CN106866996A - A kind of fast preparation method of silk fibroin matter gel - Google Patents
A kind of fast preparation method of silk fibroin matter gel Download PDFInfo
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- 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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- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0031—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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0042—Materials resorbable by the body
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/108—Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/043—Proteins; Polypeptides; Degradation products thereof
- A61L31/047—Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/145—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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
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- C—CHEMISTRY; METALLURGY
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- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
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- C25—ELECTROLYTIC OR ELECTROPHORETIC PROCESSES; APPARATUS THEREFOR
- C25B—ELECTROLYTIC OR ELECTROPHORETIC PROCESSES FOR THE PRODUCTION OF COMPOUNDS OR NON-METALS; APPARATUS THEREFOR
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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
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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Abstract
The present invention relates to a kind of fast preparation method of silk fibroin matter gel, the natural silk fiber that degumming is obtained carries out dissolution process, and regenerated silk fibroin matter solution is obtained by dialysis；Certain density silk fibroin solution is added into pH adjusting agent regulation pH value again, finally quick formation has good mechanical properties fibroin protein gel under the conditions of weak electric field.The method prepares silk extract gel without any crosslinking agent, strong adaptability, improves that traditional silk extract gel is frangible, the low deficiency such as strong, and the gel forming time is short.Silk extract gel prepared by the present invention can be applied to the fields such as organizational project, health care such as block, close, stop blooding in terms of application there are great potentiality.
Description
Technical field
It is a kind of higher force specifically the invention belongs to a kind of fast preparation method of silk fibroin matter gel
The preparation method of performance and Environmental Safety gel rubber material is learned, it can be used as drug release carrier, tissue engineering bracket, tissue repair
Etc. being applied to bio-medical field.
Background technology
Gel is a kind of spacial framework between solid and liquid, with it is soft, can keep certain shape,
The peculiar property of large quantity of moisture can be kept.Due to the effect such as the physical crosslinking of polymer molecule interchain and chemical crosslinking, gel is not
Water can be dissolved in, but can the swelling and certain shape of holding.Its property can be reduced similar to extracellular matrix components after water suction
Friction and mechanism to surrounding tissue, can improve the biology performance of material.Due to its unique water conservation and bionical spy
Property, gel rubber material can be applied to biological medicine and lead as medicinal slow release agent, tissue engineering bracket material, tissue renovation material etc.
Domain.
Tissue engineering material design should to greatest extent the 26S Proteasome Structure and Function of bionical human body cell epimatrix the characteristics of, ability
The problems such as biocompatibility and biological degradability of effective solution material.The main constituents of extracellular matrix are protein
And polysaccharide, they play respective physiological action respectively, thus jointly the regulation differentiation of cell, propagation, migration, shape, into
Living and apoptosis, the common normal configuration and function for maintaining to organize.
Fibroin albumen is a kind of special protein, with excellent physical and mechanical properties, degradation property, as biological material
, sensitization non-toxic to body and stimulation during material, catabolite will not cause inflammatory reaction, have excellent with tissue
A kind of biocompatibility, it is considered to be excellent natural polymer material that can be extensively exploited in biomedical materials field
Material.Regenerated silk solution is present in a kind of metastable condition form, that is to say, that even in the absence of any outside work
Under conditions of, silk fibroin solution can also occur from solution to gel transition phenomenon over time.Silk fibroin hydrogel
Material is not only with the shape of stabilization, while also having solute in aqueous solvent, the gel kept in gel diffusible and transmission, nothing
The poison characteristic such as non-stimulated, is a kind of biomaterial of function admirable.But under conditions of not by any external influence, silk
Plain solution greatly limit fibroin albumen and coagulate to the time that the transformation of gel needs is very long and the gel mechanics poor performance that obtains
Glue material makees the application in biomedicine field.When silk fibroin solution pH value close to fibroin albumen isoelectric point (pH=3.9)
When, fibroin albumen is susceptible to gelling, it also requires the time of 8 days or so could complete the whole process of gelation,
Silk fibroin gel pliability is equally existed poor, the problems such as brittle.Additionally, as tissue engineering bracket, although fibroin hydrogel
Preferably, but the medical science that its structure and performance adjustable extent be narrow and mechanical property is difficult to meet under different condition will for cellular affinity
Ask.
Having had various methods at present can effectively process the poor problem of silk extract gel physical mechanical strength, and mainly
Improve this class problem by being chemically crosslinked.Although these methods can effectively solve the problems, such as silk extract gel, and strength is poor,
But the introducing of organic solvent inevitably causes that the performances such as its biocompatibility, biodegradable are affected, especially
It is that Silk fibroin gel brings great resistance in the application of biomedicine field.Therefore, exigence one kind can be quick
The method for preparing the silk extract gel of the preferable mechanical property of tool；At the same time, it is desirable to which prepared silk fibroin material still has good
Good biocompatibility, good water-retaining property, biodegradable and degradation speed be slow etc., and performance can be widely applied
In biomedicine field.
The content of the invention
For above-mentioned problem, it is an object of the invention to using a kind of new quick side for preparing fibroin coagulation glue
Method, prepares and provides a kind of tissue repair engineering material for having good biocompatibility, mechanical property and good water-retaining property concurrently.
To achieve these goals, its technical solution comprises the following steps：
(1) raw silk is made the silk fibroin water solution that concentration is 1-10wt% by degumming, dissolving, dialysis, concentration；
(2) pH regulation reagents are added in the silk fibroin solution that step (1) is prepared, stirring prepares the silk that pH value is 1~5
Plain solution；
(3) silk fibroin solution processed by step (2) is placed under the conditions of weak electric field and is electrolysed；
(4) under these conditions, 50-10 minutes prepared silk fibroin matter gel.
Preferably, described fibroin protein is one or more in silkworm and wild silkworm protein.
Preferably, described fibroin protein solution concentration is 1wt%~10wt%.
Preferably, described pH regulation reagents are one or more in phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, formic acid.
Preferably, the pH value of described silk fibroin solution is 1-5.
Preferably, described electric-field intensity condition is D.C. regulated power supply.
Preferably, described D.C. regulated power supply is 12-36V.
The present invention has following obvious advantage：
(1) present invention is added compared with plural gel prepared by traditional chemical crosslinking without any organic chemical reagent, made
On the premise of possessing equal physical and mechanical properties, it has more preferable biocompatibility, biodegradable to standby gel, makes
The application that it is obtained in biomedical sector more has potentiality；
(2) between the silk fibroin molecular for compared with simple silk fibroin hydrogel, occurring in electrolytic process of the invention
Effect so that gel formation time is very of short duration, this characteristic causes it such as block in surgical operation, close, hemostasis side
The application in face has great prospect；
(3) fibroin albumen coagulation glue has inflammatory reaction small compared with other plural gels, the features such as good biocompatibility, significantly
Improve the security of material；
(4) fibroin albumen coagulation glue has similar mechanical performance and structure to some tissues and extracellular matrix in human body
Feature, these characteristics can provide standing ground for cell, and diffusible growth factor can be supplied to adhering to thin again
Cellular surface, possesses as the necessary condition of cell culture substrate.
Specific embodiment
Technical solution of the present invention is further described with reference to embodiment, following examples are explanation of the invention
And the invention is not limited in following examples.
Embodiment one
The fibroin albumen coagulation glue preparation process that the present embodiment is provided is as follows：
(1) mulberry silk raw silk is placed in degumming 3 times in the sodium carbonate boiling water that mass fraction is 0.6 ‰, it is preceding every twice
Secondary treatment 30min, third time treatment 45min, the fibroin fiber for obtaining is washed with deionized water totally, and is dried in 55 DEG C of baking ovens
It is dry；
(2) fibroin fiber that step (1) is obtained is placed in ternary solution, dissolution process 1h, obtains under the conditions of 72 DEG C
Mixed solution be fitted into bag filter (molecular weight 8000-14000) and dialysed in deionized water 4d, period every two hours changes water one
Secondary, the silk fibroin solution concentration for finally obtaining is 4wt%；
(3) the 4wt% silk fibroin solutions for preparing step (2) are added dropwise appropriate hydrochloric acid, prepare the silk fibroin solution that pH value is 2；
(4) step (3) silk fibroin solution is placed under the conditions of 24V (DC) and obtains fibroin albumen coagulation glue in 5 minutes.
Embodiment two
The fibroin albumen coagulation glue preparation process that the present embodiment is provided is as follows：
(1) mulberry silk raw silk is placed in degumming 3 times in the sodium carbonate boiling water that mass fraction is 0.6 ‰, it is preceding every twice
Secondary treatment 30min, third time treatment 45min, the fibroin fiber for obtaining is washed with deionized water totally, and is dried in 55 DEG C of baking ovens
It is dry；
(2) fibroin fiber that step (1) is obtained is placed in ternary solution, dissolution process 1h, obtains under the conditions of 72 DEG C
Mixed solution be fitted into bag filter (molecular weight 8000-14000) and dialysed in deionized water 4d, period every two hours changes water one
Secondary, the silk fibroin solution concentration for finally obtaining is 4wt%；
(3) the 4wt% silk fibroin solutions for preparing step (2) are added dropwise appropriate hydrochloric acid, prepare the silk fibroin solution that pH value is 4；
(4) electrolysis obtains fibroin albumen coagulation glue in 8 minutes under the conditions of step (3) silk fibroin solution being placed in into 24V (DC).
Embodiment three
The fibroin albumen coagulation glue preparation process that the present embodiment is provided is as follows：
(1) mulberry silk raw silk is placed in degumming 3 times in the sodium carbonate boiling water that mass fraction is 0.6 ‰, it is preceding every twice
Secondary treatment 30min, third time treatment 45min, the fibroin fiber for obtaining is washed with deionized water totally, and is dried in 55 DEG C of baking ovens
It is dry；
(2) fibroin fiber that step (1) is obtained is placed in ternary solution, dissolution process 1h, obtains under the conditions of 72 DEG C
Mixed solution be fitted into bag filter (molecular weight 8000-14000) and dialysed in deionized water 4d, period every two hours changes water one
Secondary, the silk fibroin solution concentration for finally obtaining is 5wt%；
(3) the 5wt% silk fibroin solutions for preparing step (2) are added dropwise appropriate hydrochloric acid, prepare the silk fibroin solution that pH value is 2；
(4) step (3) silk fibroin solution is placed under the conditions of 24V (DC) and obtains fibroin albumen coagulation glue in 8 minutes.
Example IV
The fibroin albumen coagulation glue preparation process that the present embodiment is provided is as follows：
(1) mulberry silk raw silk is placed in degumming 3 times in the sodium carbonate boiling water that mass fraction is 0.6 ‰, it is preceding every twice
Secondary treatment 30min, third time treatment 45min, the fibroin fiber for obtaining is washed with deionized water totally, and is dried in 55 DEG C of baking ovens
It is dry；
(2) fibroin fiber that step (1) is obtained is placed in ternary solution, dissolution process 1h, obtains under the conditions of 72 DEG C
Mixed solution be fitted into bag filter (molecular weight 8000-14000) and dialysed in deionized water 4d, period every two hours changes water one
Secondary, the silk fibroin solution concentration for finally obtaining is 5wt%；
(3) the 5wt% silk fibroin solutions for preparing step (2) are added dropwise appropriate hydrochloric acid, prepare the silk fibroin solution that pH value is 4；
(4) electrolysis obtains fibroin albumen coagulation glue in 8 minutes under the conditions of step (3) silk fibroin solution being placed in into 24V (DC).
Listed above is only some embodiments of the invention, it is clear that the invention is not restricted to above example, may be used also
To there is many deformations, all changes that one of ordinary skill in the art can directly derive from present disclosure or associate
Shape, is considered as protection scope of the present invention.
Claims (7)
1. a kind of fast preparation method of silk fibroin matter gel, it is characterised in that comprise the following steps：
(1) raw silk is made the silk fibroin water solution that concentration is 1-10wt% by degumming, dissolving, dialysis, concentration；(2) exist
PH regulation reagents are added in the silk fibroin solution that step (1) is prepared, stirring prepares the silk fibroin solution that pH value is 1~5；
(3) silk fibroin solution processed by step (2) is placed under the conditions of weak electric field and is electrolysed；
(4) under these conditions, 50-10 minutes prepared silk fibroin matter gel.
2. method according to claim 1, it is characterised in that during described fibroin protein is silkworm and wild silkworm protein
One or more.
3. method according to claim 1, it is characterised in that described fibroin protein solution concentration be 1wt%~
10wt%.
4. method according to claim 1, it is characterised in that described pH regulation reagents are phosphoric acid, hydrochloric acid, sulfuric acid, nitre
Acid, one or more in formic acid.
5. method according to claim 1, it is characterised in that the pH value of described silk fibroin solution is 1-5.
6. method according to claim 1, it is characterised in that described electric-field intensity condition is D.C. regulated power supply.
7. method according to claim 1, it is characterised in that described D.C. regulated power supply is 12-36V.
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| CN201710145572.2A CN106866996A (en) | 2017-03-13 | 2017-03-13 | A kind of fast preparation method of silk fibroin matter gel |
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| CN201710145572.2A CN106866996A (en) | 2017-03-13 | 2017-03-13 | A kind of fast preparation method of silk fibroin matter gel |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107892755A (en) * | 2017-11-17 | 2018-04-10 | 苏州大学 | A kind of method of fibroin solutions quick-gelatinizing under physiological condition |
| CN111454614A (en) * | 2020-05-28 | 2020-07-28 | 苏州大学 | 3D biological printing ink and preparation method and application thereof |
| CN112972766A (en) * | 2021-02-22 | 2021-06-18 | 苏州大学 | Silk fibroin-hydroxyapatite composite bone scaffold with high mechanical strength and preparation method thereof |
| CN113444281A (en) * | 2021-06-28 | 2021-09-28 | 武汉纺织大学 | Fabric warm-keeping filling material with down-like function and preparation method and application thereof |
| CN114539564A (en) * | 2022-03-30 | 2022-05-27 | 武汉纺织大学 | Preparation method of rapid-gelling silk fibroin-based strong hydrogel |
| CN115137009A (en) * | 2022-07-12 | 2022-10-04 | 南昌大学 | Self-assembly protein gel of globular protein under low-voltage electric field and method thereof |
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| EP2703017A1 (en) * | 2008-09-26 | 2014-03-05 | Trustees Of Tufts College | Active silk muco-adhesives, silk electrogelation process, and devices |
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| EP2703017A1 (en) * | 2008-09-26 | 2014-03-05 | Trustees Of Tufts College | Active silk muco-adhesives, silk electrogelation process, and devices |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107892755A (en) * | 2017-11-17 | 2018-04-10 | 苏州大学 | A kind of method of fibroin solutions quick-gelatinizing under physiological condition |
| CN107892755B (en) * | 2017-11-17 | 2019-04-02 | 苏州大学 | A kind of method of fibroin solutions quick-gelatinizing under physiological condition |
| WO2019095610A1 (en) * | 2017-11-17 | 2019-05-23 | 苏州大学 | Method for quick gelation of silk fibroin solution under physiological conditions |
| US11560455B2 (en) | 2017-11-17 | 2023-01-24 | Soochow University | Method for rapid gelation of silk fibroin solution under physiological conditions |
| CN111454614A (en) * | 2020-05-28 | 2020-07-28 | 苏州大学 | 3D biological printing ink and preparation method and application thereof |
| CN111454614B (en) * | 2020-05-28 | 2022-05-24 | 苏州大学 | 3D biological printing ink and preparation method and application thereof |
| CN112972766A (en) * | 2021-02-22 | 2021-06-18 | 苏州大学 | Silk fibroin-hydroxyapatite composite bone scaffold with high mechanical strength and preparation method thereof |
| CN112972766B (en) * | 2021-02-22 | 2023-02-28 | 苏州大学 | Silk fibroin-hydroxyapatite composite bone scaffold with high mechanical strength and preparation method thereof |
| CN113444281A (en) * | 2021-06-28 | 2021-09-28 | 武汉纺织大学 | Fabric warm-keeping filling material with down-like function and preparation method and application thereof |
| CN114539564A (en) * | 2022-03-30 | 2022-05-27 | 武汉纺织大学 | Preparation method of rapid-gelling silk fibroin-based strong hydrogel |
| CN114539564B (en) * | 2022-03-30 | 2023-09-12 | 武汉纺织大学 | Preparation method of rapid-gel silk fibroin-based strong hydrogel |
| CN115137009A (en) * | 2022-07-12 | 2022-10-04 | 南昌大学 | Self-assembly protein gel of globular protein under low-voltage electric field and method thereof |
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Application publication date: 20170620 |