells
Or more than one mixing；
Growth factor is VEGF, is one of VEGF121, VEGF145, VEGF165, VEGF189, VEGF206 or one
Kind or more mixing.
A kind of preparation method of above-mentioned artificial blood vessel comprising following steps:
(1), anti-coagulants, polyethylene glycol succinate, Hydrogels material, biology pottery according to parts by weight, are weighed respectively
Ceramic material, growth factor, poly butylene succinate.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent is sufficiently molten
Solution carries out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the preparation of inner stent.It is interior
Layer bracket electrostatic spinning preparation method be rotation winding, electrostatic spinning raw material solution is added in electrospinning device, using with
The identical circular metal stick of human vas internal diameter is as electrostatic spinning receiving end, metal bar circumferential direction Rotating with Uniform, a spray head left side
The right side moves horizontally, so that printing silk uniform adhesion to the outer surface of circular metal stick.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, inner stent outer surface is made in step (2) and carries out 3D printing post-crosslinking, is prepared into more than one layer
Hole fiber tubular structure completes the preparation of middle layer bracket.The 3D printing preparation method of middle layer bracket is horizontal printing and ring
Shape prints the progress that alternates, and carries out the 3D printing of horizontal direction in the outer surface of artificial blood vessel inner stent, preparing has
The rodlike layer of even distribution carries out the 3D printing of circumferencial direction in the outer surface of rodlike layer, prepares and uniformly divide with circumferencial direction
The circular layer of cloth, then along middle layer bracket in the radial direction from inside to outside successively by rodlike layer and the layer-by-layer alternating layers of circular layer
Connection is stacked, is stopped until reaching the requirement of middle layer backing thickness.After the completion of step (3), added acetum is complete
Volatilization.
(4), weighed sufficiently dissolve poly butylene succinate with appropriate volatile polar solvent of step (1) is mixed
After uniformly, the middle layer rack surface made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose structure,
Complete the preparation of outer stent；It prepares since then and completes intravascular stent.The electrostatic spinning preparation method of outer stent is rotation winding,
Electrostatic spinning raw material solution is added in electrospinning device, artificial blood vessel middle layer bracket is intermediate as electrostatic spinning receiving end
Layer bracket circumferential direction Rotating with Uniform, nozzle right move horizontally, so that printing silk uniform adhesion to the outer surface of middle layer bracket.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by the outstanding of inoculating cell
Drop is added in the hole of the middle layer bracket section in an end face of intravascular stent, completes cell inoculation, and be put into culture
In liquid, artificial blood vessel is made.Hyaluronic acid solution containing inoculating cell is suspended, suspension, the suspension are completed in preparation
The hyaluronic acid concentration of liquid be 0.1-100g/L, final concentration of the 1 × 10 of cell5-1×106；Culture solution is 10% fetal calf serum
DMEM/F12 culture solution.
Preferably, the volatile polar solvent in step (2) is one of chloroform, methylene chloride or acetone；Step (4)
In volatile polar solvent be one of chloroform, methylene chloride or acetone.Electrostatic spinning in step (2) and step (4)
After the completion, added volatile polar solvent volatilizees completely.
Preferably, when Hydrogels material selection polyvinyl alcohol hydrogel in step (3), the crosslinking after 3D printing is first
It is being freezed at least 8 hours lower than -10 DEG C, is then being placed at 20 DEG C -25 DEG C and thaws 6 hours again, above-mentioned freeze-thaw alternate treatment 4
It is secondary；When Hydrogels material selection collagen hydrogel or gelatin hydrogel, the crosslinking after 3D printing is to carry out ultraviolet light cross-linking.
The beneficial effects of the present invention are:
(1) present invention uses three-decker moulding, similar with human vas structure, and uses in middle layer backing positions
Unique printing type carries out cell adherence and proliferation, can have faster speed to carry out the generation of cell and tissue, so that people
It makes blood vessel middle layer bracket and quickly forms one layer of cells layer, have the function of artificial blood vessel, solve artificial blood vessel middle layer branch
Frame and the growth of outer stent cell and it is proliferated slow problem, is conducive to the rapid fusion of artificial blood vessel and human vas and rapid
Replacement；
(2) present invention has faster degradation speed.Using in inner stent has high-biocompatibility and speed of degrading
Spend the mixture of faster polyethylene glycol succinate and anti-coagulants, can before implantation phase anti-Hemostatic Oral Liquid in the position of artificial blood vessel
It sets and blood coagulation occurs and artificial blood vessel is caused to fail, can also be mentioned by the cell that the hole inside inner stent is middle layer bracket
For nutrition and transport waste, and can gradually degrade, so that artificial blood vessel inner stent gradually endothelialization, internal layer at this time
The cell of rack inner wall gradually forms a film, and ultimately forms the blood vessel of oneself cell, and the inoculating cell in middle layer bracket is also gradually
Film forming, when the degradable completion of inner stent, the cell of inner stent inner wall and the cell in middle layer bracket merge, significantly
Increase the growing multiplication speed of cell.Outer stent uses the poly-succinic acid-butanediol with higher stretching and bending property
Ester, degradation speed are slower than inner stent and middle layer bracket, can be in the hydrogel in inner layer scaffold material and middle layer bracket
The mechanical properties decrease area occurred after material degradation obtains certain support and protection, guarantees the adherency and fixation of cell.This hair
The degradation speed that the artificial blood vessel of bright preparation meets and cell Proliferation matches.
(3) present invention has good mechanical property.At implantation initial stage since there is no degradation behavior, artificial blood vessel warps
The structure for crossing spinning moulding guarantees it with enough mechanical properties, with the progress of degradation process, inner stent and middle layer
The implantation material of bracket gradually decreases, but the cell of inner stent and middle layer bracket gradually forms a film, so that mechanical property obtains
Compensation, therefore the stage that artificial blood vessel prepared by the present invention does not have middle layer bracket cell deficiency mechanical property to lack, Ke Yibao
It is demonstrate,proved with enough mechanical properties.
(4) characteristic that there is the present invention blood to be not easy oozing of blood.It, can since inner stent is using spinning technique
Networking is woven, diffusion of the blood to middle layer bracket and outer stent is reduced；Along with middle layer bracket is attached cell
Tight feature is presented in hydrogel structure, inside；The braiding structure of outer stent can final anti-Hemostatic Oral Liquid oozing of blood, this is artificial
Blood vessel after implanting can anti-Hemostatic Oral Liquid oozing of blood, ensure that implantation after safety.
(5) present invention has good blood seepage-proofing and mechanical property, and can gradually degrade with the extension of Implantation Time
For oneself blood vessel, there is huge advantage in cardiovascular and peripheral arterial replacement, three layers of filament can be prevented effectively
Angiorrhea, and can guarantee growth of the cell on silk thread.Middle layer bracket has faster due to the cell implantation of itself
The speed of growth.Intact cell layer is quickly formed, is engaged with vascular endothelial cell.The thicker macromolecule layer of outer stent can be
Mechanical property is kept under biggish blood pressure, and can finally be degraded to autologous vein, is not in asking for blood vessel aging
Topic makes it have significant advantage in artificial blood vessel.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of main view of the invention；
Fig. 2 is the structural schematic diagram of A-A cross-sectional view shown in FIG. 1；
Fig. 3 is the structural schematic diagram of the main view of intravascular stent of the invention；
Fig. 4 is the structural schematic diagram of B-B cross-sectional view shown in Fig. 3；
Fig. 5 is the structural schematic diagram of perspective view shown in Fig. 3；
Fig. 6 is the structural schematic diagram of cross-sectional view shown in fig. 5；
Fig. 7 is the structural schematic diagram of middle layer bracket shown in fig. 5；
Fig. 8 is the structural schematic diagram of cross-sectional view shown in Fig. 7.
Marked in the figure: 1. inner stents, 2. middle layer brackets, 3. outer stents, 4. holes, 5. inoculating cells, 6. is rodlike
Layer, 7. circular layers, 8. rod-like fibre silks, 9. circular fiber silks.
Specific embodiment
The present invention will be further explained below with reference to the attached drawings and specific examples, to help the contents of the present invention are understood.
Method used in the present invention is conventional method unless otherwise required；Used raw material and device, such as without special rule
It is fixed, it is conventional commercial product.
Embodiment 1:
As shown in FIG. 1 to FIG. 8, the present invention provides a kind of artificial blood vessel, is made of intravascular stent and inoculating cell, blood vessel
Bracket is successively made of close-connected inner stent 1, middle layer bracket 2 and outer stent 3, and inner stent 1 is by polybutadiene
One layer of porous fibre round tube shape structure that sour glycol ester and anti-coagulants are constituted；Polyethylene glycol succinate mechanical property is good, raw
Object compatibility is good, and endothelial cell quick endothelialization, anti-coagulants on the inner wall of inner stent 1 after implantation is made to greatly reduce thrombus
It is formed, improves the success rate of implantation；With the gradually degradation of polyethylene glycol succinate, anti-coagulants gradually discharges, and plays and continues
Effective anticoagulation.Middle layer bracket 2 is made of more than one layer Hydrogels material, bioceramic material, growth factor
Hole fiber round tube shape structure, middle layer bracket 2 is interior to be equipped with netted hole 4 along three-dimensional, adherency filling inoculating cell in hole 4
5；Hydrogels material and bioceramic material mechanical property are good, good biocompatibility, have good biocompatibility and can
Degradability, poly butylene succinate and polyethylene glycol succinate will form acidic environment in degradation process, be unfavorable for thin
The growth of born of the same parents can neutralize acidic environment by alkaline matter therein by the addition of bioceramic material, be formed
Be conducive to the microenvironment of cell growth.Be conducive to the growth and proliferation of cell.With Hydrogels materials, bioceramic material
It gradually degrades, growth factor gradually discharges, and plays the effect of promotion 5 growing multiplication of inoculating cell of continuous and effective；Middle layer branch
Frame 2 is equipped with rodlike layer 6 and circular layer 7, and rodlike layer 6 is equipped with as axle center annular array using the axis of middle layer bracket 2 and is spaced apart from each other
More rod-like fibre silks 8, rod-like fibre silk 8 is parallel with the axis of middle layer bracket 2, axis of the circular layer 7 along middle layer bracket 2
It is equipped at intervals with multiple circular fiber silks 9 upwards, circular fiber silk 9 and middle layer bracket 2 are coaxial；Rodlike layer 6 is with circular layer 7 in
The successively alternating layers superposition connection setting in the radial direction of interbed bracket 2, forms between rodlike layer 6 and circular layer 7 along three-dimensional side
To the hole 4 of distribution；Outer stent 3 is one layer of porous fibre round tube shape structure being made of poly butylene succinate, poly- fourth
Succinate adipate mechanical property is good, good biocompatibility.
The filament diameter of inner stent 1 is 8-20 μm, porosity 10-70%, with a thickness of 50-300 μm；Middle layer branch
The filament diameter of frame 2 is 50-300 μm, porosity 40-80%, with a thickness of 400-800 μm；The fiber filament of outer stent 3 is straight
Diameter is 10-80 μm, porosity 10-70%, with a thickness of 400-600 μm；The porosity of inner stent 1 and the hole of outer stent 3
Gap rate is respectively less than the porosity of middle layer bracket 2, is proliferated inoculating cell 5 in middle layer bracket, does not penetrate into inner stent 1
In the hole of outer stent 3；The internal diameter of artificial blood vessel is 3-46mm, with a thickness of 300-1500 μm.Meet human body different parts
Vasotransplantation demand.
Embodiment 2:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 10 parts of anti-coagulant heparin, 20 parts of polyethylene glycol succinate, Hydrogels according to parts by weight, are weighed respectively
20 parts of material collagen, 10 parts of bioceramic material tricalcium phosphate, growth factor VEGF 121 are 5 parts, poly butylene succinate 20
Part.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent chloroform are filled
Divide dissolution to carry out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of inner stent 1
It is standby；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, is made after 1 outer surface of inner stent carries out 3D printing in step (2) and carries out ultraviolet light cross-linking, prepared
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), poly butylene succinate and appropriate volatile polar solvent chloroform are sufficiently dissolved by step (1) is weighed
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose
Structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3 is rotation
Turn winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by 5 (rouge of inoculating cell
Fat stem cell) suspension be added drop-wise in the hole 4 of 2 section of middle layer bracket in an end face of intravascular stent, complete cell
Inoculation, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 8 μm, porosity 10%, with a thickness of 50 μm；In
The filament diameter of interbed bracket 2 is 50 μm, porosity 40%, with a thickness of 400 μm；The filament diameter of outer stent 3 is
10 μm, porosity 10%, with a thickness of 400 μm；The internal diameter of artificial blood vessel is 3mm.
Embodiment 3:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 20 parts of anti-coagulants sodium oxalate, 50 parts of polyethylene glycol succinate, hydrogel according to parts by weight, are weighed respectively
40 parts of class material collagen, 20 parts of bioceramic material hydroxyapatite, growth factor VEGF 145 are 10 parts, poly-succinic fourth two
30 parts of alcohol ester.
(2), by the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent dichloromethane
Alkane sufficiently dissolves carries out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes inner stent 1
Preparation；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, is made after 1 outer surface of inner stent carries out 3D printing in step (2) and carries out ultraviolet light cross-linking, prepared
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), step (1) is weighed abundant by poly butylene succinate and appropriate volatile polar methylene chloride
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning for dissolution, is prepared into one layer of porous fibre
Tubular structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3
For rotation winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, inoculating cell 5 is (more
Can stem cell) suspension be added drop-wise in the hole 4 of 2 section of middle layer bracket in an end face of intravascular stent, complete cell
Inoculation, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 20 μm, porosity 70%, with a thickness of 300 μm；
The filament diameter of middle layer bracket 2 is 300 μm, porosity 80%, with a thickness of 600 μm；The filament diameter of outer stent 3
It is 80 μm, porosity 70%, with a thickness of 600 μm；The internal diameter of artificial blood vessel is 46mm.
Embodiment 4:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 15 parts of anti-coagulants sodium citrate, 30 parts of polyethylene glycol succinate, water-setting according to parts by weight, are weighed respectively
30 parts of glue class material collagen, 15 parts of bioceramic material bio-vitric, growth factor VEGF 165 are 7 parts, poly-succinic acid-butanediol
25 parts of ester.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent acetone are filled
Divide dissolution to carry out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of inner stent 1
It is standby；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, is made after 1 outer surface of inner stent carries out 3D printing in step (2) and carries out ultraviolet light cross-linking, prepared
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), poly butylene succinate and appropriate volatile polar solvent acetone are sufficiently dissolved by step (1) is weighed
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose
Structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3 is rotation
Turn winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by inoculating cell 5 (
Mesenchymal stem cells) suspension be added drop-wise in the hole 4 of 2 section of middle layer bracket in an end face of intravascular stent, complete thin
Born of the same parents' inoculation, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 10 μm, porosity 20%, with a thickness of 80 μm；
The filament diameter of middle layer bracket 2 is 80 μm, porosity 50%, with a thickness of 500 μm；The filament diameter of outer stent 3
It is 20 μm, porosity 20%, with a thickness of 500 μm；The internal diameter of artificial blood vessel is 10mm.
Embodiment 5:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 10 parts of anti-coagulant heparin, 5 parts of sodium oxalate according to parts by weight, are weighed respectively；Polyethylene glycol succinate 40
Part；30 parts of Hydrogels material gelatin；10 parts of bioceramic material tricalcium phosphate, 5 parts of hydroxyapatite；Growth factor
VEGF121 is 4 parts, VEGF145 is 3 parts；25 parts of poly butylene succinate.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent chloroform are filled
Divide dissolution to carry out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of inner stent 1
It is standby；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, is made after 1 outer surface of inner stent carries out 3D printing in step (2) and carries out ultraviolet light cross-linking, prepared
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), step (1) is weighed abundant by poly butylene succinate and appropriate volatile polar methylene chloride
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning for dissolution, is prepared into one layer of porous fibre
Tubular structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3
For rotation winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by 5 (blood of inoculating cell
Pipe smooth muscle cell) suspension be added drop-wise in the hole 4 of 2 section of middle layer bracket in an end face of intravascular stent, complete
Cell inoculation, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 12 μm, porosity 30%, with a thickness of 100 μm；
The filament diameter of middle layer bracket 2 is 100 μm, porosity 55%, with a thickness of 600 μm；The filament diameter of outer stent 3
It is 30 μm, porosity 30%, with a thickness of 450 μm；The internal diameter of artificial blood vessel is 15mm.
Embodiment 6:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 10 parts of anti-coagulant heparin, 10 parts of sodium citrate according to parts by weight, are weighed respectively；Polyethylene glycol succinate
30 parts；20 parts of Hydrogels material gelatin；5 parts of bioceramic material tricalcium phosphate, 5 parts of bio-vitric；Growth factor VEGF 121
For 4 parts, VEGF145 be 3 parts, VEGF165 is 2 parts；30 parts of poly butylene succinate.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent chloroform are filled
Divide dissolution to carry out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of inner stent 1
It is standby；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, 1 outer surface of inner stent is made in step (2) and carries out 3D printing post-crosslinking, is prepared into one layer
Porous fibre tubulose structure completes the preparation of middle layer bracket 2.The 3D printing preparation method of middle layer bracket 2 is horizontal printing
The progress that alternates is printed with annular.When Hydrogels material selection polyvinyl alcohol hydrogel in step (3), the friendship after 3D printing
Then connection is placed at 20 DEG C -25 DEG C again and thaws 6 hours, above-mentioned freeze-thaw is handed over first to freeze at least 8 hours lower than -10 DEG C
For processing 4 times；When Hydrogels material selection collagen hydrogel or gelatin hydrogel, the crosslinking after 3D printing is that progress is ultraviolet
Photo-crosslinking.
(4), poly butylene succinate and appropriate volatile polar solvent acetone are sufficiently dissolved by step (1) is weighed
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose
Structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3 is rotation
Turn winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by 5 (rouge of inoculating cell
Fat stem cell and multipotential stem cell) suspension be added drop-wise to the hole 4 of 2 section of middle layer bracket in an end face of intravascular stent
It is interior, cell inoculation is completed, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 14 μm, porosity 40%, with a thickness of 150 μm；
The filament diameter of middle layer bracket 2 is 150 μm, porosity 60%, with a thickness of 650 μm；The filament diameter of outer stent 3
It is 40 μm, porosity 40%, with a thickness of 500 μm；The internal diameter of artificial blood vessel is 20mm.
Embodiment 7:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 10 parts of anti-coagulants sodium oxalate, 10 parts of sodium citrate according to parts by weight, are weighed respectively；Poly-succinic ethylene glycol
30 parts of ester；20 parts of Hydrogels material polyvinyl alcohol；5 parts of bioceramic material hydroxyapatite, 5 parts of bio-vitric, growth because
Sub- VEGF121 is 2.5 parts, VEGF145 is 2.5 parts, VEGF165 is 2.5 parts, VEGF189 is 2.5 parts；Poly-succinic acid-butanediol
20 parts of ester.
(2), by the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent dichloromethane
Alkane sufficiently dissolves carries out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes inner stent 1
Preparation；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Point dissolution after mixing, step (2) be made 1 outer surface of inner stent carry out 3D printing post-crosslinking, be crosslinked for elder generation lower than-
10 DEG C freeze at least 8 hours, are then placed at 20 DEG C -25 DEG C and thaw 6 hours again, above-mentioned freeze-thaw alternate treatment 4 times；Preparation
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), poly butylene succinate and appropriate volatile polar solvent acetone are sufficiently dissolved by step (1) is weighed
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose
Structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3 is rotation
Turn winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by 5 (rouge of inoculating cell
Fat stem cell, multipotential stem cell and mescenchymal stem cell) suspension be added drop-wise to the middle layer branch in an end face of intravascular stent
In the hole 4 of 2 section of frame, cell inoculation is completed, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 16 μm, porosity 50%, with a thickness of 200 μm；
The filament diameter of middle layer bracket 2 is 200 μm, porosity 70%, with a thickness of 700 μm；The filament diameter of outer stent 3
It is 50 μm, porosity 50%, with a thickness of 550 μm；The internal diameter of artificial blood vessel is 30mm.
Embodiment 8:
Prepare the preparation method of the artificial blood vessel of above-described embodiment 1 comprising following steps:
(1), 8 parts of anti-coagulant heparin, 6 parts of sodium oxalate, 6 parts of sodium citrate according to parts by weight, are weighed respectively；Poly-succinic
30 parts of glycol ester；40 parts of Hydrogels material polyvinyl alcohol；4 parts of bioceramic material tricalcium phosphate, 3 parts of hydroxyapatite,
3 parts of bio-vitric；Growth factor VEGF 121 is 2 parts, VEGF145 is 2 parts, VEGF165 is 2 parts, VEGF189 is 2 parts,
2 parts of VEGF206；20 parts of poly butylene succinate.
(2), the weighed anti-coagulants of step (1), polyethylene glycol succinate and appropriate volatile polar solvent acetone are filled
Divide dissolution to carry out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of inner stent 1
It is standby；The electrostatic spinning preparation method of inner stent 1 is rotation winding.
(3), the weighed Hydrogels material of step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Point dissolution after mixing, step (2) be made 1 outer surface of inner stent carry out 3D printing post-crosslinking, be crosslinked for elder generation lower than-
10 DEG C freeze at least 8 hours, are then placed at 20 DEG C -25 DEG C and thaw 6 hours again, above-mentioned freeze-thaw alternate treatment 4 times；Preparation
At having one layer of porous fibre tubulose structure, the preparation of middle layer bracket 2 is completed.The 3D printing preparation method of middle layer bracket 2
The progress that alternates is printed for level printing and annular.
(4), poly butylene succinate and appropriate volatile polar solvent chloroform are sufficiently dissolved by step (1) is weighed
After mixing, 2 surface of middle layer bracket made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose
Structure completes the preparation of outer stent 3.It prepares since then and completes intravascular stent；The electrostatic spinning preparation method of outer stent 3 is rotation
Turn winding.
(5), intravascular stent made from step (4) is put into after appropriate amount of deionized water cleans up, by 5 (rouge of inoculating cell
Fat stem cell, multipotential stem cell, mescenchymal stem cell and vascular smooth muscle cells) suspension be added drop-wise to one of intravascular stent end
In the hole 4 of 2 section of middle layer bracket in face, cell inoculation is completed, and be put into culture solution, artificial blood vessel is made.
The filament diameter of obtained artificial blood vessel inner stent 1 is 18 μm, porosity 60%, with a thickness of 250 μm；
The filament diameter of middle layer bracket 2 is 250 μm, porosity 75%, with a thickness of 700 μm；The filament diameter of outer stent 3
It is 60 μm, porosity 60%, with a thickness of 550 μm；The internal diameter of artificial blood vessel is 40mm.
Artificial blood vessel prepared by 2~embodiment of above-described embodiment 8 is subjected to stretch-proof experiment, thrombus test, life respectively
The detection of object compatibility, degradability detection.
(1) the stretch-proof experiment of artificial blood vessel
Extension test uses the Instron5565 type static(al) Material Testing Machine of Britain Instron company, a, and setting stretches
Speed is 5mm/min, and at 25 DEG C, relative humidity tests sample in the environment of being 50%, as a result as shown in table 1 below.
(2) thrombus is tested
New blood is taken, a small amount of anti-coagulants is added, is added dropwise in the artificial blood vessel inner wall of inventive embodiments preparation, instead
It is added dropwise 10 times again, is cast blood uniformly in stented vessel inner wall, then observe thrombosis situation, as a result as shown in table 1 below.
Tensile strength, the thrombus testing result of 1 artificial blood vessel of table
As it can be seen from table 1 artificial blood vessel tensile strength of the present invention is 35.8-46.5MPa, there is certain toughness and power
Performance is learned, is higher than existing commercially available synthetic blood vessel tensile strength 25MPa, and do not have blood or proteinosis, meets normal transplanting
It is required that.
(3) biocompatibility detection, degradability detection
It is cut along 2~embodiment of the embodiment of the present invention, 8 artificial blood vessel along its axis, is cut into the diaphragm of length and width 5mm × 5mm；
After the same method, the artificial degradable blood vessel bracket of the chitosan of existing three-decker is cut into the film of length and width 5mm × 5mm
Piece.
60 adult nude mices are chosen, first group and second group, every group of 30 nude mices are divided into；Wherein first group, divide after number
Not successively in the artificial blood vessel diaphragm of every nude mice by subcutaneous implantation 2~embodiment of the embodiment of the present invention 8, as embodiment；Second
Group, the artificial degradable blood vessel bracket film of chitosan for being successively implanted into existing three-decker in every nude mice by subcutaneous respectively after number
Piece, as a comparison case.Respectively at postoperative 1 week, 2 weeks, January, 2 months, March, April, May, June, July, August, in first group, second
Every group of group chooses 3 nude mices, takes out and is implanted into its subcutaneous artificial blood vessel diaphragm, first observes the reaction of implantation film surrounding tissue
Situation records each layer artificial blood vessel degradation situation；Then diaphragm surrounding tissue is taken, 10% formalin fixer is fixed, production
HE staining tissue slides carry out pathologic examination.
Experimental result shows that first group of artificial blood vessel diaphragm is being implanted into subcutaneous degradation in 5-6 month completely, in degradation process
In, middle layer timbering material degradation speed is most fast, and inner layer scaffold material takes second place, and outer stent degradation speed is most slow, degradation speed
It is slightly faster than the artificial degradable blood vessel bracket diaphragm of chitosan of existing three-decker；And the chitosan people of existing three-decker
The inner layer material degradation speed for making degradable blood vessel bracket diaphragm is most fast, and cladding material degradation speed takes second place, middle layer degradation speed
It is most slow.First group of artificial blood vessel diaphragm has slight tissue inflammation reaction at implantation initial stage, in the middle and later periods of implantation, with diaphragm
Degradation, inflammatory reaction fade away, and show preferable biocompatibility.It can be seen that artificial blood vessel of the present invention has well
Degeneration and histocompatbility.
Middle layer timbering material degradation speed of the present invention is most fast, matches the inoculating cell growth increment speed in middle layer bracket
Degree, and gradually form a film, inner layer scaffold material takes second place, the growth rate that the cell of inner stent inner wall is adhered in blood is matched,
And gradually form a film, with the gradually degradation of middle layer bracket and inner stent, in inner stent tapetum cell and middle layer bracket
Inoculating cell mix together and be integrally formed, there is certain intensity and toughness, outer stent degradation speed is most slow, finally completes drop
Solution, in the whole process, the growing multiplication speed of artificial blood vessel degradation speed and inoculating cell matches.The present invention has wide
Application prospect, be expected to be used in actual clinical diagnosis.
In the description of the present invention, it is to be understood that, term "left", "right", "upper", "lower", "top", "bottom", " preceding ",
The orientation or positional relationship of the instructions such as " rear ", "inner", "outside", " back ", " centre " is that orientation based on the figure or position are closed
System, is merely for convenience of description of the present invention and simplification of the description, rather than the device or element of indication or suggestion meaning must have
Specific orientation is constructed and operated in a specific orientation, therefore is not considered as limiting the invention.
Only as described above, only specific embodiments of the present invention, when the model that cannot be limited the present invention with this and implement
It encloses, therefore the displacement of its equivalent assemblies, or according to equivalent changes and modifications made by the invention patent protection scope, should still belong to this hair
The scope that bright claims are covered.
Claims (6)
1. a kind of artificial blood vessel, is made of intravascular stent and inoculating cell, the intravascular stent is successively by close-connected interior
Layer bracket, middle layer bracket and outer stent are constituted, which is characterized in that the inner stent be by polyethylene glycol succinate and
One layer of porous fibre round tube shape structure that anti-coagulants is constituted；The middle layer bracket is by Hydrogels material, bioceramic material
One layer of porous fibre round tube shape structure that material, growth factor are constituted, the middle layer bracket is interior to be equipped with reticulated cell along three-dimensional
The inoculating cell is filled in gap, the interior adherency of the hole；The middle layer branch is provided with rodlike layer and circular layer, the rodlike layer
More rod-like fibre silks being spaced apart from each other, the rod-like fibre are equipped with by axle center annular array of the axis of the middle layer bracket
Silk is parallel with the axis of the middle layer bracket, and the circular layer is equipped at intervals with multiple rings in the axial direction of the middle layer bracket
Shape fiber filament, the circular fiber silk and the middle layer bracket are coaxial；The rodlike layer and the circular layer are along the centre
The successively alternating layers superposition connection setting in the radial direction of layer bracket, forms between the rodlike layer and the circular layer along three-dimensional
The hole of directional spreding；The outer stent is one layer of porous fibre circular tube shaped knot being made of poly butylene succinate
Structure；The porosity of the inner stent and the porosity of the outer stent are respectively less than the porosity of the middle layer bracket.
2. a kind of artificial blood vessel according to claim 1, which is characterized in that the filament diameter of the inner stent is 8-
20 μm, porosity 10-70%, with a thickness of 50-300 μm；The filament diameter of the middle layer bracket is 50-300 μm, hole
Rate is 40-80%, with a thickness of 400-800 μm；The filament diameter of the outer stent is 10-80 μm, porosity 10-
70%, with a thickness of 400-600 μm.
3. a kind of artificial blood vessel according to claim 1, which is characterized in that it makes raw material mainly by following parts by weight
Component composition: 10-20 parts of anti-coagulants, 20-50 parts of polyethylene glycol succinate, 20-40 parts of Hydrogels material, bioceramic
Material 10-20, -10 parts of growth factors 5,20-30 parts of poly butylene succinate；
The Hydrogels material is one of collagen, gelatin, polyvinyl alcohol；
The bioceramic material is one of tricalcium phosphate, hydroxyapatite, bio-vitric or more than one mixing；
The anti-coagulants is one of heparin, sodium oxalate, sodium citrate or more than one mixing；
The inoculating cell is one of fat stem cell, multipotential stem cell, mescenchymal stem cell and vascular smooth muscle cells
Or more than one mixing；
The growth factor is VEGF.
4. preparing a kind of method of artificial blood vessel as claimed in claim 3, which is characterized in that itself the following steps are included:
(1), anti-coagulants, polyethylene glycol succinate, Hydrogels material, bioceramic material according to parts by weight, are weighed respectively
Material, growth factor, poly butylene succinate；
(2), the weighed anti-coagulants of the step (1), polyethylene glycol succinate and appropriate volatile polar solvent is sufficiently molten
Solution carries out electrostatic spinning after mixing, is prepared into one layer of porous fibre tubulose structure, completes the system of the inner stent
It is standby；The electrostatic spinning preparation method of the inner stent is rotation winding；
(3), the weighed Hydrogels material of the step (1), bioceramic material, growth factor and proper amount of acetic acid solution are filled
Divide dissolution after mixing, the inner stent outer surface is made in step (2) and carries out 3D printing post-crosslinking, is prepared into one
Layer porous fibre tubulose structure, completes the preparation of the middle layer bracket；The 3D printing preparation method of the middle layer bracket is
Level printing and annular print the progress that alternates；
(4), weighed sufficiently dissolve poly butylene succinate with appropriate volatile polar solvent of the step (1) is mixed
After uniformly, the middle layer rack surface made from step (2) carries out electrostatic spinning, is prepared into one layer of porous fibre tubulose knot
Structure completes the preparation of the outer stent；It prepares since then and completes the intravascular stent；It is prepared by the electrostatic spinning of the outer stent
Method is rotation winding；
(5), the intravascular stent made from the step (4) is put into after appropriate amount of deionized water cleans up, by the inoculation
The suspension of cell is added drop-wise in the hole of the middle layer bracket section in an end face of the intravascular stent, is completed cell and is connect
Kind, and be put into culture solution, the artificial blood vessel is made.
5. preparing a kind of method of artificial blood vessel as claimed in claim 4, which is characterized in that the volatilization in the step (2)
Property polar solvent be one of chloroform, methylene chloride or acetone；Volatile polar solvent in the step (4) be chloroform,
One of methylene chloride or acetone.
6. preparing a kind of method of artificial blood vessel as claimed in claim 4, which is characterized in that the water-setting in the step (3)
When glue class material selection polyvinyl alcohol hydrogel, the crosslinking after 3D printing is first to be lower than -10 DEG C of freezings at least 8 hours, then
It is placed at 20 DEG C -25 DEG C and thaws 6 hours again, the freeze-thaw alternate treatment 4 times；Hydrogels material selection collagen hydrogel
Or when gelatin hydrogel, the crosslinking after 3D printing is to carry out ultraviolet light cross-linking.
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| KR102177977B1 (en) * | 2018-12-20 | 2020-11-13 | 서강대학교산학협력단 | Manufacturing Method of Artificial Blood Vessel |
| CN109859595A (en) * | 2019-01-28 | 2019-06-07 | 深圳大学 | A kind of blood vessel network ultrasound imitates body and production method |
| CN109847103B (en) * | 2019-03-04 | 2020-11-06 | 陈汉杰 | Artificial blood vessel and preparation method thereof |
| CN110237304A (en) * | 2019-06-26 | 2019-09-17 | 上海市第六人民医院 | A kind of multilevel structure bracket and preparation method thereof |
| CN113017944A (en) * | 2019-12-25 | 2021-06-25 | 广东省人民医院(广东省医学科学院) | Artificial blood vessel stent with bioactivity, preparation method and application thereof |
| CN112603593B (en) * | 2020-12-14 | 2022-04-19 | 上海大学 | Self-anastomotic artificial blood vessel stent and preparation method thereof |
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