WO2020133668A1 - 一种镁丝蚕丝复合编织结构神经导管及其制备方法 - Google Patents

一种镁丝蚕丝复合编织结构神经导管及其制备方法 Download PDF

Info

Publication number
WO2020133668A1
WO2020133668A1 PCT/CN2019/076010 CN2019076010W WO2020133668A1 WO 2020133668 A1 WO2020133668 A1 WO 2020133668A1 CN 2019076010 W CN2019076010 W CN 2019076010W WO 2020133668 A1 WO2020133668 A1 WO 2020133668A1
Authority
WO
WIPO (PCT)
Prior art keywords
silk
catheter
nerve
magnesium
solution
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.)
Ceased
Application number
PCT/CN2019/076010
Other languages
English (en)
French (fr)
Inventor
李刚
张淑军
郑兆柱
王晓沁
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.)
Suzhou University
Nantong Textile and Silk Industrial Technology Research Institute
Original Assignee
Suzhou University
Nantong Textile and Silk Industrial Technology Research Institute
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 Suzhou University, Nantong Textile and Silk Industrial Technology Research Institute filed Critical Suzhou University
Priority to JP2021523697A priority Critical patent/JP2022506369A/ja
Publication of WO2020133668A1 publication Critical patent/WO2020133668A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

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
    • A61L27/26—Mixtures of macromolecular compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • 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
    • A61L27/02—Inorganic materials
    • A61L27/04—Metals or alloys
    • A61L27/047—Other specific metals or alloys not covered by A61L27/042 - A61L27/045 or A61L27/06
    • 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
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
    • A61L27/383—Nerve cells, e.g. dendritic cells, Schwann cells
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3839—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by the site of application in the body
    • A61L27/3878—Nerve tissue, brain, spinal cord, nerves, dura mater
    • 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
    • A61L27/52—Hydrogels or hydrocolloids
    • 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
    • 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
    • A61L27/56—Porous materials, e.g. foams or sponges
    • 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
    • A61L27/58—Materials at least partially resorbable by the body
    • 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
    • A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
    • 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
    • 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

Definitions

  • the invention belongs to the technical field of biomedical materials, and in particular relates to a magnesium-silk silk composite braided structure nerve catheter and a preparation method thereof.
  • Peripheral nerve injury is a common clinical condition.
  • the use of biomaterials and tissue engineering techniques to promote the repair and regeneration of defective nerves is a research hotspot in the field of neuroscience.
  • Ideal biological materials, appropriate physical and mechanical guidance and suitable biological microenvironment are the key factors to promote the recovery of sensory, motor and other biological functions of damaged nerves.
  • the most researched nerve catheters are the use of model casting method, electrostatic nano-spinning method, 3D bioprinting and other technologies, and the use of biodegradable materials such as silk fibroin, chitosan and collagen to produce physical guides. Hollow catheters, but the neural catheters produced by these techniques have poor mechanical mechanics and flexibility.
  • a kind of nerve catheter using silk fibroin nanofiber directional guidance function uses condensation-induced silk fibroin solution to form nano-micropores as physical guidance, but the micropores formed in this invention are difficult to control the shape size and direction, and the radial compression of the catheter General performance, bending flexibility is not considered;
  • another example is to use nano-oriented fibers to make films with different degradation concentrations and then roll them into a neural tube with a tubular structure.
  • This neural tube has excellent physical guidance to cells, but its mechanical properties are relatively good. Poor, there is less space for nerve regeneration inside;
  • another example is the use of 3D bioprinting, which has greater restrictions on materials, and the current resolution is lower, which is about 1 ⁇ 300mPa/s.
  • magnesium is one of the important elements necessary for the human body. It participates in protein synthesis in the brain and central nervous system, activates various enzymes in the body, and regulates neuromuscular activity.
  • metal magnesium ions have various biological functions such as anti-inflammatory, anti-oxidation, anti-apoptosis, and regulation of mitochondrial calcium buffer.
  • the latest research shows that magnesium ions can promote the proliferation of Schwann cells under the appropriate microenvironment, such as secretion of growth factors, extracellular matrix (ECM), etc., which play a role in supporting and protecting neurons.
  • Silkworm silk is a natural animal protein synthesized and secreted by the silkworm. It has a wide range of sources.
  • Silk fibroin is the main component of silk, accounting for 70%-80% of the quality of silk. The remaining main components are sericin and carbohydrates.
  • Silk fibroin contains a variety of amino acids, of which the more common alanine (Ala), glycine (Gly), and serine (Ser) account for about 85% of the total composition.
  • Silk fibroin itself has good physical and chemical properties, breathable moisture, biocompatibility, can be degraded into peptides and free amino acids in the body can be absorbed by the body to promote the growth of nerve cells.
  • the nerve duct made of silk fibroin has low inflammatory response in the body, is biodegradable, and can promote the functional recovery of peripheral nerve injury.
  • the present invention proposes a magnesium silk silk composite braided structure nerve catheter and a preparation method thereof.
  • the purpose of the present invention is to provide a magnesium silk silk composite braided structure neural catheter and a preparation method thereof, which can improve the order of nerve regeneration and the mechanical strength of the nerve catheter to solve the above problems.
  • a nerve conduit with a composite braided structure of magnesium silk and silk comprising an outer layer of a nerve conduit, an intermediate layer of a nerve conduit, and an inner layer of a nerve conduit, the middle layer of the nerve conduit covers the outside of the inner layer of the nerve conduit, and the outer layer of the nerve conduit
  • the outer layer of the nerve catheter is a porous sponge structure formed by mixing silk fibroin solution and chitosan solution, and the thickness of the outer layer of the nerve catheter is 0.5-1.5mm ,
  • the pore diameter is 20-150 ⁇ m, the porosity is 75%-90%
  • the middle layer of the nerve conduit is a three-dimensional braided structure, the thickness of the middle layer of the nerve conduit is 0.3-1mm, and the braid angle of the three-dimensional braided structure is 45 -60°
  • the inner layer of the nerve conduit is a porous gel structure
  • the diameter of the inner layer of the nerve conduit is 1.8-5 mm.
  • the three-dimensional braided structure is any one of a diamond braided structure, a regular braided structure and a Hergliss braided structure.
  • the three-dimensional braided structure is a three-dimensional structural catheter skeleton formed by compound braiding of magnesium wire as the shaft yarn and degummed silk as the braided yarn.
  • the diameter of the magnesium wire is 0.1-0.25 mm and the number of wires is 4-16.
  • the distance between two adjacent magnesium wires is 200-2000 ⁇ m, and the linear density of the degummed silk is 20-120D.
  • the porous gel structure is a composite gel layer composed of silk fibroin gel, Schwann cells and nerve growth factor, the silk fibroin gel is frozen to form a mesh fiber, the Schwann cell and nerve
  • the growth factor is set in the mesh fiber, the diameter of the mesh fiber is 10-1000 nm, and the pore diameter is 10-500 ⁇ m.
  • a method for preparing a nerve conduit of magnesium silk and silk composite braided structure includes the following steps:
  • a vertical spindle braiding machine with a core function is used to prepare the skeleton structure of the nerve catheter, which is composite braided with magnesium wire as the shaft yarn and degummed silk as the woven yarn;
  • a silk fibroin solution with a concentration of 0.01-0.5% cultured Schwann cells is injected into the hollow structure of the outer layer of the neural catheter, and it is left at a temperature of 37°C for 12 hours to form Gelation catheter; insert the gelation catheter vertically into a liquid nitrogen tank, freeze slowly at a speed of 0.1-1 mm/s along the axis of the gelation catheter, and then put it into the freeze dryer It was freeze-dried in medium for 24-48 hours to obtain magnesium silk silk composite braided structure neural catheter.
  • the preparation method of the silk fibroin solution in step (2) is as follows: weigh silkworm raw silk to degumme in boiling Na 2 CO 3 aqueous solution for 30 minutes, stir with a glass rod every 10 minutes, and use deionized water After the second rinse, place it in a fume hood overnight and dry it naturally to obtain dried silkworm silk fibers. Dissolve the dried silkworm silk fibers in the LiBr solution and dissolve them in an oven at 60°C for 4 hours. Ensure that the dissolution is complete, obtain the first mixed solution, centrifuge the first mixed solution after dialysis to obtain the silk fibroin solution, and place it in a refrigerator at 4 °C for use.
  • the dialysis in step (2) is: pouring the first mixed solution into a dialysis bag with a molecular weight cut-off of 3500D, dialyzing in deionized water for 36 hours, and changing the water every 4 hours.
  • step (2) centrifuging twice at 9000 r/min in a high-speed centrifuge for 20 minutes each time.
  • the preparation method of the chitosan solution in step (2) is: weigh out chitosan with a degree of deacetylation of 80-95%, dissolve in 0.5-4% acetic acid solution, and stir for 4-12 hours To make a chitosan solution with a mass percentage of 1-5%.
  • the invention provides a magnesium-silk silk composite braided structure nerve catheter and a preparation method thereof, and its advantages are:
  • the catheter has good biocompatibility, surface activity and permeability, can promote the adhesion, growth and proliferation of neuronal cells, and accelerate the regeneration of damaged nerves;
  • the degradation rate of the catheter can be adjusted to provide a suitable growth space for nerve regeneration
  • the magnesium wire in the catheter can stimulate and guide nerve growth.
  • the magnesium ions generated during degradation can help to increase the activity of neuronal cells.
  • FIG. 1 is a schematic structural view of a magnesium silk silk composite braided structure neural catheter according to the present invention in Example 1;
  • FIG. 2 is a schematic structural view of a magnesium silk silk composite braided structure neural catheter according to the present invention in Example 2;
  • Example 3 is a schematic structural diagram of a magnesium silk silk composite braided structure nerve catheter according to the present invention in Example 1;
  • FIG. 4 is a braid angle of a neural catheter with an inner diameter of 2 mm and 3 mm made of magnesium silk composite braided structure prepared with different parameters according to Example 1 of the present invention
  • FIG. 5 is a load-tensile strain curve of a neural catheter with an inner diameter of 2 mm and 3 mm prepared by using different parameters of a composite braided structure of magnesium silk and silk according to Example 1 of the present invention.
  • 1 is the outer layer of the nerve conduit
  • 2 is the middle layer of the nerve conduit
  • 3 is the inner layer of the nerve conduit
  • 4 is the shaft yarn
  • 5 is the braided yarn
  • 6 is the silk fibroin gel
  • 7 is the Schwann cell
  • 8 is the nerve Growth factor
  • 9 is titanium, nickel alloy wire.
  • the invention provides a magnesium silk silk composite braided structure neural catheter.
  • the catheter is divided into three layers, including: the outer layer 1 of the neural catheter is a porous sponge formed by mixing silk fibroin solution and chitosan solution Structure, the thickness is 0.5-1.5mm, the pore size is 20-150 ⁇ m, the porosity is 75%-90%, the middle layer 2 of the neural catheter is a three-dimensional braided structure, which can be diamond braided (1/1 interwoven), regular braided (2/ 2 interweave) and Hercules (3/3 interweave) any one of the three braided structures, the thickness is 0.3-1mm, the braid angle is 45-60°, and the inner layer 3 of the nerve conduit is a porous gel structure , Diameter is 1.8-5mm, its degradation time is 1-5 days.
  • the middle layer 2 of the nerve catheter is made of magnesium wire as the shaft yarn 4, its diameter is 0.1-0.25mm, the number is 4-16, the spacing between the magnesium wires is 200-2000 ⁇ m, and the degummed silk is woven yarn 5, and its thread Density of 20-120D, composite braided three-dimensional structure catheter skeleton; nerve catheter inner layer 3 is composed of silk fibroin gel 6, Schwann cell 7 and nerve growth factor 8 composite gel layer, which guides the directed growth of nerve cells Effect.
  • Nerve growth factor 8 is a protein molecule necessary for the growth and survival of neurons.
  • the diameter of the network fibers formed by freezing is 10-1000 nm, and the pore diameter is 10-500 ⁇ m.
  • the catheter structure has excellent flexibility and flexibility, and can well adapt to the complex stress environment of the body.
  • the preparation method of the above-mentioned magnesium silk silk composite braided structure nerve catheter includes the following steps:
  • the first mixed solution was taken out and poured into a dialysis bag (molecular weight cut off 3500D), dialyzed in deionized water for 36 hours, and the water was changed every 4 hours.
  • the dialyzed solution was centrifuged twice in a high-speed centrifuge at 9000r/min for 20 minutes each time to remove impurities, and finally a silk fibroin solution was obtained, which was placed in a 4°C refrigerator for use.
  • the "one embodiment” or “embodiments” referred to herein refer to specific features, structures, or characteristics that can be included in at least one implementation of the present invention.
  • the appearances of "in one embodiment” in different places in this specification do not all refer to the same embodiment, nor are they separate or selective embodiments mutually exclusive with other embodiments.
  • This example shows a structure and preparation method of a nerve catheter with a composite braided structure of magnesium silk and silk.
  • the nerve catheter has three layers in which the shaft yarn 4 of the middle layer 2 of the nerve catheter is magnesium wire and the middle of the nerve catheter
  • the braided yarn 5 of layer 2 is degummed silk
  • the outer layer 1 of the nerve conduit is a porous sponge layer made by mixing silk fibroin and chitosan solution
  • the mass ratio of silk fibroin to chitosan is 10:1
  • chitosan solution The concentration is 3-6 mg/mL
  • the silk fibroin aqueous solution concentration is 5-10% (w/v).
  • the inner layer 3 of the nerve conduit is a porous silk fibroin gel layer made of an aqueous solution of silk fibroin at a concentration of 5-10% (w/v).
  • the preparation process is as follows:
  • the braid angle of the magnesium wire and silk composite braided structure neural catheter varies between 40-55°.
  • the angle is stable; as can be seen in Figure 5, the first peak represents the breaking peak of the braided yarn, at this time the tensile strain is 8%, the load is up to 35N, and the second peak represents the breaking peak of the shaft yarn, when the load is greater than 65N, indicating that the shaft
  • the addition of yarn greatly enhances the tensile properties of the nerve catheter.
  • This example shows a structure and preparation method of a nerve catheter with a composite braided structure of magnesium silk and silk.
  • the nerve catheter has a total of three layers.
  • the shaft yarn 4 of the middle layer 2 of the nerve catheter is magnesium wire and braided yarn.
  • the outer layer 1 of the nerve catheter is a porous sponge made of chitosan solution with a concentration of 3-6mg/mL
  • the layer has a solution concentration of 3-6 mg/mL
  • the inner layer 3 of the neural catheter is a silk fibroin gel made of a silk fibroin aqueous solution with a concentration of 5-10% (w/v).
  • the preparation process is as follows:
  • the raw material is 20-100D degummed silk silk, magnesium wire and titanium, nickel alloy wire with a diameter of 0.05-0.2mm
  • This example shows a structure and preparation method of a nerve conduit with a composite braided structure of magnesium silk and silk.
  • the nerve conduit has three layers, in which the shaft yarn 4 of the middle layer 2 of the nerve conduit is magnesium wire and degummed silk Interweaving, braided yarn 5 is degummed silk, the outer layer 1 of the nerve conduit is a porous sponge layer prepared from an aqueous solution of silk fibroin with a concentration of 5-10% (w/v); the inner layer 3 of the nerve conduit is a concentration of 5-10 A porous silk fibroin gel layer made of% (w/v) silk fibroin aqueous solution.
  • the preparation process is as follows:
  • the present invention discloses a magnesium silk silk composite braided structure nerve catheter and a preparation method thereof.
  • the medical metal magnesium wire with strong mechanical properties is used as a shaft yarn to be added to the silk braided structure, which serves as a longitudinal guide It can also improve the radial compression performance of the braided skeleton.
  • the magnesium wire can slowly degrade, promote the growth and proliferation of Schwann cells, and is conducive to the recovery of damaged nerves; the braided yarn is degummed silk with a good biological phase Capacitive, while ensuring good mechanical properties, create a biological microenvironment that promotes nerve regeneration.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Dermatology (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Zoology (AREA)
  • Dispersion Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Botany (AREA)
  • Vascular Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Inorganic Chemistry (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Materials For Medical Uses (AREA)
  • Media Introduction/Drainage Providing Device (AREA)

Abstract

一种镁丝蚕丝复合编织结构神经导管及其制备方法。神经导管包括，丝素蛋白溶液和壳聚糖溶液混合后形成的多孔海绵结构外层，镁丝为轴纱、脱胶蚕丝为编织纱复合编织成型的三维结构骨架中间层，多孔凝胶结构内层。镁丝蚕丝复合编织结构能够提高径向抗压性能，缓慢降解促进雪旺细胞的生长和增殖，有利于损伤神经的恢复，具有良好的生物相容性。

Description

一种镁丝蚕丝复合编织结构神经导管及其制备方法 技术领域
本发明属于生物医用材料技术领域，具体涉及一种镁丝蚕丝复合编织结构神经导管及其制备方法。
背景技术
周围神经损伤是临床上常见的病症，利用生物材料和组织工程技术促进缺损神经修复与再生是神经科学领域的研究热点。理想的生物材料、合适的物理力学引导和适宜的生物微环境是促进受损神经恢复感觉、运动等生物学功能的关键因素。目前神经导管研究较多的是使用模型浇铸法、静电纳米纺丝法、3D生物打印成型等技术，以及采用丝素蛋白、壳聚糖和胶原蛋白等生物可降解材料制作出具有物理引导作用的中空导管，但是这些技术制备出来的神经导管的机械力学和柔韧性较差。如一种采用丝素纳米纤维定向引导功能的神经导管，使用冷凝诱导丝素溶液形成纳米微孔作为物理引导，但是该发明中形成的微孔难以控制形状大小和方向，而且导管的径向抗压性能一般，弯曲柔韧度未做考虑；又如先用纳米取向纤维制成不同降解浓度的薄膜然后卷成管状结构的神经导管，此种神经导管对细胞的物理导向性优良，但本身机械性能较差，内部供神经再生的空间较少；再如使用3D生物打印对材料有较大的限制，以及目前分辨率较低，大概是1～300mPa/s。
研究发现，镁元素是人体必需的重要元素之一，在大脑及中枢神经系统中参与蛋白质的合成，激活体内多种酶，调节神经肌肉的活动。在微环境下，金属镁离子具有抗炎、抗氧化、抗凋亡以及调节线粒体钙缓冲等多种生物学功能。最新研究表明镁离子在适宜的微环境的培养下可以促进雪旺细胞的增殖，如分泌生长因子、细胞外基质(Extracellular Matrix，ECM)等，对神经元起到支持和保护的作用。家蚕丝是由家蚕合成与分泌的天然动物蛋白，来源广泛，丝素蛋白是蚕丝中主要的组成部分，约占蚕丝质量的70％-80％，其余主要成分为丝胶蛋白及碳水化合物等。丝素蛋白含有多种氨基酸，其中比较常见的丙氨酸(Ala)、甘氨酸(Gly)、丝氨酸(Ser)约占总组成的85％。丝素本身具有良好的理化性质、透气透湿、生物相容性，能在体 内降解为多肽和游离的氨基酸可被机体吸收从而促进神经细胞的生长。丝素蛋白制成的神经导管体内炎症反应低，可生物降解，能够促进周围神经损伤的功能性恢复。壳聚糖又称脱乙酰甲壳素，是由自然界广泛存在的几丁质(chitin)经过脱乙酰作用得到的，结构性质与ECM中氨基多糖极为相似，吸附性良好，降解速度可调节，其降解的中间产物壳寡糖可抑制神经元凋亡，支持细胞黏附，促进受伤轴突再生。
用于制备神经导管的常见技术有：(1)浇铸-浸渍法，该方法虽然操作简便，但是支架上易残留有毒溶剂，且不易控制孔洞的大小和分布；(2)传统静电纺丝技术，能形成高孔隙率和比表面积的纳米纤维，可制造出类似细胞外基质网状结构的微环境，有利于神经再生长，但是传统静电纺制造出的神经导管机械强度不足，无法为受损神经提供稳定的力学环境；(3)编织技术，该技术采用锭子编织机，使纱线编织成管状织物，可增大导管内部的表面积，力学性能较好。但是，在诱导细胞定向迁移和组织再生方面，仍然有不足之处。
针对上述问题，本发明提出了一种镁丝蚕丝复合编织结构神经导管及其制备方法。
发明内容
本发明目的是：提供一种镁丝蚕丝复合编织结构神经导管及其制备方法，能提高神经再生有序性以及神经导管机械强度，以解决上述问题。
本发明的一种技术方案是：
一种镁丝蚕丝复合编织结构神经导管，包括神经导管外层、神经导管中间层和神经导管内层，所述神经导管中间层包覆于所述神经导管内层的外部，所述神经导管外层包覆于所述神经导管中间层的外部，所述神经导管外层为丝素蛋白溶液和壳聚糖溶液混合后形成的多孔海绵结构，所述神经导管外层的厚度为0.5-1.5mm，孔径为20-150μm，孔隙率为75％-90％，所述神经导管中间层为三维编织结构，所述神经导管中间层的厚度为0.3-1mm，所述三维编织结构的编织角为45-60°，所述神经导管内层为多孔凝胶结构，所述神经导管内层的直径为1.8-5mm。
进一步的，所述三维编织结构为菱形编织结构、规则编织结构和赫格利 斯编织结构中的任意一种。
进一步的，所述三维编织结构是以镁丝为轴纱、脱胶蚕丝为编织纱复合编织成型的三维结构导管骨架，所述镁丝的直径为0.1-0.25mm，根数为4-16根，相邻两根镁丝之间的间距为200-2000μm，所述脱胶蚕丝的线密度为20-120D。
进一步的，所述多孔凝胶结构为丝素蛋白凝胶、雪旺细胞和神经生长因子组成的复合凝胶层，所述丝素蛋白凝胶冷冻形成网状纤维，所述雪旺细胞和神经生长因子设置于所述网状纤维内，所述网状纤维的直径为10-1000nm，孔径为10-500μm。
本发明的另一种技术方案是：
一种镁丝蚕丝复合编织结构神经导管的制备方法，该方法包括如下步骤：
(1)制备神经导管中间层：采用带芯功能的立锭式编织机制备神经导管骨架结构，以镁丝为轴纱、脱胶蚕丝为编织纱复合编织成型；
(2)制备神经导管外层：按照丝素蛋白溶液:壳聚糖溶液＝1:1～1:5，将所述丝素蛋白溶液缓慢加入至所述壳聚糖溶液中，搅拌混合6-12小时，获得第二混合溶液，将所述神经导管中间层固定在模具中，再将所述第二混合溶液浇入所述模具中，然后放入-20℃冰箱冷冻4-12小时，再放入冷冻干燥机中冷冻干燥12-36小时后取出，得到具有多孔且为中空结构的神经导管外层；
(3)制备神经导管内层：将浓度为0.01-0.5％培养有雪旺细胞的丝素蛋白溶液注入到所述神经导管外层的中空结构内，在温度为37℃下放置12小时，形成凝胶化导管；将所述凝胶化导管垂直插入到液氮罐中，沿所述凝胶化导管的轴向以0.1-1mm/s的速度缓慢向下进行冷冻，再放入冷冻干燥机中冷冻干燥24-48小时，获得镁丝蚕丝复合编织结构神经导管。
进一步的，步骤(2)中所述丝素蛋白溶液的制备方式为：称取家蚕生丝在沸腾的Na 2CO 3水溶液中脱胶30分钟，每10分钟用玻璃棒进行搅拌，用去离子水多次冲洗后放在通风橱中过夜自然风干，获得干燥的家蚕丝素纤维，将所述干燥的家蚕丝素纤维溶于LiBr溶液中，在60℃的烘箱中溶解4小时，每小时轻微摇晃以保证溶解完全，获得第一混合溶液，将透析后的第 一混合溶液离心，获得丝素蛋白溶液，放置于4℃冰箱中放置待用。
进一步的，步骤(2)中所述透析为：将所述第一混合溶液倒入截留分子量为3500D的透析袋中，在去离子水中透析36小时，每4小时换一次水。
进一步的，步骤(2)中所述离心为：在高速离心机中以9000r/min离心两次，每次20分钟。
进一步的，步骤(2)中所述壳聚糖溶液的制备方式为：称取脱乙酰度为80-95％的壳聚糖，溶于0.5-4％的醋酸溶液中，搅拌4-12小时，制成质量百分比为1-5％的壳聚糖溶液。
本发明提供了一种镁丝蚕丝复合编织结构神经导管及其制备方法，其优点是：
(1)增加了导管的径向抗压和拉伸等机械性能，同时增加了神经导管的柔韧弯曲性能，更适应复杂的体内受力环境；
(2)导管生物相容性、表面活性和通透性良好，能促进神经元细胞的粘附、生长和增殖，加快受损神经的再生速度；
(3)可通过选择不同的材料和改性工艺，调控导管的降解速率，从而为神经的再生提供合适的生长空间；
(4)由于导管中的镁丝有导电性，可刺激和引导神经生长，同时降解时产生的镁离子有助于提高神经元细胞的活性。
附图说明
为了更清楚地说明本发明实施例的技术方案，下面将对实施例描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动性的前提下，还可以根据这些附图获得其它的附图。其中，
图1为本发明所述的一种镁丝蚕丝复合编织结构神经导管在实施例1中的结构示意图；
图2为本发明所述的一种镁丝蚕丝复合编织结构神经导管在实施例2中的结构示意图；
图3为本发明所述的一种镁丝蚕丝复合编织结构神经导管在实施例1中的结构示意图；
图4为本发明实施例1所述的采用不同参数制备的内径分别为2mm和3mm镁丝蚕丝复合编织结构神经导管的编织角；
图5为本发明实施例1所述的采用不同参数制备的内径分别为2mm和3mm镁丝蚕丝复合编织结构神经导管的载荷-拉伸应变曲线。
其中：1为神经导管外层、2为神经导管中间层、3为神经导管内层、4为轴纱、5为编织纱、6为丝素蛋白凝胶、7为雪旺细胞、8为神经生长因子、9为钛、镍合金丝。
具体实施方式
为使本发明的上述目的、特征和优点能够更加明显易懂，下面结合具体实施方式和附图对本发明作进一步详细的说明。
本发明提供一种镁丝蚕丝复合编织结构神经导管，如图1所示，该导管分为三层，包括：神经导管外层1为丝素蛋白溶液和壳聚糖溶液混合后形成的多孔海绵结构，厚度为0.5-1.5mm，孔径在20-150μm，孔隙率在75％-90％，神经导管中间层2为三维编织结构，可以为菱形编织(1/1交织)、规则编织(2/2交织)和赫格利斯(3/3交织)三种编织结构中的任意一种编织结构，厚度在0.3-1mm，编织角为45-60°，神经导管内层3为多孔凝胶结构，直径为1.8-5mm，其降解时间为1-5天。神经导管中间层2是由镁丝为轴纱4，其直径为0.1-0.25mm，根数为4-16根，镁丝之间的间距为200-2000μm，脱胶蚕丝为编织纱5，其线密度为20-120D，复合编织成型的三维结构导管骨架；神经导管内层3由丝素蛋白凝胶6、雪旺细胞7和神经生长因子8组成的复合凝胶层，具有引导神经细胞定向生长的效果。神经生长因子8为神经元生长和存活所必需的蛋白质分子，其冷冻形成的网状纤维直径为10-1000nm，孔径在10-500μm。导管结构具有优良的柔韧性和弯曲性，能很好地适应体内复杂的受力环境。
上述镁丝蚕丝复合编织结构神经导管的制备方法包括以下步骤：
(一)制备神经导管中间层2，采用带芯功能的立锭式编织机制备神经导管骨架结构，镁丝为轴纱4，脱胶蚕丝为编织纱5，复合编织成型。
(二)丝素蛋白溶液的制备：称取30g家蚕生丝在12L沸腾的Na 2CO 3(0.02M)水溶液中脱胶30分钟，每10分钟用玻璃棒进行搅拌，以保证脱 胶的彻底性，然后用去离水多次冲洗，将洗好的丝素蛋白纤维放在通风橱中过夜自然风干。称取干燥后的25g家蚕丝素纤维溶于100mL LiBr(9.3M)溶液中，在60℃的烘箱中溶解4小时，每小时轻微摇晃以保证溶解完全，获得第一混合溶液。取出第一混合溶液倒入透析袋(截留分子量3500D)中，在去离子水中透析36小时，每4小时换一次水。透析后的溶液在高速离心机中以9000r/min离心两次，每次20分钟，以去除杂质，最终获得丝素蛋白溶液，放置于4℃冰箱中放置待用。
(三)制备神经导管外层1：称取2-8g脱乙酰度为80-95％的壳聚糖，溶于0.5-4％的醋酸溶液中，搅拌4-12小时，制成质量百分比为1-5％的壳聚糖溶液。按照丝素蛋白溶液:壳聚糖溶液＝1:1～1:5，将丝素溶液缓慢加入到制备好的壳聚糖溶液中，搅拌混合6-12小时，获得第二混合溶液。将脱胶蚕丝编织层(神经导管中间层2)固定在模具中，再将第二混合溶液浇入模具中，然后放入-20℃冰箱冷冻4-12小时，再放入冷冻干燥机中冷冻干燥12-36小时后取出，得到具有多孔结构的海绵外层材料。
(四)制备神经导管内层3：
1)密封冷冻后的导管，将浓度为0.01-0.5％培养有雪旺细胞的丝素蛋白溶液注入到导管中空结构内，37℃下放置12小时诱导其凝胶化；2)将导管垂直插入到液氮罐中，沿导管轴向以0.1-1mm/s的速度缓慢向下进行冷冻，再放入冷冻干燥机中冷冻干燥24-48小时。
为使本发明的上述目的、特征和优点能够更加明显易懂，下面结合附图和实施例进一步说明本发明的技术方案。但是本发明不限于所列出的实施例，还应包括在本发明所要求的权利范围内其他任何公知的改变。
首先，此处所称的“一个实施例”或“实施例”是指可包含于本发明至少一个实现方式中的特定特征、结构或特性。在本说明书中不同地方出现的“在一个实施例中”并非均指同一个实施例，也不是单独的或选择性的与其他实施例互相排斥的实施例。
其次，本发明利用结构示意图等进行详细描述，在详述本发明实施例时，为便于说明，示意图会不依一般比例作局部放大，而且所述示意图只是实例，其在此不应限制本发明保护的范围。此外，在实际制作中应包含长度、宽度及深度的三维空间。
实施例1
本实施案例展示一种镁丝蚕丝复合编织结构神经导管结构及制备方法，如图1所示，该神经导管共有三层，其中，神经导管中间层2的轴纱4为镁丝，神经导管中间层2的编织纱5为脱胶蚕丝，神经导管外层1为丝素蛋白和壳聚糖溶液混合制成的多孔海绵层，丝素蛋白与壳聚糖质量比为10:1，壳聚糖溶液浓度为3-6mg/mL，丝素蛋白水溶液浓度为5-10％(w/v)。神经导管内层3是由浓度为5-10％(w/v)丝素蛋白水溶液制成的多孔丝素凝胶层。制备工艺流程如下：
(1)采用编织方法制备神经导管中间层2：原材料为20-100D脱胶蚕丝，直径在0.05-0.2mm的镁丝，编织工艺采用8-32锭立式编织机，菱形编织(1/1交织)，上机编织角为45-60°，齿轮比(大齿轮:小齿轮＝80-120:18-60)，转速设置为5-100rpm。
(2)将编织好的神经导管中间层2固定在管状聚四氟乙烯模具中，再将制备好的溶液浇入模具中，成型温度为60±2℃，成型时间15-30min。放入-20℃冰箱冷冻4-12小时，再放入冷冻干燥机中冷冻干燥12-36小时后取出后，对导管进行水蒸气处理，水蒸气处理5-10小时，处理温度为50-70℃，得到带有多孔海绵层的神经导管。
(3)密封冷冻后的导管，将含有雪旺细胞的浓度为0.01-0.5％的丝素蛋白溶液注入到导管中空结构内，37℃下放置12小时诱导其凝胶化；再将导管垂直插入到液氮罐中，沿导管轴向以0.1-1mm/s的速度缓慢向下进行液氮冷冻，最后，置于冷冻干燥机进行冷冻干燥24-48小时，得到含有雪旺细胞的丝素凝胶层。
上述方法所制得的镁丝蚕丝复合编织结构神经导管的性能请参阅图4-图5，如图4可知，镁丝蚕丝复合编织结构神经导管的编织角在40-55°之间变化，编织角稳定；如图5可知，第一个峰表示编织纱断裂峰，此时拉伸应变为8％，载荷高达35N，第二个峰表示轴纱的断裂峰，此时载荷大于65N，表明轴纱的加入大大增强了神经导管的拉伸性能。
实施例2
本实施案例展示一种镁丝蚕丝复合编织结构神经导管结构及的制备方 法，如图2所示，该神经导管共有三层，其中，神经导管中间层2的轴纱4为镁丝，编织纱5中对称加入四根可降解的钛、镍合金丝9以增强神经导管的径向抗压强度，神经导管外层1是由浓度为3-6mg/mL的壳聚糖溶液制成的多孔海绵层，其溶液浓度为3-6mg/mL，神经导管内层3是由浓度为5-10％(w/v)丝素蛋白水溶液制成的丝素凝胶。制备工艺流程如下：
(1)采用编织方法制备神经导管中间层2：原材料为20-100D脱胶蚕丝，直径在0.05-0.2mm的镁丝和钛、镍合金丝，编织工艺采用8-32锭立式编织机，规则编织(2/2交织)，上机编织角为45-60°，齿轮比(大齿轮:小齿轮＝80-120:18-60)，转速设置为5-100rpm。
(2)将神经导管中间层固定在管状聚四氟乙烯模具中，再将制备好的溶液浇入模具中，成型温度为60±2℃，成型时间15-30min。放入-20℃冰箱冷冻4-12小时，再置入冷冻干燥机中冷冻干燥12-36小时后取出后，对导管进行水蒸气处理，水蒸气处理5-10小时，处理温度为50-70℃，得到神经导管的多孔海绵层。
(3)密封冷冻后的导管，将含有雪旺细胞的浓度为0.01-0.5％的丝素蛋白溶液注入到导管中空结构内，37℃下放置12小时诱导其凝胶化；再将导管垂直插入到液氮罐中，沿导管轴向以0.1-1mm/s的速度缓慢向下进行冷冻，最后，置于冷冻干燥机进行冷冻干燥24-48小时，得到含有雪旺细胞的丝素凝胶层。
实施例3
本实施案例展示一种镁丝蚕丝复合编织结构神经导管结构及的制备方法，如图3所示，该神经导管共有三层，其中，神经导管中间层2的轴纱4为镁丝和脱胶蚕丝交织，编织纱5为脱胶蚕丝，神经导管外层1是由浓度为5-10％(w/v)的丝素蛋白水溶液制备的多孔海绵层；神经导管内层3是由浓度为5-10％(w/v)丝素蛋白水溶液制成的多孔丝素凝胶层。制备工艺流程如下：
(1)采用编织方法制备神经导管中间层2：原材料为20-100D脱胶蚕丝，直径为0.05-0.2mm的镁丝，编织工艺采用8-32锭立式编织机，赫格利斯(3/3交织)，上机编织角为45-60°，齿轮比(大齿轮:小齿轮＝80-120:18-60)，转速设置为5-100rpm。
(2)将神经导管中间层固定在管状聚四氟乙烯模具中，再将制备好的溶液浇入模具中，成型温度为60±2℃，成型时间15-30min。放入-20℃冰箱冷冻4-12小时，再置入冷冻干燥机中冷冻干燥12-36小时后取出后，对导管进行水蒸气处理5-10小时，处理温度为50-70℃，得到外层1为多孔海绵层的神经导管。
(3)密封冷冻后的导管，将含有雪旺细胞的浓度为0.01-0.5％的丝素蛋白溶液注入到导管中空结构内，37℃下放置12小时诱导其凝胶化；再将导管垂直插入到液氮罐中，沿导管轴向以0.1-1mm/s的速度缓慢向下进行冷冻，最后，置于冷冻机进行冷冻干燥24-48小时，得到含有雪旺细胞的丝素凝胶层。
综上所述，本发明公开了一种镁丝蚕丝复合编织结构神经导管及其制备方法，采用具有较强力学性能的医用金属镁丝为轴纱加入到蚕丝编织结构中，既起到纵向引导作用，又能提高编织骨架的径向抗压性能，同时，镁丝能缓慢降解，促进雪旺细胞的生长和增殖，有利于损伤神经的恢复；编织纱线为脱胶蚕丝，具有良好的生物相容性，在保证力学性能良好的同时，产生促进神经再生的生物微环境。
应说明的是，以上实施例仅用以说明本发明的技术方案而非限制，尽管参照较佳实施例对本发明进行了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围，其均应涵盖在本发明的权利要求范围当中。

Claims (9)

  1. 一种镁丝蚕丝复合编织结构神经导管，其特征在于：包括神经导管外层、神经导管中间层和神经导管内层，所述神经导管中间层包覆于所述神经导管内层的外部，所述神经导管外层包覆于所述神经导管中间层的外部，所述神经导管外层为丝素蛋白溶液和壳聚糖溶液混合后形成的多孔海绵结构，所述神经导管外层的厚度为0.5-1.5mm，孔径为20-150μm，孔隙率为75％-90％，所述神经导管中间层为三维编织结构，所述神经导管中间层的厚度为0.3-1mm，所述三维编织结构的编织角为45-60°，所述神经导管内层为多孔凝胶结构，所述神经导管内层的直径为1.8-5mm。
  2. 根据权利要求1所述的镁丝蚕丝复合编织结构神经导管，其特征在于：所述三维编织结构为菱形编织结构、规则编织结构和赫格利斯编织结构中的任意一种。
  3. 根据权利要求1所述的镁丝蚕丝复合编织结构神经导管，其特征在于：所述三维编织结构是以镁丝为轴纱、脱胶蚕丝为编织纱复合编织成型的三维结构导管骨架，所述镁丝的直径为0.1-0.25mm，根数为4-16根，相邻两根镁丝之间的间距为200-2000μm，所述脱胶蚕丝的线密度为20-120D。
  4. 根据权利要求1所述的镁丝蚕丝复合编织结构神经导管，其特征在于：所述多孔凝胶结构为丝素蛋白凝胶、雪旺细胞和神经生长因子组成的复合凝胶层，所述丝素蛋白凝胶冷冻形成网状纤维，所述雪旺细胞和神经生长因子设置于所述网状纤维内，所述网状纤维的直径为10-1000nm，孔径为10-500μm。
  5. 一种镁丝蚕丝复合编织结构神经导管的制备方法，其特征在于，该方法包括如下步骤：
    (1)制备神经导管中间层：采用带芯功能的立锭式编织机制备神经导管骨架结构，以镁丝为轴纱、脱胶蚕丝为编织纱复合编织成型；
    (2)制备神经导管外层：按照丝素蛋白溶液:壳聚糖溶液＝1:1～1:5，将所述丝素蛋白溶液缓慢加入至所述壳聚糖溶液中，搅拌混合6-12小时，获得第二混合溶液，将所述神经导管中间层固定在模具中，再将所述第二混合溶液浇入所述模具中，然后放入-20℃冰箱冷冻4-12小时，再放入冷冻干燥机中冷冻干燥12-36小时后取出，得到具有多孔且为中空结构的神经导管外层；
    (3)制备神经导管内层：将浓度为0.01-0.5％培养有雪旺细胞的丝素蛋白溶液注入到所述神经导管外层的中空结构内，在温度为37℃下放置12小时，形成凝胶化导管；将所述凝胶化导管垂直插入到液氮罐中，沿所述凝胶化导管的轴向以0.1-1mm/s的速度缓慢向下进行冷冻，再放入冷冻干燥机中冷冻干燥24-48小时，获得镁丝蚕丝复合编织结构神经导管。
  6. 根据权利要求5所述的镁丝蚕丝复合编织结构神经导管的制备方法，其特征在于，步骤(2)中所述丝素蛋白溶液的制备方式为：称取家蚕生丝在沸腾的Na 2CO 3水溶液中脱胶30分钟，每10分钟用玻璃棒进行搅拌，用去离子水多次冲洗后放在通风橱中过夜自然风干，获得干燥的家蚕丝素纤维，将所述干燥的家蚕丝素纤维溶于LiBr溶液中，在60℃的烘箱中溶解4小时，每小时轻微摇晃以保证溶解完全，获得第一混合溶液，将透析后的第一混合溶液离心，获得丝素蛋白溶液，放置于4℃冰箱中放置待用。
  7. 根据权利要求6所述的镁丝蚕丝复合编织结构神经导管的制备方法，其特征在于，步骤(2)中所述透析为：将所述第一混合溶液倒入截留分子量为3500D的透析袋中，在去离子水中透析36小时，每4小时换一次水。
  8. 根据权利要求6所述的镁丝蚕丝复合编织结构神经导管的制备方法，其特征在于，步骤(2)中所述离心为：在高速离心机中以9000r/min离心两次，每次20分钟。
  9. 根据权利要求5所述的镁丝蚕丝复合编织结构神经导管的制备方法，其特征在于，步骤(2)中所述壳聚糖溶液的制备方式为：称取脱乙酰度为80-95％的壳聚糖，溶于0.5-4％的醋酸溶液中，搅拌4-12小时，制成质量百分比为1-5％的壳聚糖溶液。
PCT/CN2019/076010 2018-12-29 2019-02-25 一种镁丝蚕丝复合编织结构神经导管及其制备方法 Ceased WO2020133668A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2021523697A JP2022506369A (ja) 2018-12-29 2019-02-25 マグネシウムワイヤー・シルク複合編組構造の神経カテーテル及びその製造方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201811638025.9A CN109513041B (zh) 2018-12-29 2018-12-29 一种镁丝蚕丝复合编织结构神经导管及其制备方法
CN201811638025.9 2018-12-29

Publications (1)

Publication Number Publication Date
WO2020133668A1 true WO2020133668A1 (zh) 2020-07-02

Family

ID=65798765

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/076010 Ceased WO2020133668A1 (zh) 2018-12-29 2019-02-25 一种镁丝蚕丝复合编织结构神经导管及其制备方法

Country Status (3)

Country Link
JP (1) JP2022506369A (zh)
CN (1) CN109513041B (zh)
WO (1) WO2020133668A1 (zh)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113634048A (zh) * 2021-09-10 2021-11-12 武汉纺织大学 天然蚕丝微纳米纤维复合多孔材料及其应用
CN114470325A (zh) * 2022-03-24 2022-05-13 合肥工业大学 一种可缝裁单面导电抗菌的甲壳素水凝胶神经导管材料的制备方法
CN114983622A (zh) * 2022-06-01 2022-09-02 奥精医疗科技股份有限公司 一种神经导管的制备方法及神经导管
CN116899014A (zh) * 2023-07-24 2023-10-20 上海工程技术大学 一种三维拓扑结构多通道神经导管及其制备方法
CN117323470A (zh) * 2023-11-08 2024-01-02 浙江大学 水凝胶复合微纤维网络的神经导管及其制备方法和应用
CN118512659A (zh) * 2024-05-10 2024-08-20 南开大学 一种管状细胞外基质纤维支架及其制备方法与应用

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109513041B (zh) * 2018-12-29 2023-10-03 南通纺织丝绸产业技术研究院 一种镁丝蚕丝复合编织结构神经导管及其制备方法
CN110251732B (zh) * 2019-06-18 2020-10-27 南通纺织丝绸产业技术研究院 一种多层复合编织的可降解神经导管结构及其制备方法
CN110786965B (zh) * 2019-11-14 2021-04-30 苏州大学 变锭编织法制备多通道抗菌神经导管的方法
CN111297513A (zh) * 2020-03-19 2020-06-19 中国海洋大学 一种负载营养因子的人工神经导管及其制备方法和应用
CN111281599A (zh) * 2020-03-19 2020-06-16 中国海洋大学 一种增强型人工神经导管及其制备方法和应用
CN116019606A (zh) * 2023-01-31 2023-04-28 重庆市畜牧科学院 一种视神经接口的制备方法

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1987850A1 (en) * 2000-12-21 2008-11-05 Ethicon, Incorporated Reinforced tissue implants and methods of manufacture and use
CN102274089A (zh) * 2011-05-24 2011-12-14 苏州大学 一种丝素蛋白管状支架及其制备方法
CN104107096A (zh) * 2014-07-18 2014-10-22 上海交通大学 可弯曲全降解镁合金神经导管及其制备方法
CN104548205A (zh) * 2014-12-29 2015-04-29 东莞颠覆产品设计有限公司 一种动物源神经支架及其制备方法
WO2015142631A1 (en) * 2014-03-17 2015-09-24 University Of Pittsburgh-Of The Commonwealth System Of Higher Education Magnesium composite-containing scaffolds to enhance tissue regeneration
CN105748171A (zh) * 2016-02-21 2016-07-13 新乡医学院 生物型神经导管
CN109513041A (zh) * 2018-12-29 2019-03-26 南通纺织丝绸产业技术研究院 一种镁丝蚕丝复合编织结构神经导管及其制备方法

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8106014B2 (en) * 2004-09-14 2012-01-31 Neurotex Limited Methods and apparatus for enhanced growth of peripheral nerves and nervous tissue
JP4982887B2 (ja) * 2007-02-20 2012-07-25 北海道曹達株式会社 神経再生チューブ及びその製造方法
KR101116237B1 (ko) * 2009-08-12 2012-03-09 서울대학교산학협력단 실크 나노섬유 신경도관 및 이의 제조방법
CN102688076B (zh) * 2011-03-25 2014-09-10 广州迈普再生医学科技有限公司 一种神经导管及其制备方法
JP2014155622A (ja) * 2013-02-15 2014-08-28 Terumo Corp 管状体
CN103083720B (zh) * 2013-02-28 2015-01-21 苏州大学 一种丝素蛋白管及其制备方法
CN103598927A (zh) * 2013-10-17 2014-02-26 上海交通大学 用于神经缺损修复的可降解镁合金神经导管及其制备方法
ES2577883B2 (es) * 2014-12-16 2016-11-21 Universitat Politècnica De València Biohíbrido para su uso en la regeneración de tractos neurales
WO2018137763A1 (en) * 2017-01-25 2018-08-02 B. Braun Melsungen Ag Endoluminal device
CN108853587B (zh) * 2017-05-10 2021-08-03 上海交通大学 镁合金与高分子丝材混编复合补片及其用途

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1987850A1 (en) * 2000-12-21 2008-11-05 Ethicon, Incorporated Reinforced tissue implants and methods of manufacture and use
CN102274089A (zh) * 2011-05-24 2011-12-14 苏州大学 一种丝素蛋白管状支架及其制备方法
WO2015142631A1 (en) * 2014-03-17 2015-09-24 University Of Pittsburgh-Of The Commonwealth System Of Higher Education Magnesium composite-containing scaffolds to enhance tissue regeneration
CN104107096A (zh) * 2014-07-18 2014-10-22 上海交通大学 可弯曲全降解镁合金神经导管及其制备方法
CN104548205A (zh) * 2014-12-29 2015-04-29 东莞颠覆产品设计有限公司 一种动物源神经支架及其制备方法
CN105748171A (zh) * 2016-02-21 2016-07-13 新乡医学院 生物型神经导管
CN109513041A (zh) * 2018-12-29 2019-03-26 南通纺织丝绸产业技术研究院 一种镁丝蚕丝复合编织结构神经导管及其制备方法

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113634048A (zh) * 2021-09-10 2021-11-12 武汉纺织大学 天然蚕丝微纳米纤维复合多孔材料及其应用
CN114470325A (zh) * 2022-03-24 2022-05-13 合肥工业大学 一种可缝裁单面导电抗菌的甲壳素水凝胶神经导管材料的制备方法
CN114983622A (zh) * 2022-06-01 2022-09-02 奥精医疗科技股份有限公司 一种神经导管的制备方法及神经导管
CN116899014A (zh) * 2023-07-24 2023-10-20 上海工程技术大学 一种三维拓扑结构多通道神经导管及其制备方法
CN116899014B (zh) * 2023-07-24 2024-05-07 上海工程技术大学 一种三维拓扑结构多通道神经导管及其制备方法
CN117323470A (zh) * 2023-11-08 2024-01-02 浙江大学 水凝胶复合微纤维网络的神经导管及其制备方法和应用
CN118512659A (zh) * 2024-05-10 2024-08-20 南开大学 一种管状细胞外基质纤维支架及其制备方法与应用

Also Published As

Publication number Publication date
JP2022506369A (ja) 2022-01-17
CN109513041A (zh) 2019-03-26
CN109513041B (zh) 2023-10-03

Similar Documents

Publication Publication Date Title
WO2020133668A1 (zh) 一种镁丝蚕丝复合编织结构神经导管及其制备方法
CN110251732B (zh) 一种多层复合编织的可降解神经导管结构及其制备方法
Han et al. A review: Current status and emerging developments on natural polymer‐based electrospun fibers
CN107952112B (zh) 一种含丝素蛋白纳米纤维定向引导功能的神经导管及其制备方法
CN103877617B (zh) 可注射蚕丝素蛋白-海藻酸盐双交联水凝胶及其制备方法和使用方法
CN103341209A (zh) 一种丝素蛋白纳米纤维膜及其制备方法
CN106237381B (zh) 一种丝素蛋白微纳米纤维多孔支架及其制备方法
CN108114315A (zh) 一种双丝素蛋白神经移植物
CN107982580A (zh) 一种基于编织和涂层技术的丝素蛋白组织工程支架及其制备方法和应用
CN106139246A (zh) 一种再生丝蛋白纤维支架及其制备方法
CN111494712B (zh) 一种融合有nt3的丝素蛋白神经移植物的制备方法
CN116139344A (zh) 一种促进成骨细胞生成的骨修复材料及其制备方法
CN106729981B (zh) 一种丝素蛋白纳米纤维及其制备方法和应用
CN109395162B (zh) 一种天然蛋白基仿生结构骨支架的制备方法
Ma et al. Application of silk fibroin-based composite films in biomedicine and biotechnology
CN106243377B (zh) 一种多孔蚕丝纤维材料及其制备方法
CN209564438U (zh) 一种镁丝蚕丝复合编织结构神经导管
CN116173306B (zh) 一种具有仿神经网络结构的复合导电水凝胶材料及其制备方法
Fang et al. Ultralight 3D silk nanofiber scaffolds with enhanced hemostatic properties and biocompatibility for rapid hemostasis
CN120329746A (zh) 仿生、高强度、生物相容的丝素蛋白水凝胶的制备方法
CN107519535B (zh) 可降解海藻酸钠/丝素双层皮肤组织工程支架材料及其制备方法
CN107096065B (zh) 含多聚唾液酸复合纳米纤维膜及制备方法和应用
CN114732954A (zh) 一种载药型人工皮肤及其制备方法
CN110464881B (zh) 具有层级结构的丝素多孔支架及其制备方法
CN102532573B (zh) 一种丝胶蛋白/羟基磷灰石复合膜的快速成型方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19903207

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2021523697

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19903207

Country of ref document: EP

Kind code of ref document: A1