CN103572507A - Preparation method for antibiosis ultraviolet prevention silk fibroin nanofiber membrane - Google Patents
Preparation method for antibiosis ultraviolet prevention silk fibroin nanofiber membrane Download PDFInfo
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Abstract
The invention relates to a preparation method for a nanofiber membrane which mixed nano materials which have an antibiosis ultraviolet prevention property and silk fibroin. The preparation method for the antibiosis ultraviolet prevention silk fibroin nanofiber membrane comprises performing surface modification on the nano materials which have the antibiosis ultraviolet prevention performance, adding into a prepared silk fibroin solution and mixing and forming into a spinning solution and obtaining the antibiosis ultraviolet prevention silk fibroin nanofiber membrane through the electrostatic spinning technique. The preparation method for the antibiosis ultraviolet prevention silk fibroin nanofiber membrane has the advantages of enabling nanoscale composite fibers obtained through the electrostatic spinning technique to have good biocompatibility as the silk fibroin, meanwhile to have the antibiosis ultraviolet prevention performance as photocatalysis type nano-antibacterial materials, to be applicable to biomedicine field and particularly to be capable of serving as cover materials for killing bacteria and treating infected wound surface and being simple in preparation technology, low in production cost and capable of satisfying production and application requirements.
Description
Technical field
The present invention relates to a kind of nanofiber and preparation method thereof, be specifically related to a kind of have antibiotic, uv resistance can nano material and the nano fibrous membrane of fibroin albumen blend and preparation method thereof.This method belongs to the technology of preparing of fiber, and prepared material can be widely used in the fields such as biological medicine.
Background technology
Fibroin albumen is a kind of protein extracting from silk, there is good biocompatibility, the material that can prepare the variforms such as film forming, gel, powder, physicochemical property due to its uniqueness, fibroin material is studied widely in field of biomedical materials at present, as immobilised enzymes material, cell culture substrate, medicinal slow release agent, artificial organs etc.In order to improve the performance of fibroin albumen, make it be applied to better technical field of biological material, in recent years, Chinese scholars has been carried out chemical modification by distinct methods to fibroin albumen, has obtained some new achievements in research.For example, the modification by fibroin albumen improves the physicochemical properties such as mechanical property, heat endurance of fibroin material; Change the rate of release of fibroin material to medicine; Give fibroin material blood-clot resistance, antibiotic property etc.Wherein, preparation antibiotic property fibroin protein film has become one of focus of research, and it is expected to become a kind of energy kill bacteria, the cladding material for the treatment of infective wound surface.
Antibiotic property fibroin protein film mainly comprises load antibiotic property medicine and loads nano anti-biotic material.Antibiotic property drug main will refer to antibiotic medicine, still, due to a large amount of uses of antibiotic medicine, makes pathogenic microorganism for hiding medicine, and in constantly variation, antibody-resistant bacterium also produces thereupon, has even occurred " superbacteria ".Its result causes antibiotic medicine result for the treatment of variation, even invalid.Nano anti-biotic material is the new material that a class possesses bacteriostasis property.Due to the high-specific surface area of antiseptic in material and the special effects of high reaction activity, greatly improved whole antibacterial effect, can make microorganism comprise that the Growth and reproduction of bacterium, fungi, saccharomycete, algae and virus etc. keeps lower level.Nano anti-biotic material can be divided into 3 classes again by Antibacterial Mechanism: the 1st class is the metal mold nano antibacterial agents such as silver (Ag); The 2nd class is quaternary ammonium salt or quaternary alkylphosphonium salt modification inorganic nanoparticles; The 3rd class is nano zine oxide (ZnO), nano titanium oxide (TiO
2) etc. photocatalyst-type nano anti-biotic material.Wherein, the 1st, 2 class anti-biotic materials all have report, and as existing antibacterial action, and the application of the photocatalyst-type nano anti-biotic material that has again an anti-ultraviolet effect in fibroin protein film has not yet to see report.
Electrostatic spinning technique is one and prepares the simple and effective technology of nanometer stage material.Polymer melt or solution, under high-pressure electrostatic effect, utilize electric field force to overcome polymer solution surface tension and form one charged injection stream, and solvent evaporates is solidified afterwards, and nano-scale fiber is arranged on collecting board disorderly, forms similar non-woven fabrics fiber film.Electrostatic spinning technique has been advantageously applied to prepares silk fibroin nano-fiber film, and prepared tunica fibrosa has the Morphological Features that fibre number is thin, surface area is large, porosity is high and can be used as the porous support of Growth of Cells, promotes migration and the propagation of cell; Also can be used as artificial graft's blood vessel, surface of a wound cladding material and pharmaceutical carrier, be widely used as organizational project and bio-medical engineering material.
Summary of the invention
The object of the invention is for the deficiencies in the prior art, provide a kind of have antibiotic, uv resistance can the preparation method-be Surface Modification of Nanosized Materials antibiotic by having, uv resistance energy of nano material and the nano fibrous membrane of fibroin albumen blend, join again in the silk fibroin protein solution of preparation, be mixed to form spinning solution, then by electrostatic spinning technique, prepare antibiotic, anti-ultraviolet silk fibroin nano-fiber film.
The preparation method who the invention provides a kind of antibiotic anti-ultraviolet silk fibroin nano-fiber film, is characterized in that, comprises the steps:
1) surface modification of nano material: by nano material, be dissolved in the aqueous solution that contains surfactant, making its mol ratio is 1:0.5～1:10, under ultrasound condition, reacts 4 hours; After reaction, solution, through centrifugal, washing, obtains the nano material of surface modification after being dried;
2) preparation of fibroin albumen: it is to stir in 0.5% sodium carbonate liquor that silk cocoon is put into mass percent concentration, boil the silk gum of removing silk cocoon surface for 60 minutes, after washing with water, at 60 ℃, be dried and prepare fibroin fiber, the calcium chloride that is 1:8:2 by dry fibroin fiber and mol ratio: water: the mixed solution of ethanol, at 75 ℃, be dissolved into silk fibroin protein solution, through dialysis, remove ethanol and the calcium chloride in solution, filtration makes silk fibroin protein solution, then obtained by freeze drying fibroin albumen;
3) preparation of electrostatic spinning raw material solution: according to weight ratio 1:0.05～0.3:0.005～0.05, the nano material of solvent, fibroin albumen and surface modification is mixed, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous;
4) electrostatic spinning: the electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare antibiotic, anti-ultraviolet silk fibroin nano-fiber film.
Described nano material is photocatalyst-type nano anti-biotic material, is specially nano titanium oxide (TiO
2), nano zine oxide (ZnO), nano-stannic oxide (SnO
2), nano tungsten trioxide (WO
3), nanometer vanadium trioxide (V
2o
3), silver ion doped nano titanium dioxide (Ag-TiO
2), copper ion doped nano titanium oxide (Cu-TiO
2), a kind of in silver ion nanometer doped zinc oxide (Ag-ZnO), copper ion doped nano zine oxide (Cu-ZnO).
Described surfactant is softex kw (CTAB), lauryl sodium sulfate (SDS), neopelex (SDBS), sodium hexadecyl sulfate, sodium stearyl sulfate, potassium stearate, potassium oleate, potassium laurate, odium stearate, enuatrol, sodium laurate, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), Tween-80 (Tu-80), ethoxylated dodecyl alcohol (Brij-35), Triton X-100 (Triton X-100), Pluronic F127 (Pluronic F127), a kind of in Pluronic P123 (Pluronic P123).
Described solvent is a kind of in water, formic acid, hexafluoroisopropanol, Hexafluoro acetone, trifluoroethanol.
Described electrostatic spinning process is 8～30 kilovolts of voltages, 0.1～2 ml/hour of liquid inventory, and receiving range is 10～20 centimetres.
The invention has the advantages that: pass through electrostatic spinning technique, prepare nano level composite fibre, make this fiber both there is the biocompatibility that fibroin albumen is good, the characteristic that simultaneously has both again antibiotic, the anti-ultraviolet of photocatalyst-type nano anti-biotic material, be more suitable for biomedical sector, can can eliminate bacterium as a kind of especially, the cladding material for the treatment of infective wound surface.Preparation technology of the present invention is simple, and production cost is low, can further meet the demand of producing and applying.
Accompanying drawing explanation
Fig. 1 is the stereoscan photograph of the prepared nano fibrous membrane of embodiment 1.
The specific embodiment
By specific embodiment, technical scheme of the present invention is further described below.Following embodiment further illustrates of the present invention, and does not limit the scope of the invention.
Embodiment 1:
1) by a certain amount of nano-ZnO, be dissolved in 20 milliliters of aqueous solution that contain PVP, making its mol ratio is 1:1, under ultrasound condition, reacts 4 hours.After reaction, solution, through centrifugal, washing, obtains the nano-ZnO of surface modification after being dried.
2) silk cocoon being put into mass percent concentration is to stir in 0.5% sodium carbonate liquor, boil the silk gum of removing silk cocoon surface for 60 minutes, after washing with water, under 60 ° of C, be dried and prepare fibroin fiber, the mixed solution of calcium chloride, water and the ethanol that is 1:8:2 by dry fibroin fiber and mol ratio, under 75 ° of C, be dissolved into silk fibroin protein solution, through dialysis, remove ethanol and the calcium chloride in solution, filter and make silk fibroin protein solution, then obtained by freeze drying fibroin albumen.
3) according to weight ratio 1:0.1:0.01, the nano-ZnO of water, fibroin albumen and PVP surface modification is mixed, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous.
4) electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare nano fibrous membrane.Electrostatic spinning process is: 10 kilovolts of voltages, and 0.1 ml/hour of liquid inventory, receiving range is 10 centimetres.
Fig. 1 is the stereoscan photograph of prepared nano fibrous membrane.As seen from the figure, the diameter of nanofiber is 200 nanometers.5 minutes is 98.7% to colibacillary bacteriostasis rate.
Embodiment 2:
1) by a certain amount of nano-TiO
2, be dissolved in 20 milliliters of aqueous solution that contain Triton X-100, making its mol ratio is 1:5, under ultrasound condition, reacts 4 hours.After reaction, solution, through centrifugal, washing, obtains the nano-TiO of surface modification after being dried
2.
2) silk cocoon being put into mass percent concentration is to stir in 0.5% sodium carbonate liquor, boil the silk gum of removing silk cocoon surface for 60 minutes, after washing with water, under 60 ° of C, be dried and prepare fibroin fiber, the mixed solution of calcium chloride, water and the ethanol that is 1:8:2 by dry fibroin fiber and mol ratio, under 75 ° of C, be dissolved into silk fibroin protein solution, through dialysis, remove ethanol and the calcium chloride in solution, filter and make silk fibroin protein solution, then obtained by freeze drying fibroin albumen.
3) according to weight ratio 1:0.3:0.05 by the nano-TiO of water, fibroin albumen and Triton X-100 surface modification
2mix, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous.
4) electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare nano fibrous membrane.Electrostatic spinning process is: 15 kilovolts of voltages, and 0.1 ml/hour of liquid inventory, receiving range is 10 centimetres.5 minutes is 95.8% to colibacillary bacteriostasis rate.
Embodiment 3:
1) by a certain amount of nano-ZnO, be dissolved in 20 milliliters of aqueous solution that contain PVA, making its mol ratio is 1:0.5, under ultrasound condition, reacts 4 hours.After reaction, solution, through centrifugal, washing, obtains the nano-ZnO of surface modification after being dried.
2) silk cocoon being put into mass percent concentration is to stir in 0.5% sodium carbonate liquor, boil the silk gum of removing silk cocoon surface for 60 minutes, after washing with water, under 60 ° of C, be dried and prepare fibroin fiber, the mixed solution of calcium chloride, water and the ethanol that is 1:8:2 by dry fibroin fiber and mol ratio, under 75 ° of C, be dissolved into silk fibroin protein solution, through dialysis, remove ethanol and the calcium chloride in solution, filter and make silk fibroin protein solution, then obtained by freeze drying fibroin albumen.
3) according to weight ratio 1:0.2:0.01, the nano-ZnO of hexafluoroisopropanol, fibroin albumen and PVA surface modification is mixed, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous.
4) electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare nano fibrous membrane.Electrostatic spinning process is: 20 kilovolts of voltages, and 0.2 ml/hour of liquid inventory, receiving range is 15 centimetres.5 minutes is 97.1% to colibacillary bacteriostasis rate.
Embodiment 4:
1) by a certain amount of nanometer Ag-TiO
2, be dissolved in 20 milliliters of aqueous solution that contain potassium oleate, making its mol ratio is 1:8, under ultrasound condition, reacts 4 hours.After reaction, solution, through centrifugal, washing, obtains the nanometer Ag-TiO of surface modification after being dried
2.
2) silk cocoon being put into mass percent concentration is to stir in 0.5% sodium carbonate liquor, boil and within 60 minutes, go out the silk gum on silk cocoon surface, extract, after washing with water, under 60 ° of C, be dried and prepare fibroin fiber, the calcium chloride that is 1:8:2 by dry fibroin fiber and mol ratio, the mixed solution of second alcohol and water are dissolved into silk fibroin protein solution under 75 ° of C, through dialysis, remove ethanol and the calcium chloride in solution, filtration makes silk fibroin protein solution, then obtained by freeze drying fibroin albumen.
3) according to weight ratio 1:0.1:0.005 by the nanometer Ag-TiO of trifluoroethanol, fibroin albumen and potassium oleate surface modification
2mix, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous.
4) electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare nano fibrous membrane.Electrostatic spinning process is: 15 kilovolts of voltages, and 0.2 ml/hour of liquid inventory, receiving range is 10 centimetres.5 minutes is 99.2% to colibacillary bacteriostasis rate.
Claims (5)
1. a preparation method for antibiotic anti-ultraviolet silk fibroin nano-fiber film, is characterized in that, comprises the steps:
1) surface modification of nano material: by nano material, be dissolved in the aqueous solution that contains surfactant, making its mol ratio is 1:0.5～1:10, under ultrasound condition, reacts 4 hours; After reaction, solution, through centrifugal, washing, obtains the nano material of surface modification after being dried;
2) preparation of fibroin albumen: it is to stir in 0.5% sodium carbonate liquor that silk cocoon is put into mass percent concentration, boil the silk gum of removing silk cocoon surface for 60 minutes, after washing with water, at 60 ℃, be dried and prepare fibroin fiber, the calcium chloride that is 1:8:2 by dry fibroin fiber and mol ratio: water: the mixed solution of ethanol, at 75 ℃, be dissolved into silk fibroin protein solution, through dialysis, remove ethanol and the calcium chloride in solution, filtration makes silk fibroin protein solution, then obtained by freeze drying fibroin albumen;
3) preparation of electrostatic spinning raw material solution: according to weight ratio 1:0.05～0.3:0.005～0.05, the nano material of solvent, fibroin albumen and surface modification is mixed, ultrasonic dispersion 0.5 hour, stirs 4 hours, forms the electrostatic spinning raw material solution of homogeneous;
4) electrostatic spinning: the electrostatic spinning raw material solution of preparation is injected to electrostatic spinning apparatus, prepare antibiotic, anti-ultraviolet silk fibroin nano-fiber film.
2. the preparation method of antibiotic anti-ultraviolet silk fibroin nano-fiber film according to claim 1, is characterized in that, described nano material is photocatalyst-type nano anti-biotic material, is specially nano titanium oxide (TiO
2), nano zine oxide (ZnO), nano-stannic oxide (SnO
2), nano tungsten trioxide (WO
3), nanometer vanadium trioxide (V
2o
3), silver ion doped nano titanium dioxide (Ag-TiO
2), copper ion doped nano titanium oxide (Cu-TiO
2), a kind of in silver ion nanometer doped zinc oxide (Ag-ZnO), copper ion doped nano zine oxide (Cu-ZnO).
3. the preparation method of antibiotic anti-ultraviolet silk fibroin nano-fiber film according to claim 1, it is characterized in that, described surfactant is softex kw (CTAB), lauryl sodium sulfate (SDS), neopelex (SDBS), sodium hexadecyl sulfate, sodium stearyl sulfate, potassium stearate, potassium oleate, potassium laurate, odium stearate, enuatrol, sodium laurate, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), Tween-80 (Tu-80), ethoxylated dodecyl alcohol (Brij-35), Triton X-100 (Triton X-100), Pluronic F127 (Pluronic F127), a kind of in Pluronic P123 (Pluronic P123).
4. the preparation method of antibiotic anti-ultraviolet silk fibroin nano-fiber film according to claim 1, is characterized in that, described solvent is a kind of in water, formic acid, hexafluoroisopropanol, Hexafluoro acetone, trifluoroethanol.
5. the preparation method of antibiotic anti-ultraviolet silk fibroin nano-fiber film according to claim 1, is characterized in that, described electrostatic spinning process is 8～30 kilovolts of voltages, 0.1～2 ml/hour of liquid inventory, and receiving range is 10～20 centimetres.
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