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 PDF

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Publication number
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
extra thin
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雷国平
王文新
朱华
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Changzhou Oman Medical Science And Technology Co Ltd
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Changzhou Oman Medical Science And Technology Co Ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14Macromolecular materials
    • A61L27/22Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/227Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14Macromolecular materials
    • A61L27/20Polysaccharides
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54Biologically active materials, e.g. therapeutic substances
    • DTEXTILES; PAPER
    • D04BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
    • D04HMAKING 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
    • D04H1/00Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
    • D04H1/40Non-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
    • D04H1/42Non-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
    • D04H1/4382Stretched reticular film fibres; Composite fibres; Mixed fibres; Ultrafine fibres; Fibres for artificial leather
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412Tissue-regenerating or healing or proliferative agents
    • A61L2300/414Growth factors
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/42Anti-thrombotic agents, anticoagulants, anti-platelet agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00Materials characterised by their function or physical properties
    • A61L2400/12Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces

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  • 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

A kind of preparation method of artificial blood vessel extra thin fabric film
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.
CN201810376666.5A 2018-04-25 2018-04-25 A kind of preparation method of artificial blood vessel extra thin fabric film Pending CN108543111A (en)

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