CN108642719B - Preparation method of integrated small-caliber artificial blood vessel stent - Google Patents
Preparation method of integrated small-caliber artificial blood vessel stent Download PDFInfo
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- CN108642719B CN108642719B CN201810448418.7A CN201810448418A CN108642719B CN 108642719 B CN108642719 B CN 108642719B CN 201810448418 A CN201810448418 A CN 201810448418A CN 108642719 B CN108642719 B CN 108642719B
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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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
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/76—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres otherwise than in a plane, e.g. in a tubular way
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- D—TEXTILES; PAPER
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- D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
- D10B2509/00—Medical; Hygiene
- D10B2509/06—Vascular grafts; stents
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Abstract
The invention discloses an electrostatic spinning integrated small-caliber artificial blood vessel stent device, which comprises a spinning device, a receiving device and a blood vessel stent forming device, wherein the spinning device is used for spinning a small-caliber artificial blood vessel stent; the intravascular stent forming device comprises a fiber winding device and a winding control device, wherein the fiber winding device is used for winding the fiber received by the receiving device into a cylindrical shape; and the winding control device is used for controlling the axial movement and the circumferential rotation of the fiber winding device to form a nanofiber tube, and the nanofiber tube is rinsed to remove the receiving roller to obtain the integrated small-caliber artificial blood vessel support. The invention also discloses a method for preparing the integrated small-caliber artificial blood vessel stent by adopting the device. The device has simple structure, low cost, easy operation of the production process and high efficiency; when the small-caliber artificial blood vessel stent is prepared, the inner diameter of the small-caliber artificial blood vessel stent can be controlled by adjusting the diameter of the cross section of the receiving roller; the small-caliber artificial blood vessel stent prepared by the device has the characteristics of high strength, high porosity, good biocompatibility and the like.
Description
Technical Field
The invention belongs to the technical field of electrostatic spinning, relates to a device for electrostatic spinning of an integrated small-caliber artificial blood vessel stent, and further relates to a method for preparing the integrated small-caliber artificial blood vessel stent by adopting the device.
Background
The human blood vessel comprises three layers of structures of intima, media and adventitia. Each layer is composed of different cells, wherein the inner layer and the outer layer are in contact with blood and have good biocompatibility and blood compatibility; the middle layer needs to maintain the vascular structure and bear the vascular pressure, and thus should have excellent mechanical properties.
In recent society, with the increase of life rhythm and irregular diet, cardiovascular diseases have become a big killer threatening human health and are important causes of high mortality of human beings. The revascularization operation is an effective method for treating cardiovascular diseases, and since the autologous blood vessels have the advantages of good biocompatibility, immunity, antigenicity and the like, the autologous blood vessels are mostly used as substitutes, but the sources of the autologous blood vessels are limited. Therefore, artificial blood vessels become a good choice for treating cardiovascular diseases. At present, large-caliber artificial blood vessels have been used clinically, but the use of small-caliber artificial blood vessels (inner diameter less than 6mm) is under further study. The reason is that when the artificial blood vessel stent with small caliber is implanted into the body, thrombus is easily formed and the intimal hyperplasia of the blood vessel is caused. Therefore, the ideal artificial blood vessel stent should have a similar hierarchical structure to that of a human blood vessel, and its inner surface should have strong adhesion to vascular endothelial cells to form a dense endothelial cell layer to prevent the formation of thrombus and the occurrence of coagulation.
With the rapid development of bioengineering technology and nanotechnology, compared with other preparation methods, the electrospun nanofiber has special advantages in the aspect of being used for the artificial blood vessel stent, because the electrospun nanofiber can form a tubular structure with a required caliber, can also simulate the composition and structure of extracellular matrix, provides a good growth environment for cells, and provides an important precondition for the regeneration and reconstruction of autologous blood vessels.
Disclosure of Invention
The invention aims to provide a device for an electrostatic spinning integrated small-caliber artificial blood vessel stent, which is used for preparing the small-caliber artificial blood vessel stent with good bionic performance.
The invention also aims to provide a method for preparing the integrated small-caliber artificial blood vessel stent by using the device.
The technical scheme adopted by the invention is that the device for the electrostatic spinning integrated small-caliber artificial blood vessel support comprises a spinning device, a receiving device and a blood vessel support forming device, wherein the receiving device and the blood vessel support forming device are arranged on a base;
the spinning device comprises an injector, one end of the injector is connected with the micro-injection pump through a liquid guide pipe, the other end of the injector is provided with a spinning needle head, the spinning needle head is connected with the direct-current high-voltage generator, the receiving device is positioned right below the spinning needle head, and the spinning needle head is vertical to the receiving device;
the receiving device is used for receiving the fibers obtained by the spinning device through electrostatic spinning;
the intravascular stent forming device comprises a fiber winding device and a winding control device, wherein the fiber winding device is used for winding the fiber received by the receiving device into a cylindrical shape; the winding control device is used for controlling the axial movement and the circumferential rotation of the fiber winding device, so that the fibers wound on the fiber winding device form a vascular stent structure.
The present invention is also characterized in that,
the receiving device comprises two receiving plates, the receiving plates are vertically and fixedly arranged on the base, the receiving plates are grounded, the spinning needle head is positioned right above a plane formed by the two receiving plates, and fibers obtained by spinning are lapped between the two receiving plates.
The two receiving plates are parallel to each other.
The fiber winding device comprises a receiving roller, the receiving roller is positioned between two receiving plates, and the receiving roller is made of terylene or polytetrafluoroethylene.
The winding control device comprises a circumferential rotation control device and an axial movement control device which are respectively used for controlling the circumferential rotation and the axial movement of the receiving roller;
the circumferential rotation control device comprises a variable frequency motor b, the variable frequency motor b is arranged on the sliding plate, and the variable frequency motor b is connected with one end of the receiving roller and used for controlling the receiving roller to rotate circumferentially;
axial displacement controlling means is including setting up track and the stroke spacing groove on the base, the slide is along track horizontal migration, the slide is kept away from receiving roller one end and is connected with the movable rod, the movable rod both ends are installed respectively in the stroke spacing inslot, the movable rod is connected with connecting rod one end, the connecting rod other end is provided with rotatory piece, inverter motor a drives rotatory piece through the dabber and is the rotation in a circumferential direction, the drive connecting rod drives the movable rod and removes in the stroke spacing inslot, thereby drive slide horizontal migration, and then drive receiving roller axial displacement.
The invention adopts another technical scheme that the preparation method of the integrated small-caliber artificial blood vessel bracket adopts a device for electrospinning the integrated small-caliber artificial blood vessel bracket, and the device comprises a spinning device, a receiving device and a blood vessel bracket forming device, wherein the receiving device and the blood vessel bracket forming device are arranged on a base;
the spinning device comprises an injector, one end of the injector is connected with the micro-injection pump through a liquid guide pipe, the other end of the injector is provided with a spinning needle head, the spinning needle head is connected with the direct-current high-voltage generator, the receiving device is positioned right below the spinning needle head, and the spinning needle head is vertical to the receiving device;
the receiving device is used for receiving the fibers obtained by electrostatic spinning of the spinning device; the receiving device comprises two receiving plates, the receiving plates are fixedly and vertically arranged on the base, the receiving plates are grounded, the spinning needle head is positioned right above a plane formed by the two receiving plates, fibers obtained by spinning are lapped between the two receiving plates, and the two receiving plates are parallel to each other;
the intravascular stent forming device comprises a fiber winding device and a winding control device, wherein the fiber winding device is used for winding the fiber received by the receiving device into a cylindrical shape; the winding control device is used for controlling the axial movement and the circumferential rotation of the fiber winding device so that the fibers wound on the fiber winding device form a vascular stent structure;
the fiber winding device comprises a receiving roller, and the receiving roller is positioned between two receiving plates;
the winding control device comprises a circumferential rotation control device and an axial movement control device which are respectively used for controlling the circumferential rotation and the axial movement of the receiving roller;
the circumferential rotation control device comprises a variable frequency motor b, the variable frequency motor b is arranged on the sliding plate, and the variable frequency motor b is connected with one end of the receiving roller and used for controlling the receiving roller to rotate circumferentially;
the axial movement control device comprises a track and a stroke limiting groove which are arranged on the base, the sliding plate horizontally moves along the track, one end, away from the receiving roller, of the sliding plate is connected with a movable rod, two ends of the movable rod are respectively installed in the stroke limiting groove, the movable rod is connected with one end of a connecting rod, the other end of the connecting rod is provided with a rotating block, a variable frequency motor a drives the rotating block to circumferentially rotate through a mandrel, and the connecting rod is driven to drive the movable rod to move in the stroke limiting groove, so that the sliding plate is driven to horizontally move, and the receiving roller is driven;
the method is implemented according to the following steps:
step 1, preparing spinning solution:
dissolving a biocompatible polymer A in a solvent A' at room temperature, and stirring until the solution is uniform to obtain a solution A with the mass fraction of the polymer A being 7-9%; dissolving a biocompatible polymer B in a solvent B', and stirring until the solution is uniform to obtain a solution B with the mass fraction of the polymer B being 10-12%;
step 2.1, preparing the fiber membrane of the inner layer of the blood vessel:
adding the prepared solution A into an injector under the environmental conditions that the temperature is 20-30 ℃ and the relative humidity is 40-60%, and setting the distance between two receiving plates; turning on a direct-current high-voltage generator, adjusting spinning voltage and spinning distance, and adjusting the flow rate of the solution controlled by the micro-injection pump; starting a variable frequency motor a, adjusting the rotating speed, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, and further enabling a receiving roller to move axially and repeatedly through a connecting rod, a movable rod and a sliding plate; meanwhile, the variable frequency motor b is started, the rotating speed is adjusted, and the receiving roller is driven to rotate in the circumferential direction; jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, and a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotates along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and a fiber membrane of the intravascular layer is obtained;
step 2.2, the solution B and the solution A are sequentially and respectively used as spinning solutions to enable fibers to be attached to the surface of the fiber membrane of the inner layer of the blood vessel in the step 2.1, so that the fiber membrane of the middle layer of the blood vessel and the fiber membrane of the outer layer of the blood vessel are respectively obtained, and the nano fiber tube with a real blood vessel structure is obtained;
and 3, rinsing the nanofiber tube with the real vascular structure obtained in the step 2, and removing the receiving roller to obtain the integrated small-caliber artificial blood vessel stent.
The present invention is also characterized in that,
the distance between the two receiving plates is 3 cm-5 cm, the spinning voltage is 10 kV-30 kV, the spinning distance is 100 mm-200 mm, the solution flow rate is 0.8 mL/h-1.5 mL/h, and the diameter of the receiving roller is 2 mm-6 mm.
The polymer A is one or more of silk fibroin, sulfated silk fibroin, polyglycolic acid, chitosan, collagen, polyethylene glycol and polyvinyl alcohol;
the polymer B is one or a mixture of more of polycaprolactone, polyurethane, poly (L-lactic acid-caprolactone), silk fibroin and polydioxanone;
the solvent A 'and the solvent B' are one or more of hexafluoroisopropanol, trichloromethane, N-dimethylformamide, dichloromethane, trifluoroacetic acid, acetone, ethanol and glycol.
The fibers forming the fiber membrane of the inner layer of the blood vessel and the receiving roller are axially arranged in parallel at an angle of less than 5 degrees; the fibers forming the fiber membrane in the middle layer of the blood vessel are axially vertical to and parallel to the receiving roller; the fibers forming the fibrous membrane on the outer layer of the blood vessel are axially vertical and parallel to the receiving roller.
The invention has the beneficial effects that:
1. the device has simple structure, low cost, easy operation of the production process and high efficiency;
2. when the small-caliber artificial blood vessel stent is prepared, the inner diameter of the small-caliber artificial blood vessel stent can be controlled by adjusting the diameter of the cross section of the receiving roller;
3. the small-caliber artificial blood vessel stent prepared by the device has the characteristics of high strength, high porosity, good biocompatibility and the like.
Drawings
FIG. 1 is a schematic structural diagram of an electrostatic spinning integrated small-caliber artificial blood vessel stent device;
fig. 2 is a structural schematic view of the integrated small-caliber artificial blood vessel stent prepared by the method of the invention.
In the figure, 1, a micro-injection pump, 2, a liquid guide tube, 3, a direct current high-voltage generator, 4, an injector, 5, a spinning needle head, 6, a variable-frequency motor a, 7, a rotating block, 8, a connecting rod, 9, a receiving roller, 10, a variable-frequency motor b, 11, a stroke limiting groove, 12, a movable rod, 13, a grounding wire, 14, a receiving plate, 15, a sliding plate, 16, a base, 17, an intravascular layer fibrous membrane, 18, an intravascular layer fibrous membrane and 19, an intravascular layer fibrous membrane.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention discloses a device for an electrostatic spinning integrated small-caliber artificial blood vessel stent, which has a structure shown in figure 1 and comprises a spinning device, a receiving device and a blood vessel stent forming device, wherein the receiving device and the blood vessel stent forming device are arranged on a base 16;
the spinning device comprises an injector 4, one end of the injector 4 is connected with the micro-injection pump 1 through a liquid guide pipe 2, the other end of the injector 4 is provided with a spinning needle head 5, the spinning needle head 5 is connected with a direct-current high-voltage generator 3, a receiving device is positioned under the spinning needle head 5, and the spinning needle head 5 is perpendicular to the receiving device.
The receiving device comprises two vertically arranged receiving plates 14 which are parallel to each other, the receiving plates 14 are fixedly installed on a base 16, the receiving plates 14 are connected with a grounding wire 13, a spinning needle head 5 is positioned right above a plane formed by the two receiving plates 14, and fibers obtained by spinning are lapped between the two receiving plates 14.
The intravascular stent forming device comprises a fiber winding device and a winding control device, wherein the fiber winding device is used for winding the fibers overlapped between the two receiving plates 14 into a cylindrical shape; the winding control device is used for controlling the axial movement and the circumferential rotation of the fiber winding device, so that the fibers wound on the fiber winding device form a vascular stent structure.
The fiber winding device comprises a receiving roller 9, and the receiving roller 9 is positioned between two receiving plates 14. The receiving roller 9 is made of terylene or polytetrafluoroethylene, and the diameter is 2 mm-6 mm.
The winding control device comprises a variable frequency motor a6 and a variable frequency motor b10, the variable frequency motor b10 is arranged on the sliding plate 15, and the variable frequency motor b10 is connected with one end of the receiving roller 9 and used for controlling the circumferential rotation of the receiving roller 9; a track and a stroke limiting groove 11 are arranged on a base 16, a sliding plate 15 horizontally moves along the track, one end, far away from a receiving roller 9, of the sliding plate 15 is connected with a movable rod 12, two ends of the movable rod 12 are respectively installed in the stroke limiting groove 11, the movable rod 12 is connected with one end of a connecting rod 8, the other end of the connecting rod 8 is provided with a rotating block 7, a variable frequency motor a6 drives the rotating block 7 to circumferentially rotate through a mandrel, and the connecting rod 8 is driven to drive the movable rod 12 to move in the stroke limiting groove 11, so that the sliding plate 15 is driven to horizontally move, and the receiving roller 9 is driven to.
The method for preparing the integrated small-caliber artificial blood vessel stent by adopting the device is implemented according to the following steps:
step 1, preparing spinning solution:
dissolving a biocompatible polymer A in a solvent A' at room temperature, and dissolving and stirring the polymer A by a constant-temperature magnetic stirrer until the solution is uniform to obtain a solution A with the mass fraction of the polymer A being 7-9%; dissolving a biocompatible polymer B in a solvent B', and dissolving and stirring the biocompatible polymer B by a constant-temperature magnetic stirrer until the solution is uniform to obtain a solution B with the mass fraction of the polymer B being 10-12%.
The polymer A is one or more of silk fibroin, sulfated silk fibroin, polyglycolic acid, chitosan, collagen, polyethylene glycol and polyvinyl alcohol;
the polymer B is one or a mixture of more of polycaprolactone, polyurethane, poly (L-lactic acid-caprolactone), silk fibroin and polydioxanone;
the solvent A 'and the solvent B' are one or more of hexafluoroisopropanol, trichloromethane, N-dimethylformamide, dichloromethane, trifluoroacetic acid, acetone, ethanol and glycol.
under the environmental conditions that the temperature is 20-30 ℃ and the relative humidity is 40-60%, the prepared solution A is added into an injector 4, and the distance between two receiving plates 14 is set to be 3-5 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller 9, a direct current high voltage generator 3 is started, the spinning voltage is adjusted to be 10kV to 30kV, the vertical distance (namely the spinning distance) between the head end of a spinning needle head 5 and the plane formed by two receiving plates 14 is adjusted to be 100mm to 200mm, and a micro-injection pump 1 is adjusted to control the solution flow rate to be 0.8mL/h to 1.5 mL/h. And starting a variable frequency motor a6, adjusting the rotating speed to be 25-35 rpm, driving a rotating block 7 to rotate in the circumferential direction by the variable frequency motor a6 through a mandrel, further enabling the receiving roller 9 to reciprocate in the axial direction through a connecting rod 8, a movable rod 12 and a sliding plate 15, and simultaneously starting a variable frequency motor b10, adjusting the rotating speed to be 300-500 rpm, and driving the receiving roller 9 to rotate in the circumferential direction. Jet flow formed by spraying the spinning solution is drawn into nano fibers under the action of an electric field force, the nano fibers are lapped between two receiving plates 14, a receiving roller 9 positioned between the two receiving plates 14 drags the fibers to axially reciprocate and simultaneously rotates along the circumferential direction, the fibers are attached to the surface of the receiving roller 9, and finally an intravascular layer fiber membrane 17 with the fibers and the receiving roller 9 in an axial direction of less than 5 degrees and parallel to each other is obtained.
under the environmental conditions that the temperature is 20-30 ℃ and the relative humidity is 40-60%, the inner layer fiber membrane prepared in the step 2 is taken as a receiving substrate, the prepared solution B is added into an injector 4, and the distance between two receiving plates 14 is set to be 3-5 cm. And opening the direct-current high-voltage generator 3, adjusting the spinning voltage to be 10 kV-30 kV, the spinning distance to be 100 mm-200 mm, and adjusting the micro-injection pump 1 to control the solution flow rate to be 0.8 mL/h-1.5 mL/h. And starting a variable frequency motor a6, adjusting the rotating speed to be 25-35 rpm, driving a rotating block 7 to rotate in the circumferential direction by a variable frequency motor a6 through a mandrel, further enabling a receiving roller 9 to reciprocate in the axial direction through a connecting rod 8, a movable rod 12 and a sliding plate 15, and simultaneously starting a variable frequency motor b10, adjusting the rotating speed to be 700-900 rpm, and driving the receiving roller 9 to rotate in the circumferential direction. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates 14, dragging the fibers to axially reciprocate between the two receiving plates 14 and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane 17 obtained in the step (2), and finally obtaining an intravascular layer fiber membrane 18 with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane on the outer layer of the blood vessel:
under the environmental conditions that the temperature is 20-30 ℃ and the relative humidity is 40-60%, the middle layer fiber membrane prepared in the step 3 is taken as a receiving substrate, the prepared solution A is added into an injector 4, and the distance between two receiving plates 14 is set to be 3-5 cm. And opening the direct-current high-voltage generator 3, adjusting the spinning voltage to be 10 kV-30 kV, the spinning distance to be 100 mm-200 mm, and adjusting the micro-injection pump 1 to control the solution flow rate to be 0.8 mL/h-1.5 mL/h. And starting a variable frequency motor a6, adjusting the rotating speed to be 25-35 rpm, driving a rotating block 7 to rotate in the circumferential direction by a variable frequency motor a6 through a mandrel, further enabling a receiving roller 9 to reciprocate in the axial direction through a connecting rod 8, a movable rod 12 and a sliding plate 15, and simultaneously starting a variable frequency motor b10, adjusting the rotating speed to be 700-900 rpm, and driving the receiving roller 9 to rotate in the circumferential direction. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates 14, dragging the fibers to axially reciprocate between the two receiving plates 14 and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane 18 obtained in the step (3), and finally obtaining a blood vessel outer layer fiber membrane 19 with the fibers basically vertical to the receiving rollers in the axial direction and arranged in parallel, namely obtaining the nano fiber tube.
And 5, removing the receiving roller:
after spinning, soaking the nanofiber tube in 75% alcohol for 3-5 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing the receiving roller 9 to finally obtain the integrated small-caliber artificial blood vessel stent, as shown in figure 2.
The invention utilizes the home-made electrostatic spinning device to prepare the small-caliber artificial blood vessel stent, and the nano-fibers are lapped between two parallel receiving plates, so that the nano-fibers in the inner layer fiber membrane are arranged along the axial direction, the nano-fibers are closely arranged, the pore diameter between the fibers is smaller, the porosity is higher, and the small-caliber artificial blood vessel stent is beneficial to the flow of blood. The nanometer fibers in the middle layer and the outer layer of fiber membrane are arranged directionally along the circumferential direction and are more regular and compact, so that the friction force between the fibers is higher, and the high strength of the intravascular stent can be ensured.
The polymers adopted in the invention have good biocompatibility and degradability, and the prepared nanofiber tissue engineering scaffold has higher specific surface area and porosity, so that the structural characteristics of extracellular matrix can be better simulated, and a more suitable environment is provided for the growth and propagation of cells.
Example 1
Step 1, preparing a spinning solution
Blending silk fibroin and sulfated silk fibroin at room temperature, dissolving in hexafluoroisopropanol, and dissolving and stirring by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer A solution with the mass fraction of 7%; dissolving polycaprolactone in chloroform/N, N-dimethylformamide (7:3), and dissolving and stirring by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer B solution with the mass fraction of 10%.
The prepared solution A was added to a syringe under ambient conditions of 20 ℃ and 45% relative humidity, with the distance between the two receiving plates set at 3 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller, a direct current high-voltage generator is started, the spinning voltage is adjusted to be 10kV, the spinning distance is adjusted to be 100mm, and the flow rate of the solution is controlled to be 0.8 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 25rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 300rpm, and driving the receiving roller to rotate circumferentially. Jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotate along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and finally the intravascular layer fiber membrane with the fibers and the receiving roller in an axial direction of less than 5 degrees and parallel to each other is obtained.
And (3) under the environmental conditions that the temperature is 20 ℃ and the relative humidity is 45%, taking the inner layer fiber membrane prepared in the step (2) as a receiving substrate, adding the prepared solution B into an injector, and setting the distance between two receiving plates to be 3 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 10kV, adjusting the spinning distance to be 100mm, and controlling the flow rate of the solution to be 0.8 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 25rpm, driving a rotating block to rotate in the circumferential direction by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 700rpm, and driving the receiving roller to rotate in the circumferential direction. And (2) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane obtained in the step (2), and finally obtaining the intravascular layer fiber membrane with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane in the middle layer of the blood vessel
And (3) under the environmental conditions that the temperature is 20 ℃ and the relative humidity is 45%, taking the middle-layer fiber membrane prepared in the step (3) as a receiving substrate, adding the prepared solution A into a syringe, and setting the distance between two receiving plates to be 3 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 10kV, adjusting the spinning distance to be 100mm, and controlling the flow rate of the solution to be 0.8 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 25rpm, driving a rotating block to rotate in the circumferential direction by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 700rpm, and driving the receiving roller to rotate in the circumferential direction. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane obtained in the step (3), and finally obtaining the blood vessel outer layer fiber membrane with the fibers basically vertical to the receiving rollers and arranged in parallel, namely obtaining the nano fiber tube.
Step 5, removing the receiving roller
And after spinning, soaking the nanofiber tube in 75% alcohol for 3 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing a receiving roller to finally obtain the integrated small-caliber artificial blood vessel stent.
Experimental results show that the artificial blood vessel stent obtained by the electrostatic spinning device has better fiber directionality and mechanical property.
Example 2
Step 1, preparing a spinning solution
At room temperature, dissolving polyglycolic acid in hexafluoroisopropanol, and dissolving and stirring the solution by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer A solution with the mass fraction of 8%; and (3) dissolving polyurethane in dichloromethane, and dissolving and stirring the solution by using a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer B solution with the mass fraction of 11%.
The prepared solution A was added to a syringe under ambient conditions of 22 ℃ and 55% relative humidity, with the distance between the two receiving plates set at 3.5 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller, a direct current high-voltage generator is started, the spinning voltage is adjusted to be 20kV, the spinning distance is adjusted to be 150mm, and the flow rate of the solution is controlled to be 1.2 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 30rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 400rpm, and driving the receiving roller to rotate circumferentially. Jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotate along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and finally the intravascular layer fiber membrane with the fibers and the receiving roller in an axial direction of less than 5 degrees and parallel to each other is obtained.
And (3) under the environmental conditions that the temperature is 22 ℃ and the relative humidity is 55%, taking the inner layer fiber membrane prepared in the step (2) as a receiving substrate, adding the prepared solution B into a syringe, and setting the distance between two receiving plates to be 3.5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 20kV, adjusting the spinning distance to be 150mm, and controlling the flow rate of the solution to be 1.2 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 30rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 800rpm, and driving the receiving roller to rotate circumferentially. And (2) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane obtained in the step (2), and finally obtaining the intravascular layer fiber membrane with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane on the outer layer of the blood vessel
Under the environmental conditions that the temperature is 22 ℃ and the relative humidity is 55%, the middle-layer fiber membrane prepared in the step 3 is taken as a receiving substrate, the prepared solution A is added into a syringe, and the distance between two receiving plates is set to be 3.5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 20kV, adjusting the spinning distance to be 150mm, and controlling the flow rate of the solution to be 1.2 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 30rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 800rpm, and driving the receiving roller to rotate circumferentially. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane obtained in the step (3), and finally obtaining the blood vessel outer layer fiber membrane with the fibers basically vertical to the receiving rollers and arranged in parallel, namely obtaining the nano fiber tube.
Step 5, removing the receiving roller
And after spinning, soaking the nanofiber tube in 75% alcohol for 4 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing a receiving roller to finally obtain the integrated small-caliber artificial blood vessel stent.
Experimental results show that the artificial blood vessel stent obtained by the electrostatic spinning device has better fiber directionality and mechanical property.
Example 3
Step 1, preparing a spinning solution
Mixing chitosan and collagen at room temperature, dissolving in trifluoroacetic acid, and dissolving and stirring by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer A solution with the mass fraction of 9%; dissolving poly (L-lactic acid-caprolactone) in acetone, and dissolving and stirring the solution by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer B solution with the mass fraction of 12%.
The prepared solution A was added to a syringe under ambient conditions of 30 ℃ and 60% relative humidity, with the distance between the two receiving plates set at 5 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller, a direct current high voltage generator is started, the spinning voltage is adjusted to be 30kV, the spinning distance is adjusted to be 200mm, and the flow rate of the solution is controlled to be 1.5 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 32rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 500rpm, and driving the receiving roller to rotate circumferentially. Jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotate along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and finally the intravascular layer fiber membrane with the fibers and the receiving roller in an axial direction of less than 5 degrees and parallel to each other is obtained.
And (3) under the environmental conditions that the temperature is 30 ℃ and the relative humidity is 60%, taking the inner layer fiber membrane prepared in the step (2) as a receiving substrate, adding the prepared solution B into an injector, and setting the distance between two receiving plates to be 5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 30kV, adjusting the spinning distance to be 200mm, and controlling the flow rate of the solution to be 1.5 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 32rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 900rpm, and driving the receiving roller to rotate circumferentially. And (2) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane obtained in the step (2), and finally obtaining the intravascular layer fiber membrane with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane on the outer layer of the blood vessel
And (3) under the environmental conditions that the temperature is 30 ℃ and the relative humidity is 60%, taking the middle-layer fiber membrane prepared in the step (3) as a receiving substrate, adding the prepared solution A into a syringe, and setting the distance between two receiving plates to be 5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 30kV, adjusting the spinning distance to be 200mm, and controlling the flow rate of the solution to be 1.5 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 32rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 900rpm, and driving the receiving roller to rotate circumferentially. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane obtained in the step (3), and finally obtaining the blood vessel outer layer fiber membrane with the fibers basically vertical to the receiving rollers and arranged in parallel, namely obtaining the nano fiber tube.
Step 5, removing the receiving roller
And after spinning, soaking the nanofiber tube in 75% alcohol for 5 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing a receiving roller to finally obtain the integrated small-caliber artificial blood vessel stent.
Experimental results show that the artificial blood vessel stent obtained by the electrostatic spinning device has better fiber directionality and mechanical property.
Example 4
Step 1, preparing a spinning solution
Dissolving polyethylene glycol in ethanol at room temperature, and dissolving and stirring the polyethylene glycol by using a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer A solution with the mass fraction of 9%; and mixing and dissolving polyurethane and silk fibroin in hexafluoroisopropanol, and dissolving and stirring the mixture by using a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer B solution with the mass fraction of 11%.
The prepared solution A was added to a syringe under ambient conditions of 25 ℃ and 40% relative humidity, with the distance between the two receiver plates set at 4 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller, a direct current high-voltage generator is started, the spinning voltage is adjusted to be 25kV, the spinning distance is adjusted to be 180mm, and the flow rate of the solution is controlled to be 1.0 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 28rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 350rpm, and driving the receiving roller to rotate circumferentially. Jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotate along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and finally the intravascular layer fiber membrane with the fibers and the receiving roller in an axial direction of less than 5 degrees and parallel to each other is obtained.
And (3) under the environmental conditions that the temperature is 25 ℃ and the relative humidity is 40%, taking the inner layer fiber membrane prepared in the step (2) as a receiving substrate, adding the prepared solution B into a syringe, and setting the distance between two receiving plates to be 4 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 25kV, adjusting the spinning distance to be 180mm, and controlling the flow rate of the solution to be 1.0 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 28rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 750rpm, and driving the receiving roller to rotate circumferentially. And (2) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane obtained in the step (2), and finally obtaining the intravascular layer fiber membrane with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane on the outer layer of the blood vessel
And (3) under the environmental conditions that the temperature is 25 ℃ and the relative humidity is 40%, taking the middle-layer fiber membrane prepared in the step (3) as a receiving substrate, adding the prepared solution A into a syringe, and setting the distance between two receiving plates to be 4 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 25kV, adjusting the spinning distance to be 180mm, and controlling the flow rate of the solution to be 1.0 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to be 28rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to be 750rpm, and driving the receiving roller to rotate circumferentially. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane obtained in the step (3), and finally obtaining the blood vessel outer layer fiber membrane with the fibers basically vertical to the receiving rollers and arranged in parallel, namely obtaining the nano fiber tube.
Step 5, removing the receiving roller
And after spinning, soaking the nanofiber tube in 75% alcohol for 4.5 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing a receiving roller to finally obtain the integrated small-caliber artificial blood vessel stent.
Experimental results show that the artificial blood vessel stent obtained by the electrostatic spinning device has better fiber directionality and mechanical property.
Example 5
Step 1, preparing a spinning solution
Dissolving polyvinyl alcohol in ethylene glycol at room temperature, and dissolving and stirring the polyvinyl alcohol by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer A solution with the mass fraction of 8%; dissolving poly (p-dioxanone) in hexafluoroisopropanol, and dissolving and stirring the solution by a constant-temperature magnetic stirrer until the solution is transparent to obtain a polymer B solution with the mass fraction of 12%.
The prepared solution A was added to a syringe under ambient conditions of 28 ℃ and 50% relative humidity, with the distance between the two receiving plates set at 4.5 cm. Before spinning, a layer of lubricant (PEG) is coated on a receiving roller, a direct current high voltage generator is started, the spinning voltage is adjusted to be 22kV, the spinning distance is adjusted to be 160mm, and the flow rate of the solution is controlled to be 1.3 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to 35rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to 450rpm, and driving the receiving roller to rotate circumferentially. Jet flow formed by spraying spinning solution is drawn into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates, a receiving roller positioned between the two receiving plates drags the fibers to axially reciprocate and simultaneously rotate along the circumferential direction, so that the fibers are attached to the surface of the receiving roller, and finally the intravascular layer fiber membrane with the fibers and the receiving roller in an axial direction of less than 5 degrees and parallel to each other is obtained.
And (3) under the environmental conditions that the temperature is 28 ℃ and the relative humidity is 50%, taking the inner layer fiber membrane prepared in the step (2) as a receiving substrate, adding the prepared solution B into a syringe, and setting the distance between two receiving plates to be 4.5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 22kV, adjusting the spinning distance to be 160mm, and controlling the flow rate of the solution to be 1.3 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to 35rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to 850rpm, and driving the receiving roller to rotate circumferentially. And (2) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between the two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the intravascular layer fiber membrane obtained in the step (2), and finally obtaining the intravascular layer fiber membrane with the fibers basically vertical to the axial direction of the receiving rollers and arranged in parallel.
Step 4, preparing the fiber membrane on the outer layer of the blood vessel
Under the environmental conditions that the temperature is 28 ℃ and the relative humidity is 50%, the middle-layer fiber membrane prepared in the step 3 is used as a receiving substrate, the prepared solution A is added into a syringe, and the distance between two receiving plates is set to be 4.5 cm. And (3) turning on the direct-current high-voltage generator, adjusting the spinning voltage to be 22kV, adjusting the spinning distance to be 160mm, and controlling the flow rate of the solution to be 1.3 mL/h. And starting a variable frequency motor a, adjusting the rotating speed to 35rpm, driving a rotating block to rotate circumferentially by the variable frequency motor a through a mandrel, further enabling the receiving roller to axially reciprocate through a connecting rod, a movable rod and a sliding plate, and simultaneously starting a variable frequency motor b, adjusting the rotating speed to 850rpm, and driving the receiving roller to rotate circumferentially. And (3) drafting jet flow formed by spraying the spinning solution into nano fibers under the action of an electric field force, overlapping the nano fibers between two receiving plates, dragging the fibers to axially reciprocate between the two receiving plates and simultaneously rotating along the circumferential direction, so that the fibers are attached to the surface of the blood vessel middle layer fiber membrane obtained in the step (3), and finally obtaining the blood vessel outer layer fiber membrane with the fibers basically vertical to the receiving rollers and arranged in parallel, namely obtaining the nano fiber tube.
Step 5, removing the receiving roller
And after spinning, soaking the nanofiber tube in 75% alcohol for 3.5 hours, rinsing with PBS (phosphate buffer solution), dissolving residual PEG on the surface of the stent, and removing a receiving roller to finally obtain the integrated small-caliber artificial blood vessel stent.
Experimental results show that the artificial blood vessel stent obtained by the electrostatic spinning device has better fiber directionality and mechanical property.
Claims (5)
1. The preparation method of an integrated small-caliber artificial blood vessel stent is characterized in that the adopted device for electrospinning the integrated small-caliber artificial blood vessel stent comprises a spinning device, a receiving device and a blood vessel stent forming device, wherein the receiving device and the blood vessel stent forming device are arranged on a base (16);
the spinning device comprises an injector (4), one end of the injector (4) is connected with the micro-injection pump (1) through a liquid guide pipe (2), the other end of the injector (4) is provided with a spinning needle head (5), the spinning needle head (5) is connected with a direct-current high-voltage generator (3), the receiving device is positioned right below the spinning needle head (5), and the spinning needle head (5) is vertical to the receiving device;
the receiving device is used for receiving the fibers obtained by the spinning device through electrostatic spinning; the receiving device comprises two receiving plates (14), the receiving plates (14) are fixedly and vertically arranged on a base (16), the receiving plates (14) are grounded, a spinning needle head (5) is positioned right above a plane formed by the two receiving plates (14), fibers obtained by spinning are lapped between the two receiving plates (14), and the two receiving plates (14) are parallel to each other;
the intravascular stent forming device comprises a fiber winding device and a winding control device, wherein the fiber winding device is used for winding the fiber received by the receiving device into a cylindrical shape; the winding control device is used for controlling the axial movement and the circumferential rotation of the fiber winding device so that the fibers wound on the fiber winding device form a vascular stent structure;
the fiber winding device comprises a receiving roller (9), and the receiving roller (9) is positioned between two receiving plates (14);
the winding control device comprises a circumferential rotation control device and an axial movement control device which are respectively used for controlling the circumferential rotation and the axial movement of the receiving roller (9);
the circumferential rotation control device comprises a variable frequency motor b (10), the variable frequency motor b (10) is installed on the sliding plate (15), and the variable frequency motor b (10) is connected with one end of the receiving roller (9) and used for controlling the circumferential rotation of the receiving roller (9);
the axial movement control device comprises a track and a stroke limiting groove (11) which are arranged on a base (16), a sliding plate (15) horizontally moves along the track, one end, far away from a receiving roller (9), of the sliding plate (15) is connected with a movable rod (12), two ends of the movable rod (12) are respectively installed in the stroke limiting groove (11), the movable rod (12) is connected with one end of a connecting rod (8), the other end of the connecting rod (8) is provided with a rotating block (7), a variable frequency motor a (6) drives the rotating block (7) to circumferentially rotate through a mandrel, and the connecting rod (8) is driven to drive the movable rod (12) to move in the stroke limiting groove (11), so that the sliding plate (15) is driven to horizontally move, and the receiving roller (9) is driven to axially move;
the method is implemented according to the following steps:
step 1, preparing spinning solution:
dissolving a biocompatible polymer A in a solvent A' at room temperature, and stirring until the solution is uniform to obtain a solution A with the mass fraction of the polymer A being 7-9%; dissolving a biocompatible polymer B in a solvent B', and stirring until the solution is uniform to obtain a solution B with the mass fraction of the polymer B being 10-12%;
step 2, preparing the nanofiber tube with the real vascular structure:
step 2.1, preparing the fiber membrane of the inner layer of the blood vessel:
under the environmental conditions that the temperature is 20-30 ℃ and the relative humidity is 40-60%, the prepared solution A is added into an injector (4), and the distance between two receiving plates (14) is set; turning on the direct-current high-voltage generator (3), adjusting spinning voltage and spinning distance, and adjusting the micro-injection pump (1) to control the flow rate of the solution; starting a variable frequency motor a (6), adjusting the rotating speed, driving a rotating block (7) to rotate circumferentially by the variable frequency motor a (6) through a mandrel, and further enabling a receiving roller (9) to reciprocate axially through a connecting rod (8), a movable rod (12) and a sliding plate (15); meanwhile, a variable frequency motor b (10) is started, the rotating speed is adjusted, and the receiving roller (9) is driven to rotate in the circumferential direction; jet flow formed by spraying spinning solution is drafted into nano fibers under the action of electric field force, the nano fibers are lapped between two receiving plates (14), and a receiving roller (9) positioned between the two receiving plates (14) drags the fibers to axially reciprocate and simultaneously rotates along the circumferential direction, so that the fibers are attached to the surface of the receiving roller (9) to obtain a fiber membrane of the inner layer of the blood vessel;
step 2.2, the solution B and the solution A are sequentially and respectively used as spinning solutions to enable fibers to be attached to the surface of the fiber membrane of the inner layer of the blood vessel in the step 2.1, so that the fiber membrane of the middle layer of the blood vessel and the fiber membrane of the outer layer of the blood vessel are respectively obtained, and the nano fiber tube with a real blood vessel structure is obtained;
and 3, rinsing the nanofiber tube with the real vascular structure obtained in the step 2, and removing the receiving roller (9) to obtain the integrated small-caliber artificial blood vessel stent.
2. The preparation method of the integrated small-caliber artificial blood vessel stent as claimed in claim 1, wherein the distance between the two receiving plates (14) is 3 cm-5 cm, the spinning voltage is 10 kV-30 kV, the spinning distance is 100 mm-200 mm, the solution flow rate is 0.8 mL/h-1.5 mL/h, and the diameter of the receiving roller (9) is 2 mm-6 mm.
3. The method for preparing the integrated small-caliber artificial blood vessel stent as claimed in claim 1, wherein the polymer A is one or more of silk fibroin, sulfated silk fibroin, polyglycolic acid, chitosan, collagen, polyethylene glycol and polyvinyl alcohol;
the polymer B is one or a mixture of more of polycaprolactone, polyurethane, poly (L-lactic acid-caprolactone), silk fibroin and polydioxanone;
the solvent A 'and the solvent B' are one or more of hexafluoroisopropanol, trichloromethane, N-dimethylformamide, dichloromethane, trifluoroacetic acid, acetone, ethanol and glycol.
4. The method for preparing the integrated small-caliber artificial blood vessel stent as claimed in claim 1, wherein in the step 2, before spinning, a layer of lubricant is coated on the surface of a receiving roller (9); the rotating speed of the variable frequency motor a (6) is 25-35 rpm; when preparing the fiber membrane of the inner layer of the blood vessel, the rotating speed of the variable frequency motor b (10) is 300 rpm-500 rpm; when the fiber membrane in the middle layer and the fiber membrane in the outer layer of the blood vessel are prepared, the rotating speed of the variable frequency motor b (10) is 700 rpm-900 rpm.
5. The method for preparing the integrated small-caliber artificial blood vessel stent as claimed in claim 1, wherein the fibers forming the fiber membrane of the intravascular layer are arranged in parallel with the receiving roller (9) in an axial direction of less than 5 degrees; the fibers forming the fiber membrane in the middle layer of the blood vessel are axially vertical and parallel to the receiving roller (9); the fibers forming the fiber membrane of the outer layer of the blood vessel are arranged in parallel and perpendicular to the axial direction of the receiving roller (9).
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| CN109700562B (en) * | 2018-12-13 | 2020-12-08 | 贵州大学 | Multilayer dual-orientation stent simulating natural vascular structure and function and preparation method thereof |
| CN109722792B (en) * | 2018-12-17 | 2021-09-07 | 河北久丰节能科技有限公司 | Automatic production equipment for fiber cotton pipe |
| CN110481019A (en) * | 2019-08-20 | 2019-11-22 | 广州帷幄生物科技有限公司 | A kind of artificial blood vessel printing device |
| CN111793899B (en) * | 2020-04-30 | 2021-06-18 | 杭州医学院 | Bionic nanofiber material and preparation method and application thereof |
| CN113737393B (en) * | 2021-09-15 | 2022-06-07 | 陕西环保产业研究院有限公司 | Electrostatic spinning nanofiber membrane and preparation method thereof |
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