CN107865981A - A kind of multilayer oriented property nano fiber artificial endocranium and preparation method thereof - Google Patents
A kind of multilayer oriented property nano fiber artificial endocranium and preparation method thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0069—Electro-spinning characterised by the electro-spinning apparatus characterised by the spinning section, e.g. capillary tube, protrusion or pin
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0076—Electro-spinning characterised by the electro-spinning apparatus characterised by the collecting device, e.g. drum, wheel, endless belt, plate or grid
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0092—Electro-spinning characterised by the electro-spinning apparatus characterised by the electrical field, e.g. combined with a magnetic fields, using biased or alternating fields
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
Abstract
The invention provides a kind of multilayer oriented property nano fiber artificial endocranium, comprising three layers, hydrophobic internal layer, hydrophilic outer layer and Weaving type intermediate layer.Weaving type intermediate layer, utilize " similar compatibility principle " that reliably inside and outside two layers is linked together.Outer layer uses hierarchy, and the second layer of outer layer is the groove structure that orientation nanofiber slice is formed, and the length direction of groove is consistent with nanofiber direction.Present invention also offers a kind of dural preparation method of multilayer oriented property nano fiber artificial, comprise the following steps:Electrospun prepares hydrophobic inner layer;Prepare braiding layer;Electrospun prepares outer layer in braiding layer, finally obtains artificial dura mater.Artificial dura mater of the present invention, the groove structure of outer layer play fixed guiding function to the brain dura mater tissue of new life, and Weaving type intermediate layer efficiently solves artificial dura mater lamination problem, and can cut randomly, and has a extensive future.
Description
Technical field
The present invention relates to a kind of multilayer oriented property nano fiber artificial endocranium and preparation method thereof, belong to electrostatic spinning skill
Art field.
Background technology
Dura defect is common in neurosurgery clinical position, open craniocerebral injury (industry, traffic, war etc.),
The erosion of tumour, congenital defect of meninges and other brain disorder reasons can trigger dura defect.Dura defect need and
When repairing it is excessive to prevent cerebrospinal fluid, prevent the bulging of brain and the compressing of atmospheric pressure, otherwise will jeopardize human life.Dura defect
The complication such as intracranial infection, brain adhesion, hypohydropses can also be triggered, often cause such as headache, brain disorder disease.
Endocranium is wrapped around the membrane structure outermost layer outside brain and spinal cord, is a kind of more firm connective tissue film,
The inner surface of natural endocranium structure is the fine and close individual layer fibroblast of a Rotating fields, thin and smooth;Contain more in intermediate layer
Fibroblast and a small amount of collagen;Outer layer is thicker, contains more collagen component and a small amount of fibroblast.Such knot
Structure inner face is smooth and overall very tough and tensile, can be good at playing protection brain and prevents the work of cerebrospinal fluid seepage
With.
Oneself has artificial dura mater made of multiple material just in Clinical practice, to can be divided mainly into two major classes at present:Biology spreads out
Green material and artificial synthesized high polymer material.Bio-derived material mainly have allogeneic human cerebral dura mater and xenogenesis pig/
Ox source pericardium, dermal matrix and biomembrane using the preparation of ox tendon forceps type i collagen etc..Artificial synthesized high polymer material is main
Including polyesters degradable macromolecule, such as PLA, polyglycolic acid, polycaprolactone and polyurethane.Additionally include poly- four
The non-degradable high polymer material such as PVF.
Meninx injury repair effect desirably multilayer artificial dura mater at present.This multilayer artificial dura mater passes through
It is layered to reach the purpose of simulation meninx physiological function.This kind of artificial dura mater is typically made up of for two layers internal layer and outer layer.Wherein
Internal layer is made up of hydrophobic material, and outer layer is made up of water wetted material.Because outer layer meninx support is using electrostatic spinning process preparation, obtain
The artificial dura mater outer layer fiber distribution arrived is disorderly and unsystematic.Fibroblast is slow-growing on such artificial dura mater, leads
Defect meninx is caused to recover excessive cycle.
2007,《NANO LETTERS》In one article of report, using Biodegradable material PLLA solution, collect
To non-woven fabrics and the electrospinning film of parallel arrangement, by cell culture, illustrate that the electrospinning film of parallel arrangement being capable of induction of neurite
Growth, strengthen the migration of Skin Cell.Kyle T. Kurpinski etc..《Biomaterials》In the article delivered, profit
Use PLLA(PLLA)3D electrospinning films have been made, the electrospinning of parallel arrangement is also demonstrated by Tube propagation and zoopery
Film enhances cell growth.
Meninx is mainly distributed the collagenous fibres of fibroblast and its secretion.General fibroblast diameter is between 20-30 μm.
Document report, when electrospun fibers footpath is between 50-1000nm, average pore size can reach 2 μm.Average pore size is at 3 μm
Hereinafter, entering in electrospun fibers film for cell, the generation of effective brain adhesion can effectively be prevented.When fibre diameter exists
Between 5-200 μm, average pore size reaches 20-100 μm.So be advantageous to fibroblastic move into, adhere to, breed and grow point
Change.
Chinese invention patent CN 103480042 A, it is entitled:" a kind of artificial endorchis and preparation method thereof and make
With method ", a kind of artificial endorchis with double-decker is disclosed, by PLA/ethanol copolymer fiber of orientation
Film inner layer, and PLA/ethanol copolymer-chitin fiber film outer layer composition of non orientation.Although the artificial endorchis
Dura mater regeneration and prevention of scar and the purpose of adhesion can be achieved, but have the disadvantage that:First, internal layer is that orientation copolymer is fine
Film is tieed up, the growth rate of meningocyte can be accelerated, but endocranium combination dynamics of the artificial dura mater with patient in itself is inadequate,
Meninx, which repairs area, the hidden danger such as the artificial meninx dislocation of generation and fold.Second, in use, first covered with internal layer impaired hard
Meninx, and the endocranium outer layer of people is mainly collagen component and a small amount of fibroblast, orientation fiber-induction cell growth
The advantages of do not give full play to.3rd, orientation tunica fibrosa preparation efficiency is low, causes artificial meninx cost too high.
Chinese invention patent CN 101507661 A, it is entitled:" it is a kind of possess multiple functional layers nano artificial it is hard
Meninx and preparation method thereof ", disclose a kind of artificial dura mater with double-decker.Although the artificial dura mater can be realized firmly
Film regenerates purpose, but has the disadvantage that:Internal layer is prepared using electrostatic spinning process, and the nanofiber in internal layer is disorderly and unsystematic,
Endocranium recovers excessive cycle.
Therefore, at present clinically there is an urgent need to one kind promote brain defect quickly to heal, will not cause again meninx dislocation and
The artificial dura mater of the defects of fold, easy to use, cheap, the wide artificial dura mater of industrialization prospect.
The content of the invention
For overcome the deficiencies in the prior art, it is an object of the present invention to provide a kind of hard brain of multilayer oriented property nano fiber artificial
Film, the artificial dura mater are with the dural difference of Traditional Man:Internal layer is formed for non-oriented property nanofiber, and outer layer is by being orientated
Property nanofiber form.Outer layer is formed by two layers, and first layer is made up of non-oriented property nanofiber;The second layer is by orientation nanometer
Fiber is formed, and it be 3mm that upper surface, which is provided with width, and depth is 5 μm, length 1000mm, and spacing is 6mm groove, nanometer
Machine direction is consistent with the direction of groove.When use, artificial dura mater need to be placed in below endocranium.
It is provided with Weaving type intermediate layer among the internal layer and outer layer of artificial dura mater, intermediate layer is using hydrophobic material and hydrophilic
Material braiding forms, according to " similar compatibility " principle, slice and artificial dura mater internal layer prepared by the hydrophobic material in braiding layer
(It is prepared by hydrophobic material Electrospun)Reliable connection, slice and artificial dura mater outer layer prepared by water wetted material(Water wetted material electrospinning
It is prepared by silk)Reliable connection, and both slices are overlapping weave ins, the slice of hydrophilic material is through hydrophobic material
The space of slice is combined with outer layer, and the slice of hydrophobic material is combined through the space of the slice of hydrophilic material with internal layer, so interior
Just reliably it is combined together for outer two layers.In a word, the artificial dura mater of this structure, the combination between internal layer and outer layer is improved
Power, it is not easy that stripping between layers occurs in the presence of external force.
It is a further object of the present invention to provide a kind of dural preparation method of multilayer oriented property nano fiber artificial.
Technical scheme is used by the present invention solves the above problems:
A kind of multilayer oriented property nano fiber artificial endocranium, comprising three layers, cerebripetal one layer of face is hydrophobic inner layer, backwards
One layer of brain is hydrophily outer layer;It is Weaving type intermediate layer between two layers.
The hydrophily outer layer is made up of electrostatic spinning, i.e. outer layer is in order to effectively induce stem cell and fibroblastic move
Enter, this layer is then using the hydrophilic material using good biocompatibility, and two layers of preparation of outer layer point, first layer is non-oriented property nanometer
Fiber, adjusted by electrospinning parameters, average pore size is reached 20-100 μm, general fibroblast diameter is at 20-30 μm
Between.So be advantageous to fibroblastic move into, adhere to, breeding and Growth and Differentiation;The second layer is orientation nanofiber,
It is 3mm that its upper surface, which is provided with width, and depth is 5 μm, length 1000mm, and spacing is 6mm groove, the direction of nanofiber and
Groove direction is consistent.For strengthening outer layer and dural bond strength, dislocation and the generation of fold are prevented.Orientation fiber can
To accelerate the growth of dura cell.
The internal layer is to use hydrophobic material, prevents moving into for cell, so as to reach the purpose to prevent adhesion;Should to preparing
The electrospinning parameters of layer material are adjusted, and make its hole aperture below nanometer, one to two orders of magnitude smaller than cell(One
As fibroblast diameter between 20-30 μm), so as to prevent cell from entering, prevent brain adhesion from producing.
The intermediate layer is formed using hydrophobic material and water wetted material braiding.
A kind of multilayer oriented property nano fiber artificial endocranium, including internal layer, intermediate layer, outer layer, its preparation method press with
Lower step is prepared:
(1) preparation of internal layer:Inner layer material composition is polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in
In hexafluoroisopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 7%(wt), stir 4 hours, Electrospun ginseng
Number be flow velocity be 0.9 ml/h, voltage 13kV, receive distance 20cm, electrostatic spinning 4h;
(2) prepared by intermediate layer.First layer material is to use polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen)Electrostatic spinning
The film of preparation, film thickness are 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Slice and any side of internal layer are placed in parallel, according to 6mm spacing, 3mmX1000mm faces inner layer, uniformly
It is booked whole internal layer.
Second layer material is to use polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen)Film prepared by electrostatic spinning,
Film thickness is 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Second layer slice and first layer slice are placed in 90 ° of angles, according to 6mm spacing, 3mmX1000mm is facing to interior
Layer, is uniformly booked whole first layer slice layer.
Especially, above-mentioned slice placement process, it should completed before internal layer solidification.
Finally, the slice that top is put is pressed down against with 50N pressure, keeps 1min.
(3) two layers of preparation of outer layer point.
From top to bottom, the first layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .4ml/h of flow velocity 1, voltage 12kV, receive distance 10cm, electrostatic spinning 2h.
From top to bottom, the second layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .4ml/h of flow velocity 0, voltage 0.9kV, distance 1mm is received, orientation electrostatic spinning process prepares 2h, obtains orientation Nanowire
Layer is tieed up, fiber direction is any parallel on one side with internal layer, and it is 3mm that width is opened in upper surface, and depth is 5 μm, and length is
1000mm grooves, ditch separation 6mm, trench length direction are consistent with fiber direction.
(4) artificial meninx takes out from culture dish by made from, is rinsed 3 times with distilled water, is vacuum-packed after freeze-dried,
- 20 DEG C of Cord bloods after the sterilizing of 25kGy cobalts -60.
The thickness for every layer of multilayer electrostatic spinning artificial dura mater being prepared is:Internal layer thickness is 80 μm ± 10 μm, outside
Thickness degree is 30 μm ± 20 μm, and intermediate layer thickness is 80 μm ± 20 μm.The two layers of thickness in intermediate layer is respectively:From top to bottom suitable
Sequence, first layer thickness are:40 μm ± 10 μm, second layer thickness is:40μm±10μm.Two layers of thickness layer by layer is respectively outside:From
Under to upper order, first layer thickness be:Non-oriented property layers of nanofibers, thickness are 20 μm, and the second layer is:Orientation Nanowire
Layer is tieed up, thickness is:10 μm, gash depth is 5 μm.
When prepared by internal layer, for electrospun fibers diameter control between 50-1000nm, average pore size is 2 μm;
When prepared by outer layer, from top to bottom, first layer, between 5-200 μm, average pore size is electrospun fibers diameter control
20-100μm.The second layer, orientation layers of nanofibers, fibre diameter are 1-3 μm.
The beneficial effects of the invention are as follows:
(1) artificial dura mater of the present invention uses three-decker, strengthens the bond strength of internal layer and outer layer by intermediate layer;
(2) the artificial exterior dura layer of the present invention uses hierarchy, and the second layer of outer layer is orientation nanofiber slice structure
Into its upper surface is provided with groove.Orientation nanofiber promotes the production of cell, brain dura mater tissue of the groove structure to new life
Play fixed guiding function;
(3) the artificial dural internal layer of the present invention is to use hydrophobic material, prevents moving into for cell, anti-so as to reach
The purpose of adhesion;The electrospinning parameters for preparing the layer material are adjusted, its hole aperture is compared cell below nanometer
Small one to two orders of magnitude(General fibroblast diameter is between 20-30 μm), so as to prevent cell from entering, prevent brain adhesion
Produce.
Brief description of the drawings
Fig. 1 is artificial dura mater structural representation, wherein 1 is internal layer, 2 be intermediate layer, and 3 be outer layer.
Fig. 2 is artificial exterior dura layer schematic diagram, and 2-1 is first layer in intermediate layer, and 2-2 is the second layer in intermediate layer.
Fig. 3 is artificial exterior dura layer schematic diagram, and 3-1 is first layer in outer layer, and 3-2 is the second layer in outer layer.
Fig. 4 is orientation nanofiber design sketch.
Embodiment
Below by specific embodiment, the present invention will be described, but the invention is not limited in this.
Experimental method used in following embodiments is conventional method unless otherwise specified:Institute in following embodiments
Reagent, material etc., unless otherwise specified, commercially obtain.
Embodiment 1
(1) preparation of internal layer:Inner layer material composition is polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in
In hexafluoroisopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 5%(wt), stir 1 hour, Electrospun ginseng
Number be flow velocity be 1 .0ml/h, voltage 12kV, receive distance 15cm, electrostatic spinning 2h;
(2) prepared by intermediate layer.First layer material is to use polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen)Electrostatic spinning
The film of preparation, film thickness are 30 μm.
Wide 4mm, long 1000mm slice is made in film.
Slice and any side of internal layer are placed in parallel, according to 8mm spacing, 4mmX1000mm faces inner layer, uniformly
It is booked whole internal layer.
Second layer material is to use polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen)Film prepared by electrostatic spinning,
Film thickness is 30 μm.
Wide 4mm, long 1000mm slice is made in film.
Slice and first layer slice are placed in 90 ° of angles, according to 8mm spacing, 4mmX1000mm faces inner layer, uniformly
It is booked whole first layer slice layer.
Especially, above-mentioned slice placement process, it should completed before internal layer solidification.
The slice that top is put is pressed down against with 50N pressure, keeps 1min.
(3) two layers of preparation of outer layer point.
From top to bottom, the first layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .2ml/h of flow velocity 1, voltage 10kV, receive distance 8cm, electrostatic spinning 2h.
From top to bottom, the second layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .3ml/h of flow velocity 0, voltage 0.7kV, distance 2mm is received, orientation electrostatic spinning process prepares 2h, obtains orientation Nanowire
Layer is tieed up, fiber direction is any parallel on one side with internal layer, and it is 3mm that width is opened in upper surface, and depth is 5 μm, and length is
1000mm grooves, ditch separation 6mm, trench length direction are consistent with fiber direction.
(4) artificial meninx takes out from culture dish by made from, is rinsed 3 times with distilled water, is vacuum-packed after freeze-dried,
- 20 DEG C of Cord bloods after the sterilizing of 25kGy cobalts -60.
Embodiment 2
(1) preparation of internal layer:Inner layer material composition is polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in
In hexafluoroisopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 7%wt, stir 4 hours, Electrospun parameter
Be 0.9 ml/h for flow velocity, voltage 13kV, receive distance 20cm, electrostatic spinning 4h;
(2) prepared by intermediate layer.First layer material is to use polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen)Electrostatic spinning
The film of preparation, film thickness are 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Slice and any side of internal layer are placed in parallel, according to 6mm spacing, 3mmX1000mm faces inner layer, uniformly
It is booked whole internal layer.
Second layer material is to use polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen)Film prepared by electrostatic spinning,
Film thickness is 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Slice and first layer slice are placed in 90 ° of angles, according to 6mm spacing, 3mmX1000mm faces inner layer, uniformly
It is booked whole first layer slice layer.
Especially, above-mentioned slice placement process, it should completed before internal layer solidification.
The slice that top is put is pressed down against with 50N pressure, keeps 1min.
(3) two layers of preparation of outer layer point.
From top to bottom, the first layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .4ml/h of flow velocity 1, voltage 12kV, receive distance 10cm, electrostatic spinning 2h.
From top to bottom, the second layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .4ml/h of flow velocity 0, voltage 0.9kV, distance 1mm is received, orientation electrostatic spinning process prepares 2h, obtains orientation Nanowire
Layer is tieed up, fiber direction is any parallel on one side with internal layer, and it is 3mm that width is opened in upper surface, and depth is 5 μm, and length is
1000mm grooves, ditch separation 6mm, trench length direction are consistent with fiber direction.
(4) artificial meninx takes out from culture dish by made from, is rinsed 3 times with distilled water, is vacuum-packed after freeze-dried,
- 20 DEG C of Cord bloods after the sterilizing of 25kGy cobalts -60.
Embodiment 3
(1) preparation of internal layer:Inner layer material composition is polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in
In hexafluoroisopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 5%wt, stir 4 hours, Electrospun parameter
Be 1 .0ml/h for flow velocity, voltage 15kV, receive distance 25cm, electrostatic spinning 5h.
(2) prepared by intermediate layer.First layer material is to use polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen)It is quiet
Film prepared by Electrospun, film thickness are 40 μm.
Wide 5mm, long 1000mm slice is made in film.
Slice and any side of internal layer are placed in parallel, according to 10mm spacing, 5mmX1000mm faces inner layer, uniformly
It is booked whole internal layer.
Second layer material is to use polycaprolactone(Bo Li biomaterials Co., Ltd of Shenzhen)Film prepared by electrostatic spinning,
Film thickness is 50 μm.
Wide 5mm, long 1000mm slice is made in film.
Slice and first layer slice are placed in 90 ° of angles, according to 10mm spacing, 5mmX1000mm faces inner layer,
It is even to be booked whole first layer slice layer.
Especially, above-mentioned slice placement process, it should completed before internal layer solidification.
The slice that top is put is pressed down against with 50N pressure, keeps 1min.
(3) two layers of preparation of outer layer point.
From top to bottom, the first layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .8ml/h of flow velocity 1, voltage 20kV, receive distance 15cm, electrostatic spinning 2h.
From top to bottom, the second layer material is polyglycolic acid(Bo Li biomaterials Co., Ltd of Shenzhen), it is dissolved in hexafluoro
In isopropanol (Suzhou Hao Fan Biological Co., Ltd.), concentration of dope 6%(wt), stir 1 hour, electrospinning parameters are:
The .8ml/h of flow velocity 0, voltage 0.9kV, distance 3mm is received, orientation electrostatic spinning process prepares 2h, obtains orientation Nanowire
Layer is tieed up, fiber direction is any parallel on one side with internal layer, and it is 3mm that width is opened in upper surface, and depth is 5 μm, and length is
1000mm grooves, ditch separation 6mm, trench length direction are consistent with fiber direction.
(4) artificial meninx takes out from culture dish by made from, is rinsed 3 times with distilled water, is vacuum-packed after freeze-dried,
- 20 DEG C of Cord bloods after the sterilizing of 25kGy cobalts -60.
Claims (10)
1. a kind of multilayer oriented property nano fiber artificial endocranium, includes three layers:The cerebripetal internal layer in face, carries on the back cerebripetal outer layer
And the intermediate layer between two layers.
A kind of 2. multilayer oriented property nano fiber artificial endocranium according to claim 1, it is characterised in that the internal layer
Thickness is 80 μm ± 10 μm, is prepared by electrostatic spinning process, and fibre diameter is controlled between 50-1000nm, average pore size
For 2 μm.
A kind of 3. multilayer oriented property nano fiber artificial endocranium according to claim 1, it is characterised in that the outer layer
Thickness is 30 μm ± 10 μm, is prepared by electrostatic spinning process and orientation electrospinning process, and fibre diameter is controlled in 5-200
Between μm, average pore size is 20-100 μm, and orientation electrospinning process fibre diameter is controlled at 1-3 μm.
4. the centripetal orientation nano fiber artificial endocranium of a kind of multilayer according to claim 1, it is characterised in that described
Intermediate layer thickness is 80 μm ± 20 μm, is formed by two layers.
5. intermediate layer according to claim 4, it is characterised in that the order of the intermediate layer from top to bottom, first layer are
Width prepared by hydrophilic material is 3mm, and length is 1000mm slice;The second layer is that width prepared by hydrophobic material is
3mm, length are 1000mm slice.
6. slice according to claim 5, it is characterised in that the first layer slice, thickness is prepared into by electrospinning process
Spend and formed for 40 μm of ± 10 μm of films, cutting;Second layer slice, it is 40 μm ± 10 μm thin to be prepared into thickness by electrospinning process
Film, cutting form.
7. slice according to claim 5, it is characterised in that the first layer slice, by slice and any side of internal layer
It is placed in parallel, according to 6mm spacing, 3mmX1000mm faces inner layer, is uniformly booked whole internal layer;Second layer slice and first
Layer slice is placed in 90 ° of angles, and according to 6mm spacing, 3mmX1000mm faces inner layer, is uniformly booked whole first layer.
8. outer layer according to claim 3, it is characterised in that the outer layer, order from top to bottom, first layer are non-
Orientation layers of nanofibers, thickness are 20 μm ± 10 μm.The second layer is orientation layers of nanofibers.Fiber direction and internal layer
Any parallel on one side, it is 3mm that width is opened in upper surface, and depth is 5 μm, and length is 1000mm grooves.
9. outer layer according to claim 8, it is characterised in that the ditch separation in the second layer in the outer layer is
6mm, trench length direction are consistent with fiber direction.
A kind of a kind of 10. dural side of multilayer oriented property nano fiber artificial prepared any one of claim 1-9
Method, comprise the steps:
(1) preparation of internal layer:Inner layer material composition is polycaprolactone, is dissolved in hexafluoroisopropanol, concentration of dope 7%
(wt), stir 4 hours, Electrospun parameter be flow velocity be 0.9 ml/h, voltage 13kV, receive distance 20cm, electrostatic spinning 4h;
(2) prepared by intermediate layer.First layer material is polyglycolic acid, is prepared into film with electrostatic spinning, film thickness is 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Slice and any side of internal layer are placed in parallel, according to 6mm spacing, 3mmX1000mm faces inner layer, is uniformly booked
Whole internal layer.
Second layer material is that polycaprolactone is quiet, is prepared into film with Electrospun, film thickness is 40 μm.
Wide 3mm, long 1000mm slice is made in film.
Second layer slice and first layer slice are placed in 90 ° of angles, according to 6mm spacing, 3mmX1000mm faces inner layer,
It is even to be booked whole first layer slice layer.
Especially, above-mentioned slice placement process, it should completed before internal layer solidification.
Finally, the slice that top is put is pressed down against with 50N pressure, keeps 1min.
(3) two layers of preparation of outer layer point.
From top to bottom, the first layer material is polyglycolic acid, is dissolved in hexafluoroisopropanol, concentration of dope 6%(wt), stirring 1
Hour, electrospinning parameters are:The .4ml/h of flow velocity 1, voltage 12kV, receive distance 10cm, electrostatic spinning 2h.
From top to bottom, the second layer material is polyglycolic acid, is dissolved in hexafluoroisopropanol, concentration of dope 6%(wt), stirring 1
Hour, electrospinning parameters are:The .4ml/h of flow velocity 0, voltage 0.9kV, distance 1mm being received, orientation electrostatic spinning process prepares 2h,
Obtain orientation layers of nanofibers, fiber direction is any parallel on one side with internal layer, and it is 3mm that width is opened in upper surface, depth
For 5 μm, length is 1000mm grooves, ditch separation 6mm, and trench length direction is consistent with fiber direction.
(4) artificial meninx takes out from culture dish by made from, is rinsed 3 times with distilled water, is vacuum-packed, passes through after freeze-dried
- 20 DEG C of Cord bloods after 25kGy cobalts -60 sterilize.
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