CN106421914B - A kind of fibroin fiber compound rest and preparation method thereof - Google Patents
A kind of fibroin fiber compound rest and preparation method thereof Download PDFInfo
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- CN106421914B CN106421914B CN201611145472.1A CN201611145472A CN106421914B CN 106421914 B CN106421914 B CN 106421914B CN 201611145472 A CN201611145472 A CN 201611145472A CN 106421914 B CN106421914 B CN 106421914B
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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
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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
- A61L27/24—Collagen
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- 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 present invention relates to a kind of fibroin fiber compound rests and preparation method thereof, disperse boiled silk in salt/formic acid mixed liquor, obtain fibroin fiber dispersion liquid；Then high molecular material and sodium chloride is added, is uniformly mixed and obtains fibroin mixed liquor；Then fibroin mixed liquor is placed in flowing water and is handled, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin fiber bracket progress freezing processing is obtained into Frozen Body, then Frozen Body is freeze-dried and obtains fibroin fiber compound rest；The high molecular material is chitosan and/or collagen.Fibroin fiber compound rest internal structure prepared by the present invention is based on fiber, in conjunction with high molecular material, with high porosity, high perforation rate, excellent mechanical property, it is very beneficial for the conveying of nutriment, the migration of cell, the growth of tissue, is ideal tissue engineering bracket.
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 technique
With the development of modern medicine and surgery operating technology, lost by tissue or organ transplant come repair function
It is widely accepted, however is but faced with huge donor notch.Tissue is formed in vivo or in vitro by regenerative medicine means body
Or organ provides new therapeutic scheme to be damaged the reparation of function.Wherein, it the selection of tissue engineering bracket material and is built into
For one of the key of the treatment method.
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.Currently, the method for preparing silk fibroin porous scaffold has very much, including freeze-drying, salting out method, gas foaming method,
3 D-printing etc., however these methods all still remain some deficiencies for being difficult to overcome.For example, freeze-drying easily forms lamella
Structure is unfavorable for cell and tissue growth, although the prior art reports one kind and film dissolution control fibroin albumen is repeated certainly
Assembling forms nanofibrous structures, and then forms porous support, but low efficiency, poor repeatability, and hole wall structure there are straight
The migration and interaction of limitation cell are connect, tissue growth is also therefore limited.
For this purpose, overcoming the above problem of existing processing technology Yu silk-fibroin supporting structure, a kind of preparation method, and structure are developed
Build out application and regeneration doctor of the fibroin bracket for being conducive to cell and tissue growth to fibroin albumen in biomedical materials field
Clinical application all has having very great significance.
Summary of the invention
The object of the present invention is to provide a kind of preparation methods of fibroin fiber compound rest, and the fibroin prepared by this method
Fibrous composite scaffold has the advantages that the large aperture high voidage of porous support, while having the inner structural features of threadiness, and
In conjunction with biocompatible polymer material, it can be greatly facilitated cell growth, it is right such as cell Proliferation and migration and tissue growth
Tissue engineering technique clinical application is highly beneficial.
In order to achieve the above objectives, the present invention provides a kind of preparation method of fibroin fiber compound rest, includes the following steps,
It disperses boiled silk in salt/formic acid mixed liquor, obtains fibroin fiber dispersion liquid；Then high molecular material and chlorination is added
Sodium is uniformly mixed and obtains fibroin mixed liquor；Then fibroin mixed liquor is placed in flowing water and is handled, obtain hygrometric state fibroin fiber
Bracket；Hygrometric state fibroin fiber bracket progress freezing processing is obtained into Frozen Body, then Frozen Body is freeze-dried and is obtained
Fibroin fiber compound rest；The high molecular material is chitosan and/or collagen.
In above-mentioned technical proposal, the silk is mulberry silk, tussah silk, ricinus silk or wild silk yarn, preferably mulberry silk.
In above-mentioned technical proposal, the mass concentration of the formic acid is 50-99%, preferably 80-98%；The 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 not dissolve, and the natural fibroin fiber internal stent structure of preparation is based on fiber, with high porosity, high perforation rate, excellent
Good mechanical property.
In above-mentioned technical proposal, in the fibroin fiber dispersion liquid, the mass concentration of fibroin fiber is 0.1-50%, preferably
1-10%。
In above-mentioned technical proposal, the partial size of the sodium chloride is 1 μm~2mm, preferably 100-600 μm.It is thus obtained more
Pore structure is more advantageous to sticking and growing for cell, and tissue is grown into.
In above-mentioned technical proposal, the mass ratio of the fibroin fiber and sodium chloride is 1: (10-500), preferably 1: (20-
200).Thus obtained bracket has more uniform porous structure, suitable porosity, is more suitable for cell and tissue growth.
In above-mentioned technical proposal, the mass ratio of the fibroin fiber and high molecular material is 1: (0.1-0.15).Macromolecule
Material has good biocompatibility, and with the compound mechanical property that porous support not only can be improved of fibroin fiber, increase can
Processability, and bracket can be kept to be used as the biological nature that institutional framework production needs.
In above-mentioned technical proposal, the temperature of freezing processing is -10~-196 DEG C, and the temperature of freeze-drying is -10~-80
DEG C, the time of freezing processing is less than the time being freeze-dried；The advantages of handling in this way takes into account energy conservation and porous structure
Stablize and obtains.
The invention also discloses the fibroin fiber compound rest prepared according to above-mentioned preparation method, inside mainly by fibroin
Fiber is constituted, and fibre diameter is 50nm-50 μm, while there is also high molecular materials between fiber；Brace aperture rate is greater than 50%,
Pore diameter range is 50 μm ~ 1mm, compression modulus 5kPa-100MPa.
Fibroin compound rest of the invention can be proliferated and organize for the adherency of cell because having three-dimensional porous structure
Regeneration provides three-dimensional space, and is conducive to the transmission of nutriment, therefore the present invention further discloses above-mentioned fibroin fibers
Compound rest is preparing the application in tissue engineering bracket.
Compared with prior art, the present invention directly constructs textile porous materials, preparation method benefit with natural fibroin fiber
Slightly soluble existing for the good dispersion of silk and a small amount of salt is acted on acid, combination processing technique, prepared fibroin fiber
Compound rest internal structure exists simultaneously high molecular material based on fiber, has high porosity, high perforation rate, excellent power
Performance is learned, is very beneficial for the conveying of nutriment, the migration of cell, the growth of tissue, is ideal tissue engineering bracket.
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 carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Fibroin fiber；It is scattered in 0.15% calcium chloride, 88% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
(2) sodium chloride particle that 200-400 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, chitosan is added, obtain fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, chitosan is 1: 100:
0.1；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -20 DEG C, and then -20 DEG C of freeze-dryings, obtain fibroin fiber compound rest, compression modulus about 11kPa, bracket
Internal fiber diameter is 880nm-50 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, pore diameter range
For 690 μm ~ 1mm.
Embodiment two
(1) natural mulberry silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 0.15% calcium chloride, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
(2) sodium chloride particle that 200-400 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, collagen is added, obtains fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen is 1: 100: 0.1；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -10 DEG C, and then -20 DEG C of freeze-dryings, obtain fibroin fiber compound rest, compression modulus about 12kPa, bracket
Internal fiber diameter is 500nm-25 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, pore diameter range
For 900 μm ~ 0.9mm.
Embodiment three
(1) natural mulberry silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.5wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 0.1% lithium bromide, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 6%；
(2) sodium chloride particle that 100-200 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, collagen is added, obtains fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen is 1: 30: 0.15；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -20 DEG C, and then -80 DEG C of freeze-dryings, obtain fibroin fiber compound rest, compression modulus about 89kPa, bracket
Internal fiber diameter is 660nm-36 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, pore diameter range
For 560 μm ~ 1mm.
Example IV
(1) natural tussah silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.5wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 0.5% calcium chloride, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 4%；
(2) sodium chloride particle that 200-250 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, collagen is added, obtains fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen is 1: 50: 0.15；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -196 DEG C, then -10 DEG C of freeze-dryings, obtains fibroin fiber compound rest, compression modulus about 38kPa, branch
Frame internal fiber diameter is 900nm-50 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, aperture model
It encloses for 520 μm ~ 0.5mm.
Embodiment five
(1) natural wild silk yarn boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 1% calcium chloride, 90% formic acid solvent, fibroin fiber dispersion liquid concentration 2%；
(2) sodium chloride particle that 200-400 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, collagen is added, obtains fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen is 1: 100: 0.12；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -100 DEG C, then -50 DEG C of freeze-dryings, obtains fibroin fiber compound rest, compression modulus about 19kPa, branch
Frame internal fiber diameter is 800nm-30 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, aperture model
It encloses for 960 μm ~ 1mm.
Embodiment six
(1) natural ricinus silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Obtain pure fibroin fiber；It is scattered in 0.5% lithium bromide, 90% formic acid solvent, fibroin fiber dispersion liquid concentration 8%；
(2) sodium chloride particle that 200-350 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, chitosan is added, obtain fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, chitosan is 1: 180:
0.15；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -10 DEG C, then -20 DEG C of freeze-dryings, obtains fibroin fiber compound rest, compression modulus about 102kPa, branch
Frame internal fiber diameter is 640nm-38 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, aperture model
It encloses for 50 μm ~ 0.5mm.
Embodiment seven
(1) natural mulberry silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 0.2% calcium chloride, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 5%；
(2) sodium chloride particle that 50-80 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and stir
It mixes uniformly, collagen and chitosan is added, obtains fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen, chitosan is
1:90:0.05:0.08；
(3) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -20 DEG C, and then -20 DEG C of freeze-dryings, obtain fibroin fiber compound rest, compression modulus about 58kPa, bracket
Internal fiber diameter is 800nm-49 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, pore diameter range
For 700 μm ~ 1mm.
Embodiment eight
(1) natural ricinus silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Obtain pure fibroin fiber；It is scattered in 0.7% calcium chloride, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 4%；
(2) sodium chloride particle that 160-220 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
It stirs evenly, collagen is added, obtain fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen, chitosan is 1: 90:
0.15；
(4) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -10 DEG C, and then -60 DEG C of freeze-dryings, obtain fibroin fiber compound rest, compression modulus about 90kPa, bracket
Internal fiber diameter is 50nm-45 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, and pore diameter range is
500μm~0.8mm。
Embodiment nine
(1) natural mulberry silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 0.6% calcium chloride, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 10%；
(3) sodium chloride particle that 200-220 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and
Stir evenly, collagen and chitosan be added, obtain fibroin mixed liquor, fibroin fiber and sodium chloride, collagen, chitosan mass ratio
It is 1: 200: 0.05: 0.05；
(4) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -80 DEG C, then -20 DEG C of freeze-dryings, obtains fibroin fiber compound rest, compression modulus about 260kPa, branch
Frame internal fiber diameter is 100nm-50 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, aperture model
It encloses for 520 μm ~ 1mm.
Embodiment ten
(1) natural tussah silk boils 30min degumming with the sodium carbonate liquor of mass fraction 0.05wt%, obtains after being repeated 3 times
Pure fibroin fiber；It is scattered in 1% lithium bromide, 98% formic acid solvent, fibroin fiber dispersion liquid concentration 6%；
(3) sodium chloride particle that 60-80 μm of diameter is obtained by sub-sieve, is then added in fibroin fiber dispersion liquid, and stir
It mixes uniformly, chitosan is added, obtain fibroin mixed liquor, the mass ratio of fibroin fiber and sodium chloride, collagen, chitosan is 1: 50:
0.15；
(4) fibroin mixed liquor is immersed in circulating water and removes sodium chloride, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold is freezed at -20 DEG C, then -20 DEG C of freeze-dryings, obtains fibroin fiber compound rest, compression modulus about 100kPa, branch
Frame internal fiber diameter is 350nm-10 μm, and there is also high molecular material between fiber, brace aperture rate is greater than 50%, aperture model
It encloses for 500 μm ~ 1mm.
Claims (3)
1. a kind of preparation method of fibroin fiber compound rest, which is characterized in that include the following steps, disperse boiled silk in
In salt/formic acid mixed liquor, fibroin fiber dispersion liquid is obtained；Then high molecular material and sodium chloride is added, is uniformly mixed and obtains silk
Plain mixed liquor；Then fibroin mixed liquor is placed in flowing water and is handled, obtain hygrometric state fibroin fiber bracket；Hygrometric state fibroin is fine
Dimensional scaffold carries out freezing processing and obtains Frozen Body, is then freeze-dried to Frozen Body and obtains fibroin fiber compound rest；
The high molecular material is chitosan and/or collagen；The mass concentration of the formic acid is 50-90%；The salt/formic acid mixed liquor
In, the mass concentration of salt is 0.1-1%；In the fibroin fiber dispersion liquid, the mass concentration of fibroin fiber is 0.1-50%；It is described
The partial size of sodium chloride is 1 μm of -2mm；The mass ratio of the fibroin fiber and sodium chloride is 1: (10-500)；The fibroin fiber with
The mass ratio of high molecular material is 1: (0.1-0.15).
2. preparation method according to claim 1, which is characterized in that the silk is mulberry silk, tussah silk, ricinus silk
Or wild silk yarn.
3. preparation method according to claim 1, which is characterized in that the temperature of the freezing processing is -10~-196 DEG C,
The temperature of freeze-drying is -10~-80 DEG C.
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| CN107812239A (en) * | 2017-09-28 | 2018-03-20 | 广东医科大学 | A kind of preparation method of tussah silk peptide collagen compound rest |
| CN113634048B (en) * | 2021-09-10 | 2022-10-14 | 武汉纺织大学 | Natural silk micro-nano fiber composite porous material and application thereof |
| CN114681682B (en) * | 2022-03-03 | 2023-04-21 | 宁波市第一医院 | Biological scaffold, preparation material and preparation method thereof |
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