CN107715178A - A kind of multilayer contains high intensity artificial dura mater of cell factor and preparation method thereof - Google Patents

A kind of multilayer contains high intensity artificial dura mater of cell factor and preparation method thereof Download PDF

Info

Publication number
CN107715178A
CN107715178A CN201711094328.4A CN201711094328A CN107715178A CN 107715178 A CN107715178 A CN 107715178A CN 201711094328 A CN201711094328 A CN 201711094328A CN 107715178 A CN107715178 A CN 107715178A
Authority
CN
China
Prior art keywords
layer
prepared
intermediate layer
slice
film
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201711094328.4A
Other languages
Chinese (zh)
Other versions
CN107715178B (en
Inventor
李瑞锋
王斐
李典
李翰林
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to CN201711094328.4A priority Critical patent/CN107715178B/en
Publication of CN107715178A publication Critical patent/CN107715178A/en
Application granted granted Critical
Publication of CN107715178B publication Critical patent/CN107715178B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • 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
    • 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
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • 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
    • 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
    • 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
    • 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
    • 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
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A61L2300/414—Growth factors
    • 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

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Textile Engineering (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Dermatology (AREA)
  • Transplantation (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Mechanical Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Materials For Medical Uses (AREA)

Abstract

The invention provides the nano fiber artificial endocranium that a kind of multilayer contains Porcine HGF, comprising three layers, the cerebripetal hydrophobic inner layer in face, carries on the back the Weaving type intermediate layer among cerebripetal hydrophily outer layer and two layers.Internal layer is non-oriented property fiber membrane, and outer layer is orientation nano-fiber film, and Porcine HGF is uniformly attached in the fiber gap of outer layer, and the slice braiding that intermediate layer is prepared by water wetted material and hydrophobic material forms.Present invention also offers the dural preparation method of nano fiber artificial that a kind of multilayer contains Porcine HGF, comprise the following steps：Electrospun prepares hydrophobic inner layer；Intermediate layer woven in advance is positioned over internal layer upper surface；Electrospun prepares outer layer；Sprinkling Porcine HGF is given birth in outer layer；Terminal disinfection obtains artificial dura mater.The problem of artificial dura mater of the present invention efficiently solves endocranium injury repair cycle length, and Porcine HGF utilization rate is low, has a extensive future.

Description

A kind of multilayer contains high intensity artificial dura mater of cell factor and preparation method thereof
Technical field
Contain high intensity artificial dura mater of cell factor and preparation method thereof the present invention relates to a kind of multilayer, belong to biology Technical field of micro-nano manufacture.
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.
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 the hydrophobic hydrophilic nmature of materials at two layers is different, cause outer layer with The coupling reliability of internal layer is not high, there is the phenomenon of stripping, the life and health of the patient of serious threat.
Meninx is mainly distributed the collagenous fibres of fibroblast and its secretion.General fibroblast diameter is at 20-30 μm Between.Document report, when electrospun fibers footpath is between 50-1000nm, average pore size can reach 2 μm.Average pore size Below 3 μm, it is possible to effectively prevent entering in electrospun fibers film for cell, the generation of effective brain adhesion.Work as fibre Diameter is tieed up between 5-200 μm, average pore size reaches 20-100 μm.So be advantageous to fibroblastic move into, adhere to, breeding And Growth and Differentiation.
Fibroblast growth factor（fibroblast growth factor,FGF）There are several isomers, it is hard in artery Change the mainly bFGF to be worked in stove（basic fibroblast growth factor）, bFGF can by endothelial cell, Smooth muscle cell, macrophages secrete.Its effect is the propagation for the migration and smooth muscle cell for promoting endothelial cell, enough to promote Neovascularization, repair the endothelial cell of infringement.
Kyle Kurpinski etc. exist《Nanomedicine》In in an article delivering, describe a kind of double-deck Electrospinning film, material therefor are PLCL/PPG solution.Article utilizes rotating shaft collection device, obtains the electrospinning film of parallel arrangement, surveys The parallel and electrospinning film of vertical fibers arragement direction mechanical strength is tried, sample size is 6 × 1cm, is measured along both direction Maximum load be respectively 12.08 ± 1.15N and 3.01 ± 0.54N.It can be seen that on direction consistent with machine direction film it is strong Spend very poor.
Chinese invention patent CN103480042A, it is entitled:" a kind of composite artificial endocranium and preparation method ", A kind of composite artificial endocranium is disclosed, is made up of gel layer and braiding layer, braiding layer is located between two layers of hydrogel layer, Contain Porcine HGF in hydrogel layer.The beneficial effects of the invention are as follows：Porcine HGF is advantageous to wound repair, accelerates The healing of endocranium damage.But have the disadvantage that:1）Meninx outer layer is colloform texture, rather than nano fiber scaffold, is unfavorable for Cell adheres to, meninx repairing efficiency length, and 2）Porcine HGF is dissolved in colloid, only the Porcine HGF and cell on top layer Contact, it is limited to the facilitation of cell production.
Chinese invention patent CN106943634A, entitled " a kind of adsorbable artificial with anti-infective function is hard Meninx and preparation method and application ", one kind is disclosed by nano fibrous membrane by degradable poly-vinegar nano fibrous membrane and chitosan Nano fibrous membrane double-decker forms.There is the adsorbable artificial endocranium of the present invention good anti-infection property can effectively prevent art Intracranial infection afterwards, certain suture power can be born, do not torn, not off-clip.But two layers of the product is using roller orientation spinning work Prepared by skill, two layers of meninx is orientation nanofiber, and this fiber membrane intensity is well below non-oriented property fiber membrane.Together When the artificial dura mater only have two layers, the bond strength of two interlayers is inadequate.
Therefore, clinically there is an urgent need to one kind at present to promote brain damage to heal, and internal layer is combined reliably with outer layer, user Just, the wide artificial dura mater of industrialization prospect.
The content of the invention
For overcome the deficiencies in the prior art, it is artificial to contain cell factor high intensity it is an object of the present invention to provide a kind of multilayer Endocranium, the artificial dura mater are with the dural difference of Traditional Man：
（1）Increase 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 silk ribbon is combined with outer layer, and the slice of hydrophobic material is combined through the space of the silk ribbon 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.
(2) artificial dura mater internal layer, prepared using electrospinning process, obtain non-oriented property nano-fiber film.It is this kind thin Film tearing toughness is far longer than orientation film, and the intensity of artificial dura mater is improved from structure.
(3) artificial dura mater outer layer, prepared using electrospinning process, obtain orientation nano-fiber film, fibre gap It is easy to fibroblast to adhere to.
(4) cell production factor is interspersed among among electricity spinning fibre, effectively increases Porcine HGF and meningocyte Contact probability, give full play to the effect of Porcine HGF.
It is a further object of the present invention to provide the preparation method that a kind of multilayer contains cell factor high intensity artificial dura mater.
Technical scheme is used by the present invention solves the above problems：
A kind of multilayer contains cell factor high intensity artificial dura mater, and comprising three layers, the cerebripetal hydrophobic inner layer in face is big backwards Weaving type intermediate layer between brain hydrophily outer layer and 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 Entering, this layer then uses the hydrophilic material using good biocompatibility, furthermore, adjusted by electrospinning parameters, make average hole Footpath reaches 20-100 μm, and general fibroblast diameter is between 20-30 μm.So be advantageous to fibroblastic move into, glue Attached, propagation and Growth and Differentiation.
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 contains cell factor high intensity artificial dura mater, including internal layer, intermediate layer, outer layer, its preparation method It is prepared according to the following steps：
(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.
Carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）Film is prepared into electrostatic spinning, film thickness is 40μm.Wide 3mm, long 1000mm slice is made in film.
Polycaprolactone（Bo Li biomaterials Co., Ltd of Shenzhen）Film, film thickness 40 are prepared into electrostatic spinning μm.Wide 3mm, long 1000mm slice is made in film.
Two kinds of slices are worked out using rhombus government establishments law, obtain Weaving type intermediate layer.
（3）Before internal layer solidification, Weaving type intermediate layer is put in internal layer upper surface.Top is put with 50N pressure Weaving type intermediate layer be pressed down against, keep 1min.
(4) prepared by outer layer：
Cladding material is carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）, it is dissolved in pure water (Jinan sea industry Work Co., Ltd) in, 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 layers of nanofibers, fiber longitudinal direction is with any while flat OK.
(5) fibroblast growth factor（Rh-bFGF dry powder, trade name：External application recombination human basic fibroblast growth The factor, Nanhai Longtime Pharmaceutical Co., Ltd.）Dissolved with water for injection (Jinan Hai Ye Chemical Co., Ltd.s), be uniformly sprayed onto brain Dura mater outer layer.Optimal dosage is 150IU/cm2。
(6) the artificial meninx by made from, it is vacuum-packed after freeze-dried, -20 DEG C of low temperature are protected after 25kGy bores -60 sterilizings Deposit.
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 ± 10 μm, and intermediate layer thickness is 80 μm ± 20 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, for electrospun fibers diameter control between 5-200 μm, average pore size is 20-100 μm.
The beneficial effects of the invention are as follows：
（1）Increase 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 silk ribbon is combined with outer layer, and the slice of hydrophobic material is combined through the space of the silk ribbon 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.
(2) artificial dura mater internal layer, prepared using electrospinning process, obtain non-oriented property nano-fiber film.It is this kind thin Film tearing toughness is far longer than orientation film, and the intensity of artificial dura mater is improved from structure.
(3) artificial dura mater outer layer, prepared using electrospinning process, obtain orientation nano-fiber film, fibre gap It is easy to fibroblast to adhere to.
(4) cell production factor is interspersed among among electricity spinning fibre, effectively increases Porcine HGF and meningocyte Contact probability, give full play to the effect of Porcine HGF.
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, and 4 be cell growth factor Son.
Fig. 2 is interlayer structure schematic diagram.2-1 is the slice of carboxymethyl chitosan material, and 2-2 is polycaprolactone material Slice.
Fig. 3 is orientation nanofiber design sketch.
Fig. 4 is non-tropism 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.
Carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）Film is prepared into electrostatic spinning, film thickness is 40μm.Wide 3mm, long 1000mm slice is made in film.
Polycaprolactone（Bo Li biomaterials Co., Ltd of Shenzhen）Film, film thickness 40 are prepared into electrostatic spinning μm.Wide 3mm, long 1000mm slice is made in film.
Two kinds of slices are worked out using rhombus government establishments law, obtain Weaving type intermediate layer.
（3）Before internal layer solidification, Weaving type intermediate layer is put in internal layer upper surface.Top is put with 50N pressure Weaving type intermediate layer be pressed down against, keep 1min.
(4) prepared by outer layer：
Cladding material is carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）, it is dissolved in pure water (Jinan sea industry Work Co., Ltd) in, concentration of dope 6%（wt）, stir 1 hour, electrospinning parameters are：The .2ml/h of flow velocity 0, voltage 0.8kV, Distance 1mm is received, orientation electrostatic spinning process prepares 3h, obtains orientation layers of nanofibers, fiber longitudinal direction is with any while flat OK.
(5) fibroblast growth factor（Rh-bFGF dry powder, trade name：External application recombination human basic fibroblast growth The factor, Nanhai Longtime Pharmaceutical Co., Ltd.）Dissolved with water for injection (Jinan Hai Ye Chemical Co., Ltd.s), be uniformly sprayed onto brain Dura mater outer layer.Optimal dosage is 150IU/cm2。
(6) the artificial meninx by made from, it is vacuum-packed after freeze-dried, -20 DEG C of low temperature are protected after 25kGy bores -60 sterilizings Deposit.
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.
Carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）Film is prepared into electrostatic spinning, film thickness is 40μm.Wide 3mm, long 1000mm slice is made in film.
Polycaprolactone（Bo Li biomaterials Co., Ltd of Shenzhen）Film, film thickness 40 are prepared into electrostatic spinning μm.Wide 3mm, long 1000mm slice is made in film.
Two kinds of slices are worked out using rhombus government establishments law, obtain Weaving type intermediate layer.
（3）Before internal layer solidification, Weaving type intermediate layer is put in internal layer upper surface.Top is put with 50N pressure Weaving type intermediate layer be pressed down against, keep 1min.
(4) prepared by outer layer：
Cladding material is carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）, it is dissolved in pure water (Jinan sea industry Work Co., Ltd) in, 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 layers of nanofibers, fiber longitudinal direction is with any while flat OK.
(5) fibroblast growth factor（Rh-bFGF dry powder, trade name：External application recombination human basic fibroblast growth The factor, Nanhai Longtime Pharmaceutical Co., Ltd.）Dissolved with water for injection (Jinan Hai Ye Chemical Co., Ltd.s), be uniformly sprayed onto brain Dura mater outer layer.Optimal dosage is 150IU/cm2。
(6) the artificial meninx by made from, it is vacuum-packed after freeze-dried, -20 DEG C of low temperature are protected after 25kGy bores -60 sterilizings Deposit.
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.
Carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）Film is prepared into electrostatic spinning, film thickness is 40μm.Wide 3mm, long 1000mm slice is made in film.
Polycaprolactone（Bo Li biomaterials Co., Ltd of Shenzhen）Film, film thickness 40 are prepared into electrostatic spinning μm.Wide 3mm, long 1000mm slice is made in film.
Two kinds of slices are worked out using rhombus government establishments law, obtain Weaving type intermediate layer.
（3）Before internal layer solidification, Weaving type intermediate layer is put in internal layer upper surface.Top is put with 50N pressure Weaving type intermediate layer be pressed down against, keep 1min.
(4) prepared by outer layer：
Cladding material is carboxymethyl chitosan（Nantong Lv Shen bioengineering Co., Ltd）, it is dissolved in pure water (Jinan sea industry Work Co., Ltd) in, concentration of dope 6%（wt）, stir 1 hour, electrospinning parameters are：The .6ml/h of flow velocity 0, voltage 1kV, connect Receipts distance 2mm, orientation electrostatic spinning process prepare 3h, obtain orientation layers of nanofibers, and fiber longitudinal direction is parallel with any one side.
(5) fibroblast growth factor（Rh-bFGF dry powder, trade name：External application recombination human basic fibroblast growth The factor, Nanhai Longtime Pharmaceutical Co., Ltd.）Dissolved with water for injection (Jinan Hai Ye Chemical Co., Ltd.s), be uniformly sprayed onto brain Dura mater outer layer.Optimal dosage is 150IU/cm2。
(6) the artificial meninx by made from, it is vacuum-packed after freeze-dried, -20 DEG C of low temperature are protected after 25kGy bores -60 sterilizings Deposit.

Claims (10)

1. a kind of multilayer contains the nano fiber artificial endocranium of Porcine HGF, three layers are included：The cerebripetal internal layer in face, the back of the body Intermediate layer between cerebripetal outer layer and two layers.
2. a kind of multilayer according to claim 1 contains the nano fiber artificial endocranium of Porcine HGF, its feature It is, the internal layer thickness is 80 μm ± 10 μm, and the fiber formed by electrostatic spinning is prepared.
3. a kind of multilayer according to claim 1 contains the nano fiber artificial endocranium of Porcine HGF, its feature It is, the outer layer thickness is 30 μm ± 10 μm, and film is prepared into by orientation electrostatic spinning process.
4. a kind of multilayer according to claim 1 contains the nano fiber artificial endocranium of Porcine HGF, its feature It is, the intermediate layer thickness is 80 μm ± 20 μm, is formed by the braiding of diamond weave method.
5. intermediate layer according to claim 4, it is characterised in that the intermediate layer have hydrophilic material prepare width be The slice braiding that width prepared by 3mm slice and hydrophobic material is 3mm forms.
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 cutting after 40 μm of ± 10 μm of films；Second layer slice, it is 40 μm ± 10 μm thin to be prepared into thickness by electrospinning process Cutting forms after film.
7. intermediate layer according to claim 5, it is characterised in that be placed on internal layer upper table before internal layer solidification Face.
8. intermediate layer according to claim 5, it is characterised in that be pressed down against, protected to whole intermediate layer with 50N pressure Hold 1min.
9. a kind of multilayer according to claim 1 contains the nano fiber artificial endocranium of Porcine HGF, its feature It is, Porcine HGF is uniformly attached in the fiber gap of mechanical brains dura mater outer layer.
A kind of 10. nanofiber people that a kind of multilayer prepared any one of claim 1-9 contains Porcine HGF The dural method of work, comprises 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.
Carboxymethyl chitosan is prepared into film with electrostatic spinning, and film thickness is 40 μm.Film is cut to wide 3mm, it is long 1000mm slice.
Polycaprolactone is prepared into film with electrostatic spinning, and film thickness is 40 μm.Film is cut to wide 3mm, long 1000mm's Slice.
By two kinds of slices, two kinds of slice 3mmx1000mm are face-down, and hydrophily slice is transversely arranged, and hydrophobicity slice is longitudinally arranged Row, worked out using rhombus government establishments law, obtain Weaving type intermediate layer.
（3）Before internal layer solidification, Weaving type intermediate layer is put in internal layer upper surface.The volume put with 50N pressure to top The type intermediate layer of knitting is pressed down against, and keeps 1min.
(4) prepared by outer layer：
Cladding material is carboxymethyl chitosan, is dissolved in pure water, concentration of dope 6%（wt）, stir 1 hour, electrospinning ginseng Number is：The .4ml/h of flow velocity 0, voltage 0.9kV, distance 1mm is received, orientation electrostatic spinning process prepares 2h, obtains orientation nanometer Fibrous layer, fiber direction are parallel with any one side.
(5) fibroblast production factor is dissolved with water for injection, is uniformly sprayed onto brain dura mater outer layer.Optimal dosage is 150IU/ cm2。
(6) by obtained artificial dura mater, it is vacuum-packed after freeze-dried, -20 DEG C of Cord bloods after 25kGy bores -60 sterilizings.
CN201711094328.4A 2017-11-09 2017-11-09 Multilayer high-strength artificial dura mater containing cell factors and preparation method thereof Active CN107715178B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711094328.4A CN107715178B (en) 2017-11-09 2017-11-09 Multilayer high-strength artificial dura mater containing cell factors and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711094328.4A CN107715178B (en) 2017-11-09 2017-11-09 Multilayer high-strength artificial dura mater containing cell factors and preparation method thereof

Publications (2)

Publication Number Publication Date
CN107715178A true CN107715178A (en) 2018-02-23
CN107715178B CN107715178B (en) 2020-10-09

Family

ID=61214081

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711094328.4A Active CN107715178B (en) 2017-11-09 2017-11-09 Multilayer high-strength artificial dura mater containing cell factors and preparation method thereof

Country Status (1)

Country Link
CN (1) CN107715178B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110368136A (en) * 2019-06-19 2019-10-25 上海市第六人民医院 A kind of bionical stndon sheath film
CN113274557A (en) * 2021-04-30 2021-08-20 上海大学 Drug-loaded vascular stent tectorial membrane with unilateral drug release capability and preparation method thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003026489A2 (en) * 2001-09-28 2003-04-03 University Of Florida Biopolymer and biopolymer-cell compositions for nerve tissue repair
WO2007021759A2 (en) * 2005-08-10 2007-02-22 Nfocus Neuromedical Inc. Anti-restenotic therapeutic device
CN103480042A (en) * 2013-10-11 2014-01-01 于凤宾 Artificial dura mater spinalis, and preparation method and use method thereof
MX2012013108A (en) * 2012-11-09 2014-05-21 Rodrigo Ramos Zuñiga Chitosan biodegradable bilayer scaffold as a substitute of dura mater.
CN104415402A (en) * 2013-08-19 2015-03-18 冯淑芹 Three-dimensional patch used for nerve regeneration and dura mater repairing
CN104414773A (en) * 2013-08-23 2015-03-18 深圳迈普再生医学科技有限公司 Anti-adhesion tissue repair membrane and preparation method thereof
CN105999419A (en) * 2016-05-05 2016-10-12 烟台正海生物科技股份有限公司 Bionical type absorbable dura mater patch and preparation method and application thereof
CN106390204A (en) * 2016-11-16 2017-02-15 浙江省人民医院 Composite type artificial dura mater and preparation method

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003026489A2 (en) * 2001-09-28 2003-04-03 University Of Florida Biopolymer and biopolymer-cell compositions for nerve tissue repair
WO2007021759A2 (en) * 2005-08-10 2007-02-22 Nfocus Neuromedical Inc. Anti-restenotic therapeutic device
MX2012013108A (en) * 2012-11-09 2014-05-21 Rodrigo Ramos Zuñiga Chitosan biodegradable bilayer scaffold as a substitute of dura mater.
CN104415402A (en) * 2013-08-19 2015-03-18 冯淑芹 Three-dimensional patch used for nerve regeneration and dura mater repairing
CN104414773A (en) * 2013-08-23 2015-03-18 深圳迈普再生医学科技有限公司 Anti-adhesion tissue repair membrane and preparation method thereof
CN103480042A (en) * 2013-10-11 2014-01-01 于凤宾 Artificial dura mater spinalis, and preparation method and use method thereof
CN105999419A (en) * 2016-05-05 2016-10-12 烟台正海生物科技股份有限公司 Bionical type absorbable dura mater patch and preparation method and application thereof
CN106390204A (en) * 2016-11-16 2017-02-15 浙江省人民医院 Composite type artificial dura mater and preparation method

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110368136A (en) * 2019-06-19 2019-10-25 上海市第六人民医院 A kind of bionical stndon sheath film
CN110368136B (en) * 2019-06-19 2021-06-08 上海市第六人民医院 Bionic tendon sheath membrane
CN113274557A (en) * 2021-04-30 2021-08-20 上海大学 Drug-loaded vascular stent tectorial membrane with unilateral drug release capability and preparation method thereof

Also Published As

Publication number Publication date
CN107715178B (en) 2020-10-09

Similar Documents

Publication Publication Date Title
Wu et al. State-of-the-art review of advanced electrospun nanofiber yarn-based textiles for biomedical applications
US20210138104A1 (en) Electrospun biocompatible fiber compositions
Sun et al. Electrospun anisotropic architectures and porous structures for tissue engineering
Yan et al. Electrospun nanofibrous membrane for biomedical application
CN104888278B (en) A kind of nano/micron fiber three-dimensional porous structure timbering material and its preparation and application
CN103263694B (en) Collagen-based dura and preparation method thereof
JP2018534113A (en) System and method for generation of gastrointestinal tissue
CN105031739B (en) A kind of GTR/GBR compound film sheets for carrying Doxycycline Hyclate and preparation method thereof
KR101387886B1 (en) Method of fabricating a scaffold capable of controlling the thickness and pore size thereof, the scaffold fabricated thereby, and a fabricating equipment used therefor
Li et al. Fabrication of multilayered nanofiber scaffolds with a highly aligned nanofiber yarn for anisotropic tissue regeneration
CN111617320B (en) Bone induction regeneration membrane and preparation method and application thereof
WO2016202281A1 (en) Reinforced electrospun fibrous membrane and preparation method thereof
CN109196152B (en) Yarn for cell culture scaffold and fabric comprising same
CN107320787A (en) A kind of periodontal reparation porous fibre membrane material and preparation method thereof
CN107715178A (en) A kind of multilayer contains high intensity artificial dura mater of cell factor and preparation method thereof
CN110917386A (en) Stem cell sponge scaffold, preparation method and application
CN106955373B (en) Multilayer nanofiber micro-caliber vascular tissue engineering scaffold material and preparation method thereof
Zhou et al. Dual release of VEGF and PDGF from emulsion electrospun bilayer scaffolds consisting of orthogonally aligned nanofibers for gastrointestinal tract regeneration
CN107875448A (en) A kind of centripetal orientation nano fiber artificial endocranium of multilayer and preparation method thereof
CN105232184B (en) A kind of artificial blood vessel and preparation method thereof and ejecting device
CN107648671B (en) Multilayer anti-infection high-strength artificial dura mater and preparation method thereof
KR101869342B1 (en) Natural silk non-woven fabric and its preparation method
CN107865981A (en) A kind of multilayer oriented property nano fiber artificial endocranium and preparation method thereof
CN107875447A (en) A kind of multi-layer braided type electrostatic spinning artificial dura mater and preparation method thereof
CN107823713A (en) A kind of multilayer fusion electrostatic spinning artificial dura mater and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant