CN106421914A - Silk fibroin fiber composite scaffold and preparation method thereof - Google Patents
Silk fibroin fiber composite scaffold and preparation method thereof Download PDFInfo
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- CN106421914A CN106421914A CN201611145472.1A CN201611145472A CN106421914A CN 106421914 A CN106421914 A CN 106421914A CN 201611145472 A CN201611145472 A CN 201611145472A CN 106421914 A CN106421914 A CN 106421914A
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- fibroin fiber
- fibroin
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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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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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
- A61L27/20—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
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
Abstract
The invention relates to a silk fibroin fiber composite scaffold and a preparation method thereof. The method comprises the following steps: spreading degummed silk in salt/formic acid mixed liquid to obtain silk fibroin fiber dispersion liquid; adding a high-molecular material which is chitosan and/or collagen and sodium chloride, and uniformly mixing to obtain silk mixed liquid; treating the silk mixed liquid in flowing water to obtain a wet silk fibroin fiber scaffold; and freezing the wet silk fibroin fiber scaffold to obtain a frozen body, and freezing and drying the frozen body to obtain the silk fibroin fiber composite scaffold. The silk fibroin fiber composite scaffold has high porosity, high connectivity and excellent mechanical performance since the inner structure is mainly prepared from fibers and combined with the high-molecular material, is very beneficial to nutrition delivery, cell migration and tissue growth, and is an ideal tissue engineering scaffold.
Description
Technical field
The present invention relates to a kind of fibroin fiber compound rest and preparation method thereof, can be used for soft tissue, hard tissue repair and
The regenerative medicine fields such as medicament slow release.
Background technology
With the development of modern medicine and surgery operating technology, lost come repair function by tissue or organ transplantation
It is widely accepted, but be but faced with huge donor breach.Tissue is formed in vivo or in vitro by regenerative medicine means body
Or the reparation that organ is impaired function provides new therapeutic scheme.Wherein, the selection of tissue engineering bracket material and being built into
One of key for this Therapeutic Method.
Fibroin is derived from the natural polymer biomaterial of nature, has excellent mechanical property, controlled
Biological degradability, workability, particularly its biocompatibility equal with collagen and become preferable regenerative medicine support
Raw material.At present, prepare silk fibroin porous scaffold method have a lot, including lyophilization, salting out method, gas foaming method,
3 D-printing etc., but all still there is the deficiency that some are difficult to overcome in these methods.For example, freeze-drying easily forms lamella
Structure, is unfavorable for cell and tissue growth, although prior art reports one kind and film dissolving control fibroin albumen is repeated certainly
Assembling forms nanofibrous structures, and then forms porous support, but efficiency is low, poor repeatability, and the presence of hole wall structure is straight
Connect the migration limiting cell and interaction, tissue growth is also therefore limited.
For this reason, overcoming the problems referred to above of existing process technology and fibroin supporting structure, develop a kind of preparation method, and structure
Build out the fibroin support being conducive to cell and tissue growth to fibroin albumen in the application of biomedical materials field and regeneration doctor
The clinical practice learned all has and has very great significance.
Content of the invention
A kind of it is an object of the invention to provide preparation method of fibroin fiber compound rest, and the fibroin prepared by the method
Fibrous composite scaffold, has the large aperture high voidage advantage of porous support, has fibrous inner structural features simultaneously, and
In conjunction with biocompatible polymer material, cell growth can be greatly facilitated, such as cell proliferation and migration and tissue growth, right
Tissue engineering technique clinical practice is highly beneficial.
For reaching above-mentioned purpose, the present invention provides a kind of preparation method of fibroin fiber compound rest, comprises the following steps,
Boiled silk is scattered in salt/formic acid mixed liquor, obtains fibroin fiber dispersion liquid；It is subsequently adding macromolecular material and chlorination
Sodium, mix homogeneously obtains fibroin mixed liquor；Then fibroin mixed liquor is placed in the water of flowing and processes, obtain hygrometric state fibroin fiber
Support；Hygrometric state fibroin fiber support is carried out freezing processing and obtains Frozen Body, then lyophilization is carried out to Frozen Body and obtain
Fibroin fiber compound rest；Described macromolecular material is shitosan and/or collagen.
In technique scheme, described silkworm silk is mulberry silk, tussah silk, ricinus silk or wild silk yarn, preferably mulberry silk.
In technique scheme, the mass concentration of described formic acid is 50-99%, preferably 80-98%；Described salt/formic acid mixing
In liquid, the mass concentration of salt is 0.1-1%, and salt is calcium chloride, lithium bromide etc..The present invention is fine with salt/formic acid dispersion fibroin albumen
Dimension, and does not dissolve, and the natural fibroin fiber internal stent structure of preparation, based on fiber, has high porosity, high insertion rate, excellent
Good mechanical property.
In technique scheme, in described fibroin fiber dispersion liquid, the mass concentration of fibroin fiber is 0.1-50%, preferably
1-10%.
In technique scheme, the particle diameter of described sodium chloride is 1 μm～2mm, preferably 100-600 μm.Thus obtained many
Pore structure is more beneficial for sticking and growth of cell, and tissue is grown into.
In technique scheme, described fibroin fiber is 1 with the mass ratio of sodium chloride:（10-500）, preferably 1:（20-
200）.Thus obtained support has the loose structure being more uniformly distributed, suitable porosity, is more suitable for cell and tissue growth.
In technique scheme, described fibroin fiber is 1 with the mass ratio of macromolecular material:（0.1-0.15）.Macromolecule
Material has good biocompatibility, is combined, with fibroin fiber, the mechanical property that not only can improve porous support, increase can
Processability, and support can be kept to be used as the biological nature that organizational structure makes needs.
In technique scheme, the temperature of freezing processing is -10～-196 DEG C, and cryodesiccated temperature is -10～-80
DEG C, the time of freezing processing is less than the cryodesiccated time；The advantage so processing takes into account energy conservation and loose structure
Stable acquisition.
The invention also discloses according to the fibroin fiber compound rest of above-mentioned preparation method preparation, mainly by fibroin inside it
Fiber is constituted, and fibre diameter is 50nm-50 μm, also there is macromolecular material between fiber simultaneously；Brace aperture rate is more than 50%,
Pore diameter range is 50 μm ~ 1mm, and modulus of compressibility is 5kPa-100MPa.
The fibroin compound rest of the present invention, because having three-dimensional porous structure, can be adhesion propagation and the tissue of cell
Regeneration provides three dimensions, and is conducive to the transmission of nutrient substance, therefore the present invention further discloses above-mentioned fibroin fiber
Application in preparing tissue engineering bracket for the compound rest.
Compared with prior art, the present invention directly builds textile porous materials with natural fibroin fiber, this preparation method profit
The slightly soluble effect good dispersion of silkworm silk and a small amount of salt being existed with acid, in conjunction with handling process, prepared fibroin fiber
Based on fiber, there is macromolecular material in compound rest internal structure, have high porosity, high insertion rate, excellent power simultaneously
Learn performance, be very beneficial for the conveying of nutrient substance, the migration of cell, the growth of tissue, be preferable tissue engineering bracket.
Specific embodiment
With reference to embodiment, the invention will be further described：
Embodiment one
（1）The sodium carbonate liquor of natural mulberry silk mass fraction 0.05wt% boils 30min degumming, obtains fibroin after being repeated 3 times
Azelon；It is scattered in 0.15% calcium chloride 88% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
（2）Obtain the sodium chloride particle of 200-400 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add shitosan, obtain fibroin mixed liquor, fibroin fiber is 1: 100: 0.1 with the mass ratio of sodium chloride, shitosan；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -20 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 11kPa, internal stent
Fibre diameter is 880nm-50 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is 690
μm~1mm.
Embodiment two
（1）The sodium carbonate liquor of natural mulberry silk mass fraction 0.05wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 0.15% calcium chloride 98% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
（2）Obtain the sodium chloride particle of 200-400 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen, obtain fibroin mixed liquor, fibroin fiber is 1: 100: 0.1 with the mass ratio of sodium chloride, collagen；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -10 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 12kPa, internal stent
Fibre diameter is 500nm-25 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is 900
μm~0.9mm.
Embodiment three
（1）The sodium carbonate liquor of natural mulberry silk mass fraction 0.5wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 0.1% lithium bromide 98% formic acid solvent, fibroin fiber dispersion liquid concentration 6%；
（2）Obtain the sodium chloride particle of 100-200 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen, obtain fibroin mixed liquor, fibroin fiber is 1: 30: 0.15 with the mass ratio of sodium chloride, collagen；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -20 DEG C, and then -80 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 89kPa, internal stent
Fibre diameter is 660nm-36 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is 560
μm~1mm.
Example IV
（1）The sodium carbonate liquor of natural tussah silk mass fraction 0.5wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 0.5% calcium chloride 98% formic acid solvent, fibroin fiber dispersion liquid concentration 4%；
（2）Obtain the sodium chloride particle of 200-250 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen, obtain fibroin mixed liquor, fibroin fiber is 1: 50: 0.15 with the mass ratio of sodium chloride, collagen；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -196 DEG C, and then -10 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 38kPa, in support
Portion's fibre diameter is 900nm-50 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is
520μm~0.5mm.
Embodiment five
（1）The sodium carbonate liquor of natural wild silk yarn mass fraction 0.05wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 1% calcium chloride 90% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
（2）Obtain the sodium chloride particle of 200-400 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen, obtain fibroin mixed liquor, fibroin fiber is 1: 100: 0.12 with the mass ratio of sodium chloride, collagen；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -100 DEG C, and then -50 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 19kPa, in support
Portion's fibre diameter is 800nm-30 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is
960μm~1mm.
Embodiment six
（1）The sodium carbonate liquor of natural ricinus silk mass fraction 0.05wt% boils 30min degumming, obtains pure after being repeated 3 times
Fibroin fiber；It is scattered in 0.5% lithium bromide 90% formic acid solvent, fibroin fiber dispersion liquid concentration 8%；
（2）Obtain the sodium chloride particle of 200-350 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add shitosan, obtain fibroin mixed liquor, fibroin fiber is 1: 180: 0.15 with the mass ratio of sodium chloride, shitosan；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -10 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 102kPa, in support
Portion's fibre diameter is 640nm-38 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is
50μm~0.5mm.
Embodiment seven
（1）The sodium carbonate liquor of natural mulberry silk mass fraction 0.05wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 0.2% calcium chloride 98% formic acid solvent, fibroin fiber dispersion liquid concentration 5%；
（2）Obtain the sodium chloride particle of 50-80 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir all
Even, add collagen and shitosan, obtain fibroin mixed liquor, fibroin fiber is 1: 90 with the mass ratio of sodium chloride, collagen, shitosan
∶0.05∶0.08；
（3）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -20 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 58kPa, internal stent
Fibre diameter is 800nm-49 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is 700
μm~1mm.
Embodiment eight
（1）The sodium carbonate liquor of natural ricinus silk mass fraction 0.05wt% boils 30min degumming, obtains pure after being repeated 3 times
Fibroin fiber；It is scattered in 0.7% calcium chloride 98% formic acid solvent, fibroin fiber dispersion liquid concentration 4%；
（2）Obtain the sodium chloride particle of 160-220 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen, obtain fibroin mixed liquor, fibroin fiber is 1: 90: 0.15 with the mass ratio of sodium chloride, collagen, shitosan；
（4）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -10 DEG C, and then -60 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 90kPa, internal stent
Fibre diameter is 50nm-45 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is 500 μ
m~0.8mm.
Embodiment nine
（1）The sodium carbonate liquor of natural mulberry silk mass fraction 0.05wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 0.6% calcium chloride 98% formic acid solvent, fibroin fiber dispersion liquid concentration 10%；
（3）Obtain the sodium chloride particle of 200-220 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir
Uniformly, add collagen and shitosan, obtain fibroin mixed liquor, fibroin fiber is 1 with the mass ratio of sodium chloride, collagen, shitosan:
200∶0.05∶0.05；
（4）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -80 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 260kPa, in support
Portion's fibre diameter is 100nm-50 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is
520μm~1mm.
Embodiment ten
（1）The sodium carbonate liquor of natural tussah silk mass fraction 0.05wt% boils 30min degumming, obtains pure silk after being repeated 3 times
Fibroin fiber；It is scattered in 1% lithium bromide 98% formic acid solvent, fibroin fiber dispersion liquid concentration 6%；
（3）Obtain the sodium chloride particle of 60-80 μm of diameter by sub-sieve, be subsequently adding in fibroin fiber dispersion liquid, and stir all
Even, add shitosan, obtain fibroin mixed liquor, fibroin fiber is 1: 50: 0.15 with the mass ratio of sodium chloride, collagen, shitosan；
（4）Fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin fiber props up
Frame freezes at -20 DEG C, and then -20 DEG C of lyophilizations obtain fibroin fiber compound rest, modulus of compressibility about 100kPa, in support
Portion's fibre diameter is 350nm-10 μm, also there is macromolecular material between fiber, and brace aperture rate is more than 50%, and pore diameter range is
500μm~1mm.
Claims (10)
1. a kind of preparation method of fibroin fiber compound rest, it is characterised in that comprising the steps, boiled silk is scattered in
In salt/formic acid mixed liquor, obtain fibroin fiber dispersion liquid；It is subsequently adding macromolecular material and sodium chloride, mix homogeneously obtains silk
Plain mixed liquor；Then fibroin mixed liquor is placed in the water of flowing and processes, obtain hygrometric state fibroin fiber support；Hygrometric state fibroin is fine
Dimensional scaffold carries out freezing processing and obtains Frozen Body, then carries out lyophilization to Frozen Body and obtains fibroin fiber compound rest；
Described macromolecular material is shitosan and/or collagen.
2. preparation method according to claim 1 is it is characterised in that described silkworm silk is mulberry silk, tussah silk, ricinus silk
Or wild silk yarn.
3. preparation method according to claim 1 is it is characterised in that the mass concentration of described formic acid is 50-99%；Described
In salt/formic acid mixed liquor, the mass concentration of salt is 0.1-1%.
4. preparation method according to claim 1 is it is characterised in that in described fibroin fiber dispersion liquid, fibroin fiber
Mass concentration is 0.1-50%.
5. preparation method according to claim 1 is it is characterised in that the particle diameter of described sodium chloride is 1 μm of -2mm.
6. preparation method according to claim 1 is it is characterised in that described fibroin fiber is 1 with the mass ratio of sodium chloride:
（10-500）.
7. preparation method according to claim 1 is it is characterised in that the mass ratio of described fibroin fiber and macromolecular material
For 1:（0.1-0.15）.
8. preparation method according to claim 1 it is characterised in that described freezing processing temperature be -10～-196 DEG C,
Cryodesiccated temperature is -10～-80 DEG C.
9. according to claim 1-8 the fibroin fiber compound rest of any one preparation method preparation it is characterised in that institute
The porosity stating fibroin fiber compound rest is more than 50%, and aperture is 50 μm of -1mm.
10. application in preparing tissue engineering bracket for the fibroin fiber compound rest described in claim 9.
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| CN113634048A (en) * | 2021-09-10 | 2021-11-12 | 武汉纺织大学 | Natural silk micro-nano fiber composite porous material and application thereof |
| CN114681682A (en) * | 2022-03-03 | 2022-07-01 | 宁波市第一医院 | Biological scaffold and preparation material and preparation method thereof |
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| CN113634048A (en) * | 2021-09-10 | 2021-11-12 | 武汉纺织大学 | Natural silk micro-nano fiber composite porous material and application thereof |
| CN114681682A (en) * | 2022-03-03 | 2022-07-01 | 宁波市第一医院 | Biological scaffold and preparation material and preparation method thereof |
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| CN106421914B (en) | 2019-07-30 |
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