CN103668782B - The electrostatic spinning preparation method of the bionical Bruch ' s of ultra-thin porous composite nano fiber film - Google Patents
The electrostatic spinning preparation method of the bionical Bruch ' s of ultra-thin porous composite nano fiber film Download PDFInfo
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Abstract
The invention belongs to bio-medical engineering material field, disclose the preparation method of the bionical Bruch ' s of a kind of ultra-thin porous composite nano fiber film, raw material is dissolved in to organic solvent and obtains electrostatic spinning solution, it is carried out to electrostatic spinning and obtain composite nano-fiber membrane; Obtain after drying the bionical Bruch ' s of described ultra-thin porous composite nano fiber film, wherein, this film comprises the bionical Bruch ' s of polycaprolactone nanofiber film, tussah silk fibroin/polycaprolactone composite nano fiber bionical Bruch ' s film or the bionical Bruch ' s of tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber film. The product that the present invention prepares can be used as the carrier of transplantation of retinal pigment epithelium cells.
Description
Technical field
The invention belongs to bio-medical engineering material field, relate to particularly a kind of prepared by electrostatic spinning technique ultra-thin, many that applyBionical Bruch ' the s of hole composite nano fiber film.
Background technology
AMD (AMD) is the great blinding illness in eye of China, and its Etiological is retinal pigment epitheliumThe layer structure of (RetinalPigmentEpithelium, RPE) or function extremely and finally cause retinal light injury photoreceptorApoptosis and then cause neural retina that irreversible sex change occurs, the radical cure of still having no idea at present. It is to treat such that RPE transplantsOne of the most promising method of disease. Preparing desirable RPE cell transplantation carrier becomes the key of this technology. Natural RPECell is positioned on Bruch ' s film, and Bruch ' s film, between neural retina and choroid, is the about 2-4 μ of thick layer m, porousStaggered network structure film. Bruch ' s film not only provides the microenvironment of growth for RPE cell, and to RPE cell functionPerformance play very important effect. Utilize external bionical structure Bruch ' the s film of biomaterial, for RPE cell provides transplantingCarrier has very tempting prospect.
PCL is in recent years at the synthetic macromolecule polyester of the wide concerned one of technical field of biological material, have good mechanical property and andPliability, food and drug administration (FDA) is because of its good biocompatibility, biodegradability, the spy such as nontoxicPoint, ratifies it as the bio-medical material that can be used for human body. But it is strong-hydrophobicity material, degraded slowly and lacked cell and knewOther site, has limited its large-scale application. Tussah silk peptide is the biodegradable natural height of one of being produced by wild silkworm-tussahMolecular material, its amino acid composition is different from bombyx mori silk fibroin, except being rich in alanine, glycine and serine (approximately 80%),The distinctive arginine of tussah silk peptide, glycine and asparatate (RGD) tripeptide sequence have been given its good cellular affinity.RGD sequence is as cell membrane integrin receptor mediated cell and material adhesion and signal transmission. Tussah silk peptide is as rare severalThe native protein that contains RGD block, is more and more subject to researcher's favor. Gelatin is the hydrolysate of collagen, is rich in hydrophilyAmino acid, has extremely strong hydrophily. In addition, compared with collagen, gelatin no antigen, be easy to absorb, often carried out modification peopleGeosynthetics.
At present, also there are some reports to utilize synthetic material, as polyurethane (PU), polycaprolactone (PCL), poly-third hands overEster is lactide (PLGA) or natural biologic material, and as collagen, bombyx mori silk fibroin, although shitosan synthetic material has wellPlasticity, the micro-structural of material, mechanical property, form etc. also can design as required and regulate and control, but lack cellRecognition site cellular affinity is poor, more difficultly meets clinical demand. Natural biologic material is rich in biogenic signaling molecule, is beneficial to cellStick and breed, but such material mechanical performance deficiency, degrading too fastly, being difficult to for a long time for cell provides support. Although there is reportClaim to improve by chemical cross-linking agent (as glutaraldehyde, carbodiimides etc.) mechanical property of support, delay the degradation rate of support,But these crosslinking agents residual tends to produce cytotoxicity.
Electrostatic spinning technique (abbreviation electrospinning) is a kind of simple and easy to do novel porous tissue engineering bracket membrane preparation method, canForm nanometer to Sub-micro Fibers, and can control tunica fibrosa thickness by controlling the electrospinning time. The fibre diameter forming and natural(50～500nm) is similar for extracellular matrix proteins. This outer fiber has high-specific surface area and high porosity and imitates extracellular baseMatter nanofiber grid structure, can provide good growing environment for cell, is beneficial to cytotrophy material, cell signal and metabolismThe transmission of refuse.
In sum, also there is no at present a kind of existing nanofiber prepared by nano bionic method porous, ultra-thin that is staggered of utilizingStructure, has again bionical Bruch ' the s film of cell recognition burst.
Summary of the invention
The present invention is intended to improve the deficiency in current RPE cell transplantation carrier, with polycaprolactone or tussah silk fibroin and poly-ownLactone or tussah silk fibroin, polycaprolactone and gelatin etc. are raw material, apply simple electrostatic spinning technique and prepareBionical Bruch ' the s of ultra-thin porous composite nano fiber film, wherein, the porosity 80%-90% of film, the thickness 3-10 μ m of film.
For realizing the object of the invention, the technical scheme of employing is as follows:
The present invention proposes the method for electrostatic spinning preparation method of the bionical Bruch ' s of a kind of ultra-thin porous gelatin-compounded nanofiber film, comprisesFollowing steps:
(1) raw material is dissolved in to organic solvent and obtains electrostatic spinning solution;
(2) described electrostatic spinning solution step (1) being obtained carries out electrostatic spinning and obtains composite nano-fiber membrane;
(3) described composite nano-fiber membrane step (2) being obtained is placed in fume hood or constant temperature blast drying oven 1-4 hour, makesResidual organic reagent volatilizees completely, obtains described ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber bionicalBruch ' s film;
Wherein, the raw material in described step (1) is: polycaprolactone or tussah silk fibroin and polycaprolactone or tussah silk peptideAlbumen, polycaprolactone and gelatin;
Wherein, the bionical Bruch ' s of described ultra-thin porous composite nano fiber film be the bionical Bruch ' s of polycaprolactone nanofiber film,Tussah silk fibroin/polycaprolactone composite nano fiber bionical Bruch ' s film or tussah silk fibroin/polycaprolactone/gelatin-compounded receivingBionical Bruch ' the s of rice fiber film.
Preferably, taking polycaprolactone as raw material, prepare the bionical Bruch ' s of polycaprolactone nanofiber film by the inventive method.Preferably, taking tussah silk fibroin and polycaprolactone as raw material, prepare polycaprolactone nanofiber by the inventive method bionicalBruch ' s film. Preferably, taking tussah silk fibroin, polycaprolactone and gelatin as raw material, prepare poly-own by the inventive methodBionical Bruch ' the s of lactone nanofiber film.
Wherein, described organic solvent is 98% formic acid or hexafluoroisopropanol.
Wherein, described raw material tussah silk fibroin, polycaprolactone, the mass ratio of gelatin in described electrostatic spinning solution are 1-5%:65-85%:10-30%. Preferably, tussah silk fibroin, polycaprolactone, the mass ratio of gelatin in described electrostatic spinning solutionFor 5%:85%:10%.
Wherein, the quality summation of described raw material tussah silk fibroin, polycaprolactone, three kinds of raw materials of gelatin and described organic solventVolume ratio is 10%-30%. Preferably, the quality summation of tussah silk fibroin, polycaprolactone, three kinds of raw materials of gelatin with described in haveThe volume ratio of machine solvent is 25%.
Wherein, when described step (1) is prepared electrospinning solution, tussah silk fibroin, polycaprolactone, gelatin are dissolved in to organic solventAfter, magnetic agitation is spent the night.
Wherein, while applying method of electrostatic spinning in described step (2), applying voltage is 10-30kV, and extruded velocity is 0.5-3mL/h,Collecting distance is 10-20cm.
Wherein, in described step (3), composite nano-fiber membrane is placed in to fume hood 1-4 hour.
Wherein, in described step (3), composite nano-fiber membrane is placed in to 40 DEG C of constant temperature blast drying oven 1-4 hour.
The preparation method of the bionical Bruch ' s of the ultra-thin porous composite nano fiber of the present invention film, comprises the steps:
1. tussah silk is come unstuck, then obtain pure tussah silk fibroin after dissolving, dialysis, purifying and freeze drying;
2. get tussah silk fibroin, polycaprolactone, gelatin, be dissolved in organic solvent, under room temperature, on magnetic stirring apparatus, stirSpend the night. Obtain tussah silk fibroin: polycaprolactone: gelatin (mass percent is 1-5:65-85:10-30) electrospinning liquid;
3. electrospinning liquid in 2 is packed in syringe, load onto the passivity syringe needle of 0.6-1.2, adjust the distance of syringe needle to receiver10-20cm, regulation voltage is at 10-30kV, and spinning solution extruded velocity is controlled at 0.5-3mL/h.
Preferably, taking tussah silk fibroin: polycaprolactone: gelatin (mass percent is as 1-5:65-85:10-30) is as raw material,Bionical Bruch ' the s of the composite nano fiber film preparing in electrostatic field, forms by the nanofiber of random arrangement is interlacedLoose structure forms, and fibre diameter can be controlled in 50-500nm, and the aperture in tunica fibrosa can regulate within the scope of 100-4000nm,Film thickness 2-200 μ m.
The present invention also provides a kind of bionical Bruch ' s of ultra-thin porous composite nano fiber film preparing by the present invention, described film bagDraw together the bionical Bruch ' s of polycaprolactone nanofiber film, the bionical Bruch ' s of tussah silk fibroin/polycaprolactone composite nano fiber film,Or the bionical Bruch ' s of tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber film.
Preferably, the fibre diameter that the bionical Bruch ' s of this ultra-thin porous composite nano fiber film comprises is 50-500nm, and aperture is500-2500nm, porosity is 80-90%.
Beneficial effect of the present invention comprises: the composite nano fiber porous membrane that the present invention utilizes electrostatic spinning technique to prepare, fiber is straightFootpath between 50-500nm, aperture 500-2500nm, porosity 80-90%, have the similar thickness of natural B ruch ' s film withNanometer topology loose structure, is beneficial to nutriment, the transmission of cell signal chemical molecular and the transport of metabolic waste. In the present invention,Tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber porous membrane can be transparent after the soaked with liquid such as ethanol, culture medium, energyDirectly observation of cell growth under inverted microscope. Preferably, select polycaprolactone, tussah silk fibroin, gelatin as raw material,Realize material advantage complementation. Tussah silk fibroin is native protein, be rich in RGD cell recognition block, can increase Biomimetic membranesBiocompatibility and cell adherence, recognition capability. The ultra-thin porous composite nano fiber of product that the present invention prepares is bionicalBruch ' s film can be used as the carrier of transplantation of retinal pigment epithelium cells, the bionical Bruch ' s of this ultra-thin porous composite nano fiber filmBe rich in cell identification sequence (RGD), can promote the adhesion of cell. After soaking in solution, this Biomimetic membranes can be transparent, is beneficial to thinBorn of the same parents plant under rear microscope and directly observe.
Brief description of the drawings
Fig. 1 represents the bionical Bruch ' s of the ultra-thin porous nanofiber of the present invention film scanning electron microscope diagram;
Fig. 2 represents the present invention's each group ultra-thin porous nanofiber bionical Bruch ' s membrane aperture and fibre diameter comparison;
Fig. 3 represents the present invention's each group ultra-thin porous nanofiber bionical Bruch ' s membrane porosity and contact angle comparison;
Fig. 4 represents the bionical Bruch ' s of the ultra-thin porous tussah silk fibroin/polycaprolactone of the present invention/gelatin-compounded nanofiber film macroscopic viewFiber pattern under pattern and inverted microscope;
Fig. 5 represent in the present invention RPE cell on the each group of bionical Bruch ' s of ultra-thin porous composite nano fiber film and control group (thinOn born of the same parents' culture plate) cultivate cell adherence figure after 24 hours.
Detailed description of the invention
In conjunction with following specific embodiments and the drawings, the present invention is described in further detail, and protection content of the present invention is not limited toIn following examples. Do not deviating under the spirit and scope of inventive concept the variation that those skilled in the art can expect and advantageAll be included in the present invention, and taking appending claims as protection domain. Implement process of the present invention, condition, examinationAgent, experimental technique etc., except the content of mentioning specially below, be universal knowledege and the common practise of this area, the present inventionBe not particularly limited content.
Bionical Bruch ' the s of the ultra-thin porous polycaprolactone nanofiber of embodiment 1 film
Preparation method comprises the following steps:
1. preparation electrospinning solution: take polycaprolactone 2.5g, in 50mL conical flask, add the formic acid solution of 10mL98%, chamberUnder temperature, magnetic agitation is spent the night, and obtains electrostatic spinning solution;
2. electrospinning process: electrospinning liquid is packed in syringe, the passivity syringe needle of syringe configuration diameter 0.6mm, at voltage 12-20kV,Collect apart from 10-20cm electrospinning under the condition of extruded velocity 1-5mL/h; Preferred voltage 18kV, collects apart from 15cm, squeezesGo out speed 2mL/h;
3. post-processed: after electrospinning completes, electrospinning film is placed on to 1-4 hour in fume hood, treats that residual organic solvents volatilizees completely;4. obtain the bionical Bruch ' s of ultra-thin porous polycaprolactone nanofiber film.
Embodiment 2 prepares the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone composite nano fiber film
Preparation method comprises the following steps:
1. preparation electrospinning solution: take tussah silk fibroin 0.125g, polycaprolactone 2.125g, in 50mL conical flask, adds 10mL98% formic acid solution, under room temperature, magnetic agitation is spent the night, and obtains electrostatic spinning solution;
2. electrospinning process: electrospinning liquid is packed in syringe, the passivity syringe needle of syringe configuration diameter 0.6mm, at voltage 12-20kV,Collect apart from 10-20cm electrospinning under the condition of extruded velocity 1-5mL/h; Preferred voltage 18kV, collects apart from 15cm, squeezesGo out speed 2mL/h;
3. post-processed: after electrospinning completes, electrospinning film is placed on to 1-4 hour in fume hood, treats that residual organic solvents volatilizees completely;
4. obtain the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone composite nano fiber film.
Embodiment 3 prepares the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber film
Preparation method comprises the following steps:
1. preparation electrospinning solution: take tussah silk fibroin 0.125g, polycaprolactone 2.125g, gelatin 0.25g is in 50mL conical flaskIn, adding the formic acid solution of 10mL98%, under room temperature, magnetic agitation is spent the night, and obtains electrostatic spinning solution;
2. electrospinning process: electrospinning liquid is packed in syringe, the passivity syringe needle of syringe configuration diameter 0.6mm, at voltage 12-20kV,Collect apart from 10-20cm electrospinning under the condition of extruded velocity 1-5mL/h; Preferred voltage 18kV, collects apart from 15cm, squeezesGo out speed 2mL/h;
3. post-processed: after electrospinning completes, electrospinning film is placed on to 1-4 hour in fume hood, treats that residual organic solvents volatilizees completely;
4. obtain the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber film.
In the present embodiment, carry out separately several specific experiment processes, by tussah silk fibroin, polycaprolactone, gelatin at Static SpinningIn the scope that the mass ratio of silk in solution is 1-5%:65-85%:10-30%, take the amount of each raw material, by tussah silk fibroin,In the scope that the quality summation of polycaprolactone, three kinds of raw materials of gelatin and the volume ratio of organic solvent are 10%-30%, take each raw materialAmount, other operating procedures are the same, and all can obtain target product is ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanometerBionical Bruch ' the s of fiber film.
Bionical Bruch ' the s of the embodiment 4 sem observations ultra-thin porous nanofiber of each group film microscopic appearance
1. get respectively the each group of bionical Bruch ' s of ultra-thin porous nanofiber film (the gathering in oneself in embodiment 1 in embodiment 1,2,3Tussah silk/polycaprolactone/gelatin film in tussah silk/polycaprolactone film, embodiment 3 in ester film, embodiment 2), be cut into 2The diaphragm of mm × 2mm size;
2. pair the bionical Bruch ' s of each group nanofiber film is placed in metal-spraying equipment and carries out metal spraying;
3. under SEM, observe the each group of bionical Bruch ' s of nanofiber film microscopic appearance, and take pictures.
As shown in Figure 1, wherein, the A in Fig. 1 represents to surpass the microscopic appearance of the bionical Bruch ' s of the ultra-thin porous nanofiber of each group filmBionical Bruch ' the s of thin porous polycaprolactone nanofiber film microscopic appearance, the B in Fig. 1 represents that ultra-thin porous tussah silk/poly-is in oneselfBionical Bruch ' the s of ester nanofiber film microscopic appearance, the C in Fig. 1 represents ultra-thin porous tussah silk fibroin/polycaprolactone/gelatinBionical Bruch ' the s of nanofiber film microscopic appearance.
Embodiment 4 is the bionical Bruch ' s of the ultra-thin porous nanofiber of each group membrane fiber diameter, aperture relatively
1. get respectively the each group of bionical Bruch ' s of the ultra-thin porous nanofiber film scanning electron microscopy that embodiment 1,2,3 preparesMirror photo, chooses at random 30 fibers and measures, and chooses immediately 30 regions simultaneously, carries out aperture calculating; Measure usedThe ImageJ (http://rsbweb.nih.gov/ij/) that software provides for America NI H;
2. the detection in the fibre diameter of the bionical Bruch ' s of the ultra-thin porous nanofiber of each group film, aperture and statistics are as Fig. 2 instituteShow. Fig. 2 shows simultaneously, and along with adding of tussah silk fibroin and gelatin, the fiber aperture of bionical Bruch ' s film enlarges markedly,Fibre diameter is along with also having occurred remarkable increase adding of gelatin.
Embodiment 5 is the bionical Bruch ' s of the ultra-thin porous nanofiber of each group membrane fiber porosity, contact angle relatively
1. under constant temperature, in 5mL specific gravity bottle, fill absolute ethyl alcohol, weigh and be designated as W1, by small-diameter intravascular supportWeigh and be designated as Ws. Respectively each group of nano bionic Bruch ' the s film preparing in embodiment 1,2,3 immersed respectively and fill secondIn the specific gravity bottle of alcohol, utilize 20mL syringe to bleed, must make absolute ethyl alcohol fill in the hole of nano bionic Bruch ' s filmIn gap. Fill it up with absolute ethyl alcohol, weigh and be designated as W2. After the sample that soaks full ethanol is taken out, remaining ethanol and specific gravity bottle claimHeavily be designated as W3. Computing formula is as follows: average, obtain nano bionic Bruch ' s membrane porosity;
2. by each group of nano bionic Bruch ' s film cutting film forming, flatten, be placed on the objective table of contact angle instrument. By approximately 5 μ LWatson distilled water splashes into nano bionic Bruch ' s film surface, utilizes automatic contact angle instrument to measure contact angle, each sampleGetting 5 different parts measurements averages. The hydrophily of the bionical Bruch ' s of the less expression of contact angle film is stronger, and contact angle is largeHydrophobic in 90 ° of expressions, be less than 90 ° of expressions hydrophilic.
3. as shown in Figure 3, the porosity of respectively organizing nano bionic Bruch ' s film is all greater than 80% to comparative result, along with adding of gelatinThe porosity that enters nano bionic Bruch ' s film significantly increases, as shown in the left figure of Fig. 3; Polycaprolactone film and tussah silk fibroin/All about 110 ° of polycaprolactone film contact angles, are strong-hydrophobicity matter, along with adding of gelatin, contact angle be significantly reduced to 70 ° withUnder, hydrophily obviously strengthens, as shown in the right figure of Fig. 3.
Bionical Bruch ' the s of the ultra-thin porous tussah silk fibroin/polycaprolactone of embodiment 6/gelatin-compounded nanofiber film transparent processing
Get the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber prepared by embodiment 3 methodsFilm, puts into 35mm culture dish, as shown in the A in Fig. 4, shows ultra-thin porous tussah silk fibroin/polycaprolactone/gelatinBionical Bruch ' the s of composite nano fiber film is opaque before processing. Add 75% ethanol, ultra-thin porous tussah silk peptide egg after soakingBionical Bruch ' the s of in vain/polycaprolactone/gelatin-compounded nanofiber film bleach, as shown in the B in Fig. 4, shows through 75% ethanolAfter immersion, this film is transparent. Add after DMEM cell culture medium, under inverted microscope, observe, as shown in the C in Fig. 4,Bionical Bruch ' the s of visible ultra-thin porous tussah silk fibroin/polycaprolactone/gelatin-compounded nanofiber film is transparent, is mutually handed over by fiberParamnesia reticulates structure, very similar with extracellular matrix.
The evaluation of embodiment 7RPE cell to the each group of bionical Bruch ' s of ultra-thin porous nanofiber film degree of adhesion
The adhesion that can bionical Bruch ' s film be beneficial to RPE cell is directly connected to the carrying out of follow-up study, for this reason according to following stepSuddenly carried out the research of cell adherence ability.
1. get the bionical Bruch ' s of the ultra-thin porous tussah silk fibroin/polycaprolactone/gelatine nano fiber film of preparation in embodiment 3;
2. bionical Bruch ' s film sterilization: three groups of ultra-thin porous films that respectively embodiment 1,2,3 prepared are cut into 2 × 2cm2The diaphragm of size, and put it in culture dish. After 75% alcohol immersion 30-120min, absorb ethanol, on super-clean bench, treatEthanol volatilizees completely, adds PBS or HBSS buffer solution to soak twice;
3. cell seeding: RPE cell is through Trypsin Induced, and counting, with 1 × 106/ mL density inoculation RPE cell 400 μ L are to filmSurface, by cell suspension coating evenly, carefully moves in CO2gas incubator and cultivates 4 hours;
4. increase culture medium: after 4 hours, supplemented medium 1-2mL, continues to cultivate;
5. cultivate after 24 hours, remove culture medium, HBSS or PBS washing 4 times, all remove the cell not adhering to or slightly adhere toRemove;
2. under inverted microscope, observation of cell adheres to, as shown in Figure 5, show RPE cell ultra-thin porous tussah silk fibroin/poly-oneselfAdhesive capacity on the bionical Bruch ' s of lactone/gelatine nano fiber film obviously more ultra-thin porous polycaprolactone nanofiber is bionicalBruch ' s film and the bionical Bruch ' s of ultra-thin porous tussah silk fibroin/polycaprolactone nanofiber film are strong, and train with cellular control unitSupport plate similar.
Claims (8)
1. a method of electrostatic spinning preparation method for the bionical Bruch ' s of ultra-thin porous composite nano fiber film, is characterized in that the methodComprise the following steps:
(1) raw material is dissolved in to organic solvent and obtains electrostatic spinning solution;
(2) described electrostatic spinning solution step (1) being obtained carries out electrostatic spinning and obtains composite nano-fiber membrane;
(3) described composite nano-fiber membrane step (2) being obtained is placed in fume hood or constant temperature blast drying oven 1-4 hour, makesResidual organic reagent volatilizees completely, obtains the bionical Bruch ' s of described ultra-thin porous composite nano fiber film;
Wherein, the raw material in described step (1) is: the combination of tussah silk fibroin and polycaprolactone or tussah silk fibroin,The combination of polycaprolactone and gelatin;
Wherein, the bionical Bruch ' s of described ultra-thin porous composite nano fiber film is tussah silk fibroin/polycaprolactone composite nano fiberBionical Bruch ' s film or the tussah silk fibroin/polycaprolactone/bionical Bruch ' s of gelatin-compounded nanofiber film;
Wherein, in the time that described raw material is the combination of tussah silk fibroin and polycaprolactone:
Tussah silk fibroin, the mass ratio of polycaprolactone in described electrostatic spinning solution are 1:17;
Wherein, in the time that described raw material is the combination of tussah silk fibroin, polycaprolactone and gelatin:
Tussah silk fibroin, polycaprolactone, the mass ratio of gelatin in described electrostatic spinning solution are 1-5%:65-85%:10-30%;Or
Tussah silk fibroin, polycaprolactone, the quality summation of three kinds of raw materials of gelatin and the volume ratio of described organic solvent are10％-30％。
2. preparation method as described in claim 1, is characterized in that the formic acid that described organic solvent is 98% or hexafluoro isopropylAlcohol.
3. preparation method as described in claim 1, is characterized in that, when described step (1) is prepared electrostatic spinning solution, and willTussah silk fibroin, polycaprolactone, gelatin are dissolved in after organic solvent, and magnetic agitation is spent the night.
4. preparation method as described in claim 1, is characterized in that, carries out electrostatic spinning and obtain compound in described step (2)When nano fibrous membrane, applying voltage is 10-30kV, and extruded velocity is 0.5-3mL/h, and collecting distance is 10-20cm.
5. preparation method as described in claim 1, is characterized in that, described step in (3) is placed in composite nano-fiber membrane1-4 hour in fume hood.
6. preparation method as described in claim 1, is characterized in that, described step in (3) is placed in composite nano-fiber membrane1-4 hour in 40 DEG C of constant temperature blast drying ovens.
7. the bionical Bruch ' s of the ultra-thin porous composite nano fiber film preparing by any one method of claim 1-6.
8. the bionical Bruch ' s of the ultra-thin porous composite nano fiber film preparing as claim 7, is characterized in that, it comprisesFibre diameter is 50-500nm, and aperture is 500-2500nm, and porosity is 80-90%.
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| CN106390196A (en) * | 2016-09-07 | 2017-02-15 | 东华大学 | Preparation method of nanofiber nerve tissue engineering scaffold |
| CN109629037B (en) * | 2018-12-10 | 2020-04-24 | 武汉大学 | Triazinized silk fibroin modified polycaprolactone nanofiber scaffold and preparation method and application thereof |
| CN111719243B (en) * | 2020-06-09 | 2022-03-01 | 紫罗兰家纺科技股份有限公司 | PGS/SF electrospun membrane and preparation method thereof |
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| WO2009052459A1 (en) * | 2007-10-19 | 2009-04-23 | University Of Medicine And Dentistry Of New Jersey | A method of using an extracellular matrix to enhance cell transplant survival and differentiation |
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| CN101036802A (en) * | 2006-12-30 | 2007-09-19 | 苏州大学 | Antheraea pernyi silk fibrion biology medicine material and the preparation method |
| CN102242463A (en) * | 2011-04-20 | 2011-11-16 | 东华大学 | Method for preparing gelatin/polycaprolactone composite nanometer fiber membrane through electrostatic spinning |
| CN102965849A (en) * | 2012-11-22 | 2013-03-13 | 天津大学 | Method for preparing medical barrier membrane by electrostatic spinning |
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