CN106751920A - Preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol silk gum Nano Silver and products thereof and application - Google Patents
Preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol silk gum Nano Silver and products thereof and application Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
Preparation method the present invention relates to the enhanced laminated film anti-biotic material of polyvinyl alcohol silk gum Nano Silver and products thereof and application, specific method is that silk gum is extracted from silk cocoon, then it is configured to silk gum solution, again to adding poly-vinyl alcohol solution in silk gum solution, it is well mixed, obtain polyvinyl alcohol silk gum laminated film by drying after freeze-thaw circulation；Then polyvinyl alcohol silk gum laminated film is sequentially placed into polyacrylic acid solution, PDDA solution and polyacrylic acid solution and is soaked, the polyvinyl alcohol silk gum laminated film of polyelectrolyte layer parcel is obtained, finally the enhanced laminated film anti-biotic material of polyvinyl alcohol silk gum Nano Silver is obtained with ultraviolet irradiation reducing process.The method enables Nano Silver to modify securely on the surface of polyvinyl alcohol silk gum laminated film, and is uniformly distributed, and makes it have the antibacterial activity of long duration；And preparation process is simple, it is with low cost, it is expected to be applied to biomedical anti-biotic material association area.
Description
Technical field
The invention belongs to biomedical anti-biotic material field, and in particular to polyvinyl alcohol-silk gum-Nano Silver is enhanced compound
The preparation method of film anti-biotic material, further relates to the application of product obtained by this method and the product.
Background technology
Fibroin is mainly made up of fibroin albumen and sericin.Wherein silk gum accounts for the 20-30% of total cocoon weight, and fibroin is accounted for
The 70-80% of total cocoon weight, fibroin is widely used in textile industry and medical field.However, for a long time, silk gum is de- in silk
Waste is taken as in glue processing procedure and is abandoned.The annual silk gum being lost in is up to 500000 tons in the world, and silk gum is in degraded
During need to consume substantial amounts of oxygen, therefore the discharge of silk gum can undoubtedly cause serious environmental pollution.If this day of silk gum
Right bioprotein can be developed reasonably, can not only produce great economic benefit, and can reduce to ring
The pollution in border.Silk gum is made up of multiple polypeptides, and containing 18 kinds of amino acid, this 18 kinds of amino acid include some of needed by human body
Must amino acid.Wherein three kinds of most amino acid serines of content, aspartic acid and glutamic acid have corresponding pharmacological use.
Serine can prevent and treat hypertension, synthesis treatment tuberculosis and antineoplastic.Glycine can reduce cholesterol, blood in blood
The content of sugar has effect well for preventing and treating hypertension and diabetes.Asparatate has protective effect to liver and cardiac muscle,
Can be used for the diseases such as treatment angina pectoris, miocardial infarction.Additionally, sericin has a good bioactivity, such as inoxidizability,
Promote propagation and differentiation of cell etc., in addition, silk gum also has biodegradability, good hydrophily, and possesses
Abundant polar group, therefore suffers from more and more paying close attention to and being applied to regenerative medicine field, but silk gum some in itself
For example frangible etc. the application for limiting it of defect.Silk gum is carried out using the method for blending, grafting macromolecule polyalcohol generally at present
It is modified, so as to widen application of the sericin in different field.
With the development of science and technology, nanometer technology is of increased attention.It is varied to possess different structure and work(
The nano material of energy is prepared out, and is widely used in different fields.Nano Silver series anti-biotic material is that a class is extensive
The nano anti-biotic material of application, with specific surface area it is big, surface reaction activity is high, Active sites are more, high catalytic efficiency, suction
The features such as attached ability is strong.Nano silver antibacterial material has the antibacterial activity of wide spectrum, for Gram-negative bacteria and Gram-positive
Bacterium is respectively provided with good resistance, and with it is heat-resist, safe, be not likely to produce the advantages such as drug resistance, but actually should
In, Nano Silver easily comes off, so as to the antibacterial activity for causing its long-term is subject to certain influence.And by Nano Silver and silk gum egg
White joint improves sericin simultaneously and the performance of Nano Silver has no report.
The content of the invention
In view of this, an object of the present invention is to provide a kind of polyvinyl alcohol-enhanced THIN COMPOSITE of silk gum-Nano Silver
The preparation method of film anti-biotic material；The second object of the present invention is to provide product obtained by this method；The purpose of the present invention
Three be that application of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver in anti-biotic material is prepared is provided.
To reach above-mentioned purpose, the present invention provides following technical scheme：
The preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, comprises the following steps：
(1) preparation of silk gum powder：Taking silk cocoon carries out degumming, collects silk gum solution, then that silk gum freezing solution is laggard
Row freeze-drying, obtains silk gum powder；
(2) preparation of polyvinyl alcohol-silk gum laminated film：Step (1) is obtained into silk gum powder to be dissolved in water, silk gum is obtained molten
Liquid；Then silk gum solution and poly-vinyl alcohol solution are blended, silk gum-polyvinyl alcohol Compound Water is obtained by freeze-thaw circulation
Gel, dries to obtain polyvinyl alcohol-silk gum laminated film；
(3) prepared by the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel：By step (2) gained polyvinyl alcohol-silk
Glue laminated film is first soaked in polyacrylic acid solution, obtains the negatively charged polyvinyl alcohol in surface-silk gum laminated film；Then
It is soaked in dimethyl diallyl ammonium chloride solution, then is soaked with polyacrylic acid solution, dried after final rinse water, obtains poly-
Polyvinyl alcohol-silk gum the laminated film of dielectric substrate parcel；
(4) preparation of the polyvinyl alcohol of nanometer modified by silver-silk gum laminated film anti-biotic material：By the poly- electricity of step (3) gained
Polyvinyl alcohol-silk gum the laminated film of solution matter layer parcel is placed in AgNO3In solution, using ultraviolet irradiation reducing process in silk gum-poly-
The surface synthesizing nano-silver of vinyl alcohol blend film, obtains the enhanced laminated film antibacterial material of polyvinyl alcohol-silk gum-Nano Silver
Material.
In the present invention, step (1) is shredded to take silk cocoon, and bath raio is 1：30 add water, the degumming under the conditions of 120 DEG C, 0.1Mpa
20~30min；Then by freeze-drying after freezing at -80 DEG C, silk gum powder is obtained.
In step (2) of the present invention, the concentration of the silk gum solution is 4% (w/t)；The poly-vinyl alcohol solution concentration is
5% (wt)；Freeze-thaw circulation is freezed 4 hours under the conditions of -20 DEG C, thaws 1 under the conditions of being then put in 18-25 DEG C
~2 hours, circulate 4 times；The temperature of the drying is 55 DEG C.
Preferably, the silk gum solution and the mass ratio of poly-vinyl alcohol solution are 3：3.
In the present invention, step (3) is that step (2) gained polyvinyl alcohol-silk gum laminated film is first soaked in into concentration for 2%
(w/v) 2~8min in polyacrylic acid solution, obtains the negatively charged polyvinyl alcohol in surface-silk gum laminated film；Then immersion
2~8min in concentration is for the dimethyl diallyl ammonium chloride solution of 2% (w/v), then be poly- the third of 2% (w/v) with concentration
Olefin(e) acid solution soaks 2~8min, is finally dried with after deionized water rinsing, obtains the polyvinyl alcohol-silk gum of polyelectrolyte layer parcel
Laminated film.
In the present invention, step (4) is by the polyvinyl alcohol-silk gum laminated film of step (3) gained polyelectrolyte layer parcel
Concentration is placed in for 20mM AgNO3In solution, ultraviolet irradiation at least 10min obtains polyvinyl alcohol-silk gum-Nano Silver enhanced multiple
Close film anti-biotic material.
2nd, the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver as obtained in the preparation method.
3rd, the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver answering in anti-biotic material is prepared
With.
Preferably, the anti-biotic material is resisting gram-positive bacteria or/and Gram-negative bacteria material.
It is furthermore preferred that the resisting gram-positive bacteria is Escherichia coli, the Gram-negative bacteria is Staphylococcus aureus
Bacterium.
The beneficial effects of the present invention are：It is anti-the invention discloses polyvinyl alcohol-enhanced laminated film of silk gum-Nano Silver
The preparation method of bacterium material, after the sericin and polyvinyl alcohol blending using the extraction from silk cocoon, is successfully prepared with good
The polyvinyl alcohol of good mechanical property-silk gum laminated film；Then by electrostatic adsorption in polyvinyl alcohol-silk gum laminated film
Area load charged polymeric nitride layer, recycles ultraviolet reducing process in polyvinyl alcohol-silk gum laminated film surface finish nano silver, carries
The load capacity of Nano Silver high, and make Nano Silver in laminated film surface modification firmly, so as to obtain a kind of there is durable antibiotic
The enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver of activity.The experimental technique letter of this self-assembled film
It is single, easily repeat；Polymer is combined together by strong electrostatic interactions between layers, improves polyelectrolyte
The stability of membrane structure；It is applied widely.In theory, any molecule with opposite charges can be by the method from group
Dress forms polyelectrolyte layer, and the method has stronger practicality.Gather electricity in the pan coating of polyvinyl alcohol-silk gum laminated film
Solution matter layer, can not only improve the load capacity of silver nanoparticle ion, and can strengthen silver nanoparticle ion by electrostatic adsorption
And the combination of polyelectrolyte layer, so as to impart the long-acting enduringly antibacterial activity of polyvinyl alcohol-silk gum laminated film.The film has
There is the anti-microbial property of good mechanical performance, biocompatibility, water imbibition and high-efficient and lasting, can be used for the related doctor such as antiseptic dressing
With technical field of biological material, have a good application prospect.
Brief description of the drawings
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below and carries out
Explanation：
Fig. 1 is the preparation principle schematic diagram of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver.
Fig. 2 shines for the surface Scanning Electron microscope of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver
Piece (A：Polyvinyl alcohol-silk gum the laminated film of polyelectrolyte layer parcel；B：Laminated film is placed in 20mM AgNO3It is ultraviolet in solution
Irradiation 10min；C：The compound silk gum film of polyelectrolyte layer parcel is placed in 20mM AgNO3Ultraviolet irradiation 10min in solution；D：
The compound silk gum film of polyelectrolyte layer parcel is placed in 20mM AgNO3Ultraviolet irradiation 30min in solution).
Fig. 3 is the XRD spectrum of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver.
Fig. 4 is the FT-IR collection of illustrative plates (a of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver：Silk gum；b：
Polyvinyl alcohol-silk gum laminated film；c：The laminated film of polyelectrolyte layer parcel；d：The polyelectrolyte layer parcel of nanometer modified by silver
Laminated film).
Fig. 5 is Escherichia coli and golden yellow after the addition enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver
Staphylococcic growth curve (A：The growth curve of Escherichia coli；B：The growth curve of staphylococcus aureus).
Fig. 6 is Escherichia coli and golden yellow after the addition enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver
(A is the antibacterial ring size figure of Escherichia coli to staphylococcic antibacterial ring size；B is the antibacterial ring size figure of staphylococcus aureus).
Specific embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
The enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, its schematic diagram are as shown in figure 1, be specifically logical
The method for crossing HTHP, extracts sericin from silkworm silk cocoon, then adds poly-vinyl alcohol solution in silk gum solution, mixes
Close uniform, polyvinyl alcohol-silk gum laminated film is obtained by drying after freeze-thaw circulation；Then electrostatic interaction is passed through
Polyelectrolyte layer is wrapped up on laminated film surface, positively charged silver ion is adsorbed the table in laminated film using Electrostatic Absorption
Face, the method for then being reduced with ultraviolet irradiation by nanometer modified by silver on laminated film surface, and then obtain polyvinyl alcohol-silk gum-
The enhanced laminated film anti-biotic material of Nano Silver.The film have good mechanical performance, biocompatibility, water imbibition and efficiently
Lasting anti-microbial property, can be used for the correlation biomaterial for medical purpose such as antiseptic dressing field.
Embodiment 1
The preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, comprises the following steps：
(1) preparation of silk gum powder：The silk cocoon of constant weight is weighed, is shredded, according to 1：30 bath raio 121 DEG C,
HTHP 30min under conditions of 0.1MPa, silk gum solution is obtained through filtration treatment, then silk gum solution is placed in -80 DEG C of bar
4h is freezed under part, then freeze-drying, obtain silk gum powder；
(2) preparation of polyvinyl alcohol-silk gum laminated film：By step (1) gained silk gum powder by concentration for 4% (wt) is molten
In hot water (90 DEG C), then the silk gum solution for obtaining be in mass ratio 3 in silk gum solution：3 add concentration for 5% (wt)
Poly-vinyl alcohol solution, be well mixed after, be put in -20 DEG C freezing 4h after, room temperature (18~25 DEG C) defrosting 1h is put in, by 4 times
After freeze-thaw circulation, the composite aquogel of shaping is taken out, polyvinyl alcohol-silk gum laminated film is obtained after 55 DEG C of drying；
(3) prepared by the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel：By step (2) gained polyvinyl alcohol-silk
Glue laminated film is soaked in 2min in 2% (w/v, g/ml) polyacrylic acid solution (PAA), is subsequently placed in 2% (w/v, g/ml's)
2min is soaked in dimethyl diallyl ammonium chloride (PDDA) solution, then is placed in 2% (w/v, g/ml) polyacrylic acid solution (PAA)
Middle immersion 2min, deionized water rinsing obtains the laminated film of polyelectrolyte layer parcel after drying, wherein polyelectrolyte layer is
PAA-PDDA-PAA。
(4) preparation of the polyvinyl alcohol of nanometer modified by silver-silk gum laminated film anti-biotic material：By the poly- electricity of step (3) gained
Polyvinyl alcohol-silk gum the laminated film of solution matter parcel is put into the AgNO of 20mM3In solution, by after ultraviolet irradiation 10min, will receive
Rice modified by silver obtains the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver on the surface of laminated film.
Embodiment 2
The preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, comprises the following steps：
(1) preparation of silk gum powder：The silk cocoon of constant weight is weighed, is shredded, according to 1：30 bath raio 121 DEG C,
HTHP 30min under conditions of 0.1MPa, silk gum solution is obtained through filtration treatment, then silk gum solution is placed in -80 DEG C of bar
4h is freezed under part, then freeze-drying, obtain silk gum powder；
(2) preparation of polyvinyl alcohol-silk gum laminated film：By step (1) gained silk gum powder by concentration for 4% (wt) is molten
In hot water (90 DEG C), then the silk gum solution for obtaining be in mass ratio 3 in silk gum solution：3 add concentration for 5% (wt)
Poly-vinyl alcohol solution, be well mixed after, be put in -20 DEG C freezing 4h after, room temperature (18~25 DEG C) defrosting 1h is put in, by 4 times
After freeze-thaw circulation, the composite aquogel of shaping is taken out, polyvinyl alcohol-silk gum laminated film is obtained after 55 DEG C of drying；
(3) prepared by the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel：By step (2) gained polyvinyl alcohol-silk
Glue laminated film is soaked in 4min in 2% (w/v, g/ml) polyacrylic acid solution (PAA), is subsequently placed in 2% (w/v, g/ml's)
4min is soaked in dimethyl diallyl ammonium chloride (PDDA) solution, then is placed in 2% (w/v, g/ml) polyacrylic acid solution (PAA)
Middle immersion 4min, deionized water rinsing obtains the laminated film of polyelectrolyte layer parcel after drying, wherein polyelectrolyte layer is
PAA-PDDA-PAA。
(4) preparation of the polyvinyl alcohol of nanometer modified by silver-silk gum laminated film anti-biotic material：By the poly- electricity of step (3) gained
Polyvinyl alcohol-silk gum the laminated film of solution matter parcel is put into the AgNO of 20mM3In solution, by after ultraviolet irradiation 10min, will receive
Rice modified by silver obtains the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver on the surface of laminated film.
Embodiment 3
The preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, comprises the following steps：
(1) preparation of silk gum powder：The silk cocoon of constant weight is weighed, is shredded, according to 1：30 bath raio 121 DEG C,
HTHP 30min under conditions of 0.1MPa, silk gum solution is obtained through filtration treatment, then silk gum solution is placed in -80 DEG C of bar
4h is freezed under part, then freeze-drying, obtain silk gum powder；
(2) preparation of polyvinyl alcohol-silk gum laminated film：By step (1) gained silk gum powder by concentration for 4% (wt) is molten
In hot water (90 DEG C), silk gum solution is obtained, be in mass ratio then 3 in silk gum solution：3 addition concentration are 5% (wt's)
Poly-vinyl alcohol solution, after being well mixed, after being put in -20 DEG C of freezing 4h, is put in room temperature (18~25 DEG C) defrosting 1h, cold by 4 times
After jelly-thaw cycles, the composite aquogel of shaping is taken out, polyvinyl alcohol-silk gum laminated film is obtained after 55 DEG C of drying；
(3) prepared by the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel：By step (2) gained polyvinyl alcohol-silk
Glue laminated film is soaked in 8min in 2% (w/v, g/ml) polyacrylic acid solution (PAA), is subsequently placed in 2% (w/v, g/ml's)
8min is soaked in dimethyl diallyl ammonium chloride (PDDA) solution, then is placed in 2% (w/v, g/ml) polyacrylic acid solution (PAA)
Middle immersion 8min, deionized water rinsing obtains the laminated film of polyelectrolyte layer parcel after drying, wherein polyelectrolyte layer is
PAA-PDDA-PAA。
(4) preparation of the polyvinyl alcohol of nanometer modified by silver-silk gum laminated film anti-biotic material：By the poly- electricity of step (3) gained
Polyvinyl alcohol-silk gum the laminated film of solution matter parcel is put into the AgNO of 20mM3In solution, by after ultraviolet irradiation 10min, will receive
Rice modified by silver obtains the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver on the surface of laminated film.
Embodiment 4, characterization test
Correlation is carried out to the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver obtained in embodiment 1~3
Characterization test, it is specific as follows：
(1) SEM (SEM) test：Each fritter of film that respectively prepared by Example 1~3 is placed in patch
On the objective table of conducting resinl, metal spraying treatment is carried out, in 10kV voltages, amplify 1300 times, under the technical parameter such as high vacuum pattern, institute
The surface sweeping sem image for obtaining is as shown in Figure 2.Result shows that the polyvinyl alcohol for wrapping up polyelectrolyte layer-silk gum strengthens THIN COMPOSITE
Film is soaked in 20mM AgNO3In solution, after ultraviolet irradiation can surface finish nano silver, and nanometer modified by silver amount with
The increase of time of ultraviolet irradiation and increase, and the laminated film for not wrapping up polyelectrolyte layer is only capable of modifying a small amount of Nano Silver, table
The bright load capacity that nano silver particles can be improved in polyvinyl alcohol-silk gum enhancing laminated film surface parcel polyelectrolyte layer.
(2) X-ray diffraction (XRD) test：The film that respectively prepared by Example 1~3 carries out XRD analysis of spectrum, as a result
As shown in Figure 3.Result shows that Nano Silver is successfully modified in the polyvinyl alcohol-silk gum laminated film surface of polyelectrolyte layer parcel.
(3) FT-IR collection of illustrative plates testing result is as shown in Figure 4.Result shown, the polyelectrolyte layer parcel of nanometer modified by silver is answered
Film is closed in 3274cm-1, 1640cm-1, 1531cm-1And 1248cm-1There is characteristic peak at place, shows the compound of polyelectrolyte layer parcel
Film surface successfully modifies Nano Silver.
(4) bacteriostatic experiment
1), growth curve experiment
The present embodiment is by the bacterium of growth naturally and addition polyvinyl alcohol-silk gum laminated film, polyelectrolyte layer bag
The bacterial growth of the polyvinyl alcohol of the polyvinyl alcohol wrapped up in-silk gum laminated film or nanometer modified by silver-silk gum enhancing laminated film is bent
Line is contrasted, so that it is determined that the polyvinyl alcohol of nanometer modified by silver-silk gum strengthens the antibacterial effect of laminated film, specific method
For：
1. the single bacterium colony for taking Escherichia coli (E.coli) and staphylococcus aureus (S.aureus) respectively is inoculated in sterilizing
100mL LB fluid nutrient mediums (pH 7.4) in, rotating speed be 220rpm, temperature be 37 DEG C under the conditions of cultivate 12 hours；
2. the Escherichia coli and the μ L of staphylococcus aureus bacteria suspension 100 for taking step (1) activation respectively are added to 10mL LB
In culture medium, every kind of bacterium solution prepares 4 groups, and one of which is blank group, and remaining 3 groups are separately added into polyvinyl alcohol-silk gum THIN COMPOSITE
The polyvinyl alcohol of film, the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel or nanometer modified by silver-silk gum enhancing THIN COMPOSITE
Film, then rotating speed be 220rpm, temperature be 37 DEG C under the conditions of cultivate, and 0h, 1h, 2h, 3h, 4h, 6h, 8h, 12h, 14h,
Bacteria suspension 0.5mL is taken when 18h, 22h, 26h and 30h, in 4 DEG C of Storage in refrigerator；
3. the bacteria suspension that different time takes out is taken out from 4 DEG C of refrigerators after after the bacteria suspension sampling of culture 30h, room temperature
Placed under (18-25 DEG C) after 20-40min using UV spectrophotometer measuring its absorbance value at 600nm, according to measuring
Absorption value draw the growth curve of Escherichia coli and staphylococcus aureus respectively, as a result as shown in Figure 5.Analysis polyethylene
The polyvinyl alcohol of alcohol-silk gum laminated film, the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel or nanometer modified by silver-
Silk gum strengthens influence of the laminated film to bacterial growth, as a result shows, the polyvinyl alcohol-silk gum enhancing for adding nanometer modified by silver is multiple
After closing film, the growth of Escherichia coli and staphylococcus aureus receives substantially suppression, and for up to more than 22h, table
Polyvinyl alcohol-silk gum the laminated film of nanometer modified by silver prepared by the bright present invention has excellent and lasting antibacterial activity.
2), Bactericidal test
To further determine that the antibacterial action of the compound silk gum film of nanometer modified by silver, tested using the method for inhibition zone
The polyvinyl alcohol of nanometer modified by silver-silk gum enhancing laminated film is to Escherichia coli and the fungistatic effect of staphylococcus aureus, tool
Body method is as follows：
1. the single bacterium colony for taking Escherichia coli and staphylococcus aureus respectively is inoculated in the 100mL LB Liquid Cultures of sterilizing
In base (pH7.4), rotating speed be 220rpm, temperature be 37 DEG C under the conditions of cultivate 10 hours；
2. the bacteria suspension for step (1) being activated takes 200~300 μ L and adds LB solid culture primary surfaces after diluting 100 times, and
Cultivated 1-2 hours in rotating speed is for the shaking table of 220rpm, dilution is evenly distributed on agar media surface；
3. polyvinyl alcohol-silk gum laminated film, the polyvinyl alcohol-silk gum of polyelectrolyte layer parcel of a diameter of 1.5cm are taken
The polyvinyl alcohol of laminated film or nanometer modified by silver-silk gum enhancing laminated film, is laid in the LB cultures that dilution is evenly distributed
Primary surface；Then 12h is cultivated under the conditions of 37 DEG C, as a result as shown in Figure 6.Result shows that polyvinyl alcohol-silk gum is multiple being placed with
Close in the LB culture mediums of film, be inoculated with no on the culture medium of the culture medium of Escherichia coli and inoculation staphylococcus aureus formation
Obvious inhibition zone, shows that polyvinyl alcohol-silk gum laminated film, in itself to the no inhibitory action of growth of Escherichia coli, is being placed
In the LB culture mediums of the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel, obvious inhibition zone is not also observed, show
Polyelectrolyte layer is being placed with the polyvinyl alcohol-silk gum of nanometer modified by silver in itself to the no too much influence of growth of bacterium
In the flat board of enhancing laminated film, it can be observed that form obvious inhibition zone, show the polyvinyl alcohol-silk of nanometer modified by silver
Glue enhancing laminated film has significant bacteriostasis, the enhanced laminated film anti-biotic material tool of polyvinyl alcohol-silk gum-Nano Silver
There is excellent anti-microbial property.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
Cross above preferred embodiment to be described in detail the present invention, it is to be understood by those skilled in the art that can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (10)
1. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver, it is characterised in that including such as
Lower step：
(1) preparation of silk gum powder：Taking silk cocoon carries out degumming, collects silk gum solution, then will be carried out after silk gum freezing solution cold
It is lyophilized dry, obtain silk gum powder；
(2) preparation of polyvinyl alcohol-silk gum laminated film：Step (1) is obtained into silk gum powder to be dissolved in water, silk gum solution is obtained；
Then silk gum solution and poly-vinyl alcohol solution are blended, silk gum-polyvinyl alcohol compound water congealing is obtained by freeze-thaw circulation
Glue, dries to obtain polyvinyl alcohol-silk gum laminated film；
(3) prepared by the polyvinyl alcohol-silk gum laminated film of polyelectrolyte layer parcel：Step (2) gained polyvinyl alcohol-silk gum is multiple
Close film to be first soaked in polyacrylic acid solution, obtain the negatively charged polyvinyl alcohol in surface-silk gum laminated film；Then immersion
In dimethyl diallyl ammonium chloride solution, then soaked with polyacrylic acid solution, dried after final rinse water, obtain poly- electrolysis
Polyvinyl alcohol-silk gum the laminated film of matter layer parcel；
(4) preparation of the polyvinyl alcohol of nanometer modified by silver-silk gum laminated film anti-biotic material：By step (3) gained polyelectrolyte
Polyvinyl alcohol-silk gum the laminated film of layer parcel is placed in AgNO3In solution, using ultraviolet irradiation reducing process in silk gum-polyethylene
The surface synthesizing nano-silver of alcohol blend film, obtains the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver.
2. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver according to claim 1,
It is characterized in that：Step (1) is shredded to take silk cocoon, and bath raio is 1：30 add water, under the conditions of 120 DEG C, 0.1Mpa degumming 20~
30min；Then by freeze-drying after freezing at -80 DEG C, silk gum powder is obtained.
3. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver according to claim 1,
It is characterized in that：In step (2), the concentration of the silk gum solution is 4% (w/t)；The poly-vinyl alcohol solution concentration is 5%
(wt)；The freeze-thaw circulation is freezed 4 hours under the conditions of -20 DEG C, defrosting 1~2 under the conditions of being then put in 18-25 DEG C
Hour, circulate 4 times；The temperature of the drying is 55 DEG C.
4. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver according to claim 3,
It is characterized in that：The silk gum solution is 3 with the mass ratio of poly-vinyl alcohol solution：3.
5. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver according to claim 1,
It is characterized in that：Step (3) is that step (2) gained polyvinyl alcohol-silk gum laminated film is first soaked in into concentration for 2% (w/v)
Polyacrylic acid solution in 2~8min, obtain the negatively charged polyvinyl alcohol in surface-silk gum laminated film；Then it is soaked in dense
2~8min in the dimethyl diallyl ammonium chloride solution for 2% (w/v) is spent, then is the polyacrylic acid of 2% (w/v) with concentration
Solution soaks 2~8min, is finally dried with after deionized water rinsing, and the polyvinyl alcohol-silk gum for obtaining polyelectrolyte layer parcel is combined
Film.
6. the preparation method of the enhanced laminated film anti-biotic material of polyvinyl alcohol-silk gum-Nano Silver according to claim 1,
It is characterized in that：Step (4) be by step (3) gained polyelectrolyte layer parcel polyvinyl alcohol-silk gum laminated film be placed in it is dense
It is 20mM AgNO to spend3In solution, ultraviolet irradiation at least 10min obtains polyvinyl alcohol-silk gum-nanometer human lymph node laminated film and resists
Bacterium material.
7. polyvinyl alcohol-silk gum obtained in the preparation method as described in any one of claim 1~6-nanometer human lymph node laminated film
Anti-biotic material.
8. polyvinyl alcohol-silk gum described in claim 7-nanometer human lymph node laminated film anti-biotic material is in anti-biotic material is prepared
Using.
9. application according to claim 8, it is characterised in that：The anti-biotic material is resisting gram-positive bacteria or/and leather
Lan Shi negative bacterium materials.
10. application according to claim 9, it is characterised in that：The resisting gram-positive bacteria is Escherichia coli, the leather
Lan Shi negative bacteriums are staphylococcus aureus.
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| CN108250478A (en) * | 2018-01-30 | 2018-07-06 | 西南大学 | Preparation method of the coated silk gum of nanometer modified by silver polyelectrolyte layer/agar composite membrane and products thereof and application |
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