CN101798756A - Method for preparing biomedical material of static self-assembly modified nano fiber - Google Patents
Method for preparing biomedical material of static self-assembly modified nano fiber Download PDFInfo
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Abstract
The invention relates to a method for preparing a biomedical material of a static self-assembly modified nano fiber, which is characterized by comprising the following concrete steps of: stirring and dissolving a water-insoluble macromolecular polymer for spinning in a solvent in a stirring kettle at room temperature to obtain an electric spinning raw material with the weight percentage of 5-30 percent; adding the electric spinning raw material into a static spinning device and statically spinning to obtain a nano fiber template material; soaking the nano fiber template material into a weak polymerized electrolyte aqueous solution which has the concentration of 0.01-10g/ml and an opposite charge, adsorbing for balance and drying; soaking the dried material into the weak polymerized electrolyte aqueous solution which has the concentration of 0.01-10g/ml and the opposite charge, adsorbing for balance, washing by distilled water for 5-30 minutes and drying; and obtaining the biomedical material of the static self-assembly modified nano fiber. The invention has simple molding and high bioactivity.
Description
Technical field
The present invention relates to a kind of method for preparing biomedical material of static self-assembly modified nano fiber, belong to macromolecular material biomedical applications field.
Background technology
Electrostatic spinning is that polymer solution or melt spray the process that forms nano-scale fiber under high voltage electric field.The electrostatic spinning process operating process is simple, and because resulting fiber is littler than the fibre fineness that conventional method obtains, so micro/nano fibrous membrane material has the specific area and the porosity of superelevation, can make the loose structure of 3 D stereo, be that preparation has the common method of surface-active tissue engineering material.Chinese patent CN1467314A and CN101172164A are by adding antiseptic ultrafine particle and curative drug in electric spinning polymer solution, then mixed solution is carried out electrostatic spinning, obtain having the nano-fiber material of biocompatibility, but the adding of antiseptic and curative drug is unfavorable for the formation of fiber in the electrostatic spinning process, and bioactive materials has restricted its deep application because the coating of polymeric material can not be brought into play its activity fully.Therefore prepare the nanofiber biomaterial that a kind of surface has good biocompatibility and become one of approach very crucial in the organizational project development.
Summary of the invention
The method for preparing biomedical material that the purpose of this invention is to provide static self-assembly modified nano fiber is to solve problems such as above-mentioned biocompatibility nanofiber electrostatic spinning difficult forming and biologically active difference.
The technical scheme of technical solution problem of the present invention is as follows:
A kind of method for preparing biomedical material of static self-assembly modified nano fiber is characterized in that, concrete steps are:
The first step: at room temperature, in stirred tank with spinning with water-insoluble macromolecular polymer with rotating speed 100-2500 rev/min stirring and dissolving in solvent, obtaining mass fraction is the electrospinning raw material of 5-30%;
Second step: the electrospinning raw material that the first step is obtained joins in the electrostatic spinning apparatus, at electrostatic pressure is the 10-40 kilovolt, the syringe pump flow velocity is 0.3-4ml/h, and spinning head for carrying out electrostatic spinning under the condition of 6-20cm, obtains the nanofiber mould material apart from the receiving screen distance;
The 3rd step: the nanofiber mould material of the second step preparation is immersed in the weak polyelectrolyte aqueous solution that has electric charge in contrast that concentration is 0.01mg/ml-10g/ml, adsorption equilibrium 560 minutes, with distilled water wash 5-30 minute, air blow drying; Dried material is immersed in the weak polyelectrolyte aqueous solution that has electric charge in contrast that concentration is 0.01mg/ml-10g/ml, adsorption equilibrium 560 minutes, with distilled water wash 5-30 minute, air blow drying; Alternately repeat above step 2-50 time, make the weak poly-zwitterion of biologically active, thereby make the bio-medical material of static self-assembly modified nano fiber alternately attached to nanofiber mould material surface.
Described water-insoluble macromolecular polymer is a cellulose acetate, cellulose, shitosan, ethylene-vinyl alcohol copolymer, nylon 6, polystyrene, polymethyl methacrylate, polyisobutene, polyacrylonitrile, polycaprolactone, polyvinyl acetate, polyethylene terephthalate, polypropylene, polyurethane, polyurethane, Kynoar, Merlon, epoxy resin, polysiloxanes, chitin, glucan, fibrin, silk-fibroin, gelatin, agar, hyaluronic acid, chondroitin sulfate, collagen, carrageenan, the mixture of one or more in mosanom and the calcium alginate.
Described solvent is an acetone, ethanol, formic acid, oxolane, N, dinethylformamide, N, the N-dimethylacetylamide, chloroform, carrene, methyl alcohol, ether, dimethyl sulfoxide (DMSO), benzene, carbon tetrachloride, 1, the 2-dichloroethanes, 1, the 1-dichloroethanes, isopropyl alcohol, acetic acid, urea, water, trichloroethanes, 2-methyl cellosolve, 1,1, the 2-trichloro-ethylene, 1, the 2-dimethoxy-ethane, 1,2,3, the 4-tetrahydronaphthalene, cellosolvo, sulfolane, pyrimidine, formamide, n-hexane, chlorobenzene, dioxane, acetonitrile, vinyl ethylene glycol, toluene, hexahydrotoluene, 1, the 2-dichloroethylene, dimethylbenzene, cyclohexane, N-Methyl pyrrolidone, pentane, acetate, methyl phenyl ethers anisole, the 1-propyl alcohol, the 2-propyl alcohol, the 1-butanols, the 2-butanols, amylalcohol, butyl acetate, three fourth MEEs, isopropyl acetate, MEK, cumene, ethyl acetate, Ethyl formate, isobutyl acetate, methyl acetate, 3-methyl isophthalic acid-butanols, methylisobutylketone, 2-methyl isophthalic acid-propyl alcohol, propyl acetate, 1, the 1-di ethyl propyl ether, 1, the 1-dimethoxymethane, 2, the 2-dimethoxy propane, isooctane, isopropyl ether, methyl isopropyl ketone, methyltetrahydrofuran, benzinum, trichloroacetic acid, the mixture of one or more in trifluoroacetic acid and the pyridine.
Described weak polyelectrolyte is weak polycation or weak polyanion; Described weak polycation is one or more the mixture in shitosan, collagen, polymine, polylysine, polypropylene-base ammonia, polypropylene-base amine hydrochlorate and the PDDA; Described weak polyanion is one or more the mixture in sodium alginate, kayexalate, dextran sulfate, chondroitin sulfate, Sodium Polyacrylate, polymethylacrylic acid, heparin, heparin sulfate, albumin, polyglutamic acid, hyaluronic acid and the polyacrylic acid.
Advantage compared with prior art of the present invention is as follows:
Technology of the present invention is simple, by simple alternate group packing technique, has realized the layer structure design of nanometer, submicron-scale at nano-material surface.The static self-assembly modified nano fiber Stability Analysis of Structures, do not destroy the activity of bioactive molecule, and this method can effectively keep the fine structure of the 3 D stereo of nanofiber mould material, this structure has the specific area and the air permeability and good of superelevation, improve the activity of bioactive molecule greatly, can be widely used in aspects such as organizational project, medical dressing, artificial skin, health-care underclothes lining, medicament slow release.
The specific embodiment
Below in conjunction with embodiment, further set forth the present invention.
Embodiment 1
Under 25 ℃ of room temperatures, in stirred tank with the 0.5g cellulose acetate fibre with rotating speed 100rpm/min stirring and dissolving at 9.5g acetone and N, in the N-dimethylacetylamide mixed solvent (weight ratio is 2: 1), obtain mass fraction and be 5% cellulose acetate fibre solution, the electrospinning raw material that obtains is joined in the electrostatic spinning apparatus, at electrostatic pressure is 10 kilovolts, syringe pump flow velocity 0.3ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 6cm, obtains cellulose acetate nanofiber mould material apart from receiving screen.
It is in the shitosan aqueous acetic acid of 0.01mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 5 minutes is used distilled water wash 5 minutes, air blow drying; It is in the 0.01mg/ml dextran sulfate aqueous solution that dried material is immersed concentration, and adsorption equilibrium 5 minutes is used distilled water wash 5 minutes, air blow drying; Alternately repeat above step 2 time, make the weak poly-zwitterion alternate group of biologically active be contained in nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified cellulose acetate nanofiber.
With 5 * 10
4The cartilage cell plant respectively in being covered with static self-assembly modified cellulose acetate nanofiber medical material and static and spin in the 50 μ lDMEM culture plates of cellulose acetate/chitosan composite nano fibre medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 2 days and be respectively 50% and 20%, this shows that the biologically active of the cellulose acetate nanofiber medical material after process is self-assembled modified obviously improves.
Embodiment 2
Under 25 ℃ of room temperatures, in stirred tank with the 1g cellulose with rotating speed 300rpm stirring and dissolving rapid dissolving cellulos after 13.3g 7wt%NaOH/12wt% aqueous solution of urea is chilled to-12 ℃ in advance, obtain mass fraction and be 7% cellulose solution, the electrospinning raw material that obtains is joined in the electrostatic spinning apparatus, at electrostatic pressure is 15 kilovolts, syringe pump flow velocity 0.5ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 8cm, obtains cellulose nano-fibrous mould material apart from receiving screen.
It is in the polyethyleneimine: amine aqueous solution of 1mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 10 minutes is used distilled water wash 7 minutes, air blow drying; It is in the 1mg/ml kayexalate aqueous solution that dried material is immersed concentration, and adsorption equilibrium 10 minutes is used distilled water wash 7 minutes, air blow drying; Alternately repeat above step 7 time, make the weak poly-zwitterion alternate group of biologically active be contained in cellulose nano-fibrous mould material surface, thereby make static self-assembly modified cellulose nano-fibrous bio-medical material.
With 5 * 10
4The human body bronchial epithelial cell plant respectively in the 100 μ lDMEM culture plates that are covered with static self-assembly modified cellulose nano-fibrous medical material and electrostatic spinning fiber element/polymine composite nano fiber medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the adhesion rate of above-mentioned two kinds of medical material pair cells cultivation after 12 hours and be respectively 95% and 50%, this shows that the biologically active of the cellulose nano-fibrous medical material after process is self-assembled modified obviously improves.
Embodiment 3
Under 25 ℃ of room temperatures, in stirred tank with the 1.5g shitosan with rotating speed 500rpm stirring and dissolving in the 15.2g acetate solvate, obtain mass fraction and be 9% chitosan-acetic acid solution, chitosan-acetic acid solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 18 kilovolts, syringe pump flow velocity 0.8ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 8cm, obtains the chitosan nano fiber mould material apart from receiving screen.
It is in the polyacrylic acid aqueous solution of 2mg/ml that the chitosan nano fiber mould material of preparation immerses concentration, and adsorption equilibrium 15 minutes is used distilled water wash 9 minutes, air blow drying; It is in the 2mg/ml collagenic aqueous solution that dried material is immersed concentration, and adsorption equilibrium 15 minutes is used distilled water wash 9 minutes, air blow drying; Alternately repeat above step 12 time, make the weak poly-zwitterion alternate group of biologically active be contained in chitosan nano fiber mould material surface, thereby make the bio-medical material of static self-assembly modified chitosan nano fiber.
With 10
5The human venous endothelial cell plant respectively in being covered with static self-assembly modified chitosan nano fiber medical material and static and spin in the 50 μ lDMEM culture plates of shitosan/collagen composite nano fiber medical material, nutrient solution is PRMI1640 and 5% calf serum, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 60% and 25%, this shows that the biologically active of the chitosan nano fiber medical material after process is self-assembled modified obviously improves.
Embodiment 4
Under 25 ℃ of room temperatures, in stirred tank with 1.5g nylon 6 with rotating speed 700rpm stirring and dissolving in the 13.5g formic acid solvent, obtain mass fraction and be 10% nylon 6 solution, nylon 6 solution are joined in the electrostatic spinning apparatus, at electrostatic pressure is 20 kilovolts, syringe pump flow velocity 1ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 10cm, obtains nylon 6/nanometer fibrous template material apart from receiving screen.
It is in the shitosan aqueous acetic acid of 3mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 20 minutes is used distilled water wash 11 minutes, air blow drying; It is in the 3mg/ml dextran sulfate aqueous solution that dried material is immersed concentration, and adsorption equilibrium 20 minutes is used distilled water wash 11 minutes, air blow drying; Alternately repeat above step 17 time, make the weak poly-zwitterion alternate group of biologically active be contained in nylon 6/nanometer fibrous template material surface, thereby make static self-assembly modified nylon 6/nanometer fiber.
With 10
5The human body bronchial epithelial cell plant respectively in being covered with static self-assembly modified nylon 6/nanometer fiber medical material and static and spin in the 100 μ lDMEM culture plates of nylon 6/ dextran sulfate composite nano fiber medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 65% and 28%, this shows that the biologically active of the nylon 6/nanometer fiber medical material after process is self-assembled modified obviously improves.
Embodiment 5
Under 25 ℃ of room temperatures, in stirred tank with the 2g ethylene-vinyl alcohol copolymer with rotating speed 900rpm stirring and dissolving in 14.7g isopropyl alcohol and water mixed solvent (weight ratio is 2: 1), obtain mass fraction and be 12% ethylene-vinyl alcohol copolymer solution, ethylene-vinyl alcohol copolymer solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 22 kilovolts, syringe pump flow velocity 1.5ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 12cm, obtains ethylene-vinyl alcohol copolymer nanofiber mould material apart from receiving screen.
It is in the shitosan aqueous acetic acid of 4mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 25 minutes is used distilled water wash 13 minutes, air blow drying; It is in the 4mg/ml polyglutamic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 25 minutes is used distilled water wash 13 minutes, air blow drying; Alternately repeat above step 21 time, make the weak poly-zwitterion alternate group of biologically active be contained in ethylene-vinyl alcohol copolymer nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified ethylene-vinyl alcohol copolymer nanofiber.
With 10
5The cerebral cortex nerve cell plant respectively in being covered with static self-assembly modified ethylene-vinyl alcohol copolymer nanofiber medical material and static and spin in the 150 μ lDMEM culture plates of ethylene-vinyl alcohol copolymer/polyglutamic acid composite nano fiber medical material, nutrient solution is 15% calf serum, 1% penicillin/streptomycin, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 65% and 25%, the adhesion rate that pair cell was cultivated after 24 hours is respectively 95% and 60%, this shows that the biologically active of the ethylene-vinyl alcohol copolymer nanofiber medical material after process is self-assembled modified obviously improves.
Embodiment 6
Under 25 ℃ of room temperatures, in stirred tank with the 3g polyethylene terephthalate with rotating speed 1000rpm stirring and dissolving at 17g1, in the 1-dichloroethanes solvent, obtain mass fraction and be 15% polyethylene terephthalate solution, polyethylene terephthalate solution is joined in the electrostatic spinning apparatus, is 25 kilovolts at electrostatic pressure, syringe pump flow velocity 1.8ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 12cm, obtains polyphenyl dioctyl phthalate second diester nanofiber mould material apart from receiving screen.
It is in the polypropylene-base ammonia spirit of 4.5mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 30 minutes is used distilled water wash 15 minutes, air blow drying; It is in the 4.5mg/ml polyacrylic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 30 minutes is used distilled water wash 15 minutes, air blow drying; Alternately repeat above step 25 time, make the weak poly-zwitterion alternate group of biologically active be contained in polyphenyl dioctyl phthalate second diester nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified polyphenyl dioctyl phthalate second diester nanofiber.
With 10
5Gegenbaur's cell plant respectively in being covered with static self-assembly modified polyethylene terephthalate nanofiber medical material and static and spin in the 100 μ lDMEM culture plates of polyethylene terephthalate/polypropylene acid composite nano fiber medical material, nutrient solution is 10% calf serum, 60IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 65% and 30%, the adhesion rate that pair cell was cultivated after 24 hours is respectively 95% and 50%, this shows that the biologically active of the polyethylene terephthalate nanofiber medical material after process is self-assembled modified obviously improves.
Embodiment 7
Under 25 ℃ of room temperatures, in stirred tank with the 3g polystyrene with rotating speed 1200rpm stirring and dissolving in the 13.7g tetrahydrofuran solvent, obtain mass fraction and be 18% polystyrene solution, polystyrene solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 25 kilovolts, syringe pump flow velocity 2ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 13cm, obtains pipe/polyhenylethylene nano fibrous template material apart from receiving screen.
It is in the polypropylene-base amine hydrochlorate aqueous solution of 5.5mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 35 minutes is used distilled water wash 17 minutes, air blow drying; It is in the 5.5mg/ml polyacrylic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 35 minutes is used distilled water wash 17 minutes, air blow drying; Alternately repeat above step 29 time, make the weak poly-zwitterion alternate group of biologically active be contained in pipe/polyhenylethylene nano fibrous template material surface, thereby make the bio-medical material of static self-assembly modified pipe/polyhenylethylene nano fiber.
With 5 * 10
5Fibroblast plant respectively in being covered with static self-assembly modified pipe/polyhenylethylene nano fiber medical material and static and spin in the 150 μ lDMEM culture plates of polystyrene/polypropylene-base amine hydrochlorate composite nano fiber medical material, nutrient solution is 20% calf serum, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 50% and 25%, the adhesion rate that pair cell was cultivated after 24 hours is respectively 90% and 50%, this shows that the biologically active of the pipe/polyhenylethylene nano fiber medical material after process is self-assembled modified obviously improves.
Embodiment 8
Under 25 ℃ of room temperatures, in stirred tank with the 3g polyacrylonitrile with rotating speed 1500rpm stirring and dissolving in the 12g dimethyl sulfoxide solvent, obtain mass fraction and be 20% polyacrylonitrile solution, polyacrylonitrile solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 30 kilovolts, syringe pump flow velocity 2.5ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 14cm, obtains polyacrylonitrile nano fibrous template material apart from receiving screen.
It is in the shitosan aqueous acetic acid of 6mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 40 minutes is used distilled water wash 19 minutes, air blow drying; It is in the 6mg/ml hyaluronic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 40 minutes is used distilled water wash 19 minutes, air blow drying; Alternately repeat above step 33 time, make the weak poly-zwitterion alternate group of biologically active be contained in polyacrylonitrile nano fibrous template material surface, thereby make the bio-medical material of static self-assembly modified polyacrylonitrile nano fiber.
With 5 * 10
5The cerebral cortex nerve cell plant respectively in being covered with static self-assembly modified polyacrylonitrile nano fiber medical material and static and spin in the 200 μ lDMEM culture plates of polyacrylonitrile/chitosan composite nano fibre medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 2 days and be respectively 70% and 30%, this shows that the biologically active of the polyacrylonitrile nano fiber medical material after process is self-assembled modified obviously improves.
Embodiment 9
Under 25 ℃ of room temperatures, in stirred tank with the 3g polycaprolactone with rotating speed 1800rpm stirring and dissolving in the 10.6g benzene solvent, obtain mass fraction and be 22% polycaprolactone solution, polycaprolactone solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 30 kilovolts, syringe pump flow velocity 3ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 16cm, obtains polycaprolactone nanofiber mould material apart from receiving screen.
It is in the PDDA aqueous solution of 7mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 45 minutes is used distilled water wash 21 minutes, air blow drying; It is in the 7mg/ml polyacrylic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 45 minutes is used distilled water wash 21 minutes, air blow drying; Alternately repeat above step 37 time, make the weak poly-zwitterion alternate group of biologically active be contained in polycaprolactone nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified polycaprolactone nanofiber.
With 5 * 10
5Lymphocyte plant respectively in being covered with static self-assembly modified polycaprolactone nanofiber medical material and static and spin in the 200 μ lDMEM culture plates of polycaprolactone/polyacrylic acid composite nano fiber medical material, nutrient solution is PRMI1640 and 10% calf serum, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 80% and 35%, the adhesion rate that pair cell was cultivated after 48 hours is respectively 95% and 50%, this shows that the biologically active of the polycaprolactone nanofiber medical material after process is self-assembled modified obviously improves.
Embodiment 10
Under 25 ℃ of room temperatures, in stirred tank with 3g polyurethane with rotating speed 2000rpm stirring and dissolving at 9g 1,1, in the 2-trichloroethylene solvent, obtain mass fraction and be 25% polyurethane solutions, polyurethane solutions is joined in the electrostatic spinning apparatus, at electrostatic pressure is 32 kilovolts, syringe pump flow velocity 3.2ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 16cm, obtains the polyurethane nanofiber mould material apart from receiving screen.
It is in the polypropylene-base ammonia spirit of 8mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 50 minutes is used distilled water wash 24 minutes, air blow drying; It is in the 8mg/ml polyacrylic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 50 minutes is used distilled water wash 24 minutes, air blow drying; Alternately repeat above step 41 time, make the weak poly-zwitterion alternate group of biologically active be contained in polyurethane nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified polyurethane nanofiber.
With 5 * 10
5The human body bronchial epithelial cell plant respectively in being covered with static self-assembly modified polyurethane nanofiber medical material and static and spin in the 200 μ lDMEM culture plates of polyurethane/polypropylene-base ammonia composite nano fiber medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 4 days and be respectively 85% and 38%, this shows that the biologically active of the polyurethane nanofiber medical material after process is self-assembled modified obviously improves.
Embodiment 11
Under 25 ℃ of room temperatures, in stirred tank with the 3g gelatin with rotating speed 2200rpm stirring and dissolving in the 7.7g formic acid solvent, obtain mass fraction and be 28% gelatin solution, gelatin solution is joined in the electrostatic spinning apparatus, at electrostatic pressure is 35 kilovolts, syringe pump flow velocity 3.6ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 18cm, obtains the gelatine nano fiber mould material apart from receiving screen.
It is in the collagen solution of 9mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 55 minutes is used distilled water wash 27 minutes, air blow drying; It is in the 9mg/ml sodium alginate aqueous solution that dried material is immersed concentration, and adsorption equilibrium 55 minutes is used distilled water wash 27 minutes, air blow drying; Alternately repeat above step 45 time, make the weak poly-zwitterion alternate group of biologically active be contained in gelatine nano fiber mould material surface, thereby make the bio-medical material of static self-assembly modified gelatine nano fiber.
With 5 * 10
5The cartilage cell plant respectively in being covered with static self-assembly modified gelatine nano fiber medical material and static and spin in the 200 μ lDMEM culture plates of gelatin/sodium alginate composite nano fiber medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 85% and 40%, the adhesion rate that pair cell was cultivated after 48 hours is respectively 95% and 60%, this shows that the biologically active of the gelatine nano fiber medical material after process is self-assembled modified obviously improves.
Embodiment 12
Under 25 ℃ of room temperatures, in stirred tank with the 3g polyurethane with rotating speed 2500rpm stirring and dissolving at 7g oxolane and N, in the dinethylformamide mixed solvent (weight ratio is 2: 1), obtain mass fraction and be 30% polyurethane solution, polyurethane solution is joined in the electrostatic spinning apparatus, is 40 kilovolts at electrostatic pressure, syringe pump flow velocity 4ml/h, spinning head apart from carrying out electrostatic spinning under the condition of 20cm, obtains polyurethane nanofiber mould material apart from receiving screen.
It is in the polypropylene-base ammonia spirit of 10mg/ml that the nanofiber mould material of preparation immerses concentration, and adsorption equilibrium 60 minutes is used distilled water wash 30 minutes, air blow drying; It is in the 10mg/ml polyacrylic acid aqueous solution that dried material is immersed concentration, and adsorption equilibrium 60 minutes is used distilled water wash 30 minutes, air blow drying; Alternately repeat above step 50 time, make the weak poly-zwitterion alternate group of biologically active be contained in polyurethane nanofiber mould material surface, thereby make the bio-medical material of static self-assembly modified polyurethane nanofiber.
With 5 * 10
5The cerebral cortex nerve cell plant respectively in being covered with static self-assembly modified polyurethane nanofiber medical material and static and spin in the 200 μ lDMEM culture plates of polyurethane/polypropylene-base ammonia composite nano fiber medical material, nutrient solution is 10% calf serum, 50IU/ml penicillin and 50 μ g/ml streptomysins, culture plate is placed 37 ℃, 5%CO
2In the incubator, change liquid every other day, record the rate of increase of above-mentioned two kinds of medical material pair cells cultivation after 3 days and be respectively 90% and 40%, the adhesion rate that pair cell was cultivated after 48 hours is respectively 95% and 70%, this shows that the biologically active of the polyurethane nanofiber medical material after process is self-assembled modified obviously improves.
Claims (4)
1. the method for preparing biomedical material of a static self-assembly modified nano fiber is characterized in that, concrete steps are:
The first step: at room temperature, in stirred tank with spinning with water-insoluble macromolecular polymer with rotating speed 100-2500 rev/min stirring and dissolving in solvent, obtaining mass fraction is the electrospinning raw material of 5-30%;
Second step: the electrospinning raw material that the first step is obtained joins in the electrostatic spinning apparatus, at electrostatic pressure is the 10-40 kilovolt, the syringe pump flow velocity is 0.3-4ml/h, and spinning head for carrying out electrostatic spinning under the condition of 6-20cm, obtains the nanofiber mould material apart from the receiving screen distance;
The 3rd step: the nanofiber mould material of the second step preparation is immersed in the weak polyelectrolyte aqueous solution that has electric charge in contrast that concentration is 0.01mg/ml-10g/ml, adsorption equilibrium 5-60 minute, with distilled water wash 5-30 minute, air blow drying; Dried material is immersed in the weak polyelectrolyte aqueous solution that has electric charge in contrast that concentration is 0.01mg/ml-10g/ml, adsorption equilibrium 5-60 minute, with distilled water wash 5-30 minute, air blow drying; Alternately repeat above step 2-50 time, make the weak poly-zwitterion of biologically active, thereby make the bio-medical material of static self-assembly modified nano fiber alternately attached to nanofiber mould material surface.
2. preparation method as claimed in claim 1, it is characterized in that described water-insoluble macromolecular polymer is a cellulose acetate, cellulose, shitosan, ethylene-vinyl alcohol copolymer, nylon 6, polystyrene, polymethyl methacrylate, polyisobutene, polyacrylonitrile, polycaprolactone, polyvinyl acetate, polyethylene terephthalate, polypropylene, polyurethane, polyurethane, Kynoar, Merlon, epoxy resin, polysiloxanes, chitin, glucan, fibrin, silk-fibroin, gelatin, agar, hyaluronic acid, chondroitin sulfate, collagen, carrageenan, the mixture of one or more in mosanom and the calcium alginate.
3. preparation method as claimed in claim 1, it is characterized in that, described solvent is an acetone, ethanol, formic acid, oxolane, N, dinethylformamide, N, the N-dimethylacetylamide, chloroform, carrene, methyl alcohol, ether, dimethyl sulfoxide (DMSO), benzene, carbon tetrachloride, 1, the 2-dichloroethanes, 1, the 1-dichloroethanes, isopropyl alcohol, acetic acid, urea, water, trichloroethanes, 2-methyl cellosolve, 1,1, the 2-trichloro-ethylene, 1, the 2-dimethoxy-ethane, 1,2,3, the 4-tetrahydronaphthalene, cellosolvo, sulfolane, pyrimidine, formamide, n-hexane, chlorobenzene, dioxane, acetonitrile, vinyl ethylene glycol, toluene, hexahydrotoluene, 1, the 2-dichloroethylene, dimethylbenzene, cyclohexane, the N methyl pyrrolidone, pentane, acetate, methyl phenyl ethers anisole, the 1-propyl alcohol, the 2-propyl alcohol, the 1-butanols, the 2-butanols, amylalcohol, butyl acetate, three fourth MEEs, isopropyl acetate, MEK, cumene, ethyl acetate, Ethyl formate, isobutyl acetate, methyl acetate, 3-methyl isophthalic acid-butanols, methylisobutylketone, 2-methyl isophthalic acid-propyl alcohol, propyl acetate, 1, the 1-di ethyl propyl ether, 1, the 1-dimethoxymethane, 2, the 2-dimethoxy propane, isooctane, isopropyl ether, methyl isopropyl ketone, methyltetrahydrofuran, benzinum, trichloroacetic acid, the mixture of one or more in trifluoroacetic acid and the pyridine.
4. preparation method as claimed in claim 1 is characterized in that, described weak polyelectrolyte is weak polycation or weak polyanion; Described weak polycation is one or more the mixture in shitosan, collagen, polymine, polylysine, polypropylene-base ammonia, polypropylene-base amine hydrochlorate and the PDDA; Described weak polyanion is one or more the mixture in sodium alginate, kayexalate, dextran sulfate, chondroitin sulfate, Sodium Polyacrylate, polymethylacrylic acid, heparin, heparin sulfate, albumin, polyglutamic acid, hyaluronic acid and the polyacrylic acid.
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