CN114732955A - Petal-shaped double-layer artificial blood vessel and preparation method thereof - Google Patents
Petal-shaped double-layer artificial blood vessel and preparation method thereof Download PDFInfo
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- 210000004204 blood vessel Anatomy 0.000 title claims abstract description 131
- 239000002473 artificial blood Substances 0.000 title claims abstract description 71
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 238000003618 dip coating Methods 0.000 claims abstract description 63
- 238000010041 electrostatic spinning Methods 0.000 claims abstract description 52
- 238000000034 method Methods 0.000 claims abstract description 18
- 239000010410 layer Substances 0.000 claims description 139
- 239000000243 solution Substances 0.000 claims description 81
- 101000691618 Homo sapiens Inactive phospholipase C-like protein 1 Proteins 0.000 claims description 59
- 102100026207 Inactive phospholipase C-like protein 1 Human genes 0.000 claims description 59
- 229920000848 poly(L-lactide-ε-caprolactone) Polymers 0.000 claims description 59
- 238000009987 spinning Methods 0.000 claims description 42
- 229910001220 stainless steel Inorganic materials 0.000 claims description 23
- 239000010935 stainless steel Substances 0.000 claims description 23
- 239000011247 coating layer Substances 0.000 claims description 17
- 239000002904 solvent Substances 0.000 claims description 14
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 9
- 238000007598 dipping method Methods 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000003517 fume Substances 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 239000012046 mixed solvent Substances 0.000 claims description 3
- 230000017531 blood circulation Effects 0.000 abstract description 9
- 230000007547 defect Effects 0.000 abstract description 3
- 230000014759 maintenance of location Effects 0.000 abstract description 3
- 230000003204 osmotic effect Effects 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 16
- 229920002451 polyvinyl alcohol Polymers 0.000 description 16
- 239000002202 Polyethylene glycol Substances 0.000 description 11
- 229920001223 polyethylene glycol Polymers 0.000 description 10
- 229920000642 polymer Polymers 0.000 description 7
- 230000006872 improvement Effects 0.000 description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 238000001523 electrospinning Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 229920001688 coating polymer Polymers 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 230000002792 vascular Effects 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000004814 polyurethane Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000004088 foaming agent Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000541 pulsatile effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
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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
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
-
- 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
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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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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- 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
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0076—Electro-spinning characterised by the electro-spinning apparatus characterised by the collecting device, e.g. drum, wheel, endless belt, plate or grid
- D01D5/0084—Coating by electro-spinning, i.e. the electro-spun fibres are not removed from the collecting device but remain integral with it, e.g. coating of prostheses
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- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
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Abstract
The application discloses a preparation method of a petal-shaped double-layer artificial blood vessel, which comprises the steps of mold manufacturing, demolding layer forming, blood vessel inner layer forming, blood vessel outer layer forming and demolding, wherein the dip-coating method is adopted for the blood vessel inner layer forming, the electrostatic spinning method is adopted for the blood vessel outer layer forming, the structural uniformity of all parts of the petal-shaped double-layer artificial blood vessel is improved, and mechanical performance indexes such as blood vessel osmotic pressure, suture retention strength and the like meet requirements. The petal-shaped double-layer artificial blood vessel prepared by the preparation method can be further stretched in the wave-shaped circumferential direction under the radial pressure of pulsating blood flow, so that the diameter of the blood vessel is enlarged, the radial expansion capability of the artificial blood vessel is improved, the defect of the compliance of the conventional artificial blood vessel can be overcome, and the petal-shaped double-layer artificial blood vessel prepared by the preparation method of the petal-shaped double-layer artificial blood vessel is also provided.
Description
Technical Field
The application relates to a petal-shaped double-layer artificial blood vessel and a preparation method thereof.
Background
Cardiovascular diseases are one of the common diseases endangering human health, and compared with more serious patients, the main and auxiliary treatment means is blood vessel transplantation, the source of autologous blood vessels is limited, so a large amount of artificial blood vessels are clinically needed as a transplant substitute. The ideal artificial blood vessel has good histocompatibility, hemocompatibility and compliance, wherein the compliance refers to the radial expansion capability of the blood vessel under blood pressure, and the blood flow dynamics abnormality at the anastomotic site after the transplantation can be caused when the compliance of the artificial blood vessel is not matched with the host blood vessel, so that the intimal hyperplasia, the thrombus and the like are caused, and the important factors influencing the patency of the blood vessel after the operation are provided.
Because the cross section of the human blood vessel is approximately circular, the cross section of most artificial blood vessels is designed to be circular or approximately circular, the compliance of most artificial blood vessels is lower than that of the human host blood vessel at present due to the limitation of materials, structures and preparation methods, and the improvement of the compliance of the artificial blood vessels from the aspect of the design of the cross section shape is a new way.
Disclosure of Invention
The application aims to provide a preparation method of a petal-shaped double-layer artificial blood vessel, which is used for preparing the double-layer artificial blood vessel with a petal-shaped section so as to improve the compliance of the artificial blood vessel.
The technical scheme adopted by the application is as follows: the preparation method of the petal-shaped double-layer artificial blood vessel comprises the following steps:
s1, preparing a mold: providing cylindrical stainless steel, enabling the diameter of the cylindrical stainless steel to be equal to that of the petal-shaped double-layer artificial blood vessel, and processing the cylindrical stainless steel to enable the cross section of the cylindrical stainless steel to be petal-shaped;
s2, forming a demolding layer: dip-coating a demolding solution on the outer side of the mold by adopting a dip-coating method to form a demolding layer;
s3, forming of the inner layer of the blood vessel: dip-coating a first PLCL solution on the outer side of the demolding layer by adopting a dip-coating method to form a dip-coating layer;
s4, forming the outer layer of the blood vessel: forming an electrostatic spinning layer on the outer side of the dip-coating layer by using a second PLCL solution as an electrostatic spinning solution by adopting an electrostatic spinning method;
s5, demolding: and separating the layered structure formed outside the mold from the mold to obtain the petal-shaped double-layer artificial blood vessel.
In step S1, the diameter of the cylindrical stainless steel is set to be 3-8 mm, the cross section of the cylindrical stainless steel includes a plurality of circular arc petals evenly distributed along the circumferential direction, and two adjacent circular arc petals are connected by a smooth curve.
As an improvement to the scheme, the cross section comprises 4-6 circular arc-shaped petals which are uniformly distributed along the circumferential direction, and the circular arc-shaped petals are semicircular.
As an improvement to the above scheme, in step S2, the demolding solution is a PVA solution, a solvent of the PVA solution is a mixed solvent of alcohol and deionized water, and a concentration of the PVA solution is 12% to 18%; dip coating a mold release solution on the outside of the mold using a dip coating method includes:
s21, fixing the die on a clamp of a dip-coating machine, and dipping the die into the PVA solution at a speed of 1000-5000 microns/s for 8-12 s;
s22, withdrawing the mold from the PVA solution at the speed of 500-5000 mu m/s;
s23, fixing the mold on a homogenizing motor, and rotating at the speed of 200-500 r/min for 5-10 min;
s24, placing the mold in an oven, and drying for 25-35 min at the temperature of 40-60 ℃;
s25, the die is reversely fixed on a clamp of the dip coater, and the steps from S21 to S24 are repeated at least once.
In a modification of the above scheme, in step S3, the solvent of the first PLCL solution is hexafluoroisopropanol, the first PLCL solution further includes PEG, and the concentration of PLCL in the first PLCL solution is 4% to 8%, and the concentration of PEG in the first PLCL solution is 2% to 4%.
As a modification to the above, dip coating the first PLCL solution outside the release layer using a dip coating method in step S3 includes:
s31, fixing the mold for forming the demolding layer on a clamp of a dip-coating machine, and dipping the mold into the first PLCL solution at the speed of 1500-5500 mu m/s for 6-10 s;
s32, withdrawing the mould from the first PLCL solution at the speed of 500-5000 mu m/s;
s33, standing the die for 8-12 min;
s34, the die is reversely fixed on a clamp of the dip coater, and the steps from S31 to S33 are repeated at least once.
As an improvement to the above scheme, in step S4, the solvent of the second PLCL solution is hexafluoroisopropanol, and the concentration of PLCL is 8% to 12%; forming an electrostatic spinning layer on the outer side of the dip-coating layer by using an electrostatic spinning method comprises the following steps:
s41, fixing the die for completing the inner layer forming of the blood vessel on a collecting device of an electrostatic spinning machine;
s42, injecting the second PLCL solution into a positive pressure injection pump and a negative pressure injection pump of the electrostatic spinning machine;
s43, starting the electrostatic spinning machine, and driving the die to rotate by the collecting device, so that the positive-pressure spinning needle head and the negative-pressure spinning needle head jet spinning preset time from two sides to the die.
As an improvement to the above scheme, the voltage of the electrostatic spinning machine is 10-15 kv, and in S41, the distance between the positive pressure spinning needle and the negative pressure spinning needle and the collecting device is 15-20 cm; and S43, the rotating speed of the collecting device is 500-800 r/min, and the spinning speeds of the positive pressure spinning needle and the negative pressure spinning needle are 0.5-1 ml/h.
As a modification of the above, the step S5 of separating the layered structure molded outside the mold from the mold includes:
s51, taking down the die for completing the outer-layer forming of the blood vessel from the electrostatic spinning machine, and drying in a fume hood for 10-20 hours;
s52, soaking the die in normal temperature-removed ionized water for 48-72 hours;
s53, the die is taken out, one end of the die is clamped, the layered structure on the outer side of the die is clamped by tweezers, and the layered structure is dragged and rotated to be separated from the die.
The technical scheme adopted by the application is as follows: the petal-shaped double-layer artificial blood vessel comprises a blood vessel inner layer and a blood vessel outer side, wherein the blood vessel inner layer is a dip coating layer of a first PLCL solution, the blood vessel outer layer is an electrostatic spinning layer of a second PLCL solution, the thickness of the dip coating layer is 60-100 mu m, and the thickness of the electrostatic spinning layer is 90-110 mu m.
The preparation method of the petal-shaped double-layer artificial blood vessel comprises the steps of mold manufacturing, demolding layer forming, blood vessel inner layer forming, blood vessel outer layer forming and demolding, wherein the dip-coating method is adopted for the blood vessel inner layer forming, the electrostatic spinning method is adopted for the blood vessel outer layer forming, the structural uniformity of all parts of the petal-shaped double-layer artificial blood vessel is improved, and mechanical performance indexes such as blood vessel osmotic pressure, suture retention strength and the like meet requirements. The petal-shaped double-layer artificial blood vessel prepared by the preparation method can be further expanded in the wave-shaped circumferential direction under the radial pressure of pulsating blood flow, so that the diameter of the blood vessel is enlarged, the radial expansion capability of the artificial blood vessel is improved, and the defect of the compliance of the conventional artificial blood vessel can be overcome.
Drawings
Fig. 1 is a schematic cross-sectional structure diagram of a petal-shaped double-layered artificial blood vessel in the present application.
Detailed Description
The application provides a preparation method of a petal-shaped double-layer artificial blood vessel, which is used for preparing the double-layer artificial blood vessel with a petal-shaped section so as to improve the compliance of the artificial blood vessel.
In the description of the present application, PVA refers to polyvinyl alcohol, PEO refers to polyethylene oxide, PLCL refers to poly L-lactide-caprolactone, and PEG refers to polyethylene glycol.
The preparation method of the petal-shaped double-layer artificial blood vessel comprises the following steps:
s1, preparing a mold: providing cylindrical stainless steel, enabling the diameter of the cylindrical stainless steel to be equal to that of the petal-shaped double-layer artificial blood vessel, and processing the cylindrical stainless steel to enable the cross section of the cylindrical stainless steel to be petal-shaped;
s2, forming a demolding layer: dip-coating a demolding solution on the outer side of the mold by adopting a dip-coating method to form a demolding layer;
s3, forming the inner layer of the blood vessel: dip-coating a first PLCL solution on the outer side of the demolding layer by adopting a dip-coating method to form a dip-coating layer;
s4, forming the outer layer of the blood vessel: forming an electrostatic spinning layer on the outer side of the dip-coating layer by using a second PLCL solution as an electrostatic spinning solution by adopting an electrostatic spinning method;
s5, demolding: and separating the layered structure formed outside the mold from the mold to obtain the petal-shaped double-layer artificial blood vessel.
Further, in step S1, the diameter of the cylindrical stainless steel is set to be 3-8 mm, the cylindrical stainless steel is machined through a lathe, so that the cross section of the cylindrical stainless steel is in a petal shape, the cross section of the cylindrical stainless steel comprises a plurality of circular arc-shaped petals which are uniformly distributed along the circumferential direction, and two adjacent circular arc-shaped petals are connected through a smooth curve, so that smooth transition between two adjacent circular arc-shaped petals of the petal-shaped double-layer vascular prosthesis formed by the die is ensured.
Furthermore, the cross section comprises 4-6 circular arc-shaped petals which are uniformly distributed along the circumferential direction, and the circular arc-shaped petals are semicircular, namely the central angle of the circular arc-shaped petals is 180 degrees.
Further, in step S2, the demolding solution is a PVA solution, a solvent of the PVA solution is a mixed solvent of alcohol and deionized water, and a concentration of the PVA solution is 12% to 18%; dip coating a mold release solution on the outside of the mold using a dip coating method includes:
s21, fixing the die on a clamp of a dip-coating machine, and dipping the die into the PVA solution at a speed of 1000-5000 microns/s for 8-12 s;
s22, withdrawing the mold from the PVA solution at the speed of 500-5000 mu m/s;
s23, fixing the mold on a homogenizing motor, and rotating at the speed of 200-500 r/min for 5-10 min;
s24, placing the mold in an oven, and drying for 25-35 min at the temperature of 40-60 ℃;
s25, the die is reversely fixed on a clamp of the dip coater, and the steps from S21 to S24 are repeated at least once.
It should be noted that the dip coater in the present application is a dip coater.
It is understood that the purpose of coating the mold surface with the release layer is to enable the molded vascular prosthesis to be separated from the mold more smoothly, and the release layer formed on the mold in the above step S2 includes at least two release base layers, each of which is molded on the mold through steps S21-S24.
Further, in step S3, the solvent of the first PLCL solution is hexafluoroisopropanol. The first PLCL solution also comprises PEG, and the first PLCL solution is used as a dip coating solution, wherein the concentration of the PLCL is 4% -8%, and the concentration of the PEG is 2% -4%.
Wherein, PEG is used as a pore-foaming agent, so that the intravascular layer is a dip-coating layer with a pore structure.
In this step, PLCL is used as a dip coating polymer, and a hexafluoroisopropanol solution of PLCL is used as a dip coating solution, where it should be noted that the dip coating polymer may also be selected from other polymers, such as polymers like Polyurethane (PU), and a solvent of the dip coating polymer may also be selected from organic solvents like chloroform and tetrahydrofuran.
Further, the dip coating of the first PLCL solution outside the release layer using a dip coating method in step S3 includes:
s31, fixing the mold for molding the demolding layer on a clamp of a dip coating machine, and dipping the mold into the first PLCL solution at the speed of 1500-5500 mu m/s for 6-10 s;
s32, withdrawing the mould from the first PLCL solution at the speed of 500-5000 mu m/s;
s33, standing the die for 8-12 min;
s34, the die is reversely fixed on a clamp of the dip coater, and the steps from S31 to S33 are repeated at least once.
The dip coating layer formed on the mold in the above step S3 includes at least two dip coating base layers, each of which is molded on the mold through steps S31-S33. It is understood that, in the actual preparation process, the number of the formed dip-coated base layer can be determined according to the thickness requirement of the inner layer of the petal-shaped double-layer artificial blood vessel, that is, the number of times of repeating the steps S31-S33 is determined.
The more the number of the layers of the dip-coating base layer is, the thicker the inner layer of the prepared petal-shaped double-layer artificial blood vessel is.
Further, in step S4, the solvent of the second PLCL solution is hexafluoroisopropanol, and the concentration of PLCL is 8% to 12%. In this step, PLCL is used as an electrospinning polymer, and a hexafluoroisopropanol solution of PLCL is used as an electrospinning solution, where the electrospinning polymer may also be selected from other polymers, such as polymers like Polyurethane (PU), and a solvent of the electrospinning polymer may also be selected from organic solvents like chloroform and tetrahydrofuran.
Forming an electrostatic spinning layer on the outer side of the dip-coating layer by adopting an electrostatic spinning method comprises the following steps:
s41, fixing the die for completing the inner layer forming of the blood vessel on a collecting device of an electrostatic spinning machine;
s42, injecting the second PLCL solution into a positive pressure injection pump and a negative pressure injection pump of the electrostatic spinning machine;
s43, starting the electrostatic spinning machine, and driving the die to rotate by the collecting device, so that the positive-pressure spinning needle head and the negative-pressure spinning needle head jet spinning preset time from two sides to the die.
It can be understood that in the actual preparation process, the spinning time can be controlled according to the thickness requirement of the outer layer of the petal-shaped double-layer artificial blood vessel. The longer the spinning time is, the thicker the electrostatic spinning layer is, and the thicker the outer layer of the prepared petal-shaped double-layer artificial blood vessel is.
The electrostatic spinning machine comprises two high-voltage direct-current power supplies, namely a positive-voltage direct-current power supply and a negative-voltage direct-current power supply, wherein the positive-voltage direct-current power supply is used for outputting positive voltage, the negative-voltage direct-current power supply is used for outputting negative voltage, a positive-voltage spinning needle head is connected with the positive-voltage direct-current power supply, and the negative-voltage spinning needle head is connected with the negative-voltage direct-current power supply.
The collecting device is arranged between the positive pressure spinning needle head and the negative pressure spinning needle head, the positive pressure spinning needle head and the negative pressure spinning needle head are arranged oppositely and are respectively positioned at two opposite sides above the collecting device, and the positive pressure spinning needle head and the negative pressure spinning needle head are obliquely downwards from two sides to jet yarns towards the die on the collecting device to form conjugated electrostatic spinning.
Furthermore, the voltage of the electrostatic spinning machine is 10-15 kv, and in S41, the distance between the positive pressure spinning needle and the collection device, and the distance between the negative pressure spinning needle and the collection device are 15-20 cm; and S43, the rotating speed of the collecting device is 500-800 r/min, and the spinning speeds of the positive pressure spinning needle and the negative pressure spinning needle are 0.5-1 ml/h.
Further, the separating the layered structure formed outside the mold from the mold in step S5 includes:
s51, taking down the die for completing the outer-layer forming of the blood vessel from the electrostatic spinning machine, and drying in a fume hood for 10-20 hours;
s52, soaking the die in normal temperature-removed ionized water for 48-72 hours;
the step is used for removing the demoulding layer formed on the mould, so that the multilayer composite pipe is convenient to separate from the mould.
S53, taking out the mold, clamping one end of the mold, clamping the layered structure outside the mold by using forceps, dragging and rotating to separate the layered structure from the mold, and obtaining the double-layer artificial blood vessel with the petal-shaped section.
In an application scenario, the preparation method of the petal-shaped double-layer artificial blood vessel further includes the step of drying and storing the petal-shaped double-layer artificial blood vessel obtained in step S5. Specifically, the petal-shaped double-layer artificial blood vessel obtained in the step S5 is naturally dried and stored in a self-sealing bag.
In the preparation method of the petal-shaped double-layer artificial blood vessel, the outer surface of the mold with the petal-shaped cross section is approximate to a wave shape, so that the structural uniformity of the positions of the wave crest and the wave trough is inconsistent when the artificial blood vessel is prepared only by adopting an electrostatic spinning method. Therefore, the dip coating method is adopted for forming the inner layer of the blood vessel, and the electrostatic spinning method is adopted for the outer layer of the blood vessel, so that the defect of insufficient uniformity of the petal-shaped blood vessel prepared by the electrostatic spinning method can be overcome, and the mechanical performance indexes of osmotic pressure, suture retention strength and the like of the artificial blood vessel can meet the requirements.
The double-layer artificial blood vessel in the shape of a petal prepared by the preparation method has a double-layer structure comprising a blood vessel inner layer with a pore structure and a blood vessel outer layer with an electrostatic spinning structure, and the inner side of the blood vessel can generate swirling flow, so that the blood flow rate and the wall shear stress close to the wall surface are improved, and the deposition of harmful substances on the inner wall of the blood vessel can be reduced; under the radial pressure of pulsating blood flow, the vessel wall can be further stretched in the wavy circumferential direction, so that the diameter of the vessel is enlarged, the radial expansion capability of the artificial vessel is enhanced, and the compliance of the artificial vessel is improved.
The present application will be described in further detail with reference to examples. It is to be noted that the following examples are only illustrative of the present application, and do not limit the scope of the present application. Likewise, the following examples are only some examples and not all examples of the present application, and all other examples obtained by a person of ordinary skill in the art without any inventive step are within the scope of the present application.
An embodiment of the application provides a preparation method of a petal-shaped double-layer artificial blood vessel, which comprises the following steps:
s1, preparing a mold:
providing cylindrical stainless steel with the length of 120mm and the diameter of 6mm, processing the cylindrical stainless steel to enable the cross section of the cylindrical stainless steel to be in a petal shape, wherein the cross section comprises 6 circular arc-shaped petals which are uniformly distributed along the circumferential direction, and the circular arc-shaped petals are arranged in a semicircular shape.
S2, forming a demolding layer:
s20, weighing 7.5g of PVA, and preparing a 15% PVA solution and a deionized water and alcohol mixed solution with a solvent of 50: 50.
S21, fixing the die on a clamp of a dip coating machine, and dipping the die into the PVA solution at a speed of 5000 microns/s for 10 s;
s22, withdrawing the mould from the PVA solution at the speed of 5000 mu m/s;
s23, fixing the mold on a homogenizing motor, and rotating for 5min at the speed of 300 r/min;
s24, placing the mold in an oven, drying for 30min at the temperature of 60 ℃, and forming a first layer of demolding base layer on the outer side of the mold;
s25, the die is reversely fixed on a clamp of the dip coating machine, and S21-S24 are repeated once, so that a second layer of demoulding base layer is formed on the outer side of the die.
S3, forming of the inner layer of the blood vessel:
s30, preparing a first PLCL solution: weighing 2g of PLCL, preparing a PLCL solution with the concentration of 6%, adding PEG (polyethylene glycol) serving as a pore-forming agent into the PLCL solution to enable the concentration of the PEG to be 3%, wherein the solvent is hexafluoroisopropanol;
s31, fixing the mold for forming the demolding layer on a clamp of a dip coating machine, and dipping the mold into the first PLCL solution at the speed of 4500 mu m/s for 8 s;
s32, withdrawing the mould from the first PLCL solution at the speed of 900 mu m/s;
s33, standing the die for 10min to form a first dip-coating base layer on the outer side of the demolding layer;
s34, the die is reversely fixed on a clamp of a dip-coating machine, and S31-S33 are repeated once, so that a second layer of dip-coating base layer is formed on the outer side of the demolding layer.
In this embodiment, the intravascular layer comprises two dip-coated base layers, and the intravascular layer has a thickness of about 60 μm.
S4, forming the outer layer of the blood vessel:
s40, preparing a second PLCL solution: weighing 1g of PLCL, and preparing a PLCL solution with the concentration of 8% by using hexafluoroisopropanol as a solvent;
s41, fixing the die for completing the inner layer forming of the blood vessel on a collecting device of an electrostatic spinning machine;
s42, injecting the second PLCL solution into a positive pressure injection pump and a negative pressure injection pump of the electrostatic spinning machine;
and S43, starting the electrostatic spinning machine, driving the die to rotate by the collecting device, and spinning for 1.5 hours from two sides of the positive pressure spinning needle and the negative pressure spinning needle towards the die to form an electrostatic spinning layer on the outer side of the inner blood vessel layer, namely the outer blood vessel layer.
In this example, the thickness of the electrospun layer was about 100 μm.
In this embodiment, the voltage of the electrostatic spinning machine is 12KV, the distances between the positive pressure spinning needle and the negative pressure spinning needle and the collecting device are 15cm, the rotation speed of the collecting device is 300rpm, and the spinning speeds of the positive pressure spinning needle and the negative pressure spinning needle are 0.8 ml/h.
(5) Demolding:
s51, taking down the die for completing the outer layer forming of the blood vessel from the electrostatic spinning machine, and drying the die in a fume hood for 12 hours;
s52, soaking the die in normal temperature deionized water for 72 hours;
s53, the mold is taken out, one end of the mold is clamped, the layered structure on the outer side of the mold is clamped by forceps, and the mold is dragged and rotated to separate the layered structure from the mold, so that the petal-shaped double-layer artificial blood vessel is obtained.
It can be understood that the cross-sectional shape of the petal-shaped double-layered artificial blood vessel is consistent with the cross-section of the mold.
The petaloid double-layer artificial blood vessel comprises a blood vessel inner layer and a blood vessel outer layer, wherein the blood vessel inner layer is a dip coating layer of a first PLCL solution, the blood vessel outer layer is an electrostatic spinning layer of a second PLCL solution, the thickness of the dip coating layer is 60-100 mu m, and the thickness of the electrostatic spinning layer is 90-110 mu m.
The petal-shaped double-layer artificial blood vessel has a double-layer structure comprising a blood vessel inner layer with a pore structure and a blood vessel outer layer with an electrostatic spinning structure, and the inner side of the blood vessel can generate swirling flow, so that the blood flow rate close to the wall surface and the wall surface shear stress are improved, and the deposition of harmful substances on the inner wall of the blood vessel can be reduced; under the radial pressure of pulsating blood flow, the vessel wall can be further stretched in the wavy circumferential direction, so that the diameter of the vessel is enlarged, the radial expansion capability of the artificial vessel is enhanced, and the compliance of the artificial vessel is improved.
Furthermore, the diameter of the petal-shaped double-layer artificial blood vessel is 3-8 mm, the cross section of the artificial blood vessel comprises a plurality of circular arc-shaped petals which are uniformly distributed along the circumferential direction, and the two adjacent circular arc-shaped petals are in smooth transition so as to reduce the radial resistance of pulsatile blood flow and further improve the compliance of the artificial blood vessel.
Under the radial pressure of a plurality of evenly distributed arc petals in pulsating blood flow, the vessel wall further extends in the circumferential direction of uniform wave shape, and the stress consistency of each position of the vessel wall is improved.
Further, the cross section of the petal-shaped double-layer artificial blood vessel comprises 4-6 circular arc petals which are uniformly distributed along the circumferential direction, please refer to fig. 1, and fig. 1 shows the structure of the cross section of the petal-shaped double-layer artificial blood vessel in the application when the cross section is 4 circular arc petals, 5 circular arc petals and 6 circular arc petals.
Through further research by the inventor of the application, the cross section of the petal-shaped artificial blood vessel is closer to the circular shape when the circular-arc petals are arranged into the circular-arc shape with the central angle smaller than 180 degrees, and the improvement degree of the compliance of the petal-shaped artificial blood vessel is not high compared with the conventional artificial blood vessel with the circular cross section; when circular arc petal sets up to the central angle and is greater than 180 circular arc, the distance of junction and the petal apex of two adjacent circular arc petals is far away, and the atress uniformity of vascular wall each position department is lower, consequently in this application, preferably, circular arc petal sets up to semi-circular, promptly circular arc petal's central angle is 180.
The above description is only an example of the present application and is not intended to limit the scope of the present application, and all modifications of equivalent structures and equivalent processes, which are made by the contents of the specification and the drawings, or which are directly or indirectly applied to other related technical fields, are intended to be included within the scope of the present application.
Claims (10)
1. A preparation method of a petal-shaped double-layer artificial blood vessel is characterized by comprising the following steps:
s1, preparing a mold: providing cylindrical stainless steel, enabling the diameter of the cylindrical stainless steel to be equal to that of the petal-shaped double-layer artificial blood vessel, and processing the cylindrical stainless steel to enable the cross section of the cylindrical stainless steel to be petal-shaped;
s2, forming a demolding layer: dip-coating a demolding solution on the outer side of the mold by adopting a dip-coating method to form a demolding layer;
s3, forming of the inner layer of the blood vessel: dip-coating a first PLCL solution on the outer side of the demolding layer by adopting a dip-coating method to form a dip-coating layer;
s4, forming the outer layer of the blood vessel: forming an electrostatic spinning layer on the outer side of the dip-coating layer by using a second PLCL solution as an electrostatic spinning solution by adopting an electrostatic spinning method;
s5, demolding: and separating the layered structure formed outside the mold from the mold to obtain the petal-shaped double-layer artificial blood vessel.
2. The method for preparing a petal-shaped double-layer artificial blood vessel according to claim 1, wherein in step S1, the diameter of the cylindrical stainless steel is set to be 3-8 mm, the cross section of the cylindrical stainless steel comprises a plurality of circular arc-shaped petals which are uniformly distributed along the circumferential direction, and two adjacent circular arc-shaped petals are connected by a smooth curve.
3. The preparation method of the petal-shaped double-layer artificial blood vessel as claimed in claim 2, wherein the cross section comprises 4-6 circular arc petals which are uniformly distributed along the circumferential direction, and the circular arc petals are arranged in a semicircular shape.
4. The method for preparing a petal-shaped double-layered artificial blood vessel according to claim 1, wherein in step S2, the mold release solution is a PVA solution, the solvent of the PVA solution is a mixed solvent of alcohol and deionized water, and the concentration of the PVA solution is 12% to 18%; dip coating a mold release solution on the outside of the mold using a dip coating method includes:
s21, fixing the die on a clamp of a dip-coating machine, and dipping the die into the PVA solution at a speed of 1000-5000 microns/s for 8-12 s;
s22, withdrawing the mold from the PVA solution at the speed of 500-5000 mu m/s;
s23, fixing the mold on a homogenizing motor, and rotating at the speed of 200-500 r/min for 5-10 min;
s24, placing the mold in an oven, and drying for 25-35 min at the temperature of 40-60 ℃;
s25, the die is reversely fixed on a clamp of the dip coater, and the steps from S21 to S24 are repeated at least once.
5. The method of claim 1, wherein in step S3, the solvent of the first PLCL solution is hexafluoroisopropanol, the first PLCL solution further comprises PEG, and the concentration of PLCL in the first PLCL solution is 4% to 8%, and the concentration of PEG in the first PLCL solution is 2% to 4%.
6. The method for preparing a petal-shaped double-layered artificial blood vessel according to claim 5, wherein the step S3 of dip-coating a first PLCL solution outside the release layer by a dip-coating method comprises:
s31, fixing the mold for forming the demolding layer on a clamp of a dip-coating machine, and dipping the mold into the first PLCL solution at the speed of 1500-5500 mu m/s for 6-10 s;
s32, withdrawing the mould from the first PLCL solution at the speed of 500-5000 mu m/s;
s33, standing the die for 8-12 min;
s34, the die is reversely fixed on a clamp of the dip coater, and the steps from S31 to S33 are repeated at least once.
7. The method for preparing a petal-shaped bilayer artificial blood vessel according to claim 1, wherein in step S4, the solvent of the second PLCL solution is hexafluoroisopropanol, and the concentration of PLCL is 8% to 12%; forming an electrostatic spinning layer on the outer side of the dip-coating layer by adopting an electrostatic spinning method comprises the following steps:
s41, fixing the die for completing the inner layer forming of the blood vessel on a collecting device of an electrostatic spinning machine;
s42, injecting the second PLCL solution into a positive pressure injection pump and a negative pressure injection pump of the electrostatic spinning machine;
and S43, starting the electrostatic spinning machine, and driving the die to rotate by the collecting device to enable the positive-pressure spinning needle and the negative-pressure spinning needle to jet spinning for a preset time from two sides to the die.
8. The preparation method of the petal-shaped double-layer artificial blood vessel as claimed in claim 7, wherein the voltage of the electrostatic spinning machine is 10-15 kv, and in S41, the distance between the positive pressure spinning needle and the negative pressure spinning needle and the collecting device is 15-20 cm; and S43, the rotating speed of the collecting device is 500-800 r/min, and the spinning speeds of the positive pressure spinning needle and the negative pressure spinning needle are 0.5-1 ml/h.
9. The method of claim 1, wherein the separating the layered structure formed outside the mold from the mold in step S5 comprises:
s51, taking down the die for completing the outer-layer forming of the blood vessel from the electrostatic spinning machine, and drying in a fume hood for 10-20 hours;
s52, soaking the die in normal temperature-removed ionized water for 48-72 hours;
s53, the die is taken out, one end of the die is clamped, the layered structure on the outer side of the die is clamped by tweezers, and the layered structure is dragged and rotated to be separated from the die.
10. A petal-shaped double-layered artificial blood vessel, which is prepared by the preparation method of the petal-shaped double-layered artificial blood vessel of claim 1, and comprises an inner blood vessel layer and an outer blood vessel layer, wherein the inner blood vessel layer is a dip coating layer of a first PLCL solution, the outer blood vessel layer is an electrostatic spinning layer of a second PLCL solution, the thickness of the dip coating layer is 60-100 μm, and the thickness of the electrostatic spinning layer is 90-110 μm.
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