CN105316933A - Preparation method of antibacterial electrospun fibrous membrane - Google Patents
Preparation method of antibacterial electrospun fibrous membrane Download PDFInfo
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- CN105316933A CN105316933A CN201510784418.0A CN201510784418A CN105316933A CN 105316933 A CN105316933 A CN 105316933A CN 201510784418 A CN201510784418 A CN 201510784418A CN 105316933 A CN105316933 A CN 105316933A
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- fiber membrane
- electrospun fiber
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- antibiotic property
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Abstract
The invention provides a preparation method of an antibacterial electrospun fibrous membrane and belongs to the technical field of biological medical materials. The preparation method is characterized in that an electrospun fibrous membrane prepared from a biodegradable synthetic polymer is taken as a base body, soluble protein is added into the base body, and then peptide bonds of the soluble protein on the surfaces of composite fibers and groups on the surfaces of synthesized metal nanoparticles can be utilized to form hydrogen bonds, so that the antibacterial metal nanoparticles to be firmly loaded on the surfaces of electrospun fibers, and the favorable biocompatibility of soluble protein can be utilized to expand application in the field of biomedical materials. The antibacterial electrospun fibrous membrane prepared by the method can be well used in not only the field of biological medicines but also other fields.
Description
Technical field
The present invention relates to a kind of preparation method of antibiotic property electrospun fiber membrane, belong to biology medical material technical field.
Background technology
Anti-biotic material occupies an important position in fields such as food, electrical equipment, medicine equipments.The anti-biotic material applied in the market still with organic antibacterial agent for many, but the general antibacterial effect of organic antibacterial agent is poor, only mould proof effect is comparatively strong, all has the organic antibacterial agent of excellent inhibition few to bacterium and mould.And the heat resistance of organic antibacterial agent, less stable, its catabolite and volatile matter may be harmful.Therefore, stronger to bacteriostasis inorganic antiseptic is shown one's talent.With organic antibacterial agent ratio, inorganic antibacterial material has continuation, persistence, broad spectrum activity, not easily produces drug resistance, good heat resistance, security high.But when inorganic antiseptic is used alone, existence and stability is poor, shortcoming not easy to be recycled.
Electrostatic spinning, as a kind of new polymers processing mode, when preparing new polymer composites, plays important effect.Due to specific area, uniform diameter, the advantage feasible to multiple polymers of electrospinning fibre, it is made to account for the ratio of can not ignore when preparing the composite containing metal nanoparticle.Electrospinning fibre can provide a stable environment (can form various chemical bond or non-bonding between metal nanoparticle and tele-release fiber) to metal nanoparticle, thus makes metal nanoparticle stable dispersion on electrospinning fibre.
Electrostatic spinning and metal nanoparticle have different combinations, roughly can be divided into three kinds, and the first slaine is blended in spinning solution, reducing metal salt (utilize Bulk reduction or separately add reductant reduction) after spinning; The second carries out spinning after metal salt back being become nano particle in spinning solution.Above-mentioned two kinds of modes all introduce metal nanoparticle, but most of Nano silver grain is positioned at electrospinning fibre intrinsic silicon, can not play the effect of Nano silver grain fully efficiently.The third method: absorption or spraying, the method can overcome the shortcoming of first two method, Nano silver grain is incorporated into the surface of electrospinning fibre, although wherein Nano silver grain can be introduced on electrospun fiber membrane surface by the method for sputtering spraying, but must depend on special equipment and strict operating condition, cost is higher and not easy to operate; And absorption is, by certain chemical bond or non-bonding, the Nano silver grain synthesized in advance is adsorbed on the surface of electrospinning fibre.Dong etc. are using natrium citricum as stabilizing agent, and the nano grain of silver sub-surface of synthesis has carboxyl, utilize carboxyl and the PA of nano grain of silver sub-surface
6hydrogen bond action between the amido link on electrospun fiber membrane surface, by the surface of Nano silver grain load at electrospun fiber membrane.But the polyamide adopted in above-mentioned experiment is not biodegradable polymers, which limits with the application at biomedicine field of the antibiotic property electrospun fiber membrane of the preparation of this material.
Summary of the invention
The object of the invention is to the preparation method proposing a kind of antibiotic property electrospun fiber membrane, make the antibiotic property electrospun fiber membrane prepared by it not only can well be used for biomedicine field, can also other field be used in.
Technical scheme of the present invention is as follows:
A preparation method for antibiotic property electrospun fiber membrane, is characterized in that described method comprises the steps:
1) biodegradable synthetic polymer and soluble protein have been dissolved in solvent have made spinning solution, then use electrostatic spinning apparatus that spinning solution is made electrospun fiber membrane, electrospun fiber membrane is dry, wherein, in electrospun fiber membrane, the mass fraction scope of soluble protein is 7%-50%; Biodegradable synthetic polymer 93%-50%;
2) utilize chemical method that the salt of the metal with antibacterial action is prepared into metal nanoparticle colloidal sol, the peptide bond on protein molecule and the group in surfaces of metal nanoparticles is utilized to form hydrogen bond, make metal nanoparticle load on electrospun fiber membrane, through washing, drying, namely obtain described antibiotic property electrospun fiber membrane.
Described biodegradable synthetic polymer refers to aliphatic polyester, polyethers or polyvinyl alcohol, or aliphatic polyester and polyamide copolymer, or the copolymer of aliphatic polyester and aromatic polyester.
Preferably, described metal nanoparticle is the nano particle of silver, copper, zinc or titanium, or titanium oxide nano particle.
Preferably, the method for described metal nanoparticle load on electrospun fiber membrane adopts filters, soaks or the method for suction filtration.
The present invention has the following advantages and high-lighting effect: the preparation method of a kind of antibiotic property electrospun fiber membrane that the present invention proposes.Because the peptide bond on protein molecule and the group in surfaces of metal nanoparticles form hydrogen bond, can ensure that metal nanoparticle disperses at electrospun fiber membrane surface-stable.Simultaneously, electrospun fiber membrane is become with soluble proteome by biodegradable synthetic polymer, wherein, soluble protein is as natural polymer, there is the advantage that many synthesis and semi-synthetic macromolecule do not have, such as: good biocompatibility etc., therefore, prepared antibiotic property electrospun fiber membrane not only can well be used for biomedicine field, can also be used in other field.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope of the antibiotic property electrospun fiber membrane that the present invention obtains by embodiment 1 preparation method, and sample film is: polycaprolactone/soluble eggshell membrane protein-Nano silver grain.
Detailed description of the invention
The preparation method of a kind of antibiotic property electrospun fiber membrane provided by the invention, the method comprises the steps:
1) biodegradable synthetic polymer and soluble protein dissolving are made spinning solution in organic solvent, then use electrostatic spinning apparatus that spinning solution is made electrospun fiber membrane, electrospun fiber membrane is dry.Wherein, in electrospun fiber membrane, the mass fraction scope of soluble protein is 7%-50%, biodegradable synthetic polymer 93%-50%, biodegradable synthetic polymer refers to aliphatic polyester, polyethers or polyvinyl alcohol, or aliphatic polyester and polyamide copolymer, or the copolymer of aliphatic polyester and aromatic polyester.
2) utilize chemical method that the salt of the metal with antibacterial action is prepared into the nano particle of silver, copper, zinc, titanium or titanyl compound.Adopt and filter, soak or the method for suction filtration, and utilize the peptide bond on protein molecule and the group in surfaces of metal nanoparticles to form hydrogen bond, make metal nanoparticle load on electrospun fiber membrane, namely obtain described
antibiotic property electricityspinning fiber film.
Embodiment 1: the preparation of electrospun fiber membrane: add hexafluoroisopropanol in sample bottle, add polycaprolactone (PCL) and soluble eggshell membrane protein (SEP), the mass ratio of three is: hexafluoroisopropanol: PCL:SEP=133:9:1, so the mass fraction of soluble eggshell membrane protein is: 10%.At room temperature be stirred to dissolving.The spinning solution prepared obtains electrospun fiber membrane through electrostatic spinning, by prepared electrospun fiber membrane desolvation 24h in 40 DEG C of vacuum drying ovens.
Synthesis Nano silver grain: by AgNO
3be dissolved in 45mL deionized water, under stirring, in this solution, add Na
3c
6h
5o
72H
2o, the mass ratio of three is AgNO
3: deionized water: Na
3c
6h
5o
72H
2o=1:5294:1.73; Take NaBH
4, be dissolved in 5mL deionized water, the mass ratio of the two is: NaBH
4: deionized water=1:526; Under rapid stirring, by NaBH
4solution is quickly poured into AgNO
3in solution.After continuing rapid stirring a period of time under room temperature, terminate reaction, final products are dark-brown dispersion liquid, are Nano silver grain colloidal sol.Regulate its PH to be 5 before using Nano silver grain colloidal sol, Nano silver grain colloidal sol is now with the current.
Because the stabilizing agent synthesizing Nano silver grain is natrium citricum, so the surface of the Nano silver grain prepared has carboxyl to exist, and carboxyl can form hydrogen bond (also can form hydrogen bond with the ester group on polycaprolactone, but quantity being few) with the peptide bond on protein.So electrospun fiber membrane can Adsorption For Ag nano particle.
The electrospun fiber membrane of the certain area of clip, under the condition of suction filtration, saturated adsorption (10ml) Nano silver grain, washs with distilled water afterwards.Take out dry at ambient temperature, obtain the electrospun fiber membrane of load Nano silver grain.
Anti-microbial property: prepared electrospun fiber membrane is measured its antibacterial ability, the contact sterilization rate measured for E.coli (DH5 α) according to following characterizing method reaches 98%.
Antibacterial assay method:
LB culture medium: add NaCl in 50mL deionized water, tryptone (tryptone), yeast extract (yeastextract), the mass ratio of four is: deionized water: NaCl:tryptone:yeastextract=200:2:2:1, be distributed in 25mL conical flask after stirring and dissolving, every bottle of 5mL, 121 DEG C of sterilizing 15min in high-pressure steam sterilizing pan.
LB-agar medium (LB is dull and stereotyped): add NaCl in 50mL deionized water, tryptone (tryptone), yeast extract (yeastextract), agar (agar) is added after stirring and dissolving, the mass ratio of five is: deionized water: NaCl:tryptone:yeastextract:agar=200:2:2:1:4,121 DEG C of sterilizing 15min in high-pressure steam sterilizing pan.Culture medium through sterilizing is distributed in the 9cm culture dish of high pressure steam sterilization before cooling curing, and each culture dish adds about 20mL culture medium, is advisable to cover culture dish completely, cooling curing in super-clean bench.
Gram-E. coli (E.coli, DH5 α) is adopted to carry out antibacterial ability test.
Bacterium preculture: bacterial classification is accessed in LB culture medium, at 37 DEG C, to hatch 18h in the shaking table of the velocity fluctuation of 230rpm, for subsequent use.The dense about 109cfu/mL of bacterium.
Anti-microbial property determination experiment uses contact sterilization method to measure.Be cut into 1.5cm × 1.5cm size by testing sample and with reference to sample, and in the ethanol of 75%, process 30min sterilizing, in super-clean bench inner drying a period of time.Through sterilizing and dried sample be positioned on LB flat board, guarantee that itself and media surface are fitted completely.Each sample is inoculated the bacterium liquid that 10 μ L precultures are good, hatch 2h in 37 DEG C of incubators after, take off sample, put into and add in the PE pipe of 1mL deionized water in advance, sustained vibration 5min, with by complete for the bacterium on sample wash-out, obtain the bacterium liquid of dilution 100 times.Bacterium liquid is through gradient dilution to 10
2, 10
3, 10
4, 10
5, 10
6, 10
7doubly, respectively get 100 μ L, be uniformly coated on LB flat board, the bacterium adopting colony counting method to measure on each sample is dense.Bacterium on test sample is dense is designated as A
1(cfu/mL) bacterium, on corresponding reference sample is dense is designated as A
0(cfu/mL), then sterilizing rate P is calculated by following formula:
Embodiment 2: change polycaprolactone in embodiment 1 into poly butylene succinate, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 97%.
Embodiment 3: change polycaprolactone in embodiment 1 into PLA, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 98%.
Embodiment 4: change saturated adsorption (10ml) Nano silver grain in embodiment 1 into unsaturated absorption (5ml) Nano silver grain, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 92%.
Embodiment 5: be 10% become 7% by the mass concentration mark of soluble protein in electrospun fiber membrane in embodiment 1, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 90%.
Embodiment 6: be 10% become 50% by the mass fraction of soluble protein in electrospun fiber membrane in embodiment 1, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 7: change soluble eggshell membrane protein in embodiment 1 into soluble collagen, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 8: change the E.coli (DH5 α) in embodiment 1 into B.Subtilis (168), preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 90%.
Embodiment 9: the pH of Nano silver grain in embodiment 1 is adjusted to 3, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 10: change the mode of Adsorption For Ag nano particle in embodiment 1 into filtration, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 11: the mass concentration of soluble eggshell membrane protein in embodiment 1 is adjusted to 30%, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 12: the mass concentration of soluble eggshell membrane protein in embodiment 1 is adjusted to 50%, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 99%.
Embodiment 13: change Nano silver grain in embodiment 1 into copper nano-particle, preparation method is with embodiment 1, and the 2h contact sterilization rate for E.coli (DH5 α) reaches 82%.
Comparative example 1: if directly use Nano silver grain to be used for sterilization, so Nano silver grain cannot effectively reclaim, and can be blended in bacteria samples, and cost is high, and there is potential danger.
Claims (4)
1. a preparation method for antibiotic property electrospun fiber membrane, is characterized in that described method comprises the steps:
1) biodegradable synthetic polymer and soluble protein have been dissolved in solvent have made spinning solution, then use electrostatic spinning apparatus that spinning solution is made electrospun fiber membrane, electrospun fiber membrane is dry, wherein, in electrospun fiber membrane, the mass fraction of soluble protein is 7%-50%; The mass fraction of biodegradable synthetic polymer is 93%-50%;
2) utilize chemical method that the salt of the metal with antibacterial action is prepared into metal nanoparticle colloidal sol, the peptide bond on protein molecule and the group in surfaces of metal nanoparticles is utilized to form hydrogen bond, make metal nanoparticle load on electrospun fiber membrane, through washing, drying, namely obtain described antibiotic property electrospun fiber membrane.
2. by the preparation method of a kind of antibiotic property electrospun fiber membrane described in claim 1, it is characterized in that: biodegradable synthetic polymer refers to aliphatic polyester, polyethers or polyvinyl alcohol, or aliphatic polyester and polyamide copolymer, or the copolymer of aliphatic polyester and aromatic polyester.
3., by the preparation method of a kind of antibiotic property electrospun fiber membrane described in claim 1 or 2, it is characterized in that: described metal nanoparticle is the nano particle of silver, copper, zinc or titanium, or titanium oxide nano particle.
4. by the preparation method of a kind of antibiotic property electrospun fiber membrane described in claim 1 or 2, it is characterized in that: the method for metal nanoparticle load on electrospun fiber membrane adopts filters, soaks or the method for suction filtration.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105803777A (en) * | 2016-06-02 | 2016-07-27 | 耿云花 | Method for manufacturing antibacterial materials |
| CN105821678A (en) * | 2016-06-07 | 2016-08-03 | 耿云花 | Preparation technology of easy-to-dye synthetic leather |
| CN106049716A (en) * | 2016-06-02 | 2016-10-26 | 耿云花 | Construction process for anti-mould anti-cracking outer wall |
| CN106037128A (en) * | 2016-06-07 | 2016-10-26 | 耿云花 | Breathable and antibacterial leather shoes |
| CN106073089A (en) * | 2016-06-07 | 2016-11-09 | 耿云花 | A kind of easy dyeing processing suitcase |
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| DE19811900A1 (en) * | 1998-03-18 | 1999-09-23 | Feinchemie Gmbh Sebnitz | Biocompatible composite material useful for making bone or tooth prostheses, for coating metal, ceramic, silicon or polymer implants, or for coating textile fabrics |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105803777A (en) * | 2016-06-02 | 2016-07-27 | 耿云花 | Method for manufacturing antibacterial materials |
| CN106049716A (en) * | 2016-06-02 | 2016-10-26 | 耿云花 | Construction process for anti-mould anti-cracking outer wall |
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| CN109537281A (en) * | 2016-06-02 | 2019-03-29 | 耿云花 | A kind of manufacturing method of antibacterial fabric |
| CN105821678A (en) * | 2016-06-07 | 2016-08-03 | 耿云花 | Preparation technology of easy-to-dye synthetic leather |
| CN106037128A (en) * | 2016-06-07 | 2016-10-26 | 耿云花 | Breathable and antibacterial leather shoes |
| CN106073089A (en) * | 2016-06-07 | 2016-11-09 | 耿云花 | A kind of easy dyeing processing suitcase |
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