CN108543111A - A kind of preparation method of artificial blood vessel extra thin fabric film - Google Patents
A kind of preparation method of artificial blood vessel extra thin fabric film Download PDFInfo
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- CN108543111A CN108543111A CN201810376666.5A CN201810376666A CN108543111A CN 108543111 A CN108543111 A CN 108543111A CN 201810376666 A CN201810376666 A CN 201810376666A CN 108543111 A CN108543111 A CN 108543111A
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- blood vessel
- silk
- artificial blood
- thin fabric
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- 239000004744 fabric Substances 0.000 title claims abstract description 28
- 210000004204 blood vessel Anatomy 0.000 title claims abstract description 27
- 239000002473 artificial blood Substances 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 108010022355 Fibroins Proteins 0.000 claims abstract description 39
- 229920001872 Spider silk Polymers 0.000 claims abstract description 32
- 238000009987 spinning Methods 0.000 claims abstract description 28
- 229920001661 Chitosan Polymers 0.000 claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims description 33
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 20
- 239000012528 membrane Substances 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 12
- 229910019142 PO4 Inorganic materials 0.000 claims description 12
- 239000000706 filtrate Substances 0.000 claims description 12
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 12
- 239000010452 phosphate Substances 0.000 claims description 12
- 229910052751 metal Inorganic materials 0.000 claims description 11
- 239000002184 metal Substances 0.000 claims description 11
- 230000008569 process Effects 0.000 claims description 10
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 10
- 235000019441 ethanol Nutrition 0.000 claims description 9
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 8
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 6
- 239000004471 Glycine Substances 0.000 claims description 6
- 238000009835 boiling Methods 0.000 claims description 6
- 238000010041 electrostatic spinning Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 239000011521 glass Substances 0.000 claims description 6
- 238000007711 solidification Methods 0.000 claims description 6
- 230000008023 solidification Effects 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 claims description 4
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000008363 phosphate buffer Substances 0.000 claims 2
- 239000000835 fiber Substances 0.000 abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 9
- 239000001257 hydrogen Substances 0.000 abstract description 7
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 7
- 239000012620 biological material Substances 0.000 abstract description 3
- 239000007943 implant Substances 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 230000009471 action Effects 0.000 abstract description 2
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- 230000001737 promoting effect Effects 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 230000003993 interaction Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 31
- 238000003756 stirring Methods 0.000 description 12
- 238000007605 air drying Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 210000001772 blood platelet Anatomy 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- -1 disodium hydrogen Chemical class 0.000 description 5
- 238000004108 freeze drying Methods 0.000 description 5
- 239000003102 growth factor Substances 0.000 description 5
- 239000012475 sodium chloride buffer Substances 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 4
- 125000005909 ethyl alcohol group Chemical group 0.000 description 4
- 230000035699 permeability Effects 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
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- 206010033892 Paraplegia Diseases 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
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- 230000011132 hemopoiesis Effects 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 206010002329 Aneurysm Diseases 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 208000009087 False Aneurysm Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 206010048975 Vascular pseudoaneurysm Diseases 0.000 description 1
- PTDQUWYFMZSJJM-UHFFFAOYSA-K [Na+].[Na+].[Na+].[Cl-].OP([O-])([O-])=O Chemical compound [Na+].[Na+].[Na+].[Cl-].OP([O-])([O-])=O PTDQUWYFMZSJJM-UHFFFAOYSA-K 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- 206010002895 aortic dissection Diseases 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
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- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 230000007786 learning performance Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002070 nanowire Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000009864 tensile test Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000006453 vascular barrier function Effects 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4382—Stretched reticular film fibres; Composite fibres; Mixed fibres; Ultrafine fibres; Fibres for artificial leather
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/42—Anti-thrombotic agents, anticoagulants, anti-platelet agents
-
- 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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
Landscapes
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Textile Engineering (AREA)
- Vascular Medicine (AREA)
- Materials For Medical Uses (AREA)
- Prostheses (AREA)
Abstract
The present invention relates to a kind of artificial blood vessel preparation methods of extra thin fabric film, belong to technical field of biological materials.The present invention utilizes excellent in mechanical performance, flexility and elasticity are far superior to staple fibre, spider silk with good biomechanical property and cell compatibility is raw material, it is small using the proportion of spider silk fiber, hydrogen bond action combination fibroin between the high molecular weight and macromolecular of spider silk fibroin, enhance the tensile property of fibroin, water stability, solubility property, tridimensional network is unified into chitosan again and spinning solution is made, it is formed and is closely spaced between fiber, Interaction enhanced between fiber, effectively improve fiber and the tensile property of spinning, it can be made into extra thin fabric film；The present invention is conducive to implant survival and the reparation for promoting body defect, and the biocompatibility of material can be greatly improved.
Description
Technical field
The present invention relates to a kind of artificial blood vessel preparation methods of extra thin fabric film, belong to technical field of biological materials.
Background technology
The progression of disease such as dissecting aneurysm of aorta, false or true aneurysm are rapid, lethality is high, very dangerous.It passes
Open the chest open surgery death rate, the paraplegia rate of system are up to 10% or more.In recent years the Endovascular intervention method implantation overlay film branch occurred
The operation that frame is repaired has unrivaled minimally invasive advantage, and the death rate, paraplegia rate drop 3% hereinafter, substituted original substantially
Open surgery.So the research of endovascular exclusion will will produce larger science, social value, and can promote
Represent the raising of the weaving technology of high-caliber unique fabric.
Can artificial blood vessel be the chief component of endoluminal vascular barrier systems, be that isolate from lesion blood vessel normally
The key point except blood flow is recycled, so the application benefit of artificial blood vessel is high, each company is exhausted to its processing technology in the world
To secrecy.Implant system of the China used in endovascular graft is imported product at present, and it is expensive, supply of material week
Phase is long, limited amount, and clinical application has certain limitation, is unfavorable for the smooth treatment of many patients.Fully to meet patient's need
It asks, breaks external monopolization, we meet the people required by endovascular graft it is necessary to develop production domesticization inner cavity insulation utensil, production
Hematopoiesis pipe.
As endovascular exclusion, thickness is generally required to be no more than 0.12mm, permeability rate is no more than 300mL/
cm2/ min, while other performances will also meet certain requirement.But since Asian's aorta pectoralis diameter is relatively small, add
The limitations such as the thickness of upper conveyer, the rigidity of holder generally require the thickness of the covered fabric membrane of holder in 0.06mm or so, even
It is thinner.But the overlay film thickness of external overlay film frame product can not fully meet most of China in 0.8mm or so at present
Human body matter characteristic requirements.So China is it is necessary to carry out the development of this special artificial blood vessel, in order to which endovascular graft is in state
Interior popularization.
Invention content
The technical problems to be solved by the invention：For the extra thin fabric film developed at present in thickness and water penetration side
Face can not fully meet the problem of most people constitutional nature requirement, provide a kind of system of artificial blood vessel extra thin fabric film
Preparation Method.
In order to solve the above technical problems, the technical solution adopted by the present invention is：
A kind of preparation method of artificial blood vessel extra thin fabric film, specific preparation process are：
（1）It is immersed in after spider silk alcohol is washed in anhydrous ether 2~3 days, obtains pretreatment spider silk, be immersed in after silk alcohol is washed
2~3 days in anhydrous ether, pretreatment silk is obtained；
（2）It takes pretreatment spider silk, pretreatment silk that mass fraction is added to be in 0.5% sodium carbonate liquor, and it is heated to 95~
100 DEG C of 30~40min of boiling, filter to obtain filter residue, by filter residue washing and drying, obtain compound degumming fibroin albumen；
（3）Compound degumming fibroin albumen is taken, it is to be mixed in 10% calcium chloride solution that mass fraction, which is added, adds absolute ethyl alcohol, and
It is fitted into the bag filter that molecular weight is 1000~5000 and dialyses, filter to get filtrate, filtrate is freeze-dried 10~20h, is obtained compound
Fibroin albumen；
（4）Compound fibroin albumen, chitosan are taken, it is to be mixed in 1% acetum that mass fraction, which is added, obtains spinning solution；
（5）Spinning solution is added in glass syringes of the 10mL with metal needle, and is spun on electrostatic spinning apparatus
Film, and it is 1~2h in 1% glutaraldehyde solution that film forming matter, which is immersed in mass fraction, film forming matter is taken out in filtering, then uses mass fraction
Dry nano fibrous membrane after being washed 3~5 times for 1% glycine solution；
（6）Platelet derived growth factor is taken, it is in 7.4 disodium hydrogen phosphates-sodium chloride buffer, 36~37 that acid-base value, which is added,
It is cultivated 1~2 day at DEG C, phosphate culture solution is obtained, then nano fibrous membrane is immersed in phosphate culture solution, at 36~37 DEG C
Culture growth 7~10 days takes out nano fibrous membrane and drains rear high-temperature sterilization, obtains artificial blood vessel extra thin fabric film.
Step（2）It is described pretreatment spider silk, pretreatment silk, sodium carbonate liquor parts by weight be 10~20 parts pretreatment
Spider silk, 20 ~ 30 parts of pretreatment silks, 2000 ~ 3000 parts of sodium carbonate liquors.
Step（3）The mass ratio of the compound degumming fibroin albumen and calcium chloride solution is 1：5~1：20, the anhydrous second
Alcohol dosage is 1 ~ 4 times of compound degumming fibroin protein amount.
Step（4）The compound fibroin albumen, chitosan, acetum parts by weight be 5 ~ 10 parts of compound fibroin albumens, 1
~ 2 parts of chitosans, 100 ~ 200 parts of acetums.
Step（5）The spinning film-forming process is that high-voltage DC power supply connects on front end of the syringe needle metal needle, and control pushes away
It is 0.3~1.0mL/h into speed, spinning voltage is 15~18kV, and solidification distance is 15~18cm.
Step（6）The mass ratio of the platelet derived growth factor and disodium hydrogen phosphate-sodium chloride buffer is 1：40~
1：100.
Compared with other methods, advantageous effects are the present invention：
（1）The present invention utilizes excellent in mechanical performance, flexility and elasticity to be far superior to staple fibre, have good Biological Strength
The spider silk for learning performance and cell compatibility is raw material, the high molecular weight of spider silk fibroin small using the proportion of spider silk fiber
And the hydrogen bond action combination fibroin between macromolecular, enhance the tensile property, water stability, dissolubility of fibroin
Can, then be unified into tridimensional network with chitosan and spinning solution is made, it is formed and is closely spaced between fiber, between fiber mutually
Effect enhancing, effectively improves fiber and the tensile property of spinning, can be made into extra thin fabric film；
（2）The active site tactophily blood platelet that the effective extra thin fabric film of the present inventor's hematopoiesis passes through compound fibroin albumen surface
Derivative growth factor, effectively pre- preventing thrombosis generate, and promote vascular endothelial cell proliferation, stimulation improve surrounding tissue angiogenic growth and
Promote blood vessel to grow into biomaterial from surrounding tissue, is conducive to implant survival and the reparation for promoting body defect, energy
The biocompatibility of material is enough greatly improved.
Specific implementation mode
It is immersed in anhydrous ether 2~3 days after washing spider silk 3 ~ 5 times with absolute ethyl alcohol, takes out spider silk and natural wind
It is dry, pretreatment spider silk is obtained, is immersed in anhydrous ether 2~3 days after wash silk 3 ~ 5 times with absolute ethyl alcohol, taking-up silk is simultaneously
Natural air drying obtains pretreatment silk, and 10~20g is taken to pre-process spider silk, and 20 ~ 30g pre-processes silk, and 2~3L mass point is added
Number is and to be heated to 95~100 DEG C of 30~40min of boiling in 0.5% sodium carbonate liquor, filter residue is filtered to obtain after being cooled to room temperature, and is used
Deionized water washs filter residue 2~3 times, then filter residue is placed in drying box, and dry 2~3h, obtains compound degumming at 70~80 DEG C
Fibroin albumen, takes the compound degumming fibroin albumens of 10~20g, and 100~200g mass fractions are added and are in 10% calcium chloride solution, with
300~400r/min stirs 20~30min, adds 20~40g absolute ethyl alcohols, molecule is packed into after continuing 20~30min of stirring
For amount to dialyse 3~5 days in 1000~5000 bag filter, every 4~8h replaces deionized water, filters to get filtrate after dialysis,
Filtrate is placed in 10~20h of freeze-drying in freeze drying box, compound fibroin albumen is obtained, takes the compound fibroin albumens of 5 ~ 10g, 1 ~ 2g
Chitosan, it is to stir 20 ~ 30min in 1% acetum with 300 ~ 400r/min, obtain spinning that 100 ~ 200g mass fractions, which are added,
Spinning solution is added in glass syringes of the 10mL with metal needle liquid, and on electrostatic spinning apparatus, high voltage direct current
Source connects on front end of the syringe needle metal needle, and control fltting speed is 0.3~1.0mL/h, and spinning voltage is 15~18kV, solidification
Distance is 15~18cm, and film forming matter is simultaneously immersed in mass fraction as 1~2h in 1% glutaraldehyde solution by spinning film forming, and filtering is taken out
Film forming matter, then wash 3~5 times for 1% glycine solution with mass fraction and be placed in vacuum drying chamber, it is dry at 60 ~ 70 DEG C
Nano fibrous membrane is obtained, it is 7.4 disodium hydrogen phosphates-chlorine to take 3~5g platelet derived growth factors, addition 200~300g acid-base values
Change in sodium buffer solution, is cultivated 1~2 day at 36~37 DEG C, obtain phosphate culture solution, then nano fibrous membrane is immersed in phosphate
It in culture solution, is placed in climatic chamber, culture growth 7~10 days at 36~37 DEG C, after taking-up nano fibrous membrane drains
High-temperature sterilization obtains artificial blood vessel extra thin fabric film.
Example 1
It is immersed in anhydrous ether 2 days after washing spider silk 3 times with absolute ethyl alcohol, takes out spider silk and natural air drying, obtain pre- place
Spider silk is managed, is immersed in anhydrous ether 2 days after wash silk 3 times with absolute ethyl alcohol, silk simultaneously natural air drying is taken out, obtains pre- place
Manage silk, take 10g pre-process spider silk, 20g pre-process silk, be added 2L mass fractions be 0.5% sodium carbonate liquor in, and add
Heat filters to obtain filter residue to 95 DEG C of boiling 30min after being cooled to room temperature, filter residue is washed with deionized 2 times, then filter residue is placed in dry
In dry case, dry 2h, obtains compound degumming fibroin albumen at 70 DEG C, takes the compound degumming fibroin albumens of 10g, and 100g mass point is added
Number is to stir 20min in 10% calcium chloride solution with 300r/min, add 20g absolute ethyl alcohols, continues to be packed into after stirring 20min
It dialyses 3 days in the bag filter that molecular weight is 1000, deionized water is replaced per 4h, filters to get filtrate after dialysis, filtrate is set
It is freeze-dried 10h in freeze drying box, obtains compound fibroin albumen, takes the compound fibroin albumens of 5g, 1g chitosans that 100g matter is added
It is to stir 20min in 1% acetum with 300r/min, obtain spinning solution to measure score, and 10mL band metal needles are added in spinning solution
Glass syringe in, and on the electrostatic spinning apparatus, high-voltage DC power supply connects on front end of the syringe needle metal needle, control
Fltting speed processed is 0.3mL/h, spinning voltage 15kV, and solidification distance is 15cm, and spinning forms a film and film forming matter is immersed in matter
It is 1h in 1% glutaraldehyde solution to measure score, and film forming matter is taken out in filtering, then washs 3 postpositions with mass fraction for 1% glycine solution
In vacuum drying chamber, dry the nano fibrous membrane at 60 DEG C takes 3g platelet derived growth factors, 200g acid-base values is added
In 7.4 disodium hydrogen phosphates-sodium chloride buffer, to be cultivated 1 day at 36 DEG C, phosphate culture solution is obtained, then by nano fibrous membrane
It is immersed in phosphate culture solution, is placed in climatic chamber, nano fibrous membrane drip is taken out in culture growth 7 days at 36 DEG C
High-temperature sterilization after dry, obtains artificial blood vessel extra thin fabric film.
Example 2
It is immersed in anhydrous ether 2 days after washing spider silk 4 times with absolute ethyl alcohol, takes out spider silk and natural air drying, obtain pre- place
Spider silk is managed, is immersed in anhydrous ether 2 days after wash silk 4 times with absolute ethyl alcohol, silk simultaneously natural air drying is taken out, obtains pre- place
Manage silk, take 15g pre-process spider silk, 25g pre-process silk, be added 2L mass fractions be 0.5% sodium carbonate liquor in, and add
Heat filters to obtain filter residue to 98 DEG C of boiling 35min after being cooled to room temperature, filter residue is washed with deionized 2 times, then filter residue is placed in dry
In dry case, dry 2h, obtains compound degumming fibroin albumen at 75 DEG C, takes the compound degumming fibroin albumens of 15g, and 150g mass point is added
Number is to stir 25min in 10% calcium chloride solution with 350r/min, add 30g absolute ethyl alcohols, continues to be packed into after stirring 25min
It dialyses 4 days in the bag filter that molecular weight is 3000, deionized water is replaced per 6h, filters to get filtrate after dialysis, filtrate is set
It is freeze-dried 15h in freeze drying box, obtains compound fibroin albumen, takes the compound fibroin albumens of 8g, 1g chitosans that 150g matter is added
It is to stir 25min in 1% acetum with 350r/min, obtain spinning solution to measure score, and 10mL band metal needles are added in spinning solution
Glass syringe in, and on the electrostatic spinning apparatus, high-voltage DC power supply connects on front end of the syringe needle metal needle, control
Fltting speed processed is 0.7mL/h, spinning voltage 16kV, and solidification distance is 16cm, and spinning forms a film and film forming matter is immersed in matter
It is 1h in 1% glutaraldehyde solution to measure score, and film forming matter is taken out in filtering, then washs 4 postpositions with mass fraction for 1% glycine solution
In vacuum drying chamber, dry the nano fibrous membrane at 65 DEG C takes 4g platelet derived growth factors, 250g acid-base values is added
In 7.4 disodium hydrogen phosphates-sodium chloride buffer, to be cultivated 1 day at 36 DEG C, phosphate culture solution is obtained, then by nano fibrous membrane
It is immersed in phosphate culture solution, is placed in climatic chamber, nano fibrous membrane drip is taken out in culture growth 8 days at 36 DEG C
High-temperature sterilization after dry, obtains artificial blood vessel extra thin fabric film.
Example 3
It is immersed in anhydrous ether 3 days after washing spider silk 5 times with absolute ethyl alcohol, takes out spider silk and natural air drying, obtain pre- place
Spider silk is managed, is immersed in anhydrous ether 3 days after wash silk 5 times with absolute ethyl alcohol, silk simultaneously natural air drying is taken out, obtains pre- place
Manage silk, take 20g pre-process spider silk, 30g pre-process silk, be added 3L mass fractions be 0.5% sodium carbonate liquor in, and add
Heat filters to obtain filter residue to 100 DEG C of boiling 40min after being cooled to room temperature, filter residue is washed with deionized 3 times, then filter residue is placed in
In drying box, dry 3h, obtains compound degumming fibroin albumen at 80 DEG C, takes the compound degumming fibroin albumens of 20g, and 200g mass is added
Score is to stir 30min in 10% calcium chloride solution with 400r/min, add 40g absolute ethyl alcohols, continues to fill after stirring 30min
Enter and dialyse 5 days in the bag filter that molecular weight is 5000, deionized water is replaced per 8h, is filtered to get filtrate after dialysis, by filtrate
It is placed in freeze drying box and is freeze-dried 20h, obtain compound fibroin albumen, the compound fibroin albumens of 10g, 2g chitosans is taken to be added
200g mass fractions are to stir 30min in 1% acetum with 400r/min, obtain spinning solution, and 10mL band gold is added in spinning solution
In the glass syringe for belonging to syringe needle, and on electrostatic spinning apparatus, high-voltage DC power supply connects front end of the syringe needle metal needle
On head, control fltting speed is 1.0mL/h, spinning voltage 18kV, and solidification distance is 18cm, and spinning forms a film and soaks film forming matter
Bubble 2h in mass fraction is 1% glutaraldehyde solution, film forming matter is taken out in filtering, then washs 5 with mass fraction for 1% glycine solution
Secondary to be placed in vacuum drying chamber, dry the nano fibrous membrane at 70 DEG C takes 5g platelet derived growth factors, 300g is added
Acid-base value is to be cultivated 2 days at 37 DEG C in 7.4 disodium hydrogen phosphates-sodium chloride buffer, obtain phosphate culture solution, then by nanometer
Tunica fibrosa is immersed in phosphate culture solution, is placed in climatic chamber, and culture growth 10 days, take out Nanowire at 37 DEG C
Dimension film drains rear high-temperature sterilization, obtains artificial blood vessel extra thin fabric film.
Reference examples：The artificial blood vessel extra thin fabric film of Dongguan company production.
Example and the artificial blood vessel of reference examples are detected with extra thin fabric film, specific detection is as follows：
Sample rate is tested：The fabric photo within the scope of 2mm is first shot by digital 3 D video microscope, is then passed through
Captured photo counts sample in the line radical of 2mm, is then scaled standard density again, same sample measures 5 averagings
Value.
Thickness of sample is tested：Using YG141D type automatic thickness testers, in grade III Standard atmospheric conditions（Temperature 20 ± 2
DEG C, relative humidity be 65 ± 5%）Relative humidity and test.Instrument Foot area 2000mm2, pressing force 400cN.In view of sample
The uniformity of product and the influence of test error, each sample are tested 5 times in different parts, are then averaged.
Sample permeability rate is tested：Requirement of the test equipment using Jia Lixia with reference to ISO7198 standards to this test index
And specially design the water penetration instrument for the artificial blood vessel built.Test result reference standard ISO7198：1998（E）, with permeability rate come
It indicates.
Tensile property test method：Tension failure experiment is surveyed respectively using the universal tensile testing machine of model WDW-20
Try tensile property.
Specific testing result such as table 1.
1 performance characterization contrast table of table
As shown in Table 1, the artificial blood vessel that prepared by the present invention has good density, thickness, permeability rate and drawing with extra thin fabric film
Performance is stretched, comprehensive performance is preferable.
Claims (6)
1. a kind of artificial blood vessel preparation method of extra thin fabric film, which is characterized in that specifically preparation process is：
（1）It is immersed in after spider silk alcohol is washed in anhydrous ether 2~3 days, obtains pretreatment spider silk, be immersed in after silk alcohol is washed
2~3 days in anhydrous ether, pretreatment silk is obtained；
（2）It takes pretreatment spider silk, pretreatment silk that mass fraction is added to be in 0.5% sodium carbonate liquor, and it is heated to 95~
100 DEG C of 30~40min of boiling, filter to obtain filter residue, by filter residue washing and drying, obtain compound degumming fibroin albumen；
（3）Compound degumming fibroin albumen is taken, it is to be mixed in 10% calcium chloride solution that mass fraction, which is added, adds absolute ethyl alcohol, and
It is fitted into the bag filter that molecular weight is 1000~5000 and dialyses, filter to get filtrate, filtrate is freeze-dried 10~20h, is obtained compound
Fibroin albumen；
（4）Compound fibroin albumen, chitosan are taken, it is to be mixed in 1% acetum that mass fraction, which is added, obtains spinning solution；
（5）Spinning solution is added in glass syringes of the 10mL with metal needle, and is spun on electrostatic spinning apparatus
Film, and it is 1~2h in 1% glutaraldehyde solution that film forming matter, which is immersed in mass fraction, film forming matter is taken out in filtering, then uses mass fraction
Dry nano fibrous membrane after being washed 3~5 times for 1% glycine solution；
（6）Take platelet derived growth factor, acid-base value be added be in 7.4 phosphate buffers cultivate 1 at 36~37 DEG C~
2 days, obtain phosphate culture solution, then nano fibrous membrane is immersed in phosphate culture solution, at 36~37 DEG C culture growth 7~
It 10 days, takes out nano fibrous membrane and drains rear high-temperature sterilization, obtain artificial blood vessel extra thin fabric film.
2. a kind of preparation method of artificial blood vessel extra thin fabric film as described in claim 1, which is characterized in that step（2）
It is described pretreatment spider silk, pretreatment silk, sodium carbonate liquor parts by weight be 10~20 parts pretreatment spider silks, 20 ~ 30 parts
Pre-process silk, 2000 ~ 3000 parts of sodium carbonate liquors.
3. a kind of preparation method of artificial blood vessel extra thin fabric film as described in claim 1, which is characterized in that step（3）
The mass ratio of the compound degumming fibroin albumen and calcium chloride solution is 1：5~1：20, the absolute ethyl alcohol dosage is compound degumming
1 ~ 4 times of fibroin albumen quality.
4. a kind of preparation method of artificial blood vessel extra thin fabric film as described in claim 1, which is characterized in that step（4）
The compound fibroin albumen, chitosan, acetum parts by weight be 5 ~ 10 parts of compound fibroin albumens, 1 ~ 2 part of chitosan, 100 ~
200 parts of acetums.
5. a kind of preparation method of artificial blood vessel extra thin fabric film as described in claim 1, which is characterized in that step（5）
The spinning film-forming process be high-voltage DC power supply connect front end of the syringe needle metal needle on, control fltting speed be 0.3~
1.0mL/h, spinning voltage are 15~18kV, and solidification distance is 15~18cm.
6. a kind of preparation method of artificial blood vessel extra thin fabric film as described in claim 1, which is characterized in that step（6）
The mass ratio of the platelet derived growth factor and phosphate buffer is 1：40~1：100.
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