CN106633927A - Preparation method of antibacterial composite silver nanoparticle-silk fibroin film - Google Patents
Preparation method of antibacterial composite silver nanoparticle-silk fibroin film Download PDFInfo
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- CN106633927A CN106633927A CN201611260464.1A CN201611260464A CN106633927A CN 106633927 A CN106633927 A CN 106633927A CN 201611260464 A CN201611260464 A CN 201611260464A CN 106633927 A CN106633927 A CN 106633927A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
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- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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Abstract
The invention discloses a preparation method of an antibacterial composite silver nanoparticle-silk fibroin film. The preparation method includes: 1) preparing a silver nanoparticle solution, 2) preparing degummed silk, 3) preparing composite silver nanoparticle-silk fibroin powder, 4) preparing the composite silver nanoparticle-silk fibroin film. According to the preparation, the silk fibroin is utilized as a material matrix, the composite silver nanoparticle-silk fibroin material uniform in size is prepared from the silver nanoparticles which are synthetized by means of a wet chemical method, and then the composite silver nanoparticle-silk fibroin film high in mechanical property, antimicrobial property and water-insolubility is prepared.
Description
Technical field
The present invention relates to a kind of preparation method, more particularly, it relates to a kind of Nano silver grain-fibroin albumen antibacterial is compound
The preparation method of film.
Background technology
Fibroin albumen is a kind of natural polymer albumen, due to its have excellent mechanical property, biocompatibility and
Biological degradability, and it is nontoxic to human body, harmless, the neck such as biomedicine, cosmetics, drug delivery is widely used in recent years
Domain.In recent years, with the development of technology, more and more the biomaterial with fibroin albumen as substrate is by innovation and creation and answers
With.For example, fibroin albumen film can be as the substrate for preparing biology sensor, while can also be as the fresh-keeping of food antiseptic
Film；Silk fibroin powder can be as the additive of skin care item increasing the water-retaining property and biocompatibility of mutual powder product；Fibroin egg
White nanometer bead can be applied to biologic medical field as the carrier of drug delivery and release.
It is a critically important neck that metal nanoparticle-fibroin protein functional composite is prepared using fibroin albumen
Domain.Basic ideas are to put method by specific physics, chemistry metal nanoparticle and fibroin albumen are effectively combined one
Rise so that the functional composite material for preparing has the advantage of metal nanoparticle and fibroin albumen simultaneously.Metal nano
The preparation method of particle-fibroin protein functional composite is broadly divided into two big class：One is first to prepare metal nanoparticle, so
Afterwards fibroin albumen mixed with metal nano again, compound is obtained (such as American invention through methods such as Electrospun, masking, dryings
Patent US20130310908A1；Chinese invention patent CN201210258495)；Two is using fibroin material elder generation and metal
Ions binding, recycles chemical reduction method and compound is obtained (such as Chinese invention patent CN101044848；Chinese invention is special
Sharp CN102618043), former approach fibroin albumen and metal nanoparticle interphase interaction it is weak, the compound for preparing
Homogeneity is poor；The morphology controllable of later approach metal nano is poor, and due to fibroin albumen participate in chemical reaction so that
Its structural change, affects the bio-compatible performance and mechanical property of composite.
Nano silver grain is a kind of function metal nano material being widely studied, not only with specific to nano material
Skin effect and quantum size effect, while also having excellent optical property, catalytic performance and anti-microbial property.Therefore, its quilt
It is widely used as optical material, catalyst and biology sensor material etc..In addition, because Nano silver grain toxicity is low, and
There is different degrees of inhibitory action to the bacterium of numerous species, fungi and virus in life, therefore apply also for medical row
Industry and cosmetic industry.The preparation method of Nano silver grain is a lot, there is chemical reduction method, electrochemical process and photoreduction met hod etc..This
Outward, by changing specific experiment condition, the such as species of surface stabilizer, experimental temperature etc., silver nanoparticle can effectively be regulated and controled
The pattern and size of particle.
The excellent biocompatibility of good anti-microbial property and fibroin albumen based on Nano silver grain and mechanical property,
Prepare a kind of Nano silver grain with antibiotic property-fibroin albumen laminated film and have become one of focus of research, it is expected to
Become a kind of functional material for suppressing bacterium, treating infective wound surface.Fibroin albumen in-situ reducing Nano silver grain can typically be utilized
Method prepared Nano silver grain-silk-fibroin composite, but fibroin albumen take part in chemical reaction in the method,
Its original protein structure receives destruction, and the molecular weight of albumen is reduced, so as to reduce composite mechanical property and its
Allogene performance；In addition, the Nano silver grain dimensional homogeneity for preparing is poor, pattern is also uncontrollable.
The content of the invention
1. the invention technical problem to be solved
It is an object of the invention to overcome the shortcomings of above-mentioned, there is provided a kind of Nano silver grain-fibroin albumen antibacterial is combined
The preparation method of film, using technical scheme, using natural fibroin albumen as material collective, by mixing
The Nano silver grain of Moist chemical synthesis is learned so as to prepare the Nano silver grain-fibroin albumen compound of size uniformity, and then is made
Nano silver grain-fibroin albumen laminated film, Nano silver grain-fibroin albumen laminated film have good mechanical property,
Anti-microbial property and water-insoluble.
2. technical scheme
To reach above-mentioned purpose, the technical scheme that the present invention is provided is：
A kind of preparation method of Nano silver grain-fibroin albumen antibacterial laminated film of the present invention, its step is：
1) preparation of silver nano-particle solution：(1-1) mixed aqueous solution of silver nitrate and sodium citrate, wherein nitric acid are prepared
Silver and sodium citrate are mixed according to mol ratio for 1: 1；(1-2) above-mentioned mixed solution is quickly stirred, while will be pre-cooling to
The sodium borohydride solution of zero degree is rapidly injected, and the color of liquid quickly becomes yellow from water white transparency；(1-3) persistently stirring 2 is little
When, you can obtain silver nano-particle solution；
2) preparation of boiled silk：(2-1) mixed aqueous solution of the Marsellies Soap of the sodium carbonate and 1wt% of 0.5wt% is utilized
Silk cocoon is boiled, boiling time is 30 minutes, while requiring to be stirred continuously in boiling part；(2-2) by degumming after the completion of boiling
Silk pull out and cleaned repeatedly with water purification；(2-3) repeat the above steps (2-1) and step (2-2) carry out secondary degumming；(2-
4) boiled silk for completing to be obtained after step (2-3) is placed in and is dried ventilation dried overnight；
3) preparation of Nano silver grain-fibroin albumen composite powder：(3-1) it is molten with the trifluoroacetic acid that concentration is 99.9%
Solution step 2) obtained in boiled silk；(3-2) solution obtained in step (3-1) is injected into into step 1) obtained in Nano silver grain
In solution, and it is slowly stirred state homogeneous to solution presentation；(3-3) solution obtained in sodium hydroxide solution regulating step (3-2) is used
PH value is to neutrality；(3-4) solution obtained in quick whipping step (3-3) is to precipitating generation, and it is clear to continue stirring until upper solution
Clearly；(3-5) precipitation in step (3-4) is filtered out, and clear water is washed, drying, you can obtain Nano silver grain-fibroin albumen and be combined
Thing powder；
4) preparation of Nano silver grain-fibroin albumen complex thin film：(4-1) it is molten with the trifluoroacetic acid that concentration is 99.9%
Solution step 3) obtained in Nano silver grain-fibroin albumen composite powder；(4-2) by solution-cast obtained in step (4-1) in
In masking container, and it is dried at room temperature for film forming；(4-3) by film clear water and 95% ethanol water obtained in step (4-2)
Solution is dried at room temperature for after washing successively.
Further, described step 1) in obtained Nano silver grain ultraviolet-visible absorption spectroscopy figure in,
There is at 408nm characteristic absorption peak in the range of 300nm to 800nm；Described step 1) in obtained Nano silver grain transmission
Scale is 20nm in electron microscope figure, and particle size is homogeneous, and in spherical, diameter is 10nm.
Further, described step 2) in obtained boiled silk FTIR spectrum figure in, in 1400cm-1
To 1800cm-1Spectral region in 1512cm-1、1622cm-1、1698cm-1There is characteristic peak at place；Described step 4) in be obtained
Nano silver grain-fibroin albumen laminated film FTIR spectrum figure in, in 1400cm-1To 1800cm-1Spectrum model
In 1512cm in enclosing-1、1531cm-1、1622cm-1、1643cm-1、1698cm-1There is characteristic peak at place.
3. beneficial effect
The technical scheme provided using the present invention, compared with existing known technology, with following remarkable result：
(1) preparation method of a kind of Nano silver grain-fibroin albumen antibacterial laminated film of the invention, it adopts natural
Fibroin albumen as material collective, by the silver for mixing the Nano silver grain of wet chemical synthesis so as to prepare size uniformity
Nano-particle-fibroin albumen compound, operation is simple, environmental protection；
(2) preparation method of a kind of Nano silver grain-fibroin albumen antibacterial laminated film of the invention, what it was prepared
The uniformity of Nano silver grain is good in Nano silver grain-fibroin albumen laminated film, spherical in rule, and Nano silver grain-silk
Fibroin laminated film has good mechanical property and water-insoluble, can bend, and stretches, and extends；Prepare in addition
Nano silver grain-fibroin albumen laminated film can suppress the growth of bacterium, with good anti-microbial property.
Description of the drawings
Fig. 1 is the ultraviolet-visible absorption spectroscopy figure of Nano silver grain in the present invention；
Fig. 2 is the transmission electron microscope picture of Nano silver grain in the present invention；
Fig. 3 is boiled silk in the present invention, the FTIR spectrum figure of Nano silver grain-fibroin albumen laminated film.
Fig. 4 is the anti-microbial property test schematic diagram of Nano silver grain in the present invention-fibroin albumen laminated film.
Specific embodiment
To further appreciate that present disclosure, in conjunction with the accompanying drawings and embodiments the present invention is described in detail.
A kind of preparation method of Nano silver grain of the present embodiment-fibroin albumen antibacterial laminated film, its step is：
1) preparation of silver nano-particle solution：(1-1) mixed aqueous solution of silver nitrate and sodium citrate, wherein nitric acid are prepared
Silver and sodium citrate are mixed according to mol ratio for 1: 1；(1-2) above-mentioned mixed solution is quickly stirred, while will be pre-cooling to
The sodium borohydride solution of zero degree is rapidly injected, and the color of liquid quickly becomes yellow from water white transparency；(1-3) persistently stirring 2 is little
When, you can obtain silver nano-particle solution；The mixed aqueous solution of 200 milliliters of silver nitrates and sodium citrate is specially prepared, wherein
Silver nitrate is 0.25mM, sodium citrate 0.25mM；Described step (1-2) is specially obtained in quick whipping step (1-1) and mixes
Solution is closed, while 6.0 milliliters of 10mM sodium borohydride solutions for being pre-cooling to zero degree are rapidly injected, the color of solution is from colourless
It is bright to quickly become yellow；
Obtain to step 1) in the ultraviolet-visible absorption spectroscopy figure (participate in Fig. 1 shown in) of Nano silver grain for preparing,
There is characteristic absorption peak at 408nm in the range of 300nm to 800nm in figure, show that Nano silver grain size is less and homogeneous；
To step 1) in the Nano silver grain for preparing carry out
Obtaining step 1) in the transmission electron microscope picture (participate in Fig. 2 shown in) of Nano silver grain for preparing, scale is in figure
20nm, particle size is homogeneous, and in spherical, diameter is 10nm；
2) preparation of boiled silk：(2-1) mixed aqueous solution of the Marsellies Soap of the sodium carbonate and 1wt% of 0.5wt% is utilized
Silk cocoon is boiled, boiling time is 30 minutes, be specially to shear 14 silk cocoons while requiring to be stirred continuously in boiling part
Into little fine strip shape, above-mentioned silk cocoon is boiled using the mixed aqueous solution of the Marsellies Soap of the sodium carbonate and 1wt% of 1 liter of 0.5wt%, boiled
The boiling time is 30 minutes, while requiring to be stirred continuously in boiling part；(2-2) silk of degumming is pulled out after the completion of boiling is used in combination
Water purification is cleaned repeatedly；(2-3) repeat the above steps (2-1) and step (2-2) carry out secondary degumming；(2-4) step (2- will be completed
3) boiled silk obtained after is placed in and is dried ventilation dried overnight；
3) preparation of Nano silver grain-fibroin albumen composite powder：(3-1) it is molten with the trifluoroacetic acid that concentration is 99.9%
Solution step 2) obtained in boiled silk, be specially with the trifluoroacetic acid dissolving step 2 that 400 milliliters of concentration is 99.9%) be obtained
Boiled silk；(3-2) solution obtained in step (3-1) is injected into into step 1) obtained in silver nano-particle solution, and delay
Slowly stir to solution and homogeneous state be presented, specially solution obtained in step (3-1) is injected into into 200 milliliters of steps 1) obtained in silver
In nano-particle solution, and it is slowly stirred state homogeneous to solution presentation；(3-3) made with sodium hydroxide solution regulating step (3-2)
Solution ph to neutrality, specially with solution ph obtained in 1M sodium hydroxide solution regulating steps (3-2) to neutrality；
(3-4) solution obtained in quick whipping step (3-3) is to precipitating generations, and continues stirring until upper solution and clarify；(3-5) filter
Go out precipitation in step (3-4), and clear water is washed, drying, you can obtain Nano silver grain-fibroin albumen composite powder；Using
Natural fibroin albumen is equal so as to prepare size by the Nano silver grain for mixing wet chemical synthesis as material collective
One Nano silver grain-fibroin albumen compound, operation is simple, environmental protection；
4) preparation of Nano silver grain-fibroin albumen complex thin film：(4-1) it is molten with the trifluoroacetic acid that concentration is 99.9%
Solution step 3) obtained in Nano silver grain-fibroin albumen composite powder；(4-2) by solution-cast obtained in step (4-1) in
In masking container, and it is dried at room temperature for film forming；(4-3) by film clear water and 95% ethanol water obtained in step (4-2)
Solution is dried at room temperature for after washing successively；Nano silver grain in the Nano silver grain for preparing-fibroin albumen laminated film
Uniformity it is good, spherical in rule, and Nano silver grain-fibroin albumen laminated film has good mechanical property and non-aqueous
Property, can bend, stretch, extend, the Nano silver grain for preparing in addition-fibroin albumen laminated film can suppress bacterium
Growth, with good anti-microbial property.
Obtaining step 4) in obtained Nano silver grain-fibroin albumen laminated film and step 2) in obtained boiled silk
In 1400cm-1To 1800cm-1Spectral region in FTIR spectrum figure (participate in Fig. 3 shown in), curve A is in figure
Boiled silk, the infrared signature peak of boiled silk occurs in 1512cm-1、1622cm-1、1698cm-1Place, shows that fibroin is tied
Structure is mainly Silk II structures (beta sheet structure), shows water-insoluble；Curve B is Nano silver grain-fibroin albumen in figure
Laminated film, the infrared signature peak of Nano silver grain-fibroin albumen laminated film occurs in 1512cm-1、1531cm-1、1622cm-1、1643cm-1、1698cm-1, wherein 1531cm-1、1643cm-1Silk I structures (random coil and α-helixstructure) are represented,
Step 4 is understood according to analysis) in Nano silver grain-fibroin albumen laminated film for preparing simultaneously containing beta sheet structure,
α-helixstructure and random coil structure, silk structure is not destroyed in Nano silver grain-fibroin albumen laminated film, alpha-helix
The presence of structure and random coil structure causes Nano silver grain-fibroin albumen laminated film to show the good spy of ductility
Levy, but from the point of view of relative amount, beta sheet structure be primary structure, α-helixstructure and random coil structure content compared with
It is low, so as to Nano silver grain-fibroin albumen laminated film show it is water-insoluble.
By step 4) in obtained Nano silver grain-fibroin albumen laminated film be placed in colibacillary culture dish, put
In 37 degrees Celsius about 12 hours, then take out colibacillary life at observation Nano silver grain-fibroin albumen laminated film position
Long situation (participating in shown in Fig. 4), surface Nano silver grain-fibroin albumen laminated film can effectively suppress colibacillary in figure
Growth, and common fibroin albumen film then can not effectively suppress colibacillary growth, further relate to Nano silver grain-fibroin
Albumen laminated film has excellent anti-microbial property.
The preparation method of a kind of Nano silver grain-fibroin albumen antibacterial laminated film of the present invention, using natural fibroin
Albumen as material collective, by the silver nanoparticle for mixing the Nano silver grain of wet chemical synthesis so as to prepare size uniformity
Particle-fibroin albumen compound, and then Nano silver grain-fibroin albumen laminated film is obtained, Nano silver grain-fibroin albumen is multiple
Close film and there is good mechanical property, anti-microbial property and water-insoluble.
Below schematically to the present invention and embodiments thereof be described, the description does not have restricted, institute in accompanying drawing
What is shown is also one of embodiments of the present invention, and actual structure is not limited thereto.So, if the common skill of this area
Art personnel enlightened by it, in the case of without departing from the invention objective, is designed and the technical scheme without creative
Similar frame mode and embodiment, all should belong to protection scope of the present invention.
Claims (3)
1. the preparation method of a kind of Nano silver grain-fibroin albumen antibacterial laminated film, its step is：
1) preparation of silver nano-particle solution：(1-1) prepare the mixed aqueous solution of silver nitrate and sodium citrate, wherein silver nitrate and
Sodium citrate is mixed according to mol ratio for 1: 1；(1-2) above-mentioned mixed solution is quickly stirred, while zero degree will be pre-cooling to
Sodium borohydride solution be rapidly injected, the color of liquid quickly becomes yellow from water white transparency；(1-3) 2 hours are persistently stirred, i.e.,
Silver nano-particle solution is obtained；
2) preparation of boiled silk：(2-1) using 0.5wt% sodium carbonate and 1wt% Marsellies Soap mixed aqueous solution by silkworm
Cocoon boils, and boiling time is 30 minutes, while requiring to be stirred continuously in boiling part；(2-2) by the silkworm of degumming after the completion of boiling
Silk is pulled out and cleaned repeatedly with water purification；(2-3) repeat the above steps (2-1) and step (2-2) carry out secondary degumming；(2-4) will
The boiled silk for completing to be obtained after step (2-3) is placed in and is dried ventilation dried overnight；
3) preparation of Nano silver grain-fibroin albumen composite powder：(3-1) with the trifluoroacetic acid dissolving step that concentration is 99.9%
It is rapid 2) obtained in boiled silk；(3-2) solution obtained in step (3-1) is injected into into step 1) obtained in silver nano-particle solution
In, and it is slowly stirred state homogeneous to solution presentation；(3-3) solution ph obtained in sodium hydroxide solution regulating step (3-2) is used
To neutrality；(3-4) solution obtained in quick whipping step (3-3) is to precipitating generations, and continues stirring until upper solution and clarify；
(3-5) precipitation in step (3-4) is filtered out, and clear water is washed, drying, you can obtain Nano silver grain-fibroin albumen compound
Powder；
4) preparation of Nano silver grain-fibroin albumen complex thin film：(4-1) with the trifluoroacetic acid dissolving step that concentration is 99.9%
It is rapid 3) obtained in Nano silver grain-fibroin albumen composite powder；(4-2) by solution-cast obtained in step (4-1) in masking
In container, and it is dried at room temperature for film forming；(4-3) by film clear water and 95% ethanol water obtained in step (4-2)
It is dried at room temperature for after washing successively.
2. the preparation method of a kind of Nano silver grain according to claim 1-fibroin albumen antibacterial laminated film, its feature
It is：Described step 1) in obtained Nano silver grain ultraviolet-visible absorption spectroscopy figure in, in 300nm to 800nm scopes
It is interior to have characteristic absorption peak at 408nm；Described step 1) in obtained Nano silver grain transmission electron microscope picture figure in scale be
20nm, particle size is homogeneous, and in spherical, size is 10nm.
3. the preparation method of a kind of Nano silver grain according to claim 2-fibroin albumen antibacterial laminated film, its feature
It is：Described step 2) in obtained boiled silk FTIR spectrum figure in, in 1400cm-1To 1800cm-1Light
In 1512cm in spectral limit-1、1622cm-1、1698cm-1There is characteristic peak at place；Described step 4) in obtained Nano silver grain-
In the FTIR spectrum figure of fibroin albumen laminated film, in 1400cm-1To 1800cm-1Spectral region in 1512cm-1、1531cm-1、1622cm-1、1643cm-1、1698cm-1There is characteristic peak at place.
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| CN112451748A (en) * | 2020-11-25 | 2021-03-09 | 西北有色金属研究院 | Method for preparing silk fibroin-based bimetal antibacterial coating |
| CN113647409A (en) * | 2021-08-11 | 2021-11-16 | 武汉纺织大学 | Silver-loaded silk fibroin nanoparticle antibacterial agent, preparation method and application |
| CN114316315A (en) * | 2020-10-09 | 2022-04-12 | 苏州大学 | Method for forming nano-pores on silk fibroin membrane |
| CN115105634A (en) * | 2022-06-27 | 2022-09-27 | 西北有色金属研究院 | Process for preparing composite coating with timeliness for orthopedics department |
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| CN114316315A (en) * | 2020-10-09 | 2022-04-12 | 苏州大学 | Method for forming nano-pores on silk fibroin membrane |
| CN114316315B (en) * | 2020-10-09 | 2023-06-27 | 苏州大学 | Method for forming nano-holes on silk fibroin film |
| CN112451748A (en) * | 2020-11-25 | 2021-03-09 | 西北有色金属研究院 | Method for preparing silk fibroin-based bimetal antibacterial coating |
| CN113647409A (en) * | 2021-08-11 | 2021-11-16 | 武汉纺织大学 | Silver-loaded silk fibroin nanoparticle antibacterial agent, preparation method and application |
| CN113647409B (en) * | 2021-08-11 | 2023-03-07 | 武汉纺织大学 | Silver-loaded silk fibroin nanoparticle antibacterial agent, preparation method and application |
| CN115105634A (en) * | 2022-06-27 | 2022-09-27 | 西北有色金属研究院 | Process for preparing composite coating with timeliness for orthopedics department |
| CN115105634B (en) * | 2022-06-27 | 2023-09-19 | 西北有色金属研究院 | Process for preparing composite coating with timeliness for orthopedics |
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