CN111388760A - Small blood vessel stent with nano fibers oriented along circumferential direction and preparation method - Google Patents

Small blood vessel stent with nano fibers oriented along circumferential direction and preparation method Download PDF

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Publication number
CN111388760A
CN111388760A CN202010220104.9A CN202010220104A CN111388760A CN 111388760 A CN111388760 A CN 111388760A CN 202010220104 A CN202010220104 A CN 202010220104A CN 111388760 A CN111388760 A CN 111388760A
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bacterial cellulose
section
vessel stent
blood vessel
circumferential direction
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CN111388760B (en
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罗红林
周建业
万怡灶
杨志伟
张全超
王捷
甘德强
欧阳晨曦
邓小燕
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East China Jiaotong University
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East China Jiaotong University
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/28Materials for coating prostheses
    • A61L27/34Macromolecular materials
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/52Hydrogels or hydrocolloids
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2400/00Materials characterised by their function or physical properties
    • A61L2400/12Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces

Abstract

The invention discloses a small blood vessel stent with nanofibers oriented along the circumferential direction, which is composed of circumferentially arranged bacterial cellulose nanofibers, wherein the diameters of the nanofibers are smaller than 100 nanometers, the diameter of the small blood vessel stent is 2-6 millimeters, the thickness of a tube wall is controllable, and the minimum thickness can be as small as 10 micrometers. The preparation process comprises the following steps: preparing a bacterial cellulose hydrogel film with the length of 10-30 mm, the film thickness of 0.01-1 mm and the inner diameter of 2-6 mm on the outer rotary surface of a round rod with the diameter changing by adopting a film liquid interface culture technology, then moving the hydrogel film from the thin section end to the thick section end of the round rod to gradually increase the pipe diameter of the tubular bacterial cellulose hydrogel, forcing the bacterial cellulose fibers to be oriented in situ along the circumferential direction, and finally, taking out the round rod after heating, drying and shaping to obtain the bacterial cellulose hydrogel. The structure of the small blood vessel stent has similarity with the fiber arrangement structure of natural blood vessels, the performance of the small blood vessel stent is favorable for cell adhesion, proliferation and final endothelialization, and a better treatment effect is expected to be obtained.

Description

Small blood vessel stent with nano fibers oriented along circumferential direction and preparation method
Technical Field
The invention relates to a biomedical engineering product, in particular to a small blood vessel stent.
Background
Cardiovascular disease is the leading enemy of human health today, with about 1/3 total deaths worldwide, and is growing rapidly. Vascular surgery is performed and replacement of diseased blood vessels is a recognized and effective clinical treatment. Autologous blood vessel transplantation is the clinical first choice, but the source is limited, and secondary trauma can be caused; the allogeneic blood vessels easily cause rejection reaction, blood coagulation and other defects; it is widely believed that the artificial blood vessel (stent) constructed by the tissue engineering method has the most development prospect. Since the first completion of artificial vascular artery transplantation surgery in 1952, human beings have made great progress and succeeded in large-caliber blood vessel transplantation. However, small-bore (less than 6mm) artificial blood vessels (hereinafter simply referred to as "small blood vessels") are a worldwide problem that has not been overcome so far because thrombi are very easy to form (rapid endothelialization is not easy). Therefore, the development of high-performance small vessel stents is a common pursuit goal of domestic and foreign scientists.
In vivo blood vessels are composed of circumferentially arranged fibers, and it is particularly important that the size of the fibers is in the range of nanometer (1-100 nm) to submicron (100-1000 nm), and that such nanostructures are likely to have a direct relationship with their antithrombotic formation. It follows that from a biomimetic perspective, the fibers of the small vessel scaffold should be composed of circumferentially arranged nanofibers. However, current techniques do not allow for small vessel stents with nanofibers oriented in the circumferential direction.
Disclosure of Invention
In view of the above problems, the present invention is directed to provide a small vessel stent with nanofibers oriented in the circumferential direction, which has a structure similar to the fiber arrangement structure of natural vessels, facilitates cell adhesion, proliferation and final endothelialization, and is expected to achieve a better therapeutic effect.
In order to solve the technical problems, the invention provides a small blood vessel stent with nanofibers oriented along the circumferential direction, which is formed by uniformly surrounding bacterial cellulose nanofibers into a circular tube shape, wherein the bacterial cellulose nanofibers are arranged along the circumferential direction, and the small blood vessel stent has an inner diameter of 2-6 mm, a wall thickness of 0.01-1 mm and a length of 10-30 mm. The preparation method mainly comprises the following steps: firstly, forming bacterial cellulose hydrogel on the rotary surface of a round bar in situ, gradually increasing the inner diameter of the tubular bacterial cellulose hydrogel by a displacement method to achieve the in-situ circumferential orientation of the nano fibers, heating, drying and shaping, and then removing the round bar to obtain the small vessel stent with the bacterial cellulose nano fibers oriented along the circumferential direction and the controllable tube wall thickness. The method comprises the following specific steps:
step one, preparing a culture medium by adopting a conventional method, and placing the culture medium in a sterilization pot for high-temperature and high-pressure sterilization for 30-60 minutes;
step two, preparing a bacterial cellulose base membrane: inoculating the strain into a culture medium in an aseptic environment; statically culturing the inoculated culture medium at 30 ℃ for 3 days to obtain a bacterial cellulose base membrane;
taking a round bar, wherein the round bar comprises an A section, a middle section and a B section which are coaxially connected in sequence; the diameter of the section A is Da, the Da is 1.5-4.5 mm, and the length of the section A is 10-30 mm; the diameter of the section B is Db, Db is 2-6 mm, Db is 1.3-1.4 Da, and the length of the section B is 10-30 mm; the diameter of the connecting end of the middle section and the section A is Da, the diameter of the connecting end of the middle section and the section B is Db, the length of the middle section is 100-200 mm, the middle section is a diameter gradually-changing section, and the diameter gradually changes from Da to Db along the length of the middle section; after ultraviolet sterilization, the round rod is fixed in an oxygen-permeable silica gel tube with a sampling hole at the upper end downwards according to the section A, and is vertically placed on the surface of the bacterial cellulose base membrane obtained in the step two;
step four, preparing a tubular bacterial cellulose hydrogel film with the length of 10-30 mm and the film thickness of 0.01-1 mm on the outer rotating surface of the section A of the round bar by adopting a film liquid interface culture technology;
step five, slowly pushing the tubular bacterial cellulose hydrogel film to the section B from the section A of the round rod through the middle section with the taper, so that the inner diameter of the tubular bacterial cellulose hydrogel film is gradually increased, and the bacterial cellulose nanofibers are oriented in situ along the circumferential direction;
and sixthly, putting the round bar with the bacterial cellulose hydrogel film into deionized water, cleaning for 2 times, then putting the round bar into the deionized water, heating and boiling, after the bacterial cellulose hydrogel film turns white from light yellow, putting the round bar into 0.5 mol/L NaOH solution, boiling for 20 minutes, finally boiling in the deionized water, repeating the boiling for 3-4 times, and washing with the deionized water until the round bar is neutral.
And seventhly, drying the cleaned round bar with the bacterial cellulose hydrogel film in an oven at 60 ℃ for 6-12 hours, and then taking down the tubular bacterial cellulose dry film to obtain the small blood vessel stent with the bacterial cellulose nanofibers oriented along the circumferential direction, wherein the inner diameter of the small blood vessel stent is 2-6 millimeters, the wall thickness of the small blood vessel stent is 0.01-1 millimeter, and the length of the small blood vessel stent is 10-30 millimeters.
Further, the preparation method of the small blood vessel stent with the nano-fiber oriented along the circumferential direction is provided, wherein,
in the second step, the strains at least comprise acetobacter xylinum, acetobacter aceti, acetobacter gluconicum, acetobacter acetogenes and acetobacter pasteurianus.
The round bar in the third step at least comprises a glass round bar, a ceramic round bar, an alkali-boiling resistant metal round bar and an alkali-boiling resistant high polymer material round bar.
The specific process of the fourth step is that the mixture is in a mist form and is according to 0.012-0.025 m L/cm2Spraying amount of the obtained productSpraying a culture medium on the contact surface of the round bar and the bacterial cellulose base membrane from a sample inlet hole at the upper end of the oxygen permeable silicone tube, spraying again after the culture medium on the bacterial cellulose base membrane is completely consumed, and repeatedly spraying for 50-200 times; thus, the tubular bacterial cellulose hydrogel film with the length of 10-30 mm and the film thickness of 0.01-1 mm is prepared on the outer surface of the section A of the round bar.
Compared with the prior art, the invention has the beneficial effects that:
the diameter of the small vessel stent prepared by the invention is 2-6 mm, the thickness of the vessel wall is controllable, and the minimum thickness can be as small as 10 microns. The small blood vessel stent has a bionic structure, wherein fibers with the diameter smaller than 100 nanometers are oriented in situ along the circumferential direction, the pores of the stent are small, blood seepage can be prevented, and rapid endothelialization is easy to realize. In addition, the diameter of the stent is controllable, and the thickness of the stent is adjustable, so that the stent becomes a small blood vessel stent or graft with great development prospect.
Drawings
FIG. 1(a) is a schematic cross-sectional structure diagram of a small vessel stent with nanofibers oriented in the circumferential direction;
FIG. 1(b) is a schematic diagram of the overall structure of a small vessel stent;
FIG. 2(a) is a photograph 1 of a small vessel stent prepared in example 1 of the present invention;
FIG. 2(b) is a photograph 2 of a small vessel stent prepared in example 1 of the present invention;
FIG. 3(a) is a scanning electron micrograph of the oriented arrangement of bacterial cellulose fibers of the small vessel stent prepared in example 1 of the present invention;
FIG. 3(b) is a partial enlarged view of the directional arrangement of bacterial cellulose fibers of the small vessel stent of FIG. 3 (a).
Detailed Description
The design idea of the invention is that the structure of the prepared small vessel stent has similarity with the fiber arrangement structure of natural vessels, so that the small vessel stent has the performance of facilitating cell adhesion, proliferation and final endothelialization. The small blood vessel stent is composed of bacterial cellulose nanofibers which are circumferentially arranged, wherein the diameter of the nanofibers is smaller than 100 nanometers, the diameter of the small blood vessel stent is 2-6 millimeters, the thickness of the tube wall is controllable, and the minimum thickness can be as small as 10 micrometers. The preparation method mainly comprises the following steps: the method comprises the steps of forming a bacterial cellulose hydrogel membrane with the length of 10-30 mm, the membrane thickness of 0.01-1 mm and the inner diameter of 2-6 mm in situ on the outer rotary surface of a round rod with the diameter changing (the outer diameter of a thick section of the round rod is 1.3-1.4 times of the outer diameter of a thin section, and an intermediate transition section is arranged between the thin section and the thick section) by adopting a membrane-liquid interface culture technology, then moving the hydrogel membrane from the thin section end to the thick section end of the round rod, gradually increasing the inner diameter of a tubular bacterial cellulose hydrogel by a displacement method, forcing bacterial cellulose fibers to be oriented in situ along the circumferential direction, taking out the round rod after heating, drying and shaping, and finally obtaining the small vessel stent with the bacterial cellulose nanofibers oriented along the circumferential direction and controllable tube wall thickness.
The invention is further illustrated with reference to the following figures and specific examples, which are not intended to limit the invention in any way.
Example 1, preparation of a small vessel stent, comprising the steps of:
step one, preparing a culture medium, namely sequentially adding glucose (25 g/L), yeast powder (7.5 g/L), peptone (10 g/L) and disodium hydrogen phosphate (10 g/L) into a beaker filled with 1000 ml of ultrapure water, stirring until the components are completely dissolved, adjusting the pH value of a system to be 4-5 by using glacial acetic acid to prepare the culture medium, and then placing the culture medium into a sterilization pot to sterilize for 40 minutes under the conditions of the temperature of 115 ℃ and the pressure of 0.1 MPa.
Step two, preparing a bacterial cellulose base membrane: the strain is inoculated into the culture medium under a sterile environment. The strain in this example is acetobacter xylinum. The strain can also be selected from one of acetobacter aceti, acetobacter gluconicum, acetobacter acetogenes and acetobacter pasteurianus; and statically culturing the inoculated culture medium at the temperature of 30 ℃ for 3 days to obtain a bacterial cellulose hydrogel membrane with the thickness of about 2.5 mm, wherein the bacterial cellulose hydrogel membrane is used as a basement membrane for membrane-liquid interface culture.
And step three, taking a round rod, wherein the round rod in the embodiment is a glass round rod, and the round rod can also be one of a ceramic round rod, an alkali-boiling-resistant metal round rod and an alkali-boiling-resistant high polymer material round rod. The total length of the glass round rod is 260 mm, the glass round rod is sequentially provided with a thin section, a middle section and a thick section from one end to the other end, the length of the thin section is 30 mm, and the diameter of the thin section is 2.5 mm; the length of the thick section is 30 mm, and the diameter is 3.5 mm; the middle section has a length of 200 mm and a diameter that increases gradually (equal slope) from 2.5 mm at the end of the thin section to 3.5 mm at the end of the thick section; and (3) after ultraviolet sterilization, fixing the thin section in an oxygen-permeable silica gel tube with a sample inlet at the upper end, and vertically placing the thin section on the surface of the bacterial cellulose base membrane obtained in the second step.
Step four, preparing a tubular bacterial cellulose hydrogel membrane with the length of 30 mm and the membrane thickness of 1.8 mm on the outer rotating surface of the thin section of the glass round rod by adopting the membrane liquid interface culture technology disclosed in the published patent document (a through membrane bracket and a preparation method thereof, application number 201910715079.9); the specific process is as follows:
in the form of a mist and at a rate of 0.018m L/cm2Spraying the culture medium obtained in the step one on the contact surface of the glass round bar and the bacterial cellulose base membrane from a sample inlet hole at the upper end of the oxygen permeable silicone tube, spraying again after the culture medium on the bacterial cellulose base membrane is completely consumed, and repeatedly spraying for 200 times; a tubular bacterial cellulose hydrogel film with the length of 30 mm and the film thickness of 1.8 mm is formed on the outer rotating surface of the thin section of the glass round rod.
And step five, slowly pushing the tubular bacterial cellulose hydrogel film on the round glass rod from the thin section to the thick section through the middle section with the taper, and gradually increasing the inner diameter of the tubular bacterial cellulose hydrogel film to the outer diameter (namely 3.5 mm) of the thick section so as to enable the bacterial cellulose nanofibers in the tubular bacterial cellulose hydrogel film to be oriented in the circumferential direction.
And sixthly, putting the glass round bar with the bacterial cellulose hydrogel film on the surface into deionized water, cleaning for 2 times, then putting the glass round bar into the deionized water, heating and boiling, putting the glass round bar into 0.5 mol/L NaOH solution, heating and boiling for 20 minutes after the film turns white from light yellow, and finally boiling for 4 times in the deionized water, and washing the glass round bar to be neutral by the deionized water.
And step seven, drying the cleaned glass round bar with the bacterial cellulose hydrogel film on the surface in an oven at 60 ℃ for 10 hours, and then removing the bacterial cellulose dry film to obtain the small blood vessel stent with the bacterial cellulose diameter smaller than 100 nanometers and in-situ circumferential orientation, wherein as can be seen from fig. 1(a) and 1(b), the small blood vessel stent is uniformly surrounded by the bacterial cellulose nanofibers and axially extends to form a round tube shape, the length of the small blood vessel stent is about 30 millimeters, the thickness of the small blood vessel stent is about 0.5 millimeters, the inner diameter of the small blood vessel stent is about 3.5 millimeters, macroscopic photographs of the small blood vessel stent are shown in fig. 2(a) and 2(b), and fig. 3(a) and 3(b) show SEM photographs in which the bacterial cellulose nanofibers are directionally arranged.
Example 2, preparation of small vessel stents comprising the following steps:
step one, the same as example 1.
Step two, the same as example 1.
Step three, different from the embodiment 1, the total length of the glass round rod in the embodiment is 200 mm, the length of the thin section is 20 mm, and the diameter is 3.5 mm; the length of the thick section is 20 mm, and the diameter is 4.6 mm; the middle section has a length of 160 mm and a diameter that gradually increases (equal in slope) from 3.5 mm at the end of the thin section to 4.6 mm at the end of the thick section; otherwise, the same procedure as in example 1 was repeated.
Step four, which is different from example 1 in that the number of spraying was 140, thereby forming a bacterial cellulose tubular hydrogel having a length of 20 mm and a thickness of 2.0 mm, which was attached to the surface of revolution of a thin section of a glass rod having an outer diameter of about 3.5 mm.
And step five, slowly pushing the tubular bacterial cellulose hydrogel film on the round glass rod from the thin section to the thick section through the middle section with the taper, and gradually increasing the inner diameter of the tubular bacterial cellulose hydrogel film to the outer diameter (namely 4.6 millimeters) of the thick section so as to enable the bacterial cellulose nanofibers in the tubular bacterial cellulose hydrogel film to be oriented in the circumferential direction.
Step six, the same as example 1.
Seventhly, as in the embodiment 1, the small blood vessel stent with the bacterial cellulose nanofibers oriented along the circumferential direction is finally obtained, wherein the length of the small blood vessel stent is about 20 millimeters, the thickness of the small blood vessel stent is about 0.6 millimeter, and the inner diameter of the small blood vessel stent is about 4.6 millimeters.
Example 3, preparation of small vessel stents comprising the following steps:
step one, the same as example 1.
Step two, the same as example 1.
Step three, different from the embodiment 1, the total length of the glass round rod in the embodiment is 214 mm, the length of the thin section is 17 mm, and the diameter is 4.0 mm; the length of the thick section is 17 mm, and the diameter is 5.6 mm; the middle section has a length of 180 mm and a diameter that gradually increases (equal in slope) from 4.0 mm at one end of the thin section to 5.6 mm at one end of the thick section; otherwise, the same procedure as in example 1 was repeated.
Step four, which is different from example 1 in that the number of spraying was 110, thereby forming a bacterial cellulose tubular hydrogel having a length of 17 mm and a thickness of 1.8 mm, which was attached to the surface of revolution of a thin section of a glass rod having an outer diameter of about 4.0 mm.
And step five, slowly pushing the tubular bacterial cellulose hydrogel film on the round glass rod from the thin section to the thick section through the middle section with the taper, and gradually increasing the inner diameter of the tubular bacterial cellulose hydrogel film to the outer diameter (namely 5.6 millimeters) of the thick section B so as to enable the bacterial cellulose nanofibers in the tubular bacterial cellulose hydrogel film to be oriented in the circumferential direction.
Step six, the same as example 1.
Seventhly, as in the embodiment 1, the small blood vessel stent with the bacterial cellulose nanofibers oriented along the circumferential direction is finally obtained, wherein the length of the small blood vessel stent is about 17 millimeters, the thickness of the small blood vessel stent is about 0.4 millimeter, and the inner diameter of the small blood vessel stent is about 5.6 millimeters.
Although the present invention has been described in detail with reference to the drawings, the present invention is not limited to the above embodiments, which are only illustrative and not restrictive, and those skilled in the art can make many modifications without departing from the spirit of the present invention, which falls within the protection of the present invention.

Claims (6)

1. The small blood vessel stent with the nanofibers oriented in the circumferential direction is characterized in that the small blood vessel stent is formed by uniformly surrounding bacterial cellulose nanofibers into a circular tube shape, the bacterial cellulose nanofibers are arranged in the circumferential direction, and the small blood vessel stent is 2-6 mm in inner diameter, 0.01-1 mm in wall thickness and 10-30 mm in length.
2. The method for preparing the small vessel stent with the nanofibers oriented in the circumferential direction according to claim 1, wherein a bacterial cellulose hydrogel is formed in situ on a rotating surface of a round rod, the inner diameter of the tubular bacterial cellulose hydrogel is gradually increased by a displacement method to achieve the in-situ circumferential orientation of the nanofibers, and after heating, drying and shaping, the round rod is removed to obtain the small vessel stent with the nanofibers oriented in the circumferential direction and the controllable wall thickness of the tube.
3. The method for preparing a small vessel stent with nanofibers oriented in the circumferential direction according to claim 2, characterized by the following steps:
step one, preparing a culture medium by adopting a conventional method, and placing the culture medium in a sterilization pot for high-temperature and high-pressure sterilization for 30-60 minutes;
step two, preparing a bacterial cellulose base membrane: inoculating the strain into a culture medium in an aseptic environment; statically culturing the inoculated culture medium at 30 ℃ for 3 days to obtain a bacterial cellulose base membrane;
taking a round bar, wherein the round bar comprises an A section, a middle section and a B section which are coaxially connected in sequence; the diameter of the section A is Da, the Da is 1.5-4.5 mm, and the length of the section A is 10-30 mm; the diameter of the section B is Db, Db is 2-6 mm, Db is 1.3-1.4 Da, and the length of the section B is 10-30 mm; the diameter of the connecting end of the middle section and the section A is Da, the diameter of the connecting end of the middle section and the section B is Db, the length of the middle section is 100-200 mm, the middle section is a diameter gradually-changing section, and the diameter gradually changes from Da to Db along the length of the middle section; after ultraviolet sterilization, the round rod is fixed in an oxygen-permeable silica gel tube with a sampling hole at the upper end downwards according to the section A, and is vertically placed on the surface of the bacterial cellulose base membrane obtained in the step two;
step four, preparing a tubular bacterial cellulose hydrogel film with the length of 10-30 mm and the film thickness of 0.01-1 mm on the outer rotating surface of the section A of the round bar by adopting a film liquid interface culture technology;
step five, slowly pushing the tubular bacterial cellulose hydrogel film from the section A of the round rod to the section B through the middle section with the taper, so that the inner diameter of the tubular bacterial cellulose hydrogel film is gradually increased, and the bacterial cellulose nanofibers are oriented along the circumferential direction;
and sixthly, putting the round bar with the bacterial cellulose hydrogel film into deionized water, cleaning for 2 times, then putting the round bar into the deionized water, heating and boiling, after the bacterial cellulose hydrogel film turns white from light yellow, putting the round bar into 0.5 mol/L NaOH solution, boiling for 20 minutes, finally boiling in the deionized water, repeating the boiling for 3-4 times, and washing with the deionized water until the round bar is neutral.
And seventhly, drying the cleaned round bar with the bacterial cellulose hydrogel film in an oven at 60 ℃ for 6-12 hours, and then taking down the tubular bacterial cellulose dry film to obtain the small blood vessel stent with the bacterial cellulose nanofibers oriented along the circumferential direction, wherein the inner diameter of the small blood vessel stent is 2-6 millimeters, the wall thickness of the small blood vessel stent is 0.01-1 millimeter, and the length of the small blood vessel stent is 10-30 millimeters.
4. The method for preparing the small blood vessel stent with the nanofibers oriented in the circumferential direction according to claim 3, wherein the specific process of the fourth step is to spray the nanofibers at 0.012-0.025 m L/cm2Spraying the culture medium obtained in the step one on a contact surface of the round bar and the bacterial cellulose base membrane from a sample inlet hole at the upper end of the oxygen permeable silicone tube, spraying again after the culture medium on the bacterial cellulose base membrane is completely consumed, and repeatedly spraying for 50-200 times; thus, the tubular bacterial cellulose hydrogel film with the length of 10-30 mm and the film thickness of 0.01-1 mm is prepared on the outer surface of the section A of the round bar.
5. The method for preparing a small vessel stent with nanofibers oriented in the circumferential direction according to claim 3, wherein the round rods at least comprise glass round rods, ceramic round rods, alkali-boiling resistant metal round rods and alkali-boiling resistant high polymer material round rods.
6. The method for preparing a small vessel stent with nanofibers oriented in a circumferential direction according to claim 3, wherein in the second step, the bacterial species at least include Acetobacter xylinum, Acetobacter aceti, Acetobacter xylinum, Acetobacter acetogenium and Acetobacter pasteurianus.
CN202010220104.9A 2020-03-25 2020-03-25 Small blood vessel stent with nano fibers oriented along circumferential direction and preparation method Expired - Fee Related CN111388760B (en)

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