CN117582561A - Preparation method of silk fibroin vascular stent and silk fibroin vascular stent - Google Patents
Preparation method of silk fibroin vascular stent and silk fibroin vascular stent Download PDFInfo
- Publication number
- CN117582561A CN117582561A CN202311559198.2A CN202311559198A CN117582561A CN 117582561 A CN117582561 A CN 117582561A CN 202311559198 A CN202311559198 A CN 202311559198A CN 117582561 A CN117582561 A CN 117582561A
- Authority
- CN
- China
- Prior art keywords
- silk fibroin
- solution
- silk
- vascular stent
- drying
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010022355 Fibroins Proteins 0.000 title claims abstract description 294
- 230000002792 vascular Effects 0.000 title claims abstract description 103
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 130
- 238000001035 drying Methods 0.000 claims abstract description 64
- 239000000843 powder Substances 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 238000002791 soaking Methods 0.000 claims abstract description 10
- 238000009966 trimming Methods 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims description 123
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 47
- 239000012498 ultrapure water Substances 0.000 claims description 47
- 229910003002 lithium salt Inorganic materials 0.000 claims description 43
- 159000000002 lithium salts Chemical class 0.000 claims description 43
- 239000007864 aqueous solution Substances 0.000 claims description 36
- 238000000034 method Methods 0.000 claims description 28
- 238000003756 stirring Methods 0.000 claims description 28
- 238000000502 dialysis Methods 0.000 claims description 27
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 26
- 238000010438 heat treatment Methods 0.000 claims description 25
- 238000004321 preservation Methods 0.000 claims description 19
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 14
- 238000003760 magnetic stirring Methods 0.000 claims description 14
- 239000011259 mixed solution Substances 0.000 claims description 14
- 238000009423 ventilation Methods 0.000 claims description 14
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 13
- AMXOYNBUYSYVKV-UHFFFAOYSA-M lithium bromide Chemical compound [Li+].[Br-] AMXOYNBUYSYVKV-UHFFFAOYSA-M 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 12
- 239000011148 porous material Substances 0.000 claims description 9
- 239000008176 lyophilized powder Substances 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 7
- 238000003698 laser cutting Methods 0.000 claims description 7
- 238000010147 laser engraving Methods 0.000 claims description 7
- -1 polypropylene Polymers 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 229910052751 metal Inorganic materials 0.000 claims description 5
- 239000002184 metal Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000004743 Polypropylene Substances 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 229920001155 polypropylene Polymers 0.000 claims description 4
- 241000255789 Bombyx mori Species 0.000 claims description 3
- 108010013296 Sericins Proteins 0.000 claims description 3
- ZJZXSOKJEJFHCP-UHFFFAOYSA-M lithium;thiocyanate Chemical compound [Li+].[S-]C#N ZJZXSOKJEJFHCP-UHFFFAOYSA-M 0.000 claims description 3
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 3
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 2
- 238000007710 freezing Methods 0.000 claims description 2
- 230000008014 freezing Effects 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 210000004204 blood vessel Anatomy 0.000 abstract description 12
- 238000004090 dissolution Methods 0.000 abstract description 5
- 238000006065 biodegradation reaction Methods 0.000 abstract description 3
- 239000002861 polymer material Substances 0.000 abstract description 3
- 238000011112 process operation Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 208000007536 Thrombosis Diseases 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000003892 spreading Methods 0.000 description 4
- 230000007480 spreading Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 2
- 238000003754 machining Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000010907 mechanical stirring Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 208000037803 restenosis Diseases 0.000 description 2
- 241000382353 Pupa Species 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000005297 material degradation process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
- C07K14/43586—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects from silkworms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/043—Proteins; Polypeptides; Degradation products thereof
- A61L31/047—Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/146—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Vascular Medicine (AREA)
- Surgery (AREA)
- Heart & Thoracic Surgery (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Insects & Arthropods (AREA)
- Dispersion Chemistry (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention relates to the technical field of organic high polymer materials, in particular to a preparation method of a silk fibroin vascular stent and the silk fibroin vascular stent. Preparing a silk fibroin solution by adopting silk fibroin freeze-dried powder, drying to form a film and trimming to obtain a silk fibroin wet film, immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and drying to obtain a target silk fibroin film; processing the target silk fibroin film to obtain an initial silk fibroin vascular stent; and (3) carrying out water soaking softening treatment on the initial silk fibroin vascular stent, embedding the initial silk fibroin vascular stent on a preset die, drying and demolding to obtain the silk fibroin vascular stent. The invention has simple process operation, can realize biodegradation, can regulate and control the dissolution rate of the silk fibroin vascular stent and regulate and control the mechanical property and the mechanical property of the silk fibroin vascular stent, and can better play a role in supporting blood vessels.
Description
Technical Field
The invention relates to the technical field of organic high polymer materials, in particular to a preparation method of a silk fibroin vascular stent and the silk fibroin vascular stent.
Background
The acute thrombus can be quickly removed by mechanical thrombus suction and catheter thrombolysis, so as to achieve the aim of restoring blood flow, and the vascular stent is an indispensable consumable. The vascular stents used in clinic comprise degradable vascular stents and non-degradable vascular stents. The degradable vascular stent comprises a degradable metal vascular stent and a high polymer vascular stent, mechanical support is provided within a certain period of time, and the vascular stent can pass through the restenosis after the expansion for 6 months after the operation, but the material degradation controllability of the degradable vascular stent is difficult to control, the mechanical performance is difficult to meet the requirement, and the cost is high. The main material of the nondegradable vascular stent is metal, has good plasticity and geometric stability, high mechanical strength, and can better play a role in supporting blood vessels when being delivered to lesion sites, but the undegraded vascular stent is easy to expand excessively when being expanded to cause rupture, and the metal material has thrombus source, so that thrombus formation and intimal hyperplasia are possibly caused, and restenosis of blood vessels is caused.
Disclosure of Invention
In order to solve the technical problems, the invention provides a preparation method of a silk fibroin vascular stent, which comprises the following steps:
preparing a silk fibroin solution based on silk fibroin freeze-dried powder; the average molecular weight of silk fibroin in the silk fibroin freeze-dried powder is 10 KDa-350 KDa;
drying the silk fibroin solution to form a film and trimming to obtain a silk fibroin wet film;
immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and drying to obtain a target silk fibroin film;
processing the target silk fibroin film to obtain an initial silk fibroin vascular stent; the initial silk fibroin vascular stent has a grid pore structure;
soaking the initial silk fibroin vascular stent in ultrapure water for 1-50 times, and performing softening treatment;
and (3) embedding the softened initial silk fibroin tubular stent on a preset die, drying and demolding to obtain the target silk fibroin vascular stent.
Optionally, preparing the silk fibroin solution based on the silk fibroin lyophilized powder comprises:
placing the clean and dry cocoons in a sodium carbonate solution, and performing heating degumming treatment to form silk;
washing silk, drying, dissolving in lithium salt water solution, stirring and mixing uniformly, and preserving heat under preset heat preservation conditions to obtain silk fibroin lithium salt water solution;
dialyzing and centrifuging the silk fibroin lithium salt aqueous solution to obtain supernatant after centrifuging to obtain silk fibroin aqueous solution; the concentration of the aqueous solution of silk fibroin is 0.05-0.25 g/ml;
freeze-drying the aqueous silk fibroin solution to obtain freeze-dried silk fibroin powder;
and dissolving the silk fibroin freeze-dried powder by using hexafluoroisopropanol to prepare a silk fibroin solution.
Optionally, placing the clean and dry cocoons in a sodium carbonate solution, and performing heat degumming treatment to form silk comprises:
the acquisition area is 1-1000 mm 2 Placing the cocoons in a sodium carbonate solution with the concentration of 0.1-3 g/L to obtain a first mixed solution; the volume ratio of the mass of the silkworm cocoons to the sodium carbonate solution is (5:2) g/L;
and (3) heating the first mixed solution at the heating temperature of 60-100 ℃, the heating pressure of 0.5-1 Mpa and the heating time of 1 s-1000 h, and stirring to remove sericin, thereby obtaining silk.
Optionally, after washing silk and drying, dissolving the silk in lithium salt water solution, stirring and mixing uniformly, and preserving heat under preset heat preservation conditions to obtain silk fibroin lithium salt water solution, wherein the step of obtaining silk fibroin lithium salt water solution comprises the following steps:
placing silk in ultrapure water to obtain a second mixed solution; the mass ratio of silk to ultrapure water is 1:20-1:50;
stirring the second mixed solution by mechanical stirring or magnetic stirring; the stirring speed of the magnetic stirring is 1-2000 r/min, and the stirring time is 60 s-5 h;
taking out silk and squeezing, repeatedly placing the silk into ultrapure water and stirring, wherein the repetition time is 1-50 times;
the silk washed by natural drying treatment or heating ventilation drying treatment is heated and ventilated to be dried at the drying temperature of 50-70 ℃ for 1-20 h;
dissolving the dried silk in lithium salt aqueous solution, wherein the lithium salt aqueous solution is lithium thiocyanate solution or lithium bromide solution, and the concentration of the lithium salt aqueous solution is 0.01-2 g/ml; the volume ratio of the mass of the silk after the drying treatment to the lithium salt water solution is (1-100): 100g/ml;
preserving heat under a preset heat preservation condition to obtain a silk fibroin lithium salt aqueous solution; the preset heat preservation condition is that the heat preservation temperature is 30-80 ℃ and the heat preservation time is 0.5-5 h.
Optionally, dialyzing and centrifuging the silk fibroin lithium salt aqueous solution comprises:
pouring the silk fibroin lithium salt aqueous solution into a dialysis bag, placing the dialysis bag into 1 mL-1000L of ultrapure water, and performing dialysis by standing or magnetic stirring; the specification of the dialysis bag is 10-1000000 Da; the stirring speed of magnetic stirring dialysis is 1-2000 r/min; the total dialysis time is 24-72 h, and the ultrapure water is replaced at intervals of 0.1-100 h;
transferring the silk fibroin lithium salt water solution subjected to dialysis treatment into a centrifuge tube, and performing centrifugal treatment; the rotational speed of the centrifugal treatment is 10-20000 r/min, the time is 6 s-2 h, and the temperature is-3-5 ℃.
Optionally, freeze-drying the aqueous silk fibroin solution to obtain silk fibroin freeze-dried powder comprises:
freezing the aqueous solution of the silk fibroin at the temperature of-80 to 0 ℃, and then performing freeze drying treatment for 1 to 500 hours under the vacuum condition that the vacuum pressure is 0.001 to 1mBar and the temperature is-80 to 0 ℃ to obtain the silk fibroin freeze-dried powder.
Optionally, dissolving the silk fibroin freeze-dried powder by using hexafluoroisopropanol, and preparing the silk fibroin solution comprises the following steps:
at the temperature of 5-80 ℃, the silk fibroin freeze-dried powder and hexafluoroisopropanol are placed into a sealed container to be dissolved for 0.1-100 h, so as to obtain silk fibroin solution; the mass ratio of the silk fibroin freeze-dried powder to hexafluoroisopropanol is 1: (1-20).
Optionally, drying the silk fibroin solution to form a film and finishing, wherein the obtaining of the silk fibroin wet film comprises the following steps:
loading the silk fibroin solution into an injection device with the specification of 1-200 ml, and standing for 1 min-100 h until bubbles are dissipated;
uniformly injecting a silk fibroin solution into a kit body, spreading, ventilating and drying to form a film, and trimming the edge to obtain a silk fibroin wet film; wherein,
the reagent material of the kit body comprises one of polypropylene, polytetrafluoroethylene and a metal kit; the area of the bottom of the kit body is 1-1000 cm < 2 >;
the tiling thickness of the silk fibroin solution is 1-1000 mm; the ambient temperature of the ventilation drying treatment is 10-80 ℃, and the drying time is 1 h-20 days.
Optionally, immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and performing drying treatment to obtain a target silk fibroin film, wherein the method comprises the following steps:
immersing the silk fibroin wet film in 1-50L of methanol solution for 1-500 h; changing the methanol solution at intervals of 1-50 h, wherein the number of times of changing the methanol solution is 1-50 times;
taking out the silk fibroin wet film from the methanol solution, and soaking in 1-5L of ultrapure water; the ultrapure water is replaced at intervals of 1-100 h, and the times of replacing the ultrapure water is 1-50 times, so that the methanol solution is removed;
and (3) carrying out ventilation drying treatment on the silk fibroin wet film from which the methanol solution is removed at the environment temperature of 10-80 ℃ for 1-200 days to obtain the target silk fibroin film.
Optionally, processing the target silk fibroin film to obtain a processing mode in the initial silk fibroin vascular stent, including but not limited to laser engraving, laser cutting or machine tool processing; the laser power of the laser engraving and the laser cutting is 1-30W;
the initial silk fibroin vascular stent has the length of 0.1-10 cm and the diameter of 1-15 mm.
Optionally, immersing the initial silk fibroin vascular scaffold in ultrapure water 1-50 times comprises:
soaking the initial silk fibroin vascular stent in 1-50L of ultrapure water, and replacing the ultrapure water at intervals of 1-50 h for 1-50 times.
Optionally, the preset die is a bar-shaped die with the diameter of 0.1-20 mm;
and demolding after drying to obtain the target silk fibroin vascular stent, wherein the drying time is 1-1000 h.
In another aspect, the present invention provides a silk fibroin vascular stent made by the silk fibroin vascular stent preparation method described above.
The technical scheme provided by the embodiment of the invention has the following technical effects:
the silk fibroin vascular stent prepared by the invention has simple technological operation and is suitable for industrialization; the silk is used as a raw material, so that the cost is low, and the biodegradation can be realized; the average molecular weight of the silk fibroin can be controlled by the heating time length, so that the mechanical property of the silk fibroin vascular stent can be regulated and controlled; the methanol can change the protein molecular structure in the silk fibroin wet film so as to regulate and control the mechanical property and the mechanical property of the silk fibroin vascular stent, can better play a role in supporting blood vessels, and in addition, the silk fibroin wet film is soaked in the methanol to perform insoluble treatment so as to regulate and control the dissolution rate of the silk fibroin vascular stent; the size, shape and the like of the grid pore structure can be adjusted according to actual requirements so as to ensure that the silk fibroin vascular stent is firmly combined with a target entity such as a blood vessel.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions and advantages of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are only some embodiments of the invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic flow chart of a method for preparing a silk fibroin vascular stent according to an embodiment of the present invention;
FIGS. 2 a-2 i are schematic structural flow diagrams of a method for preparing a silk fibroin vascular stent according to an embodiment of the present invention;
fig. 3 is a diagram of an example of a silk fibroin blood vessel according to an embodiment of the present invention.
Reference numerals illustrate:
1-silk fibroin aqueous solution; 2-silk fibroin solution; 3-an injection device; 4-silk fibroin wet film; 5-methanol solution; 6-insoluble silk fibroin film; 7-a target silk fibroin membrane; 8-initial silk fibroin vascular stent; 9-presetting a die; 10-target silk fibroin vascular stent.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that reference throughout this specification to "one embodiment" or "an embodiment" of an embodiment of the present invention means that a particular feature, structure, or characteristic may be included in at least one implementation of the present invention. It should be understood that the terms "first" and "second" in the description and claims of embodiments of the invention and in the above-described figures are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may include one or more of the feature, either explicitly or implicitly. Moreover, the terms "first," "second," and the like, are used for distinguishing between similar objects and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used may be interchanged where appropriate such that the embodiments of the invention described herein may be implemented in sequences other than those illustrated or otherwise described herein. In addition, in the description of the present embodiment, unless otherwise specified, the meaning of "a plurality" is two or more. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, or article that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed but may include other steps or elements not expressly listed or inherent to such process, method, article, or article.
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings and examples. It should be understood that the detailed description and specific examples, while indicating the embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
Where numerical ranges are provided in the examples, it is understood that unless otherwise stated herein, both endpoints of each numerical range and any number between the two endpoints are significant both in the numerical range. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, materials used in the embodiments, any methods, devices, and materials of the prior art similar or equivalent to those described in the embodiments of the present invention may be used to practice the present invention according to the knowledge of one skilled in the art and the description of the present invention.
Fibroin material is one of the earliest natural protein materials developed by human beings as a biological material with a long history of application. The silk fibroin is a main constituent material of silk (accounting for about 70% of the mass of silk), and is a novel green bio-based material with modern technological characteristics. Compared with other protein materials and organic polymer materials, the silk fibroin has unique and excellent biological, optical and mechanical properties, including high light transmittance, no human rejection, good biocompatibility, adjustable dissolution and degradation rate, high mechanical strength, wide sources and the like, is very suitable for being used as a material of a vascular stent, is low in price, and can reduce the expenditure of cardiovascular medical aspects in China.
Referring to fig. 1, fig. 1 is a schematic flow chart of a preparation method of a silk fibroin vascular stent according to an embodiment of the present invention, as illustrated in fig. 1, the preparation method includes S101-S106:
s101, preparing a silk fibroin solution based on silk fibroin freeze-dried powder; the average molecular weight of silk fibroin in the silk fibroin freeze-dried powder is 10 KDa-350 KDa.
First, a silk fibroin lyophilized powder is prepared, and in one possible embodiment, the process of preparing silk fibroin lyophilized powder can include the steps of:
(1) And (3) placing the clean and dry cocoons into a sodium carbonate solution, and performing heating degumming treatment to form silk.
In the embodiment of the invention, the cocoons are clean and dry cocoons after the pupa is removed; the cocoons may be rectangular or square in shape, in one possible embodiment, the acquisition area is 1-1000 mm 2 Placing cocoons in a sodium carbonate solution with the concentration of 0.1-3 g/L to obtain a first mixed solution; wherein the volume ratio of the mass of the silkworm cocoons to the sodium carbonate solution is (5:2) g/L. For example, in an exemplary embodiment, 10g portions of cocoons are weighed and sheared and placed in 4L of sodium carbonate solution at a concentration of 2.12 g/L.
And (3) heating the first mixed solution at the heating temperature of 60-100 ℃, the heating pressure of 0.5-1 Mpa and the heating time of 1 s-1000 h, and stirring to remove sericin, thereby obtaining silk. The average molecular weight of silk fibroin of the silk fibroin freeze-dried powder can influence the mechanical property of the finally prepared silk fibroin vascular stent, and in the embodiment of the invention, the average molecular weight of silk fibroin can be controlled by the heating time. For example, in the exemplary embodiment, the first mixed solution is heated to boiling by an electric furnace under normal pressure and magnetically stirred at a rotation speed of 200r/min for 30 minutes.
(2) And (3) washing silk, drying, dissolving in lithium salt water solution, stirring and mixing uniformly, and preserving heat under the preset heat preservation condition to obtain the silk fibroin lithium salt water solution.
The silk may be washed by stirring or soaked by standing, and in one possible embodiment, the silk is placed in ultrapure water to obtain a second mixed solution; the mass ratio of silk to ultrapure water is 1:20-1:50; stirring the second mixed solution by mechanical stirring or magnetic stirring; the stirring speed of the magnetic stirring is 1-2000 r/min, and the stirring time is 60 s-5 h; taking out the silk and squeezing, and repeatedly placing the silk into ultrapure water and stirring for 1-50 times.
The silk is washed for many times by adopting the steps, in each washing process, the mass ratio of the silk to the ultrapure water is 1:20-1:50, for example, in an exemplary embodiment, each part of silk is placed into the ultrapure water according to the mass ratio of 1:20, stirred, washed and kneaded to dryness, magnetic stirring is adopted, the stirring speed is 200r/min, the stirring time is 20min, the silk is taken out and squeezed to dryness, the ultrapure water with the same amount is replaced each time, and the washing is repeated for 3 times.
The silk washed by natural drying treatment or heating ventilation drying treatment is heated and ventilated to be dried at the drying temperature of 50-70 ℃ for 1-20 h.
Dissolving the dried silk in lithium salt aqueous solution, wherein the lithium salt aqueous solution is lithium thiocyanate solution or lithium bromide solution, and the concentration of the lithium salt aqueous solution is 0.01-2 g/ml; the volume ratio of the mass of the silk after the drying treatment to the lithium salt water solution is (1-100): 100g/ml.
Preserving heat under a preset heat preservation condition to obtain a silk fibroin lithium salt aqueous solution; the preset heat preservation condition is that the heat preservation temperature is 30-80 ℃ and the heat preservation time is 0.5-5 h.
For example, in an exemplary embodiment, the washed silk is subjected to a heating and ventilating drying process, the temperature of the heating and ventilating drying process is 50 ℃, and the drying time is 2 hours. And (3) dissolving the dried silk in a lithium bromide solution with the concentration of 0.98g/mL, wherein the relation between the volume of the lithium bromide solution and the mass of the dried silk is 4 mL/1 g, and preserving the temperature at 60 ℃ for 4 hours to obtain a silk fibroin lithium salt aqueous solution, namely a mixed solution of fibroin and lithium bromide.
(3) Dialyzing and centrifuging the silk fibroin lithium salt aqueous solution to obtain supernatant after centrifuging to obtain silk fibroin aqueous solution; the concentration of the aqueous solution of silk fibroin is 0.05-0.25 g/ml.
In one possible embodiment, the aqueous solution of silk fibroin lithium salt is poured into a dialysis bag, placed in 1mL to 1000L of ultrapure water, dialyzed by standing or magnetic stirring; the specification of the dialysis bag is 10-1000000 Da; the stirring speed of magnetic stirring dialysis is 1-2000 r/min; the total dialysis time is 24-72 h, and the ultrapure water is replaced at intervals of 0.1-100 h; transferring the silk fibroin lithium salt water solution subjected to dialysis treatment into a centrifuge tube, and performing centrifugal treatment; the rotational speed of the centrifugal treatment is 10-20000 r/min, the time is 6 s-2 h, and the temperature is-3-5 ℃.
For example, in an exemplary embodiment, the silk fibroin lithium salt aqueous solution is filled into a dialysis bag with the molecular weight cutoff of 3500Da, and is placed into 4L of ultrapure water for dialysis, wherein the dialysis mode is magnetic stirring dialysis, and the stirring speed of the magnetic stirring dialysis is 1-2000 r/min; the ultrapure water is replaced once at intervals of 5 hours, the volume of the ultrapure water is 4L each time, and the total dialysis time is 48 hours. Transferring the silk fibroin lithium salt water solution subjected to dialysis treatment into a centrifuge tube, and performing centrifugal treatment; the rotational speed of the centrifugal treatment is 12000r/min, the time is 30min, the temperature is 4 ℃, and the supernatant after the centrifugal treatment is obtained to obtain the silk fibroin aqueous solution.
(4) And freeze-drying the aqueous silk fibroin solution to obtain freeze-dried silk fibroin powder.
In one possible embodiment, the silk fibroin aqueous solution is frozen at the temperature of-80 to 0 ℃, and then the silk fibroin aqueous solution is frozen and dried for 1 to 500 hours under the vacuum condition that the vacuum pressure is 0.001 to 1mBar and the temperature is-80 to 0 ℃ to obtain the silk fibroin freeze-dried powder.
For example, in an exemplary embodiment, an aqueous silk fibroin solution is frozen in an environment of-80 ℃ and then vacuum frozen in a freeze dryer at a vacuum pressure of 1mBar and a temperature of-80 ℃ for 1 hour.
Next, a silk fibroin solution is prepared based on silk fibroin lyophilized powder, and in one possible embodiment, the silk fibroin solution is obtained by dissolving silk fibroin lyophilized powder with hexafluoroisopropanol. At the temperature of 5-80 ℃, the silk fibroin freeze-dried powder and hexafluoroisopropanol are placed into a sealed container to be dissolved for 0.1-100 h, so as to obtain silk fibroin solution; the mass ratio of the silk fibroin freeze-dried powder to hexafluoroisopropanol is 1: (1-20).
For example, in an exemplary embodiment, at a temperature of 25 ℃, the mass ratio of the silk fibroin lyophilized powder to hexafluoroisopropanol is 1:10 is dissolved in a 2L beaker for 1h to obtain a silk fibroin solution, and sealing is carried out by a sealing film during dissolution to prevent volatilization of hexafluoroisopropanol.
S102, drying the silk fibroin solution to form a film and trimming to obtain a silk fibroin wet film.
In one possible embodiment, the silk fibroin solution is loaded into an injection device of 1-200 ml in size, and left to stand for 1 minute-100 hours until bubbles dissipate; uniformly injecting a silk fibroin solution into a kit body, spreading, ventilating and drying to form a film, and trimming the edge to obtain a silk fibroin wet film; wherein, the reagent material of the reagent box body comprises one of polypropylene, polytetrafluoroethylene and a metal box; the area of the bottom of the kit body is 1-1000 cm < 2 >; the tiling thickness of the silk fibroin solution is 1-1000 mm; the ambient temperature of the ventilation drying treatment is 10-80 ℃, and the drying time is 1 h-20 days.
For example, in the exemplary embodiment, the silk fibroin solution is filled into a 200ml injection device, and is kept stand for 1h, and after the bubbles are completely dissipated, the silk fibroin solution is uniformly coated on a box bottom with an area of 200cm 2 Uniformly spreading in the polypropylene kit body, ventilating and drying for 12 hours at the temperature of 10-50 ℃, taking off the silk fibroin wet film, and trimming the edge with uneven thickness.
S103, immersing the silk fibroin wet film in a methanol solution, removing the methanol solution, and drying to obtain the target silk fibroin film.
Specifically, the methanol can change the protein molecular structure in the silk fibroin wet film so as to regulate and control the mechanical property and the mechanical property of the silk fibroin vascular stent, and can better play a role in supporting blood vessels.
In one possible embodiment, the silk fibroin wet film is immersed in 1-50L of methanol solution for 1-500 h; changing the methanol solution 1-50 times at intervals of 1-50 h to obtain an insoluble silk fibroin film; taking out the silk fibroin wet film from the methanol solution, and soaking in 1-5L of ultrapure water; the ultrapure water is replaced at intervals of 1-100 h, and the times of replacing the ultrapure water is 1-50 times so as to remove the methanol solution; and (3) carrying out ventilation drying treatment on the silk fibroin wet film from which the methanol solution is removed at the environment temperature of 10-80 ℃ for 1-200 days to obtain the target silk fibroin film.
For example, in an exemplary embodiment, a trimmed silk fibroin wet film is filled into a 2L beaker, methanol is poured into the 2L beaker, the 2L beaker is sealed by a sealing film, the methanol solution is replaced once at intervals of 24 hours, and the total soaking time is 500 hours, so that an insoluble silk fibroin wet film is obtained. Then taking out the insoluble silk fibroin wet film, filling into a 2L beaker, pouring ultrapure water to 2L scale marks, sealing by using a sealing film, replacing the ultrapure water at intervals of 24 hours for 35 times, and removing the methanol solution. And then carrying out ventilation drying treatment for 100 days in a ventilation environment at the temperature of between 10 and 80 ℃ to obtain the target silk fibroin film.
S104, processing a target silk fibroin film to obtain an initial silk fibroin vascular stent; the initial silk fibroin vascular stent has a lattice pore structure.
In one possible embodiment, the machining means includes, but is not limited to, laser engraving, laser cutting or machine tool machining; the laser power of the laser engraving and the laser cutting is 1-30W; the initial silk fibroin vascular stent has the length of 0.1-10 cm and the diameter of 1-15 mm. According to the embodiment, the target silk fibroin film is engraved to form a grid pore structure, and the size, shape and the like of the grid pore structure can be adjusted according to actual requirements, so that the silk fibroin vascular stent is firmly combined with a target entity such as a blood vessel.
For example, in an exemplary embodiment, laser engraving and laser cutting with a laser power of 1-30W is used to obtain an initial silk fibroin vascular stent with a length of 10cm and a diameter of 15mm.
S105, soaking the initial silk fibroin vascular stent in ultrapure water for 1-50 times, and performing softening treatment.
In one possible embodiment, the initial silk fibroin vascular stent is immersed in 1 to 50L of ultrapure water to be softened, and the ultrapure water is replaced at intervals of 1 to 50 hours, wherein the number of times of replacement of the ultrapure water is 1 to 50 times. For example, in an exemplary embodiment, the initial silk fibroin vascular stent is placed in a 2L beaker, ultrapure water is added to the 2L graduation line, water is changed once a day, and after water is changed for 4 times, the softened initial silk fibroin vascular stent is taken out. By means of the above embodiment, the mechanical properties of the silk fibroin vascular stent are regulated, and the shape thereof is regulated, so that the silk fibroin vascular stent can be better combined with a target entity such as a blood vessel.
S106, embedding the softened initial silk fibroin tubular stent on a preset die, drying and demolding to obtain the target silk fibroin vascular stent.
In one possible embodiment, the preset die is a bar-shaped die with a diameter of 0.1-20 mm; and demolding after drying to obtain the target silk fibroin vascular stent, wherein the drying time is 1-1000 h. For example, in an exemplary embodiment, the softened initial silk fibroin tubular stent is nested on a rod-shaped mold with the diameter of 6mm, moved into a ventilation drying cabinet, naturally dried for 6 days, and then demolded to obtain the target silk fibroin vascular stent. The silk fibroin vascular stents of interest obtained by this embodiment can be better adapted to bind to a target entity such as a blood vessel.
In another aspect, the present invention provides a silk fibroin vascular stent made by the silk fibroin vascular stent preparation method described above.
According to the invention, through the embodiments, the silk fibroin freeze-dried powder is adopted to prepare the silk fibroin solution, and the silk fibroin solution is dried to form a film and is trimmed to obtain a silk fibroin wet film; immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and drying to obtain a target silk fibroin film; processing the target silk fibroin film to obtain an initial silk fibroin vascular stent; the initial silk fibroin vascular stent has a grid pore structure; soaking the initial silk fibroin vascular stent in ultrapure water for softening treatment; the initial silk fibroin tubular stent after softening treatment is nested on a preset die, and the silk fibroin vascular stent obtained by demolding after drying is simple in technological operation and suitable for industrialization; the silk is used as a raw material, so that the cost is low, and the biodegradation can be realized; the average molecular weight of the silk fibroin can be controlled by the heating time length, so that the mechanical property of the silk fibroin vascular stent can be regulated and controlled; the methanol can change the protein molecular structure in the silk fibroin wet film so as to regulate and control the mechanical property and the mechanical property of the silk fibroin vascular stent, can better play a role in supporting blood vessels, and in addition, the silk fibroin wet film is soaked in the methanol to perform insoluble treatment so as to regulate and control the dissolution rate of the silk fibroin vascular stent; the size, shape and the like of the grid pore structure can be adjusted according to actual requirements so as to ensure that the silk fibroin vascular stent is firmly combined with a target entity such as a blood vessel.
Based on the above description of a method for preparing a silk fibroin vascular stent, the above preparation method will be further described with reference to a structural flow chart, so as to facilitate understanding of structural changes in a silk fibroin vascular stent preparation method. It should be understood that the following figures are merely exemplary illustrations and that other figures may be obtained by those skilled in the art from the foregoing description without the inventive effort. In addition, the structural changes shown in the following drawings are only one possible example of the production method according to the present invention, and do not represent that the production method according to the embodiment of the present invention is limited to the following examples.
Fig. 2 a-fig. 2i are schematic structural flow diagrams of a method for preparing a silk fibroin vascular stent according to an embodiment of the present invention.
Referring to fig. 2a, a clean and dry cocoon is put into a sodium carbonate solution and is subjected to heating degumming treatment to form silk; washing silk, drying, dissolving in lithium salt water solution, stirring and mixing uniformly, and preserving heat under preset heat preservation conditions to obtain silk fibroin lithium salt water solution; dialyzing and centrifuging the silk fibroin lithium salt aqueous solution to obtain supernatant after centrifuging to obtain silk fibroin aqueous solution 1;
referring to fig. 2b, the aqueous silk fibroin solution 1 is freeze-dried to obtain silk fibroin freeze-dried powder; dissolving silk fibroin freeze-dried powder by using hexafluoroisopropanol to prepare silk fibroin solution 2;
referring to fig. 2c, the silk fibroin solution 2 is loaded into the injection device 3 and allowed to stand until bubbles dissipate; uniformly injecting the silk fibroin solution 2 into a kit body, spreading, carrying out ventilation drying treatment to form a film, and trimming the edge to obtain a silk fibroin wet film 4;
referring to fig. 2d, the wet silk fibroin film 4 is immersed in a methanol solution 5 to obtain an insoluble silk fibroin film 6;
referring to FIG. 2e, the insoluble silk fibroin film 6 is taken out of the methanol solution 5, immersed in ultrapure water, and the methanol solution 5 is removed; removing the silk fibroin wet film 4 of the methanol solution 5, and carrying out ventilation drying treatment to obtain a target silk fibroin film 7;
referring to fig. 2f, processing the target silk fibroin film 7 to obtain an initial silk fibroin vascular stent 8, wherein the initial silk fibroin vascular stent 8 has a grid pore structure;
referring to FIG. 2g, the initial silk fibroin vascular stent 8 is immersed in ultrapure water and subjected to softening treatment;
referring to fig. 2h, the initial silk fibroin tubular stent after softening treatment is nested on a preset mold 9;
referring to fig. 2i, the silk fibroin vascular stent of interest 10 is obtained by demolding after drying.
The specific exemplary flow of the steps performed in fig. 2 a-2 i may be referred to above and will not be described again here.
Further, referring to fig. 3, a diagram of an example of a silk fibroin vascular stent prepared according to an embodiment of the present invention is shown.
It should be noted that: the sequence of the embodiments of the present invention is only for description, and does not represent the advantages and disadvantages of the embodiments. And the foregoing description has been directed to specific embodiments of this specification. Other embodiments are within the scope of the following claims. In some cases, the actions or steps recited in the claims can be performed in a different order than in the embodiments and still achieve desirable results. In addition, the processes depicted in the accompanying figures do not necessarily require the particular order shown, or sequential order, to achieve desirable results. In some embodiments, multitasking and parallel processing are also possible or may be advantageous.
In this specification, each embodiment is described in a progressive manner, and identical and similar parts of each embodiment are all referred to each other, and each embodiment mainly describes differences from other embodiments. In particular, for the apparatus embodiments, since they are substantially similar to the method embodiments, the description is relatively simple, and reference is made to the description of the method embodiments in part.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (13)
1. A method for preparing a silk fibroin vascular stent, comprising the steps of:
preparing a silk fibroin solution based on silk fibroin freeze-dried powder; the average molecular weight of silk fibroin in the silk fibroin freeze-dried powder is 10-350 kDa;
drying the silk fibroin solution to form a film and trimming to obtain a silk fibroin wet film;
immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and drying to obtain a target silk fibroin film;
processing the target silk fibroin film to obtain an initial silk fibroin vascular stent; the initial silk fibroin vascular stent has a grid pore structure;
soaking the initial silk fibroin vascular stent in ultrapure water for 1-50 times, and performing softening treatment;
and embedding the softened initial silk fibroin tubular stent on a preset die, drying and demolding to obtain the target silk fibroin vascular stent.
2. The method for preparing a silk fibroin vascular stent according to claim 1, wherein the preparation of silk fibroin solution based on silk fibroin lyophilized powder comprises:
placing the clean and dry cocoons in a sodium carbonate solution, and performing heating degumming treatment to form silk;
washing the silk, drying, dissolving in lithium salt water solution, stirring and mixing uniformly, and preserving heat under preset heat preservation conditions to obtain silk fibroin lithium salt water solution;
dialyzing and centrifuging the silk fibroin lithium salt aqueous solution to obtain supernatant after centrifuging to obtain silk fibroin aqueous solution; the concentration of the silk fibroin aqueous solution is 0.05-0.25 g/ml;
freeze-drying the silk fibroin aqueous solution to obtain silk fibroin freeze-dried powder;
and dissolving the silk fibroin freeze-dried powder by adopting hexafluoroisopropanol to prepare the silk fibroin solution.
3. The method of preparing silk fibroin vascular stent according to claim 2, wherein the heating degumming treatment of the clean and dry cocoons in sodium carbonate solution to form silk comprises:
the acquisition area is 1-1000 mm 2 Placing the cocoons in a sodium carbonate solution with the concentration of 0.1-3 g/L to obtain a first mixed solution; the volume ratio of the mass of the silkworm cocoons to the sodium carbonate solution is (5:2) g/L;
and heating the first mixed solution at the heating temperature of 60-100 ℃, the heating pressure of 0.5-1 Mpa and the heating time of 1 s-1000 h, and stirring to remove sericin, thereby obtaining the silk.
4. The method for preparing a silk fibroin vascular stent according to claim 2, wherein the steps of washing and drying the silk, dissolving the silk in a lithium salt water solution, stirring and mixing uniformly, and performing heat preservation under a preset heat preservation condition to obtain the silk fibroin lithium salt water solution comprise:
placing the silk in ultrapure water to obtain a second mixed solution; the mass ratio of the silk to the ultrapure water is 1:20-1:50;
stirring the second mixed solution mechanically or magnetically; the stirring speed of the magnetic stirring is 1-2000 r/min, and the stirring time is 60 s-5 h;
taking out the silk and squeezing, and repeatedly placing the silk into ultrapure water and stirring for 1-50 times;
the silk washed by natural drying treatment or heating ventilation drying treatment is heated to be ventilated and dried at a drying temperature of 50-70 ℃ for 1-20 h;
dissolving the silk after the drying treatment in a lithium salt water solution, wherein the lithium salt water solution is a lithium thiocyanate solution or a lithium bromide solution, and the concentration of the lithium salt water solution is 0.01-2 g/ml; the volume ratio of the mass of the silk after the drying treatment to the lithium salt aqueous solution is (1-100): 100g/ml;
preserving heat under a preset heat preservation condition to obtain a silk fibroin lithium salt aqueous solution; the preset heat preservation condition is that the heat preservation temperature is 30-80 ℃ and the heat preservation time is 0.5-5 h.
5. The method of preparing a silk fibroin vascular stent according to claim 2, wherein the dialysis and centrifugation of the silk fibroin lithium salt aqueous solution comprises:
pouring the silk fibroin lithium salt aqueous solution into a dialysis bag, placing the dialysis bag into 1 mL-1000L of ultrapure water, and performing dialysis by standing or magnetic stirring; the specification of the dialysis bag is 10-1000000 Da; the stirring speed of the magnetic stirring dialysis is 1-2000 r/min; the total dialysis time is 24-72 h, and the ultrapure water is replaced at intervals of 0.1-100 h;
transferring the silk fibroin lithium salt water solution subjected to dialysis treatment into a centrifuge tube, and performing centrifugal treatment; the rotational speed of the centrifugal treatment is 10-20000 r/min, the time is 6 s-2 h, and the temperature is-3-5 ℃.
6. The method for preparing a silk fibroin vascular stent according to claim 2, wherein the freeze-drying the silk fibroin aqueous solution to obtain the silk fibroin freeze-dried powder comprises:
freezing the silk fibroin aqueous solution at the temperature of-80-0 ℃, and then performing freeze drying treatment for 1-500 h under the vacuum condition that the vacuum pressure is 0.001-1 mBar and the temperature is-80-0 ℃ to obtain the silk fibroin freeze-dried powder.
7. The method of preparing a silk fibroin vascular stent according to claim 2, wherein the dissolving the silk fibroin lyophilized powder with hexafluoroisopropanol to prepare the silk fibroin solution comprises:
under the temperature condition of 5-80 ℃, the silk fibroin freeze-dried powder and hexafluoroisopropanol are placed into a sealed container to be dissolved for 0.1-100 h, and the silk fibroin solution is obtained; the mass ratio of the silk fibroin freeze-dried powder to the hexafluoroisopropanol is 1: (1-20).
8. The method of preparing a silk fibroin vascular stent according to claim 1, wherein drying the silk fibroin solution to form a film and trimming the film to obtain a silk fibroin wet film comprises:
filling the silk fibroin solution into an injection device with the specification of 1-200 ml, and standing for 1 min-100 h until bubbles are dissipated;
uniformly injecting the silk fibroin solution into a kit body, tiling, ventilating and drying to form a film, and trimming the edge to obtain the silk fibroin wet film; wherein,
the reagent material of the kit body comprises one of polypropylene, polytetrafluoroethylene and a metal kit; the area of the bottom of the kit body is 1-1000 cm 2 ；
The tiling thickness of the silk fibroin solution is 1-1000 mm; the ambient temperature of the ventilation drying treatment is 10-80 ℃, and the drying time is 1 h-20 days.
9. The method for preparing a silk fibroin vascular stent according to claim 1, wherein immersing the silk fibroin wet film in a methanol solution, removing the methanol solution and drying to obtain a target silk fibroin film comprises:
immersing the silk fibroin wet film in 1-50L of the methanol solution for 1-500 h; the methanol solution is replaced at intervals of 1-50 h, and the times of replacing the methanol solution is 1-50 times;
taking out the silk fibroin wet film from the methanol solution, and soaking in 1-5L of ultrapure water; replacing the ultrapure water at intervals of 1-100 h for 1-50 times to remove the methanol solution;
and (3) carrying out ventilation drying treatment on the silk fibroin wet film from which the methanol solution is removed at the environment temperature of 10-80 ℃ for 1-200 days to obtain the target silk fibroin film.
10. The method of claim 1, wherein the processing of the target silk fibroin film to obtain an initial silk fibroin stent includes, but is not limited to, laser engraving, laser cutting, or machine tool processing; the laser power of the laser engraving and the laser cutting is 1-30W;
the initial silk fibroin vascular stent has the length of 0.1-10 cm and the diameter of 1-15 mm.
11. The method of preparing a silk fibroin vascular stent according to claim 1, wherein immersing the initial silk fibroin vascular stent in ultrapure water 1 to 50 times comprises:
immersing the initial silk fibroin vascular stent in 1-50L of ultrapure water, and replacing the ultrapure water at intervals of 1-50 h for 1-50 times.
12. The method for preparing a silk fibroin vascular stent according to claim 1, wherein the preset mold is a bar-shaped mold with a diameter of 0.1-20 mm;
and demolding after drying to obtain the target silk fibroin vascular stent, wherein the drying time is 1-1000 h.
13. A silk fibroin vascular stent prepared by the method of any one of claims 1 to 12.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311559198.2A CN117582561A (en) | 2023-11-21 | 2023-11-21 | Preparation method of silk fibroin vascular stent and silk fibroin vascular stent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311559198.2A CN117582561A (en) | 2023-11-21 | 2023-11-21 | Preparation method of silk fibroin vascular stent and silk fibroin vascular stent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117582561A true CN117582561A (en) | 2024-02-23 |
Family
ID=89914578
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311559198.2A Pending CN117582561A (en) | 2023-11-21 | 2023-11-21 | Preparation method of silk fibroin vascular stent and silk fibroin vascular stent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117582561A (en) |
-
2023
- 2023-11-21 CN CN202311559198.2A patent/CN117582561A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105169483B (en) | A kind of preparation method and its acellular matrix gel of acellular matrix gel | |
| CN109851819B (en) | Injectable silk fibroin porous hydrogel and preparation method thereof | |
| CN1622836A (en) | Scaffolds for tissue engineered hair | |
| CN110201223A (en) | A kind of synthesis macromolecule and natural extracellular matrix composite material, artificial blood vessel and preparation method thereof | |
| CN106668956B (en) | Fibroin bone nail and preparation method thereof | |
| US10730928B2 (en) | Biofabrication techniques for the implementation of intrinsic tissue geometries to an in vitro collagen hydrogel | |
| WO2008130068A1 (en) | Method for preparing a porous polymer scaffold using dry ice | |
| CN110841112A (en) | Method for producing decellularized tissue product, and graft having decellularized tissue product | |
| CN108409992B (en) | Silk fibroin block and preparation method thereof | |
| CN106667595B (en) | Fibroin dental implant and preparation method thereof | |
| CN107149702A (en) | A kind of preparation of poly-dopamine modified lithium porous support | |
| CN104548200A (en) | Method of preparing highly branched polysaccharide-fibroin hydrogel bracket | |
| CN104548196B (en) | A kind of tissue engineering bracket material being crosslinked based on vinyl sulfydryl and preparation method thereof | |
| CN113663130A (en) | Artificial blood vessel and preparation method thereof | |
| JPS6389165A (en) | Electrostatically spun product and its production | |
| CN117582561A (en) | Preparation method of silk fibroin vascular stent and silk fibroin vascular stent | |
| CN115554467B (en) | Skull repairing material and preparation method thereof | |
| CN110078880B (en) | Isocyanate cross-linked polyethylene glycol-polysebacic acid glyceride biological elastomer and preparation method and application thereof | |
| CN109867811B (en) | Porous-solid composite silk material and preparation method thereof | |
| CN115105631A (en) | Photopolymerization artificial exosome blood vessel prepared by cold casting method, and preparation method and application thereof | |
| CN117462768A (en) | Silk fibroin vascular stent and preparation method thereof | |
| CN113694253A (en) | Preparation method of small-caliber artificial blood vessel | |
| CN204840394U (en) | Preparation and application of biological activity support with chemotactic function | |
| CN106667964B (en) | Fibroin capsule and preparation method thereof | |
| CN106668939B (en) | Fibroin dental implant and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |