CN103993426A - Preparing method of polyethylene terephthalate-keratin composite nano fiber tubular materials - Google Patents
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Abstract
The invention relates to a preparing method of polyethylene terephthalate-keratin composite nano fiber tubular materials. The preparing method comprises the steps that polyethylene terephthalate is dissolved in a solvent, dissolution by stirring is carried out, and a polyethylene terephthalate solution is obtained; keratin is dissolved in a solvent, dissolution by stirring is carried out, and a keratin solution is obtained; and the polyethylene terephthalate solution and the keratin solution are mixed according to the weight ratio of 95 : 5-5 : 95, blending by stirring is carried out, a spinning solution is obtained, then electrostatic spinning is carried out, a cylindrical collecting roller is used as a receiver, and the polyethylene terephthalate-keratin composite nano fiber tubular materials are obtained. According to the polyethylene terephthalate-keratin composite nano fiber tubular materials, the good biological activity of the keratin is maintained, the good tenacity and the good mechanical strength of the polyethylene terephthalate are achieved, meanwhile, the materials have the advantages that inner diameter size can be adjusted, the preparing method is simple, and the materials are expected to be used in fields such as surgical dressing.
Description
Technical field
The invention belongs to the preparation field of nano-fiber tubular material, particularly a kind of preparation method of polyethylene terephthalate-keratin composite nano fiber tubular material.
Background technology
Keratin is a kind of insoluble fibrous animal protein, is the structural proteins of ectoderm cell, is extensively present in animal skin and cutaneous appendage, as hair, hoof, shell, pawl, angle, scale etc.Large quantity research in recent years shows, keratin is a kind of good biocompatibility and the high-quality bio-medical material that do not repelled by immunity of organism, has broad application prospects.The most outstanding, through the keratin in the sources such as wool is carried out to determined amino acid sequence discovery, it contains same Arg-Gly-Asp (RGD) tripeptide sequence.This tripeptide sequence is acknowledged as effective binding site of realizing Cell binding in extracellular matrix, has the function that promotes cell absorption.Therefore, a large amount of basic research and animal experiment studies about keratin based biomaterial have been carried out both at home and abroad, and at wound dressing [Wound Repair and Regeneration, 2012,20:236-242], artifical bone [Journal of Bioactive and Compatible Polymers, 2013,28:141-153] and the neural aspects such as [Biomaterials34 (2013) 5907-5914] of repairing all obtained good result, existing portioned product is applied to clinical.
But existing studies have shown that, because keratic molecular weight is lower, the membrane material that single keratin material is made is conventionally more crisp, and mechanical strength is not high, and its application is restricted.Therefore, at present, most keratin based biomaterial often adopts keratin and natural polymer or artificial macromolecule compound, so both can keep improving the poor deficiency of single keratin mechanical property, as fibroin albumen [Biomacromolecules, 2008, 9, 1299 – 1305], PVA[Advances in Materials Science and Engineering, 2014, Article ID163678], PLGA[Journal of Bioactive and Compatible Polymers, 2013, 28:141-153], PLLA[Biomed.Mater.2013, 8:1-9] etc.
Polyethylene terephthalate, be commonly called as dacron, terylene, reach gram synthetic fibre etc., it is common a kind of resin in life, have that physical and mechanical properties is strong, creep-resistant property is high, endurance and the advantage such as crocking resistance is good, good stability of the dimension, be applied to clinically at present artificial blood vessel's preparation, for example adopt the artificial blood vessel of polyster fibre braiding can treat aortic aneurysm, art etc. is changed in aortostenosis and superior and inferior vena cava excision.But the inanimate object of polyethylene terephthalate materials own is active, lacks cell recognition signal site, is unfavorable for histiocytic adhesion and the growths such as endothelial cell.
Electrostatic spinning technique refers to and utilizes high voltage electric field environment to make polymer dope form charged injection stream, and this injection stream is elongated under electric field action, and solvent evaporates finally forms the nanofiber of certain form on receiving system.In recent ten years, this technology has become one of effective way of preparing nano-fiber material.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of preparation method of polyethylene terephthalate-keratin composite nano fiber tubular material, the tubular material that this invention prepares has kept the biologically active that keratin is good, possess again the good toughness of polyethylene terephthalate and mechanical strength, there is the advantages such as internal diameter size is adjustable, preparation method is simple simultaneously, be expected to for fields such as artificial blood vessels.
The preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material of the present invention, comprising:
(1) polyethylene terephthalate is dissolved in solvent, stirring and dissolving, obtains polyethylene terephthalate solution;
(2) keratin is dissolved in solvent, stirring and dissolving, obtains keratin solution;
(3) by polyethylene terephthalate solution with keratin solution in mass ratio for 95:5-5:95 mixes, stir and evenly mix, obtain spinning solution, then carry out electrostatic spinning, and take cylindrical collecting drum as receiver, obtain polyethylene terephthalate-keratin composite nano fiber tubular material.
In described step (1), the concentration of polyethylene terephthalate solution is 5-20mg/ml.
In described step (2), keratin molecule amount is 3-300kDa.
Described keratin can be to adopt the whole bag of tricks of open report at present to extract the keratin of preparation, comprises reducing process, and oxidizing process and Hydrolyze method etc., can be also above-mentioned keratic derivative, as carboxymethyl keratin.
Described keratin can extract and obtain from human or animal's hairs such as human hair, wool, poultry feather, ox hairs, can be also to derive from the animal body that other have been reported.
In described step (2), the concentration of keratin solution is 10-20mg/ml.
In described step (1), (2), solvent is one or more in hexafluoroisopropanol, DMF, DMA, 1-METHYLPYRROLIDONE, carrene, dichloroethanes, chloroform, oxolane.
The preferred hexafluoroisopropanol of described solvent.
In described step (3), spinning solution gross mass percentage concentration is 5-20%.
In described step (3), in spinning solution, add other reagent that can be dissolved in this mixed solution system, comprising: one or more in medicine, inorganic antiseptic, organic antiseptic, anticoagulant.
In described step (3), electrostatic spinning process parameter is: voltage 13-35kV, and receiving range 8-22cm, spinning speed is 0.3-1.5ml/h, collecting drum slewing rate 60-130rpm, spinneret orifice internal diameter is 0.7-0.9mm, spinning temperature 20-30 ℃, spinning humidity 45-65%.
Columniform collecting drum in described step (3), diameter dimension can be 0.6-2 centimetre, or regulates according to the internal diameter size demand of required tubular material; The axle of collecting drum is perpendicular to spinning-nozzle opening direction, and does uniform rotation.
beneficial effect
The present invention proposes keratin and polyethylene terephthalate is compound, and the method that adopts electrostatic spinning prepares nanofiber type tubular material, to improving the deficiency that single keratin fiber material mechanical performance is lower and polyethylene terephthalate biologically active is low;
The tubular material porosity that the present invention obtains is high, has good biologically active, and good toughness and mechanical strength have the advantages such as internal diameter size is adjustable, preparation method is simple simultaneously, are expected to for fields such as artificial blood vessels.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read the content of the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
The wool of take extracts keratin as raw material adopts reducing process, and concrete grammar is: take 5g wool, the benzinum of take is removed wool surfaces grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans wool, air-dry.Above-mentioned wool is added in the mercaptoethanol of 100ml, 1mol/l, with NaOH, regulate pH approximately 10.0, then at 40 ℃ of conditioned response 12h.After reaction finishes, reactant liquor is collected to (being designated as filtrate 1), will react remaining wool solid filtering simultaneously and collect, continue under 40 ℃ of conditions, to process 2h with the Tris alkali lye of 100ml, 0.1mol/l, then solids removed by filtration, collects filtrate (being designated as filtrate 2).. filtrate 1 and 2 is merged, in 6000rpm centrifugal 5 minutes, then use the bag filter dialysis 36h of molecular cut off 40,000, finally dislysate is added liquid nitrogen to carry out fast frozen, then insert the dry keratin powder that obtains dry state of freeze drier.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.15g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 15mg/ml.0.2g keratin solid is dissolved in to 10ml hexafluoroisopropanol, under room temperature, carries out magnetic agitation to dissolving completely, obtain the keratin solution that concentration is 20mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 50:50 with keratin solution, and spinning solution total concentration is 15%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 15kV, receiving range 12cm, and spinning speed is 0.5ml/h, spinneret orifice internal diameter is 0.7mm, 25 ℃ of spinning temperatures, spinning humidity 50%.The cylindrical collecting drum of diameter 1.0cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 100rpm, through the collection of 4 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 2
The human hair of take is extracted keratin as raw material adopts reducing process, and concrete grammar is: take 5g human hair, the benzinum of take is removed human hair surface grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans human hair, air-dry.Above-mentioned human hair is added in the solution that 200ml water mixes with 5g sodium pyrosulfite, with NaOH, regulate pH approximately 10.0, then at 60 ℃ of conditioned response 5h.After reaction finishes, reactant liquor is collected to (being designated as filtrate 1), will react remaining human hair solid filtering simultaneously and collect, continue under 40 ℃ of conditions, to process 2h with the Tris alkali lye of 100ml, 0.1mol/l, then solids removed by filtration, collects filtrate (being designated as filtrate 2).. filtrate 1 and 2 is merged, in 6000rpm centrifugal 5 minutes, then use the bag filter dialysis 36h of molecular cut off 8,000, finally dislysate is added liquid nitrogen to carry out fast frozen, then insert the dry keratin powder that obtains dry state of freeze drier.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.2g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 20mg/ml.0.15g keratin solid is dissolved in hexafluoroisopropanol, under room temperature, carries out magnetic agitation to dissolving completely, obtain the keratin solution that concentration is 15mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 20:80 with keratin solution, and spinning solution total concentration is 20%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 13kV, receiving range 10cm, and spinning speed is 0.8ml/h, spinneret orifice internal diameter is 0.7mm, 30 ℃ of spinning temperatures, spinning humidity 65%.The cylindrical collecting drum of diameter 0.6cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 60rpm, through the collection of 4 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 3
The wool of take extracts keratin as raw material adopts oxidizing process, and concrete grammar is: take 5g wool, the benzinum of take is removed wool surfaces grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans wool, air-dry.To adding 100ml deionized water and 5ml, 30% hydrogen peroxide in above-mentioned wool, then at 100 ℃ of conditioned response 2h.After reaction finishes, reactant liquor is collected to (being designated as filtrate 1), will react remaining wool solid filtering simultaneously and collect, continue under 40 ℃ of conditions, to process 2h with the Tris alkali lye of 100ml, 0.1mol/l, then solids removed by filtration, collects filtrate (being designated as filtrate 2).. filtrate 1 and 2 is merged, in 6000rpm centrifugal 5 minutes, then use the bag filter dialysis 36h of molecular cut off 8,000, finally dislysate is added liquid nitrogen to carry out fast frozen, then insert the dry keratin powder that obtains dry state of freeze drier.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.05g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 5mg/ml.0.1g keratin solid is dissolved in 10ml hexafluoroisopropanol, under room temperature, carries out magnetic agitation to dissolving completely, obtain the keratin solution that concentration is 10mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 95:5 with keratin solution, and spinning solution total concentration is 5%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 13kV, receiving range 8cm, and spinning speed is 1.5ml/h, spinneret orifice internal diameter is 0.9mm, 20 ℃ of spinning temperatures, spinning humidity 45%.The cylindrical collecting drum of diameter 1.5cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 110rpm, through the collection of 3 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 4
The wool of take extracts keratin as raw material adopts oxidizing process, and concrete grammar is: take 5g wool, the benzinum of take is removed wool surfaces grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans wool, air-dry.To adding 100ml, 4% Peracetic acid in above-mentioned wool, then at 60 ℃ of conditioned response 5h.After reaction finishes, reactant liquor is collected to (being designated as filtrate 1), will react remaining wool solid filtering simultaneously and collect, continue under 40 ℃ of conditions, to process 2h with the Tris alkali lye of 100ml, 0.1mol/l, then solids removed by filtration, collects filtrate (being designated as filtrate 2).. filtrate 1 and 2 is merged, in 6000rpm centrifugal 5 minutes, then use the bag filter dialysis 36h of molecular cut off 8,000, finally dislysate is added liquid nitrogen to carry out fast frozen, then insert the dry keratin powder that obtains dry state of freeze drier.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.15g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 15mg/ml.0.2g keratin solid is dissolved in 10ml hexafluoroisopropanol, under room temperature, carries out magnetic agitation to dissolving completely, obtain the keratin solution that concentration is 20mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 50:50 with keratin solution, and spinning solution total concentration is 15%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 35kV, receiving range 22cm, and spinning speed is 1.5ml/h, spinneret orifice internal diameter is 0.9mm, 20 ℃ of spinning temperatures, spinning humidity 55%.The cylindrical collecting drum of diameter 1.0cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 130rpm, through the collection of 3 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 5
The wool of take extracts keratin as raw material adopts reducing process, and keratin is carried out to derivatization modification.Concrete grammar is: take 5g wool, the benzinum of take is removed wool surfaces grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans wool, air-dry.Above-mentioned wool is added in the mercaptoethanol of 100ml, 1mol/l, with NaOH, regulate pH approximately 10.0, then at 40 ℃ of conditioned response 12h.Then, with acetic acid, above-mentioned reactant liquor pH is adjusted to 7.5, add wherein the NaBrO3 solution of 100ml, 0.33mol/l, continue at ambient temperature to stir 24h.After reaction finishes, solids removed by filtration thing, collects filtrate and use the bag filter of molecular cut off 8,000 36h that dialyses, and finally dislysate is added liquid nitrogen to carry out fast frozen, then inserts freeze drier and is dried the carboxymethyl keratin powder that obtains dry state.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.15g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 15mg/ml.0.1g carboxymethyl keratin solid is dissolved in 10ml hexafluoroisopropanol, carries out magnetic agitation to dissolving completely under room temperature, obtaining concentration is the carboxymethyl keratin solution of 10mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 70:30 with keratin solution, and spinning solution total concentration is 10%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 15kV, receiving range 12cm, and spinning speed is 1.0ml/h, spinneret orifice internal diameter is 0.7mm, 15 ℃ of spinning temperatures, spinning humidity 55%.The cylindrical collecting drum of diameter 1.0cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 100rpm, through the collection of 4 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 6
The human hair of take is extracted keratin as raw material adopts reducing process, and keratin is carried out to derivatization modification.Concrete grammar is: take 5g human hair, the benzinum of take is removed human hair surface grease as solvent adopts Soxhlet extraction process, then with ethanol, cleans human hair, air-dry.Above-mentioned human hair is added in the solution that 200ml water mixes with 5g sodium pyrosulfite, with NaOH, regulate pH approximately 10.0, then at 60 ℃ of conditioned response 5h.Then, with acetic acid, above-mentioned reactant liquor pH is adjusted to 8.5, add wherein 8.0g iodoacetic acid, continue at ambient temperature to stir 6h.After reaction finishes, solids removed by filtration thing, collects filtrate and use the bag filter of molecular cut off 8,000 36h that dialyses, and finally dislysate is added liquid nitrogen to carry out fast frozen, then inserts freeze drier and is dried the carboxymethyl keratin powder that obtains dry state.
The preparation of polyethylene terephthalate-keratin composite nano-fiber membrane: the polyethylene terephthalate that is 0.15g by quality is dissolved in 10ml hexafluoroisopropanol, under room temperature, carry out magnetic agitation to dissolving completely, obtaining concentration is the polyethylene terephthalate solution of 15mg/ml.0.2g carboxymethyl keratin solid is dissolved in to 10ml hexafluoroisopropanol, carries out magnetic agitation to dissolving completely under room temperature, obtaining concentration is the carboxymethyl keratin solution of 20mg/ml.Above-mentioned polyethylene terephthalate is mixed according to the ratio of mass ratio 50:50 with keratin solution, and spinning solution total concentration is 12%, and magnetic agitation is to mixing.Above-mentioned gained polyethylene terephthalate/keratin solution is carried out to electrostatic spinning, and electrostatic spinning parameter is voltage 15kV, receiving range 12cm, and spinning speed is 1.2ml/h, spinneret orifice internal diameter is 0.7mm, 15 ℃ of spinning temperatures, spinning humidity 55%.The cylindrical collecting drum of diameter 1.0cm of take is that receiver receives and sprays the nano silk producing, and the slewing rate of collecting drum is 100rpm, through the collection of 4 hours, obtains keratin-polyethylene terephthalate nano-fiber tubular material.
Embodiment 7
According to the preparation method described in embodiment 1, prepare keratin-polyethylene terephthalate nano-fiber tubular material, measure by analysis, the fracture strength of nano-fiber material under drying regime is 5.2MPa, and elongation at break is 30%.Take vascular endothelial cell as cell model, adopt sem observation cell to stick form on material, and by mtt assay, the propagation behavior of cell is evaluated, result shows, endothelial cell shows good stick form and propagation behavior on nano-fiber material, compares and has significant difference with single polyethylene terephthalate nano-fiber material.According to the preparation method described in embodiment 1, the nano-fiber material of preparing with single keratin under drying regime fragility by force, easily broken.
Claims (10)
1. a preparation method for polyethylene terephthalate-keratin composite nano fiber tubular material, comprising:
(1) polyethylene terephthalate is dissolved in solvent, stirring and dissolving, obtains polyethylene terephthalate solution;
(2) keratin is dissolved in solvent, stirring and dissolving, obtains keratin solution;
(3) by polyethylene terephthalate solution with keratin solution in mass ratio for 95:5-5:95 mixes, stir and evenly mix, obtain spinning solution, then carry out electrostatic spinning, and take cylindrical collecting drum as receiver, obtain polyethylene terephthalate-keratin composite nano fiber tubular material.
2. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: in described step (1), the concentration of polyethylene terephthalate solution is 5-20mg/ml.
3. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: in described step (2), keratin molecule amount is 3-300kDa.
4. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: in described step (2), the concentration of keratin solution is 10-20mg/ml.
5. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, it is characterized in that: in described step (1), (2), solvent is hexafluoroisopropanol, N, one or more in dinethylformamide, DMA, 1-METHYLPYRROLIDONE, carrene, dichloroethanes, chloroform, oxolane.
6. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 5, is characterized in that: described solvent is hexafluoroisopropanol.
7. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: in described step (3), spinning solution gross mass percentage concentration is 5-20%.
8. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: in described step (3), in spinning solution, add one or more in medicine, inorganic antiseptic, organic antiseptic, anticoagulant.
9. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, it is characterized in that: in described step (3), electrostatic spinning process parameter is: voltage 13-35kV, receiving range 8-22cm, spinning speed is 0.3-1.5ml/h, collecting drum slewing rate 60-130rpm, spinneret orifice internal diameter is 0.7-0.9mm, spinning temperature 20-30 ℃, spinning humidity 45-65%.
10. the preparation method of a kind of polyethylene terephthalate-keratin composite nano fiber tubular material according to claim 1, is characterized in that: columniform collecting drum in described step (3), and diameter dimension is 0.6-2 centimetre; The axle of collecting drum is perpendicular to spinning-nozzle opening direction, and does uniform rotation.
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Application publication date: 20140820 |