CN107875447B - Multilayer woven electrostatic spinning artificial dura mater and preparation method thereof - Google Patents
Multilayer woven electrostatic spinning artificial dura mater and preparation method thereof Download PDFInfo
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- 238000010041 electrostatic spinning Methods 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
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- 238000000034 method Methods 0.000 claims abstract description 15
- 230000008569 process Effects 0.000 claims abstract description 13
- 210000004556 brain Anatomy 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims description 64
- 229920000954 Polyglycolide Polymers 0.000 claims description 14
- 229920001610 polycaprolactone Polymers 0.000 claims description 14
- 239000004632 polycaprolactone Substances 0.000 claims description 14
- 239000004633 polyglycolic acid Substances 0.000 claims description 14
- 238000009987 spinning Methods 0.000 claims description 14
- 239000000835 fiber Substances 0.000 claims description 13
- 239000011148 porous material Substances 0.000 claims description 11
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 10
- GUTLYIVDDKVIGB-OUBTZVSYSA-N Cobalt-60 Chemical compound [60Co] GUTLYIVDDKVIGB-OUBTZVSYSA-N 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000010408 film Substances 0.000 claims 8
- 239000010409 thin film Substances 0.000 claims 2
- 238000004108 freeze drying Methods 0.000 claims 1
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- 230000001954 sterilising effect Effects 0.000 claims 1
- 238000009461 vacuum packaging Methods 0.000 claims 1
- 238000001523 electrospinning Methods 0.000 abstract description 13
- 238000009941 weaving Methods 0.000 abstract description 5
- 239000010410 layer Substances 0.000 description 133
- 210000002950 fibroblast Anatomy 0.000 description 11
- 230000007547 defect Effects 0.000 description 8
- 210000002418 meninge Anatomy 0.000 description 8
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- 238000013508 migration Methods 0.000 description 7
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 238000010146 3D printing Methods 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 208000032544 Cicatrix Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000012422 Collagen Type I Human genes 0.000 description 1
- 108010022452 Collagen Type I Proteins 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 1
- 206010019196 Head injury Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
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- 230000000735 allogeneic effect Effects 0.000 description 1
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- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
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- 229920000728 polyester Polymers 0.000 description 1
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- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 230000037387 scars Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
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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
- A61L27/14—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
-
- 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
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4326—Condensation or reaction polymers
- D04H1/435—Polyesters
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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
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- Oral & Maxillofacial Surgery (AREA)
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Abstract
The invention provides a multilayer weaving type electrostatic spinning artificial dura mater, which comprises three layers, a hydrophobic inner layer facing the brain, a hydrophilic outer layer facing away from the brain, and a weaving type middle layer between the two layers. The invention also provides a preparation method of the multilayer weaving type electrostatic spinning artificial dura mater, which comprises the following steps: preparing a hydrophobic inner layer by electrospinning; preparing hydrophilic thin strips; preparing a hydrophobic fine strip; preparing a woven intermediate layer; and preparing an outer layer on the middle layer by using an electrospinning process, and finally obtaining the artificial dura mater. The artificial dura mater effectively solves the layering problem of the artificial dura mater, can be cut randomly according to the needs, and has wide application prospect.
Description
Technical Field
The invention relates to a multilayer braiding electrostatic spinning artificial dura mater and a preparation method thereof, belonging to the technical field of biological 3D printing.
Background
The dural defect is common in neurosurgery clinical work, and can be caused by open craniocerebral injury (industry, traffic, war, and the like), tumor erosion, congenital meningal defect and other craniocerebral disease reasons. The dural defect needs to be repaired in time to prevent the cerebrospinal fluid from overflowing, and prevent the brain from bulging and the atmospheric pressure from being pressed, otherwise, the life of a human body is endangered. The dural defect can also cause complications such as intracranial infection, cerebral adhesion, subcutaneous effusion and the like, and often causes diseases such as headache, cerebral dysfunction and the like.
At present, artificial dura mater made of various materials is in clinical use, and the artificial dura mater can be mainly divided into two main types, namely biological derivative materials and artificial synthetic high polymer materials. The biological derivative material mainly comprises allogeneic human dura mater, xenogeneic porcine/bovine pericardium, dermal matrix, biomembrane prepared by utilizing bovine muscle bond type I collagen, and the like. The synthetic polymer material mainly comprises polyester degradable polymers such as polylactic acid, polyglycolic acid, polycaprolactone, polyurethane and the like. In addition, the material also comprises non-degradable high polymer materials such as polytetrafluoroethylene and the like.
At present, the repair effect of the meninges injury is ideal to be a multilayer artificial dura mater. The multilayer artificial dura mater achieves the purpose of simulating the physiological function of the dura mater by layering. Such artificial dura mater generally consists of an inner layer and an outer layer. Wherein the inner layer is made of a hydrophobic material and the outer layer is made of a hydrophilic material. Because the two layers of materials have different hydrophobic and hydrophilic properties, the connection reliability of the outer layer and the inner layer is not high, the peeling phenomenon occurs, and the life and the health of patients are seriously threatened.
The meninges are mainly distributed by fibroblasts and collagen fibers secreted by them. Typically, the fibroblast diameter is between 20-30. Mu.m. The literature reports that when the diameter of the electrospun fiber is between 50 and 1000nm, the average pore diameter can reach 2 μm. The average pore diameter is below 3 mu m, so that cells can be effectively prevented from entering the electrostatic spinning fiber film, and the brain adhesion is effectively generated. When the fiber diameter is between 5 and 200 mu m, the average pore diameter reaches 20 to 100 mu m. This facilitates the migration, adhesion, proliferation and growth differentiation of fibroblasts.
The Chinese patent CN 103480042A is named as "an artificial dura mater, a preparation method and a use method thereof", and discloses an artificial dura mater with a double-layer structure, which consists of an oriented polylactic acid/glycollic acid copolymer fiber film inner layer and a non-oriented polylactic acid/glycollic acid copolymer-chitosan fiber film outer layer. Although the artificial dura mater can realize the purposes of dura mater regeneration and prevention of scars and adhesion, the artificial dura mater has the following defects that the outer layer fiber and the inner layer fiber are required to be separately placed at the position of the meninges defect in the clinical use process, and meanwhile, the adhesive is required to be coated between the two layers, so that the inconvenience in the clinical use process is greatly increased.
The Chinese patent CN 101507661A is named as nano artificial dura mater with multiple functional layers and a preparation method thereof, and discloses an artificial dura mater with a double-layer structure. Although the artificial dura mater can realize the purpose of dura mater regeneration, the artificial dura mater has the following defects that the outer layer prepared from the hydrophilic material is directly subjected to electrostatic spinning on the inner layer prepared from the hydrophobic material, and the connection between the outer layer and the inner layer is not firm and is easy to peel off due to the different hydrophobic and hydrophilic properties of the two layers of materials, so that the life safety of a meninges repair operation patient is directly threatened.
Therefore, the artificial dura mater with reliable connection between the inner layer and the outer layer, convenient use and wide industrialization prospect is urgently needed in clinic at present.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a multilayer braiding type electrostatic spinning artificial dura mater, which is different from the traditional artificial dura mater in that: the weaving type middle layer is additionally arranged between the inner layer and the outer layer of the artificial dura mater, the middle layer is woven by adopting a hydrophobic material and a hydrophilic material, the thin strips prepared by the hydrophobic material in the weaving layer are reliably combined with the inner layer of the artificial dura mater (prepared by electrospinning of the hydrophobic material), the thin strips prepared by the hydrophilic material are reliably combined with the outer layer of the artificial dura mater (prepared by electrospinning of the hydrophilic material), the two thin strips are overlapped and interweaved together, the thin strips of the hydrophilic material are combined with the outer layer through gaps of the thin strips of the hydrophobic material, and the thin strips of the hydrophobic material are combined with the inner layer through gaps of the thin strips of the hydrophilic material, so that the inner layer and the outer layer are reliably combined together. In a word, the artificial dura mater of this kind of structure has improved the cohesion between inlayer and the extexine, is difficult for taking place the peeling off between the layer under the effect of external force.
The invention further aims to provide a preparation method of the multilayer woven electrostatic spinning artificial dura mater.
The invention solves the problems by adopting the following technical scheme:
a multilayer braiding electrostatic spinning artificial dura mater comprises three layers, wherein one layer facing the brain is a hydrophobic inner layer, and the other layer facing away from the brain is a hydrophilic outer layer; a braiding type middle layer is arranged between the two layers.
The hydrophilic outer layer is made of electrostatic spinning, namely, the outer layer is made of hydrophilic materials with good biocompatibility for effectively inducing the migration of stem cells and fibroblasts, and the average pore diameter is 20-100 mu m and the diameter of the fibroblasts is 20-30 mu m generally through electrostatic spinning parameter adjustment. This facilitates the migration, adhesion, proliferation and growth differentiation of fibroblasts.
The inner layer is made of a hydrophobic material, so that the migration of cells is prevented, and the anti-adhesion purpose is achieved; the electrostatic spinning parameters for preparing the layer material are adjusted to ensure that the pore diameter is below nanometer and is one to two orders of magnitude smaller than that of cells (generally, the diameter of fibroblast is between 20 and 30 mu m), thereby preventing the cells from entering and preventing the brain adhesion.
The middle layer is woven by adopting a hydrophobic material and a hydrophilic material.
The multilayer woven electrostatic spinning artificial dura mater comprises an inner layer, an intermediate layer and an outer layer, and the preparation method comprises the following steps:
(1) Preparation of the inner layer: the inner layer material comprises polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and is dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) to obtain a spinning solution with a concentration of 7% (wt), and stirring for 4 hours, wherein the electrospinning parameters are that the flow rate is 0.9 ml/h, the voltage is 13kV, the receiving distance is 20cm, and the electrospinning is performed for 4 hours;
(2) And (5) preparing an intermediate layer. The first layer material is a film prepared by electrostatic spinning of polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 40 μm.
The film was made into strips 3mm wide and 1000mm long.
The strips are placed in parallel with any side of the inner layer, and 3mmx1000mm faces towards the inner layer according to the interval of 6mm, so that the whole inner layer is uniformly arranged.
The second layer material is a film prepared by electrostatic spinning of polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 40 μm.
The film was made into strips 3mm wide and 1000mm long.
The second layer of thin strips and the first layer of thin strips are placed at an angle of 90 degrees, and 3mmx1000mm faces towards the inner layer according to the interval of 6mm, so that the whole first layer of thin strips are uniformly distributed.
In particular, the strand placement process described above should be completed prior to curing the inner layer.
Finally, the thin strips placed on the upper side are pressed downwards by using 50N pressure and kept for 1min.
(3) The outer layer material is polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) and the concentration of spinning solution is 6% (wt.), and the spinning parameters are: the flow rate is 1.4 ml/h, the voltage is 12kV, the receiving distance is 10cm, and the electrostatic spinning is carried out for 4h.
(4) The artificial meninges are taken out of the culture dish, rinsed 3 times with distilled water, freeze-dried, vacuum-packed, sterilized by 25kGy cobalt-60 and stored at-20 ℃.
The thickness of each layer of the prepared multilayer electrostatic spinning artificial dura mater is as follows: the thickness of the inner layer is 80 mu m +/-10 mu m, the thickness of the outer layer is 30 mu m +/-10 mu m, and the thickness of the middle layer is 80 mu m +/-20 mu m. The thickness of the three layers in the fusion layer is respectively as follows: from bottom to top, the first layer thickness is: 40 μm.+ -. 10 μm, the second layer thickness is: 40 μm.+ -. 10 μm.
When the inner layer is prepared, the diameter of the electrostatic spinning fiber is controlled between 50 nm and 1000nm, and the average pore diameter is 2 mu m;
when the outer layer is prepared, the diameter of the electrostatic spinning fiber is controlled between 5 and 200 mu m, and the average pore diameter is 20 to 100 mu m.
The beneficial effects of the invention are as follows:
(1) The artificial dura mater adopts a three-layer structure, and the bonding strength of the inner layer and the outer layer is enhanced through the middle layer.
(2) According to the artificial dura mater, a braiding type middle layer is additionally arranged between an inner layer and an outer layer of the artificial dura mater, the middle layer is formed by braiding a hydrophobic material and a hydrophilic material, the thin strips prepared by the hydrophobic material in the braiding layer are reliably combined with the inner layer of the artificial dura mater (prepared by electrospinning of the hydrophobic material), the thin strips prepared by the hydrophilic material are reliably combined with the outer layer of the artificial dura mater (prepared by electrospinning of the hydrophilic material), the thin strips of the hydrophilic material are overlapped and interweaved together, the thin strips of the hydrophilic material are combined with the outer layer through gaps of the thin strips of the hydrophilic material, and the thin strips of the hydrophobic material are combined with the inner layer through gaps of the thin strips of the hydrophilic material, so that the inner layer and the outer layer are reliably combined together.
(3) In order to effectively induce the migration of stem cells and fibroblasts, the outer layer of the artificial dura mater adopts hydrophilic materials with good biocompatibility, and the average pore diameter reaches 20-100 mu m through electrostatic spinning parameter adjustment, and the diameter of the fibroblasts is generally 20-30 mu m. This facilitates the migration, adhesion, proliferation and growth differentiation of fibroblasts.
(4) The inner layer of the artificial dura mater adopts a hydrophobic material to prevent the migration of cells, thereby achieving the purpose of preventing adhesion; the electrostatic spinning parameters for preparing the layer material are adjusted to ensure that the pore diameter is below nanometer and is one to two orders of magnitude smaller than that of cells (generally, the diameter of fibroblast is between 20 and 30 mu m), thereby preventing the cells from entering and preventing the brain adhesion.
Drawings
Fig. 1 is a schematic diagram of the structure of the artificial dura mater, wherein 1 is an inner layer, 2 is an intermediate layer, and 3 is an outer layer.
Fig. 2 is a schematic three-dimensional structure of artificial dura mater, 4 is a thin strip cut from a polycaprolactone film, and 5 is a thin strip cut from a polyglycolic acid film.
FIG. 3 is a schematic illustration of the structure of the middle layer of artificial dura mater.
Detailed Description
The present invention will be described with reference to the following specific examples, but the present invention is not limited thereto.
The experimental methods used in the following examples are conventional methods, and the reagents, materials, etc. used in the following examples are commercially available, unless otherwise specified.
Example 1
(1) Preparation of the inner layer: the inner layer material comprises polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and is dissolved in hexafluoroisopropanol (Suzhou Kazakh Sail biological Co., ltd.) to obtain a spinning solution with a concentration of 5% (wt), stirring for 1 hr, and electrospinning for 2 hr at a flow rate of 1.0 ml/h, a voltage of 12kV, a receiving distance of 15 cm;
(2) And (5) preparing an intermediate layer. The first layer material is a film prepared by electrostatic spinning of polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) and the thickness of the film is 30 μm.
The film was made into thin strips 4mm wide and 1000mm long.
The strips are placed in parallel with any side of the inner layer, and 4mmx1000mm faces towards the inner layer according to the interval of 8mm, so that the whole inner layer is uniformly arranged.
The second layer material is a film prepared by electrostatic spinning of polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 30 μm.
The film was made into thin strips 4mm wide and 1000mm long.
The strips are placed at an angle of 90 degrees with the first layer of strips, and the whole first layer of strips are uniformly arranged according to the interval of 8mm and the surface of 4mmx1000mm facing the inner layer.
In particular, the strand placement process described above should be completed prior to curing the inner layer.
The thin strips placed on the upper side are pressed downwards by using 50N pressure and kept for 1min.
(3) The outer layer material is polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) and the concentration of spinning solution is 6% (wt.), and the spinning parameters are: the flow rate is 1.2 ml/h, the voltage is 14kV, the receiving distance is 15cm, and the electrostatic spinning is carried out for 6h.
(4) The artificial meninges are taken out of the culture dish, rinsed 3 times with distilled water, freeze-dried, vacuum-packed, sterilized by 25kGy cobalt-60 and stored at-20 ℃.
Example 2
(1) Preparation of the inner layer: the inner layer material is polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and is dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) to obtain a spinning solution with a concentration of 7% wt, and stirring for 4 hours, wherein the electrospinning parameters are that the flow rate is 0.9 ml/h, the voltage is 13kV, the receiving distance is 20cm, and the electrospinning is performed for 4 hours;
(2) And (5) preparing an intermediate layer. The first layer material is a film prepared by electrostatic spinning of polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 40 μm.
The film was made into strips 3mm wide and 1000mm long.
The strips are placed in parallel with any side of the inner layer, and 3mmx1000mm faces towards the inner layer according to the interval of 6mm, so that the whole inner layer is uniformly arranged.
The second layer material is a film prepared by electrostatic spinning of polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 40 μm.
The film was made into strips 3mm wide and 1000mm long.
The strips and the first layer of strips are placed at an angle of 90 degrees, and 3mmx1000mm faces towards the inner layer according to the interval of 6mm, and the whole first layer of strips is uniformly arranged.
In particular, the strand placement process described above should be completed prior to curing the inner layer.
The thin strips placed on the upper side are pressed downwards by using 50N pressure and kept for 1min.
(3) The outer layer material is polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) and the concentration of spinning solution is 6% (wt.), and the spinning parameters are: the flow rate is 1.4 ml/h, the voltage is 12kV, the receiving distance is 10cm, and the electrostatic spinning is carried out for 4h.
(4) The artificial meninges are taken out of the culture dish, rinsed 3 times with distilled water, freeze-dried, vacuum-packed, sterilized by 25kGy cobalt-60 and stored at-20 ℃.
Example 3
(1) Preparation of the inner layer: the inner layer material is polycaprolactone (Shenzhen Boli biological materials Co., ltd.) dissolved in hexafluoroisopropanol (Suzhou Kazakh Sail biological Co., ltd.) and stirred for 4 hr, and the electrospinning parameters are flow rate of 1.0 ml/h, voltage of 15kV, receiving distance of 25cm, and electrospinning for 5 hr.
(2) And (5) preparing an intermediate layer. The first layer material is a film prepared by electrostatic spinning of polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 40 μm.
The film was formed into strips 5mm wide and 1000mm long.
The strips are placed in parallel with any side of the inner layer, and the whole inner layer is uniformly arranged according to the interval of 10mm and the surface of 5mmx1000mm facing the inner layer.
The second layer material is a film prepared by electrostatic spinning of polycaprolactone (Shenzhen Boli biological materials Co., ltd.) and has a film thickness of 50 μm.
The film was formed into strips 5mm wide and 1000mm long.
The strips are placed at an angle of 90 degrees with the first layer of strips, and the whole first layer of strips are uniformly arranged according to a distance of 10mm and a surface of 5mmx1000mm facing the inner layer.
In particular, the strand placement process described above should be completed prior to curing the inner layer.
The thin strips placed on the upper side are pressed downwards by using 50N pressure and kept for 1min.
(3) The outer layer material is polyglycolic acid (Shenzhen Boli biological materials Co., ltd.) dissolved in hexafluoroisopropanol (Suzhou Kaisha Sail biological Co., ltd.) and the concentration of spinning solution is 6% (wt.), and the spinning parameters are: the flow rate is 1.2 ml/h, the voltage is 14kV, the receiving distance is 10cm, and the electrostatic spinning is carried out for 4h.
(4) The artificial meninges are taken out of the culture dish, rinsed 3 times with distilled water, freeze-dried, vacuum-packed, sterilized by 25kGy cobalt-60 and stored at-20 ℃.
Claims (4)
1. A multilayer woven electrospun artificial dura mater comprising: an inner layer facing the brain, an outer layer facing away from the brain, and an intermediate layer between the inner and outer layers, wherein:
the thickness of the inner layer is 80 mu m plus or minus 10 mu m, the inner layer is prepared by an electrostatic spinning process, the diameter of the electrostatic spinning fiber is controlled between 50 nm and 1000nm, the average pore diameter is 2 mu m, and the inner layer material is polycaprolactone;
the thickness of the outer layer is 30 mu m plus or minus 10 mu m, the fiber is prepared by an electrostatic spinning process, the diameter of the electrostatic spinning fiber is controlled between 5 mu m and 200 mu m, the average pore diameter is 20 mu m to 100 mu m, and the outer layer material is polyglycolic acid;
the thickness of the middle layer is 80 mu m plus or minus 20 mu m, and the middle layer is composed of two layers, wherein the first layer is a thin strip with the width of 3mm and the length of 1000mm prepared from hydrophilic materials, and the second layer is a thin strip with the width of 3mm and the length of 1000mm prepared from hydrophobic materials;
preparing a film with the thickness of 40 mu m plus or minus 10 mu m by using polyglycolic acid through an electrostatic spinning process, and then cutting the film into a first layer of thin strips; preparing a thin film with the thickness of 40 mu m plus or minus 10 mu m by using polycaprolactone through an electrostatic spinning process, and then cutting the thin film into a second layer of thin strips;
the first layer of thin strips are placed in parallel with any side edge of the inner layer, 3mm multiplied by 1000mm faces towards the inner layer according to the interval of 6mm, and the whole inner layer is uniformly filled; the second layer of thin strips are placed at an angle of 90 degrees with the first layer of thin strips, and 3mm multiplied by 1000mm faces the inner layer according to the interval of 6mm, and the whole first layer of thin strips are uniformly distributed.
2. The multilayer knitted electrospun artificial dura mater of claim 1, wherein the two-layer strand placement process is completed before the inner layer is cured.
3. The multilayer knitted electrospun artificial dura mater according to claim 1, wherein the laid sliver is pressed vertically downward with a pressure of 50N for 1min.
4. A method for preparing a multilayer woven electrospun artificial dura mater according to any one of claims 1 to 3, comprising the steps of:
(1) Preparation of the inner layer: the inner layer material comprises polycaprolactone, and is dissolved in hexafluoroisopropanol, the concentration of spinning solution is 7wt%, and the spinning solution is stirred for 4 hours, and the electrostatic spinning parameters are as follows: the flow rate is 0.9 mL/h, the voltage is 13kV, the receiving distance is 20cm, and the electrostatic spinning is carried out for 4h;
(2) Intermediate layer preparation: the first layer of material is a film prepared by electrostatic spinning of polyglycolic acid, and the thickness of the film is 40 mu m;
making the film into a first layer of thin strips with the width of 3mm and the length of 1000 mm;
the first layer of thin strips are placed in parallel with any side edge of the inner layer, 3mm multiplied by 1000mm faces towards the inner layer according to the interval of 6mm, and the whole inner layer is uniformly filled;
the second layer of material is a film prepared by using polycaprolactone electrostatic spinning, and the thickness of the film is 40 mu m;
making the film into a second layer of thin strips with the width of 3mm and the length of 1000 mm;
placing the second layer of thin strips at an angle of 90 degrees with the first layer of thin strips, and uniformly arranging the whole first layer of thin strips according to a spacing of 6mm and a surface of 3mm multiplied by 1000mm facing the inner layer;
the two-layer thin strip placing process is completed before the inner layer is solidified;
finally, the placed thin strips are pressed downwards by using 50N pressure and kept for 1min;
(3) The outer layer material is polyglycolic acid, dissolved in hexafluoroisopropanol, the concentration of the spinning solution is 6wt%, and the mixture is stirred for 1 hour, and the electrostatic spinning parameters are as follows: the flow rate is 1.4 mL/h, the voltage is 12kV, the receiving distance is 10cm, and the electrostatic spinning is carried out for 4h;
(4) Taking out the prepared artificial dura mater from the culture dish, rinsing with distilled water for 3 times, freeze-drying, vacuum packaging, sterilizing with 25kGy cobalt-60, and preserving at-20deg.C.
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