CN110028685A - A kind of preparation method of fibroin/chitosan/porous oxidation graphene complex three-dimensional bracket - Google Patents
A kind of preparation method of fibroin/chitosan/porous oxidation graphene complex three-dimensional bracket Download PDFInfo
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Abstract
The present invention relates to a kind of fibroin/chitosan/porous oxidation graphene complex three-dimensional bracket preparation methods, comprising the following steps: (1) takes porous oxidation graphene solution to be added in silk fibroin protein solution, obtain mixed solution A;(2) chitosan is taken to be added in acetic acid aqueous solution, heating stirring is made into chitosan aqueous solution;(3) it will be added in the mixed solution A in step (1) after the chitosan aqueous solution centrifugation in step (2), obtain mixed solution B;(4) mixed solution B obtained by step (3) is subjected to freeze-drying process to get purpose product is arrived.Compared with prior art, simple process of the invention, commercial viability is strong, and the three-dimensional stent material of acquisition has the advantages that good mechanical performance, bioactivity are high, degradability is strong.
Description
Technical field
The invention belongs to technical field of composite preparation, and it is compound to be related to a kind of fibroin/chitosan/porous oxidation graphene
The preparation method of three-dimensional rack.
Background technique
Silk fibroin protein is a kind of by the way that silk is dissolved purification, is obtained by aminoacetic acid, alanine, serine etc. 18
Natural macromolecular material composed by kind of amino acid, abundance have good biocompatibility, breathable moisture permeability and can
The advantages that degradability.Fibroin material is removed for industries such as surgical sewing thread, cosmetics and food additives, in recent years
More and more researchers have started to inquire into it applied to cell culture medium, the biologies such as drug controlled release carrier and artificial skin
A possibility that medical material.Though natural silk has excellent mechanical strength and toughness, due to the change of its molecular secondary structure
Change, after dissolution regeneration, the timbering material that fibroin albumen is formed usually becomes very crisp under dry state, it is difficult to process.Lead in industry
It crosses outside the measure for adding different inducers, also solves the problems, such as this by mixing with other natural or synthetic macromolecules.In addition,
Freeze-drying is to prepare one of three-dimensional porous rack material simple effective method, but to simple fibroin albumen, freezing
In drying process, fibroin molecule can be self-assembled into lamellar structure, this will affect its pore structure macroscopically to a certain extent,
It can improve the interaction between silk fibroin molecular by outer-penetrating agent is added, inhibit being self-assembled into for silk fibroin molecular
One of sheet and critical issue.
Chitosan (CS) is a kind of natural biological polysaccharide of the absorption that can degrade in vivo generated after deacetylated by chitin, is
The only positively charged polysaccharide high molecular material of nature.Due to its with good biocompatibility, biodegradable and
Polyfunctional group, chitosan cause extensively in wide range of areas such as water process, seperation film, food packaging, organizational project and drug conveyings
General concern.
Graphene has excellent mechanical property, big specific surface area and good biocompatibility, thus cures in biology
Field possesses huge application potential.Porous oxidation graphene (HGO) is a kind of derivative of graphene, porous oxidation graphite
There are many oxygen-containing functional groups, such as carbonyl, carboxyl, hydroxyl on alkene lamella, allow to carry out with a variety of silk-fibroin peptide chains more
Weight covalent reaction, while there is also the noncovalent interactions such as ionic bond and hydrogen bond.
Therefore, if three kinds of material advantages can be combined for three kinds of fibroin albumen, chitosan and graphene Material cladding preparations
Composite material must can be with a wide range of applications.
Summary of the invention
It is an object of the present invention to overcome the above-mentioned drawbacks of the prior art and provide a kind of fibroin/chitosan/
The preparation method of porous oxidation graphene complex three-dimensional bracket, is mixed by solution and the measures such as freeze-drying, acquisition have more
The fibroin albumen three-dimensional compound rest of good mechanical property and bioactivity.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of preparation method of fibroin/chitosan/porous oxidation graphene complex three-dimensional bracket, comprising the following steps:
(1) it takes porous oxidation graphene solution to be added in silk fibroin protein solution, obtains mixed solution A;
(2) chitosan is taken to be added in acetic acid aqueous solution, heating stirring is made into chitosan aqueous solution;
(3) it will be added in the mixed solution A in step (1), mixed after the chitosan aqueous solution centrifugation in step (2)
Close solution B;
(4) mixed solution B obtained by step (3) is subjected to freeze-drying process to get purpose product is arrived.
Further, in step (1), the additive amount of porous oxidation graphene solution and silk fibroin protein solution meets: porous
The mass ratio of graphene oxide and fibroin albumen is 1-2:1.
Further, in step (1), silk fibroin protein solution is the silk fibroin water solution of mass fraction 3.5%;
The concentration of porous oxidation graphene solution is 0.1-0.3g/L.
Further, in step (1), the process for preparation of silk fibroin protein solution are as follows: the threadiness that will be obtained after degumming silkworm cocoons
Fibroin is placed in LiBr solution, and Na is added2CO3Solution, so that fibrous fibroin dissolution, then carry out dialysis processing, that is, it completes.
Further, the concentration of LiBr solution is 9M, Na2CO3The concentration of solution is 0.02M, and the temperature that solution is prepared is
60℃。
Further, in step (2), the mass fraction of acetic acid aqueous solution is 2%, the condition of heating stirring are as follows: at 100 DEG C
1h is stirred with 600r/min revolving speed in water-bath, the mass fraction for the chitosan aqueous solution being made into is 2%.
Further, in step (3), the mass ratio of chitosan aqueous solution and mixed solution A is 0.1-1:1.
Further, in step (4), the temperature of freeze-drying is -20 DEG C~-60 DEG C, drying time 10-15h.
Fibroin albumen has good biocompatibility as natural polymer, but its crystallinity is higher, and brittleness is big, is added
Chitosan can improve the crystallinity of fibroin albumen, and porous oxidation graphene can be well in conjunction with the two, and is three-dimensional
Bracket provides a better porous network structure.For silk fibroin protein solution in configuration process, it is effective to carry out to need LiBr
Degumming compares and is dissolved with ternary solvent, and degumming rate convenient using single lithium bromide is higher.If fibroin albumen amount was added
More, the laminated structure that will lead to bracket increases;If not adding HGO, SF/CS forms the chip architecture of mainly aperture, seriously affects
The overall structure of three-dimensional rack has an impact subsequent absorption etc..
Compared with prior art, the invention has the following advantages that
1) fibroin albumen, contact crosslinks and Supramolecular self assembly between chitosan and porous oxidation graphene, nothing
Crosslinking agent must be added and adjust PH, and the presence of graphene oxide can be such that fibroin albumen is tied by random conformation steering rule
Structure, it is possible to reduce the addition of the chemical reagent such as methanol, simple process can industrialized production;
2) the three-dimensional rack good mechanical performance, bioactivity of acquisition are high, biodegradability is strong;
3) it is with a wide range of applications: can be used as tissue biomaterial, man-made organ, facial mask is separated by filtration material etc.
The application of other field.
Detailed description of the invention
Fig. 1 is preparation technology flow chart of the invention;
Fig. 2 is the scanning electron microscope (SEM) photograph of three-dimensional rack prepared by embodiment 1
Fig. 3 is fibroin/chitosan compound rest scanning electron microscope (SEM) photograph prepared by embodiment 4.
Specific embodiment
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings.The present embodiment is with technical solution of the present invention
Premised on implemented, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to
Following embodiments.
Embodiment 1
A kind of preparation method of fibroin, chitosan and porous oxidation graphene complex three-dimensional bracket, the specific steps of which are as follows:
1) 0.02mol/L Na is utilized2CO3Solution will obtain fibroin albumen after degumming silkworm cocoons, it is dissolved at 60 DEG C
4h in 9mol/L LiBr solution, after with dialysis membrane dialyse 72h, obtain quality volume fraction be 6~7w/v% fibroin albumen water
Solution is added appropriate water and is configured to the silk fibroin protein solution that mass fraction is 3.5%, is placed in 4 DEG C of refrigerators and saves for use;
2) the porous oxidation graphene solution of 0.1g/l is added in the silk fibroin solution that step 1) obtains, obtains mixing molten
Liquid A, wherein graphene oxide: the mass ratio of fibroin albumen is 2:1;
3) 2.5g chitosan is placed in 2% acetic acid aqueous solution, the 600r/min magnetic agitation 1h in 100 DEG C of water-baths,
The chitosan solution that mass fraction is 2% is prepared, uses preceding centrifugation;
4) it will be added in the mixed solution A of step 2) acquisition after the centrifugation of step 3) chitosan solution, obtain mixed solution
B, wherein chitosan and the mass ratio of mixed solution A are 0.1:1,15~20min are swung in mixed solution B ultrasound acoustic shock, so that porous
Graphene is cross-linked to each other between fibroin albumen and chitosan, sufficiently combines;
5) by freeze overnight under conditions of obtained in step 4) -20 DEG C of warp of mixed solution B, then it is dry with freeze drier
10h to get arrive fibroin, chitosan and porous oxidation graphene complex three-dimensional timbering material.
Embodiment 2
A kind of preparation method of fibroin, chitosan and porous oxidation graphene complex three-dimensional bracket, the specific steps of which are as follows:
1) silk fibroin protein solution that mass fraction is 3.5% is prepared according to 1 method of embodiment, is placed in 4 DEG C of refrigerators and saves
For use;
2) the porous oxidation graphene solution of 0.2g/l is added in the silk fibroin solution that step 1) obtains, obtains mixing molten
Liquid A, wherein graphene oxide: the mass ratio of fibroin albumen is 1.5:1;
3) chitosan solution that mass fraction is 2% is prepared according to the method for embodiment 1, uses preceding centrifugation;
4) it will be added in the mixed solution A of step 2) acquisition after the centrifugation of step 3) chitosan solution, obtain mixed solution
B, wherein chitosan and the mass ratio of mixed solution A are 0.6:1,15~20min are swung in mixed solution B ultrasound acoustic shock, so that porous
Graphene is cross-linked to each other between fibroin albumen and chitosan, sufficiently combines;
5) by freeze overnight under conditions of obtained in step 4) -40 DEG C of warp of mixed solution B, then it is dry with freeze drier
12h to get arrive fibroin, chitosan and porous oxidation graphene complex three-dimensional timbering material.
Embodiment 3
A kind of preparation method of fibroin, chitosan and porous oxidation graphene complex three-dimensional bracket, the specific steps of which are as follows:
1) silk fibroin protein solution that mass fraction is 3.5% is prepared according to 1 method of embodiment, is placed in 4 DEG C of refrigerators and saves
For use;
2) the porous oxidation graphene solution of 0.3g/l is added in the silk fibroin solution that step 1) obtains, obtains mixing molten
Liquid A, wherein graphene oxide: the mass ratio of fibroin albumen is 1:1;
3) chitosan solution that mass fraction is 2% is prepared according to the method for embodiment 1, uses preceding centrifugation;
4) it will be added in the mixed solution A of step 2) acquisition after the centrifugation of step 3) chitosan solution, obtain mixed solution
B, wherein chitosan and the mass ratio of mixed solution A are 1:1,15~20min are swung in mixed solution B ultrasound acoustic shock, so that porous stone
Black alkene, is cross-linked to each other between fibroin albumen and chitosan, sufficiently combines;
5) by freeze overnight under conditions of obtained in step 4) -60 DEG C of warp of mixed solution B, then it is dry with freeze drier
15h to get arrive fibroin, chitosan and porous oxidation graphene complex three-dimensional timbering material.
Shape is carried out to fibroin obtained in the present embodiment 1, chitosan and porous oxidation graphene complex three-dimensional timbering material
Looks characterization.Attached drawing 1 is complex three-dimensional timbering material preparation technology flow chart, if Fig. 2 is the scanning electron microscope (SEM) photograph of three-dimensional stent material,
Complex three-dimensional bracket prepared by the present invention as shown in the figure has highly porous, and aperture is about between 20um~40um, and hole with
Hole is interconnected, which has good porous structure and bioactivity, can be used as tissue biomaterial, artificial device
Official, is separated by filtration material at facial mask.
Embodiment 4
A kind of preparation method of fibroin, chitosan compound rest, the specific steps of which are as follows:
1) silk fibroin protein solution that mass fraction is 3.5% is prepared according to 1 method of embodiment, is placed in 4 DEG C of refrigerators and saves
For use;
2) chitosan solution that mass fraction is 2% is prepared according to the method for embodiment 1, uses preceding centrifugation;
3) it will be added in the silk fibroin protein solution of step 1) acquisition after the centrifugation of step 2) chitosan solution, obtain mixing molten
Liquid A, wherein the mass ratio of chitosan and fibroin albumen is 1:1, by mixed solution A 15~20min of ultrasonic vibration, so that fibroin egg
It is white to be cross-linked to each other between chitosan, sufficiently combine;
4) by freeze overnight under conditions of -60 DEG C of mixed solution A warp obtained in step 3), then it is dry with freeze drier
15h to get arrive fibroin/Chitosan Composite Scaffold Materials.
Shape is carried out to fibroin obtained in the present embodiment 1, chitosan and porous oxidation graphene complex three-dimensional timbering material
Looks characterization.Attached drawing 1 is complex three-dimensional timbering material preparation technology flow chart, if Fig. 2 is the scanning electron microscope (SEM) photograph of three-dimensional stent material,
Complex three-dimensional bracket prepared by the present invention as shown in the figure has highly porous, and aperture is about between 20um~40um, and hole with
Hole is interconnected, which has good porous structure and bioactivity, can be used as tissue biomaterial, artificial device
Official, is separated by filtration material at facial mask.
Fibroin/chitosan compound rest progress morphology characterization obtained in the present embodiment 4.Attached drawing 3 is sweeping for compound rest
Electron microscope is retouched, the present embodiment as shown in the figure is in the case where no addition porous oxidation graphene, main slabbing small structure,
And brittleness is larger, poor mechanical property.
The above description of the embodiments is intended to facilitate ordinary skill in the art to understand and use the invention.
Person skilled in the art obviously easily can make various modifications to these embodiments, and described herein general
Principle is applied in other embodiments without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments, ability
Field technique personnel announcement according to the present invention, improvement and modification made without departing from the scope of the present invention all should be of the invention
Within protection scope.
Claims (8)
1. a kind of fibroin/chitosan/porous oxidation graphene complex three-dimensional bracket preparation method, which is characterized in that including with
Lower step:
(1) it takes porous oxidation graphene solution to be added in silk fibroin protein solution, obtains mixed solution A;
(2) chitosan is taken to be added in acetic acid aqueous solution, heating stirring is made into chitosan aqueous solution;
(3) it will be added in the mixed solution A in step (1) after the chitosan aqueous solution centrifugation in step (2), obtain mixing molten
Liquid B;
(4) mixed solution B obtained by step (3) is subjected to freeze-drying process to get purpose product is arrived.
2. a kind of fibroin/chitosan according to claim 1/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that in step (1), the additive amount of porous oxidation graphene solution and silk fibroin protein solution meets: porous oxidation
The mass ratio of graphene and fibroin albumen is 1-2:1.
3. a kind of fibroin/chitosan according to claim 1/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that in step (1), silk fibroin protein solution is the silk fibroin water solution of mass fraction 3.5%;
The concentration of porous oxidation graphene solution is 0.1-0.3g/L.
4. a kind of preparation of fibroin/chitosan according to claim 1 or 3/porous oxidation graphene complex three-dimensional bracket
Method, which is characterized in that in step (1), the process for preparation of silk fibroin protein solution are as follows: the fibrous silk that will be obtained after degumming silkworm cocoons
Element is placed in LiBr solution, and Na is added2CO3Solution, so that fibrous fibroin dissolution, then carry out dialysis processing, that is, it completes.
5. a kind of fibroin/chitosan according to claim 4/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that the concentration of LiBr solution is 9M, Na2CO3The concentration of solution is 0.02M, and the temperature that solution is prepared is 60 DEG C.
6. a kind of fibroin/chitosan according to claim 1/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that in step (2), the mass fraction of acetic acid aqueous solution is 2%, the condition of heating stirring are as follows: in 100 DEG C of water
1h is stirred with 600r/min revolving speed in bath, the mass fraction for the chitosan aqueous solution being made into is 2%.
7. a kind of fibroin/chitosan according to claim 1/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that in step (3), the mass ratio of chitosan aqueous solution and mixed solution A is 0.1-1:1.
8. a kind of fibroin/chitosan according to claim 1/porous oxidation graphene complex three-dimensional bracket preparation side
Method, which is characterized in that in step (4), the temperature of freeze-drying is -20 DEG C~-60 DEG C, drying time 10-15h.
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Cited By (3)
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CN110452397A (en) * | 2019-08-19 | 2019-11-15 | 中国科学院苏州纳米技术与纳米仿生研究所 | Three-dimensional graphene foam/natural polysaccharide based aquagel compound rest and its preparation method |
CN111214710A (en) * | 2020-01-08 | 2020-06-02 | 广州贝奥吉因生物科技股份有限公司 | Composite scaffold for promoting skin regeneration and preparation method thereof |
CN113244463A (en) * | 2021-05-31 | 2021-08-13 | 杭州凤喆凰生物科技有限公司 | Method for manufacturing nano microcapsule acellular biological scaffold |
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