CN106310380B - A kind of nanofiber Silk fibroin gel and preparation method thereof - Google Patents
A kind of nanofiber Silk fibroin gel and preparation method thereof Download PDFInfo
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- CN106310380B CN106310380B CN201610692291.4A CN201610692291A CN106310380B CN 106310380 B CN106310380 B CN 106310380B CN 201610692291 A CN201610692291 A CN 201610692291A CN 106310380 B CN106310380 B CN 106310380B
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- 108010022355 Fibroins Proteins 0.000 title claims abstract description 70
- 239000002121 nanofiber Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 238000001879 gelation Methods 0.000 title description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Substances OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000012460 protein solution Substances 0.000 claims abstract description 15
- 238000002347 injection Methods 0.000 claims abstract description 3
- 239000007924 injection Substances 0.000 claims abstract description 3
- 239000000017 hydrogel Substances 0.000 claims description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 10
- 235000019253 formic acid Nutrition 0.000 claims description 10
- 239000006166 lysate Substances 0.000 claims description 10
- 240000000249 Morus alba Species 0.000 claims description 8
- 235000008708 Morus alba Nutrition 0.000 claims description 8
- AMXOYNBUYSYVKV-UHFFFAOYSA-M lithium bromide Chemical group [Li+].[Br-] AMXOYNBUYSYVKV-UHFFFAOYSA-M 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- 239000001110 calcium chloride Substances 0.000 claims description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 5
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 4
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 claims description 4
- 239000000835 fiber Substances 0.000 claims description 4
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 3
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 claims description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 2
- 235000003846 Ricinus Nutrition 0.000 claims description 2
- 241000322381 Ricinus <louse> Species 0.000 claims description 2
- 239000003292 glue Substances 0.000 claims description 2
- 229910052744 lithium Inorganic materials 0.000 claims description 2
- 229910052749 magnesium Inorganic materials 0.000 claims description 2
- 239000011777 magnesium Substances 0.000 claims description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 2
- VGTPCRGMBIAPIM-UHFFFAOYSA-M sodium thiocyanate Chemical compound [Na+].[S-]C#N VGTPCRGMBIAPIM-UHFFFAOYSA-M 0.000 claims description 2
- 229910001631 strontium chloride Inorganic materials 0.000 claims description 2
- AHBGXTDRMVNFER-UHFFFAOYSA-L strontium dichloride Chemical compound [Cl-].[Cl-].[Sr+2] AHBGXTDRMVNFER-UHFFFAOYSA-L 0.000 claims description 2
- 239000011592 zinc chloride Substances 0.000 claims description 2
- 235000005074 zinc chloride Nutrition 0.000 claims description 2
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N thiocyanic acid Chemical compound SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 claims 2
- 239000000499 gel Substances 0.000 abstract description 24
- 239000000243 solution Substances 0.000 abstract description 20
- 238000000034 method Methods 0.000 abstract description 9
- 239000003960 organic solvent Substances 0.000 abstract description 8
- 239000007864 aqueous solution Substances 0.000 abstract description 6
- 239000011664 nicotinic acid Substances 0.000 abstract description 4
- 230000006835 compression Effects 0.000 description 11
- 238000007906 compression Methods 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 9
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 235000013601 eggs Nutrition 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- 239000003814 drug Substances 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 235000017557 sodium bicarbonate Nutrition 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 230000001172 regenerating effect Effects 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010016654 Fibrosis Diseases 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000004761 fibrosis Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002070 nanowire Substances 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229960000502 poloxamer Drugs 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- -1 stirring Chemical compound 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- 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
- A61L27/52—Hydrogels or hydrocolloids
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- 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
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
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Abstract
The present invention relates to a kind of nanofiber Silk fibroin gels and preparation method thereof, and salt-formic acid is dissolved in after natural silk degumming and obtains silk fibroin protein solution；It will be in fibroin albumen-salt-formic acid solution injection mold；Mold equipped with silk fibroin protein solution is immersed into organic solvent or aqueous solution, standing a period of time is to form Silk fibroin gel.Preparation method of the present invention is simple, process is short, facilitates operation, is easy to produce in batches.Silk fibroin gel prepared by the present invention has natural bionic nano fibrillar structure, adjustable mechanical property and good biocompatibility.
Description
Technical field
The present invention relates to a kind of nanofiber Silk fibroin gels and preparation method thereof, and prepared material can be applied to
The regenerative medicine fields such as organizational project, medicament slow release.
Background technique
The organ or tissue as caused by disease and accident damage and afunction patient have every year it is as many as millions of,
Only the U.S. needs more than 800 ten thousand operations to give treatment to this kind of patient every year, and economic costs are at 400,000,000,000 dollars or more.With
The development of modern medicine and surgery operating technology, it is widely accepted come repair function loss by tissue or organ transplant,
However but it is faced with huge donor notch.Tissue is formed in vivo or in vitro by regenerative medicine means body or organ is impaired
The reparation of function provides new therapeutic scheme.Wherein, the selection of tissue engineering bracket material and it is configured to the treatment method
One of key.The gel (hydrogel) formed using water as medium nature and it is artificial synthesized in the case where be all widely present,
It is small in organism to arrive organ greatly to cell, tissue, it can regard complicated aquogel system as.In biomedical materials field, tool
There is the polym eric gel of good biocompatibility and degradability that there is high application value and prospect, can be widely applied to
Tissue recovery support material, medicine controlled release carrier, the loading of cell and bioactive molecule and transmitting etc..
Hydrogel for loading and transmitting cell must have biocompatibility, suitable substance conveying capacity, enough
Mechanical stability and controllable biological degradability.Hydrogel is prepared using synthesis macromolecule, its advantage is that choosing can be passed through
Gel process and hydrogel properties, but hydrogel are accurately controlled with specific molecular weight, molecular structure and crosslinking method etc.
Biocompatibility it is poor；It is preferable with tissue, the biocompatibility of cell with hydrogel prepared by natural macromolecular material,
But controllability and less reproducible.
Fibroin is derived from the natural polymer biomaterial of nature, with excellent mechanical property, controllably
Biological degradability, workability, especially its biocompatibility same with collagen and become ideal regenerative medicine bracket
Raw material.Fibroin aqueous solution is being self-assembly of gel in its natural state, but the time is longer, needs a couple of days
It can complete.It, can be by adjusting solution parameter (such as solution concentration, pH value, temperature), application quickly to prepare silk fibroin hydrogel
Extrinsic factor (such as stirring, ultrasonication) and addition other materials (such as poloxamer) are realized.Although fibroin hydrogel can
To produce easily, but still face some urgent problems to be solved.Existing critical issue first is that the dissolution process of existing fibroin is made
It greatly degrades at fibroin, seriously affects the mechanical property of fibroin hydrogel, keep its application limited；Problem second is that fibroin water-setting
The reparation technology of glue is complex, including following three key steps: dialysis obtains for a long time for the dissolution of 1. high concentration neutral salt, 2.
Pure silk fibroin water solution, 3. by interior extrinsic factor induction silk fibroin aqueous solution agglomerate to form gel；Problem third is that at present
Fibroin hydrogel inside be still similar sponge porous structure, do not have bionic nano fibre structure, therefore its biology
Characteristic still needs to further be promoted, and biologic applications needs are not achieved.
Therefore overcome the above problem in the prior art, develop a kind of simple, easy-operating preparation method, and construct knot
The silk fibroin nano-fiber hydrogel that structure is bionical, mechanical property is controllable to fibroin albumen biomedical materials field application
It has very important significance.
Summary of the invention
The object of the present invention is to provide a kind of preparation sides of simple, easy-operating nanofiber silk fibroin hydrogel
Method, and the silk fibroin hydrogel with bionic nano fibrosis formation and excellent in mechanical performance by this method preparation.
In order to achieve the above objectives, the technical solution adopted by the present invention is that: a kind of nanofiber silk fibroin hydrogel
Preparation method, comprising the following steps:
(1) it is dissolved in after natural silk degumming in salt-formic acid lysate and obtains silk fibroin protein solution；
It (2) will be in the silk fibroin protein solution injection mold of step (1)；
(3) step (2) are immersed in the aqueous solution or aqueous solution of organic solvent equipped with the mold of silk fibroin protein solution, is stood
Form nanofiber silk fibroin hydrogel.
In above-mentioned technical proposal, step (1) silk is mulberry silk, tussah silk, ricinus silk or wild silk yarn.
In above-mentioned technical proposal, salt in step (1) salt-formic acid lysate be lithium bromide, calcium chloride, zinc chloride,
Magnesium chloride, strontium chloride, lithium rhodanate, sodium sulfocyanate, magnesium rhodanate or calcium nitrate.
In above-mentioned technical proposal, in step (1) salt-formic acid lysate, salinity is 1~20 w/v%；Formic acid concn
For 98wt%；The concentration of fibroin albumen is 5~50 w/v% in silk fibroin protein solution.
In above-mentioned technical proposal, the dissolution of silk can not only be guaranteed by limiting formic acid, protect the fibrillar structure of silk, simultaneously
Excessive degradation silk is avoided, so that regenerated fibroin material has good structure and performance；Avoid other acid as phosphoric acid,
Hydrochloric acid can seriously degrade silk, thus the problem of losing use value；Also avoid other acid as acetic acid cannot dissolve silk, no
It is able to achieve the defect of the regeneration preparation of fibroin.So that formic acid of the invention is conducive to, silk fibroin protein solution is uniform, it is molten to guarantee
Xie Du provides good basis to form nanofiber silk fibroin hydrogel, especially excellent mechanical property and bionical
Nanofibrillar structures.
In above-mentioned technical proposal, organic solvent described in step (2) is methanol, ethyl alcohol, isopropanol；Organic solvent it is water-soluble
Liquid concentration is 1~99wt%, i.e., the mass content of organic solvent is 1~99wt%.
In above-mentioned technical proposal, in step (2), the shape of product that mold is prepared as needed is designed.
The invention also discloses manufactured nanofiber silk fibroin hydrogels according to the above technical scheme；Gel master
It to be made of the fiber of 10~100nm, hygrometric state compression modulus is 10kPa~100MPa；It can be applicable to and prepare tissue engineering bracket
In.
The molecular weight and multistage self-assembled structures, especially nanofibrillar structures of the excellent mechanical property of silk and its superelevation
It is directly related.In the present invention, salt-formic acid lysate can dissolve silk in nanometer fibril level, and it is molten to obtain fibroin nanometer fibril
Liquid, because silk still saves its fibrillar structure, therefore the solution does not dissolve each other with water and organic solvent；It is received when by the fibroin
When rice fibril solution is immersed in water and organic solvent, the concentration decline of formic acid, pH value of solution are reduced, to induce silk fibroin molecular
Occur to change from amorphous to beta sheet crystalline texture, while being cross-linked between nanometer fibril, final cohesion forms gel；
It is main to assign Silk fibroin gel excellent mechanical property and bionic nano fibrillar structure, gel for original fibrillar structure in solution
It is made of 10 ~ 100nm fiber, hygrometric state compression modulus is 10kPa ~ 100MPa, achieves unexpected technical effect.
Due to the above technical solutions, the present invention has the following advantages over the prior art:
(1) preparation method of the present invention is simple, controllability is strong, after silk dissolves in salt-formic acid, is directly immersed in organic molten
Gel can be formed in agent or water, is mainly made of 10 ~ 100nm fiber, and hygrometric state compression modulus is 10kPa ~ 100MPa, is achieved
Unexpected technical effect.
(2) silk fibroin solution that preparation method of the present invention may be selected that a step is promoted to dissolve using pure water as inducer directly occurs
Gel-forming bracket can avoid organic solvent post-processing and thus caused bracket toxicity problem；And water process is for fibroin egg
White gel is mainly made of nanometer fibril, and fibril diameter is between 10 ~ 100 nanometers；Prepare the compression modulus of Silk fibroin gel
Between 10kPa ~ 100MPa, soft tissue healing such as skin, cornea cannot be only used for, and can be used for sclerous tissues' damage
It repairs, such as bone；Good basis is provided as regeneration medical material application for silk.
Detailed description of the invention
Fig. 1 is the camera and scanning electron microscope (SEM) photograph of fibroin egg nanofiber hydrogels made from embodiment one；
Fig. 2 is the x-ray diffraction pattern of fibroin egg nanofiber hydrogels made from embodiment two；
Fig. 3 is the scanning electron microscope (SEM) photograph after the freeze-drying of fibroin egg nanofiber hydrogels made from embodiment two；
Fig. 4 is the camera and scanning electron microscope (SEM) photograph of fibroin egg nanofiber hydrogels made from embodiment three.
Specific embodiment
The present invention will be further described below with reference to examples:
Embodiment one
(1) natural mulberry silk boils 30min degumming with the sodium bicarbonate solution of mass fraction 0.5wt%, obtains after being repeated 3 times
Obtain degummed mulberry silk；
(2) boiled silk is dissolved in 2w/v calcium chloride -98wt% formic acid solution and obtains 10 w/v % silk protein solutions；
(3) above-mentioned fibroin albumen lysate is directly injected into plastic tube, is then immersed in deionized water, 8 hours i.e. shape
At nanofiber silk fibroin hydrogel.
The camera photos and the scanning electron microscope (SEM) photograph after freeze-drying that attached drawing 1 is fibroin egg nanofiber hydrogels obtained above.
Gel is milky as seen from the figure, and surface is smooth, is mainly made of fibroin albumen nanometer fibril inside gel.It compresses and surveys through mechanics
Examination, the compression modulus of the gel are 523.2KPa.
Embodiment two:
(1) natural mulberry silk boils 30min degumming with the sodium bicarbonate solution of mass fraction 0.05wt%, obtains after being repeated 3 times
Obtain degummed mulberry silk；
(2) boiled silk is dissolved in the silk protein solution that 4w/v% calcium chloride -98wt% formic acid solution obtains 25 w/v%；
(3) above-mentioned fibroin albumen lysate is injected in 24 orifice plates, is then immersed in deionized water, formed within 6 hours and received
Rice fibrosis silk fibroin hydrogel.
Attached drawing 2, attached drawing 3 are respectively X-ray diffraction spectrogram and the freeze-drying of fibroin egg nanofiber hydrogels obtained above
Scanning electron microscopic picture afterwards, silk-fibroin secondary structure is mainly beta sheet crystalline texture as seen from the figure, and internal stent is by Nanowire
Dimension composition.Through mechanics compression verification, the compression modulus of the gel is 16.2MPa.
Embodiment three:
(1) natural tussah silk boils 30min degumming with the sodium bicarbonate solution of mass fraction 0.5wt%, obtains after being repeated 3 times
Obtain degumming tussah silk；
(2) degumming tussah silk is dissolved in and obtains the silk that concentration is 50w/v% in 10w/v% lithium bromide -98wt% formic acid solution
Protein solution；
(3) above-mentioned silk fibroin protein solution is injected in 6 orifice plates, is then immersed in aqueous solution, form tussah silk within 20 hours
Nanofiber silk fibroin hydrogel.
Attached drawing 4 is the scanning electron microscope (SEM) photograph after fibroin egg nanofiber hydrogels obtained above freeze-drying.The water as seen from the figure
Nanofibrillar structures are similarly inside gel.Through mechanics compression verification, the compression modulus of the gel is 98.2MPa.
Example IV:
(1) natural mulberry silk boils 30min degumming with the sodium bicarbonate solution of mass fraction 0.05wt%, obtains after being repeated 3 times
Obtain degummed mulberry silk；
(2) boiled silk is dissolved in and obtains the silk egg that concentration is 6 w/v% in 10w/v% calcium chloride -98wt% formic acid solution
White solution；
(3) above-mentioned fibroin albumen lysate is injected in 6 orifice plates, is then immersed in 50 wt% deionized waters, 12 hours i.e.
Form nanofiber silk fibroin hydrogel.
Through mechanics compression verification, the compression modulus of the gel is 213.2KPa.
Claims (3)
1. a kind of preparation method of nanofiber silk fibroin hydrogel, which comprises the following steps:
(1) it is dissolved in after natural silk degumming in salt-formic acid lysate and obtains silk fibroin protein solution；In the salt-formic acid lysate, salt
Concentration is 2~10 w/v%；Formic acid concn is 98wt%；The concentration of fibroin albumen is 5~50 w/ in the silk fibroin protein solution
v%；
It (2) will be in the silk fibroin protein solution injection mold of step (1)；
(3) step (2) are immersed in the water equipped with the mold of silk fibroin protein solution, standing forms nanofiber fibroin albumen water-setting
Glue；The nanofiber silk fibroin hydrogel includes the fiber of 10 ~ 100nm of diameter.
2. the preparation method of nanofiber silk fibroin hydrogel according to claim 1, it is characterised in that: the silk
For one or more of mulberry silk, tussah silk, ricinus silk, wild silk yarn.
3. the preparation method of nanofiber silk fibroin hydrogel according to claim 1, it is characterised in that: the salt-
Salt in formic acid lysate is lithium bromide, calcium chloride, zinc chloride, magnesium chloride, strontium chloride, lithium rhodanate, sodium sulfocyanate, thiocyanic acid
One or more of magnesium, calcium nitrate.
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| CN107687088A (en) * | 2017-09-04 | 2018-02-13 | 西南大学 | Conductive silk fabric is prepared with zinc chloride/formic acid solution |
| EP3706712A1 (en) * | 2017-11-10 | 2020-09-16 | Givaudan SA | Silk alcohol formulations |
| CN109096501B (en) * | 2018-07-16 | 2022-11-01 | 武汉纺织大学 | Silk fibroin three-dimensional porous scaffold and preparation method thereof |
| CN111358706A (en) * | 2020-04-13 | 2020-07-03 | 杭州万事利丝绸文化股份有限公司 | Preparation method of alcohol-fibroin gel washing-free disinfectant |
| CN111603607B (en) * | 2020-05-21 | 2021-01-05 | 武汉理工大学 | Fibroin-calcium hydrogen phosphate compound obtained by induction with fibroin as template and preparation method thereof |
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| CN112940293A (en) * | 2021-02-25 | 2021-06-11 | 中国科学院上海微系统与信息技术研究所 | Preparation method of hydrogel based on biological protein |
| CN113444281B (en) * | 2021-06-28 | 2022-11-11 | 武汉纺织大学 | Fabric warm-keeping filling material with down-like function and preparation method and application thereof |
| CN113754898A (en) * | 2021-09-13 | 2021-12-07 | 复旦大学 | Full-fibroin-protein-based conductive gel sensor and preparation method thereof |
| CN114163684B (en) * | 2021-12-31 | 2023-09-05 | 浙江理工大学 | Method for directly extracting silk fibroin nanofibers from waste cocoons and recovering hydrolyzed silk proteins and extracting solution |
| CN114395141B (en) * | 2022-01-18 | 2023-09-12 | 苏州大学 | Preparation method of high-strength silk fibroin nanofiber hydrogel |
| CN115137883B (en) * | 2022-08-03 | 2023-12-29 | 尧舜泽生物医药(南京)有限公司 | Bionic composite mineralization bracket and preparation method thereof |
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