WO2020237785A1 - 一种引导牙周硬软组织再生梯度材料及其制备方法 - Google Patents

一种引导牙周硬软组织再生梯度材料及其制备方法 Download PDF

Info

Publication number
WO2020237785A1
WO2020237785A1 PCT/CN2019/095661 CN2019095661W WO2020237785A1 WO 2020237785 A1 WO2020237785 A1 WO 2020237785A1 CN 2019095661 W CN2019095661 W CN 2019095661W WO 2020237785 A1 WO2020237785 A1 WO 2020237785A1
Authority
WO
WIPO (PCT)
Prior art keywords
fiber membrane
gradient material
regeneration
printing
layer
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/CN2019/095661
Other languages
English (en)
French (fr)
Inventor
李吉东
金蜀鄂
李玉宝
左奕
袁琛
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.)
Sichuan University
Original Assignee
Sichuan University
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 Sichuan University filed Critical Sichuan University
Priority to US16/771,221 priority Critical patent/US11696974B2/en
Publication of WO2020237785A1 publication Critical patent/WO2020237785A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • D01D5/0038—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion the fibre formed by solvent evaporation, i.e. dry 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
    • 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
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/12—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L31/125—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L31/127—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix containing fillers of phosphorus-containing inorganic 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/146—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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B32—LAYERED PRODUCTS
    • B32B—LAYERED PRODUCTS, i.e. PRODUCTS BUILT-UP OF STRATA OF FLAT OR NON-FLAT, e.g. CELLULAR OR HONEYCOMB, FORM
    • B32B37/00—Methods or apparatus for laminating, e.g. by curing or by ultrasonic bonding
    • B32B37/14—Methods or apparatus for laminating, e.g. by curing or by ultrasonic bonding characterised by the properties of the layers
    • B32B37/144—Methods or apparatus for laminating, e.g. by curing or by ultrasonic bonding characterised by the properties of the layers using layers with different mechanical or chemical conditions or properties, e.g. layers with different thermal shrinkage, layers under tension during bonding
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y80/00—Products made by additive manufacturing
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L67/00—Compositions of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Compositions of derivatives of such polymers
    • C08L67/04—Polyesters derived from hydroxycarboxylic acids, e.g. lactones
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
    • C08L89/04—Products derived from waste materials, e.g. horn, hoof or hair
    • C08L89/06—Products derived from waste materials, e.g. horn, hoof or hair derived from leather or skin, e.g. gelatin
    • 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
    • D01D1/00—Treatment of filament-forming or like material
    • D01D1/02—Preparation of spinning solutions
    • 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
    • D01D5/0076—Electro-spinning characterised by the electro-spinning apparatus characterised by the collecting device, e.g. drum, wheel, endless belt, plate or grid
    • D01D5/0084—Coating by electro-spinning, i.e. the electro-spun fibres are not removed from the collecting device but remain integral with it, e.g. coating of prostheses
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F1/00—General methods for the manufacture of artificial filaments or the like
    • D01F1/02—Addition of substances to the spinning solution or to the melt
    • D01F1/10—Other agents for modifying properties
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F6/00—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof
    • D01F6/58—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from homopolycondensation products
    • D01F6/62—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from homopolycondensation products from polyesters
    • D01F6/625—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from homopolycondensation products from polyesters derived from hydroxy-carboxylic acids, e.g. lactones
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F6/00—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof
    • D01F6/78—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from copolycondensation products
    • D01F6/84—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from copolycondensation products from copolyesters
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F6/00—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof
    • D01F6/88—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from mixtures of polycondensation products as major constituent with other polymers or low-molecular-weight compounds
    • D01F6/92—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from mixtures of polycondensation products as major constituent with other polymers or low-molecular-weight compounds of polyesters
    • 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
    • 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
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
    • 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
    • A61L2430/12—Materials or treatment for tissue regeneration for dental implants or prostheses
    • 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
    • A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2005/00—Use of polysaccharides or derivatives as moulding material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2067/00—Use of polyesters or derivatives thereof, as moulding material
    • B29K2067/04—Polyesters derived from hydroxycarboxylic acids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2067/00—Use of polyesters or derivatives thereof, as moulding material
    • B29K2067/04—Polyesters derived from hydroxycarboxylic acids
    • B29K2067/043—PGA, i.e. polyglycolic acid or polyglycolide
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2067/00—Use of polyesters or derivatives thereof, as moulding material
    • B29K2067/04—Polyesters derived from hydroxycarboxylic acids
    • B29K2067/046—PLA, i.e. polylactic acid or polylactide
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2073/00—Use of other polymers having oxygen as the only hetero atom in the main chain, as moulding material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29K—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES B29B, B29C OR B29D, RELATING TO MOULDING MATERIALS OR TO MATERIALS FOR MOULDS, REINFORCEMENTS, FILLERS OR PREFORMED PARTS, e.g. INSERTS
    • B29K2075/00—Use of PU, i.e. polyureas or polyurethanes or derivatives thereof, as moulding material
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
    • C08K3/00—Use of inorganic substances as compounding ingredients
    • C08K3/32—Phosphorus-containing compounds
    • C08K2003/321—Phosphates
    • C08K2003/325—Calcium, strontium or barium phosphate
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
    • D—TEXTILES; PAPER
    • D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B2331/00—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products
    • D10B2331/04—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products polyesters, e.g. polyethylene terephthalate [PET]
    • D—TEXTILES; PAPER
    • D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B2331/00—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products
    • D10B2331/04—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products polyesters, e.g. polyethylene terephthalate [PET]
    • D10B2331/041—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products polyesters, e.g. polyethylene terephthalate [PET] derived from hydroxy-carboxylic acids, e.g. lactones
    • D—TEXTILES; PAPER
    • D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B2331/00—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products
    • D10B2331/10—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products polyurethanes
    • D—TEXTILES; PAPER
    • D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
    • D10B2331/00—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products
    • D10B2331/30—Fibres made from polymers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polycondensation products polycondensation products not covered by indexing codes D10B2331/02 - D10B2331/14

Definitions

  • the invention belongs to the technical field of biomedical materials, and specifically relates to a gradient material for guiding the regeneration of periodontal hard and soft tissues and a preparation method thereof.
  • GBR Guided Bone Regeneration
  • the traditional non-degradable GBR membrane needs to be removed by a second operation, which will increase the risk of infection and patient suffering, so it is gradually eliminated; the collagen-based degradable membrane has the risk of disease transmission because most of its collagen components are derived from mammalian tissues, and There is the problem of excessive degradation; the biosynthetic polymer-based degradable membrane has the problem of poor biocompatibility.
  • the present invention constructs a gradient composite material integrating the dual functions of guiding tissue regeneration membrane and bone filling.
  • the ideal tissue repair material can simulate the local microenvironment of natural tissues in terms of composition and structure and has specific biological functions.
  • Electrospinning technology as a simple, cost-effective and effective method for preparing ultrafine fibers, can prepare micro-nano-level fibers.
  • the fiber structure mimics the extracellular matrix structure well, and the prepared fiber membrane has a large specific surface area. , High porosity and other characteristics, so it is widely used in the field of biomedical materials.
  • 3D printing can produce high-porosity structures with good interconnectivity, and can print tissue engineering scaffold materials with specific and complex shapes. The process is simple and the prepared material structure is stable and controllable.
  • biopolymer materials commonly used in electrospinning or bio 3D printing include L-polylactic acid, polycaprolactone, polylactic-glycolic acid, chitosan, polyurethane, etc.
  • a single polymer fiber often cannot meet the needs of clinical applications due to low mechanical properties or poor biocompatibility.
  • researchers have modified polymer fibers in order to obtain excellent performance tissue engineering scaffold materials, and have made a series of breakthroughs.
  • a guided tissue regeneration membrane composed of chitosan and calcium phosphate salt.
  • the membrane structure is a dense layer and a loose porous layer, and has good bone-promoting activity, but chitosan-based biological materials exist The disadvantage of poor mechanical properties.
  • Brown et al. prepared a composite fiber membrane of type I collagen modified polylactic acid-glycolic acid by electrospinning, but the collagen was derived from rat tail.
  • commercial collagen-based guided tissue regeneration membranes (Bio-Gide, Bio-Mend, etc.) in clinical use also have defects: (1) Due to their rapid degradation behavior, they lose the ability to maintain space; (2) Most of the collagen is derived from mammals Organs or tissues, such as pig skin, cattle Achilles tendon, etc., have the risk of potential disease transmission and religious restrictions.
  • Qingqiang Yao et al. used PCL as a matrix to graft amino acids and prepared a scaffold material by 3D printing for the integrated repair of soft and hard bones.
  • the materials used for periodontal hard and soft tissue regeneration and repair should also have a certain mechanical barrier effect to prevent gingival epithelium. Cells migrate to the root surface, so the scaffold material should have a denser structural layer to act as a physical barrier.
  • the purpose of the present invention is: based on the above research status and defects, the present invention combines the traditional electrospinning technology and the emerging bio-3D printing technology to prepare an integrated repair gradient material that can simultaneously guide the regeneration of periodontal hard and soft tissues.
  • a gradient material for guiding the regeneration of periodontal hard and soft tissues comprising a 3D printing stent layer and an electrospinning fiber membrane layer, wherein the content of hydroxyapatite in the 3D printing stent layer is higher than that of the electrospinning fiber membrane layer, the 3D printing stent
  • the pore size of the layer is larger than the electrospun fiber membrane layer, the pore size of the 3D printing scaffold layer is 100-1000 ⁇ m, the fiber diameter of the electrospun fiber membrane layer is 300-5000 nm, and the electrospun fiber membrane layer
  • the structure is random distribution or oriented arrangement or grid structure, and the thickness of the electrospun fiber membrane layer is 0.08-1mm.
  • the pore structure changes gradually, from 300-10000nm of the fiber membrane to a few hundred microns of the scaffold; the composition of the gradient changes, the content of apatite in the fiber membrane is lower, while the content of apatite in the 3D scaffold is higher: fish collagen is in the fiber membrane The content of medium is higher, and it is less in the porous scaffold, or even not added. This reflects the gradient changes in the pore structure and composition content of the fiber membrane and the porous scaffold.
  • Fish collagen can promote the adhesion and growth of cells derived from soft tissues; scaffolds with high apatite content can promote osteogenic differentiation of osteoblasts.
  • the gradient material can be obtained by compounding the electrospun fiber membrane and the biological 3D printed scaffold.
  • the specific composite method can be the ABAB structure formed by the fiber membrane-3D printed scaffold. The structure alternates in three-dimensional directions, and the pore structure and porosity are repeated Alternate.
  • the gradient material can be an electrospun fiber membrane wrapped on the surface of the biological 3D printing scaffold to form a hard-soft gradient structure and promote the interface combination of the scaffold material and the host tissue.
  • the gradient material can be a composite scaffold material formed by cutting the electrospun fiber membrane into fragments and filling the hole structure of the 3D printing scaffold.
  • the upper layer of the gradient material is composed of an electrospun fiber membrane with a small porosity, which can effectively prevent the migration of gingival epithelial cells and gingival connective tissue cells to the root surface, and the lower layer is composed of a 3D printed scaffold with a large porosity.
  • Gradient materials have excellent mechanical properties, adjustable degradation rates, low immunogenicity and good biological activity in vivo, and can have gradient changes in structure and composition.
  • the method for preparing the gradient material for guiding the regeneration of periodontal hard and soft tissue includes the following steps:
  • step S2 Stirring the spinning solution obtained in step S1 to volatilize the solvent to obtain a 3D printing ink; wherein the solvent is partially volatilized to obtain a 3D printing ink of the desired concentration;
  • step S3 The spinning solution obtained in step S1 is prepared by an electrostatic spinning method to obtain an electrostatic spinning fiber membrane layer;
  • step S4 Place the electrospun fiber membrane layer obtained in step S3 on the biological 3D printer platform, and use the 3D printing ink obtained in step S2 to print on the electrospun fiber membrane layer by the biological 3D printer to construct the electrospun fiber membrane layer and 3D printing the composite gradient material of the scaffold layer.
  • the 3D printing stent layer can also be printed with a 3D printing ink through a biological 3D printer, and then an electrostatic spinning fiber membrane layer is prepared on the 3D printing stent layer by an electrostatic spinning method, thereby constructing an electrostatic spinning fiber membrane layer Gradient material compounded with 3D printed support layer.
  • Polylactic acid-glycolic acid is a biopolymer approved by the US FDA for use in the field of biomedicine.
  • Fish collagen is introduced as a protein molecule into polylactic acid-glycolic acid to interact with its molecular chain to form a molecular network structure, thereby improving the mechanics of the material performance.
  • Nano-hydroxyapatite as the main inorganic component of human bone, is introduced into polylactic acid-glycolic acid to give the material significant biological activity and bone-inducing ability.
  • the gradient material can be loaded with biological factors or drugs through nano-hydroxyapatite, such as dexamethasone, BMP, TGF, FGF, baicalin, etc., to achieve sustained release of factors or drugs, and endow the material with versatility.
  • nano-hydroxyapatite such as dexamethasone, BMP, TGF, FGF, baicalin, etc.
  • the content of nano-hydroxyapatite in the electrospun fiber membrane layer is 5-40% by weight, and the content of fish collagen is 1-30% by weight; preferably, the content of nano-hydroxyapatite in the electrospun fiber membrane layer is 10%. -25wt%, and the content of fish collagen is 5-15wt%.
  • the content of nano-hydroxyapatite in the 3D printing support layer is 10-70 wt%.
  • the hydroxyapatite in step S1 includes short rod-shaped, needle-shaped, microspherical and mesoporous hydroxyapatite, and the hydroxyapatite can be replaced by calcium phosphate or calcium silicate.
  • polylactic acid-glycolic acid can be replaced by polycaprolactone, polylactic acid, polyurethane, or chitosan.
  • step S1 the fish collagen is derived from fish skin or scales, and the fish is cod or tilapia or grass carp or silver carp.
  • the solvent in step S1 is trifluoroethanol or hexafluoroisopropanol or dichloromethane or acetone or N,N-dimethylformamide or trifluoroethanol/N with a volume ratio of 7-9:1-3.
  • step S3 is: collecting randomly distributed, oriented, and grid-like fiber membranes by using a flat-plate receiver, an orientation receiver, and a grid-like receiver, respectively, and the electrospinning process parameter is set to voltage 7-12kV, receiving distance 12-18cm, bolus injection speed 0.3-0.6mL/h; roller speed of orientation receiver is 2000-4000r/min, grid aperture size of grid receiver is 400-800 ⁇ m; among them, preferred The process parameters are voltage 7-9kV, receiving distance 15cm, bolus speed 0.4-0.5mL/h, the preferred rotation speed of the orientation receiver roller is 2500-3000r/min, and the preferred mesh aperture size of the grid receiver is 500- 600 ⁇ m.
  • step S4 the 3D printed stent layer is prepared by the biological 3D printing technology, the diameter of the extruded filament is 0.1-0.4 mm, and the shape of the stent is cube, cylinder or rhombus or other customized shapes according to clinical needs.
  • the gradient material is a functionally gradient material composed of an electrospun fiber membrane layer and a biological 3D printing scaffold layer, which can realize the simultaneous repair of periodontal hard and soft tissues.
  • the composition, pore structure, fiber membrane and 3D printing of the gradient material The arrangement of extruded filaments can present a gradient change. Compared with a single fiber membrane or a 3D printing scaffold material, the gradient material has a better application prospect;
  • the fibrous membrane in the gradient material of the present invention has a relatively dense microporous structure, which can act as a mechanical barrier to prevent gingival fibroblasts from migrating to the root surface, and at the same time guide periodontal soft tissue repair; the 3D printing scaffold material in the gradient material can guide Alveolar bone regeneration, thereby realizing the integrated restoration of periodontal hard and soft tissues;
  • the fish collagen used in the present invention is derived from marine organisms, and its amino acid composition is similar to that of mammals. It has higher biocompatibility, lower immunogenicity, higher cell affinity and biodegradability, and its amino acid sequence Unlike mammals, it has different immune epitopes, can avoid the risk of disease transmission, and has no religious, cultural, and ethical issues;
  • the present invention introduces a small amount of fish collagen into polylactic acid-glycolic acid. Because fish collagen molecules and polylactic acid-glycolic acid molecular chains form a molecular network structure through hydrogen bonding, the mechanical strength of the material is significantly improved, and the The introduction changed the main degradation behavior of the polylactic acid-glycolic acid matrix, resulting in porous degradation, significantly accelerating the degradation rate of gradient materials, and regulating the degradation rate of materials by adjusting the amount of fish collagen added;
  • the present invention uses electrospinning to prepare fiber membranes, which simulates the structure of extracellular matrix, and has simple operation, mature technology, and stable process.
  • the preparation of stents through biological 3D printing technology has strong designability and repeatability, and can be based on defects.
  • the shape and size of the part meets the customized needs of patients and realizes personalized clinical treatment.
  • Figure 1 shows the morphology of nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane
  • Figure 2 is the fiber diameter distribution diagram of nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane
  • Figure 3 shows the morphology of nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane after being degraded in phosphate buffer for 8 weeks;
  • Figure 4 shows the effect of micro-CT reconstruction of the bone repair effect of the nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane (PFC5H15) implanted in the rat bilateral skull defect model for 4 weeks;
  • Figure 5 shows the topography of the grid-like fiber membrane
  • Figure 6 is a stress-strain curve diagram
  • Fig. 7 is a diagram of gradient materials constructed by composite construction of fiber membranes and 3D printed scaffolds with different pore structures.
  • first and “second” and other relational terms are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply one of these entities or operations. There is any such actual relationship or order between.
  • the terms “include”, “include” or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or device that includes a series of elements includes not only those elements, but also includes Other elements of, or also include elements inherent to this process, method, article or equipment. If there are no more restrictions, the element defined by the sentence “including a" does not exclude the existence of other same elements in the process, method, article, or equipment including the element.
  • a preferred embodiment of the present invention provides a method for preparing a gradient material for guiding the regeneration of periodontal hard and soft tissues.
  • the raw materials include: fish collagen from Shenggong Biological Co., Ltd., hexafluoroisopropanol from Aladdin, Shandong Jinan Daigang Biology
  • the specific steps of polylactic acid-glycolic acid of Technology Co., Ltd. are as follows:
  • Step 1 Weigh 0.06 g of nano-hydroxyapatite and disperse it in 2 mL of hexafluoroisopropanol, and use an ultrasonic cell pulverizer for ultrasonic dispersion for 1 hour;
  • Step 2 Weigh 0.02g fish collagen into the dispersion obtained in step 1, and shake it with a constant temperature oscillator for 10 minutes;
  • Step 3 Weigh 0.4 g of polylactic acid-glycolic acid and add it to the mixed solution obtained in step 2, and shake it in a constant temperature oscillator at 25°C for 2 hours to obtain a spinning solution;
  • Step 4 Disperse the spinning solution obtained in step 3 again with ultrasonic cell crusher for 30 minutes;
  • Step 5 Use the spinning solution obtained in step 4 to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane through an electrospinning machine.
  • the needle used is a flat-headed needle 23G, a flat receiver is used, and the spinning parameters are set It is: voltage 8kV, bolus injection speed 0.5mL/h, receiving distance 16cm;
  • Step 6 Use a grid-like receiver in step 5, take off the fiber membrane after continuous collection for 1 hour, and place it in a vacuum drying oven for 3 days to obtain an electrospun fiber membrane layer;
  • Step 7 Weigh 0.6 g of nano-hydroxyapatite and disperse it in 10 mL of dichloromethane, and pre-disperse it for 20 minutes with an ultrasonic cleaner;
  • Step 8 Disperse the dispersion liquid obtained in step 7 with an ultrasonic cell pulverizer for 30 minutes;
  • Step 9 Weigh 0.2g of fish collagen and add it to the dispersion obtained in step 8, and shake it with a constant temperature oscillator for 20 minutes to obtain a mixed suspension of fish collagen evenly dispersed in the solution;
  • Step 10 Weigh 4g of polylactic acid-glycolic acid and add it to the mixed suspension obtained in Step 9, and shake it in a constant temperature shaker at 25°C for 2 hours;
  • Step 11 Stir the mixed solution obtained in Step 10 in a fume hood, and measure its viscosity to be 40 ⁇ 10mPa ⁇ s to obtain 3D printing ink;
  • Step 12 Place the electrospun fiber membrane layer obtained in step 6 on the bio-3D printer platform, take the ink obtained in step 11 and print on the composite fiber membrane by the bio-3D printer to prepare nano-hydroxyapatite/fish collagen/polylactic acid- Glycolic acid composite holder, the needle used is a tapered needle with an inner diameter of 0.16-0.41mm, and the 3D printing parameters are set to: holder size 10 ⁇ 10 ⁇ 5mm 3 , push out air pressure 5bar, needle temperature 30°C, receiving platform temperature 20°C, needle speed 4mm/s, the initial needle tip platform spacing is 0.208mm, the layer spacing is 0.208mm, the initial needle tip platform spacing is 0.208mm, and the printing line spacing is 0.3-0.8mm.
  • Step 13 Place the printing support in a vacuum drying oven for 3 days to get it.
  • a preferred embodiment of the present invention provides a method for preparing a gradient material for guiding the regeneration of periodontal hard and soft tissues.
  • the raw materials include: fish collagen from Shenggong Biological Co., Ltd., hexafluoroisopropanol from Aladdin, Shandong Jinan Daigang Biological
  • the specific steps of polylactic acid-glycolic acid of Technology Co., Ltd. are as follows:
  • Step 1 Weigh 0.06 g of nano-hydroxyapatite and disperse it in 2 mL of hexafluoroisopropanol, and use an ultrasonic cell pulverizer to ultrasonically disperse for 1 hour;
  • Step 2 Weigh 0.02g fish collagen and add it to the dispersion obtained in Step 1, and shake it with a constant temperature oscillator for 10 minutes;
  • Step 3 Weigh 0.4 g of polylactic acid-glycolic acid and add it to the mixed solution obtained in step 2, and shake it in a constant temperature oscillator at 25°C for 2 hours to obtain a spinning solution;
  • Step 4 Disperse the spinning solution obtained in step 3 again with ultrasonic cell crusher for 30 minutes;
  • Step 5 Use the spinning solution obtained in step 4 to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane through an electrospinning machine.
  • the needle used is a flat-head needle 23G, a flat receiver is used, and the spinning parameters are set It is: voltage 8kV, bolus injection rate 0.5mL/h, receiving distance 16cm;
  • Step 6 Use a grid-like receiver in step 5, take off the fiber membrane after continuous collection for 1 hour, and place it in a vacuum drying oven for 3 days to obtain an electrospun fiber membrane layer;
  • Step 7 Weigh 0.6 g of nano-hydroxyapatite and disperse it in 10 mL of dichloromethane, and use an ultrasonic cleaner to pre-disperse ultrasonically for 20 minutes;
  • Step 8 Disperse the dispersion liquid obtained in step 7 with an ultrasonic cell crusher for 30 minutes;
  • Step 9 Weigh 0.2 g of fish collagen and add it to the dispersion obtained in step 8, and shake it with a constant temperature oscillator for 20 minutes to obtain a mixed suspension of fish collagen evenly dispersed in the solution;
  • Step 10 Weigh 4g of polylactic acid-glycolic acid and add it to the mixed suspension obtained in step 9, and shake it in a constant temperature shaker at 25°C for 2 hours;
  • Step 11 Stir the mixed solution obtained in Step 10 in a fume hood, and measure its viscosity to be 40 ⁇ 10mPa ⁇ s to obtain 3D printing ink;
  • Step 12 Place the electrospun fiber membrane layer obtained in step 6 on the bio 3D printer platform, take the ink obtained in step 11 to print on the composite fiber membrane by the bio 3D printer, and pause after printing a layer, and spin the electrospinning fiber film obtained in step 6
  • the fiber membrane layer is placed on the first layer of 3D printing, and the above operation is repeated ten times to prepare the nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite formed by alternating layers of electrospun fiber membrane and biological 3D printing scaffold.
  • the needle used is a tapered needle with an inner diameter of 0.16-0.41mm
  • the 3D printing parameters are set to: holder size 10 ⁇ 10 ⁇ 5mm 3 , push air pressure 5bar, needle temperature 30°C, receiving platform temperature 20°C, needle speed 4mm /s, the initial needle tip platform spacing is 0.208mm, the layer spacing is 0.2-0.3mm, the initial needle tip platform spacing is 0.208mm, and the printing line spacing is 0.3-0.8mm;
  • Step 13 Place the printing support in a vacuum drying oven for 3 days to get it.
  • Step 1 Weigh 0.06 g of nano-hydroxyapatite and disperse it in 2 mL of hexafluoroisopropanol, and use an ultrasonic cell pulverizer to ultrasonically disperse for 1 hour;
  • Step 2 Weigh 0.02g fish collagen and add it to the dispersion obtained in Step 1, and shake it with a constant temperature oscillator for 10 minutes;
  • Step 3 Weigh 0.4 g of polylactic acid-glycolic acid and add it to the mixed solution obtained in step 2, and shake it in a constant temperature oscillator at 25°C for 2 hours to obtain a spinning solution;
  • Step 4 Disperse the spinning solution obtained in step 3 again with an ultrasonic cell crusher for 30 minutes;
  • Step 5 Use the spinning solution obtained in step 4 to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane through an electrospinning machine.
  • the needle used is a flat-head needle 23G, a flat receiver is used, and the spinning parameters are set It is: voltage 8kV, bolus injection rate 0.5mL/h, receiving distance 16cm;
  • Step 6 After continuous collection for 2 hours, remove the fiber membrane from the receiver and place it in a vacuum drying oven to dry for 3 days to fully evaporate the solvent;
  • Step 7 Electrospinning to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid fiber membrane
  • the electrospinning method was used to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane.
  • the specific steps are as follows:
  • Step 1 Weigh 0.06 g of nano-hydroxyapatite and disperse it in 2 mL of hexafluoroisopropanol, and use an ultrasonic cell pulverizer for ultrasonic dispersion for 1 hour;
  • Step 2 Weigh 0.02g fish collagen and add it to the dispersion obtained in Step 1, and shake it with a constant temperature oscillator for 10 minutes;
  • Step 3 Weigh 0.4 g of polylactic acid-glycolic acid and add it to the mixed solution obtained in step 2, and shake it in a constant temperature oscillator at 25°C for 2 hours to obtain a spinning solution;
  • Step 4 Disperse the spinning solution obtained in step 3 again with ultrasonic cell crusher for 30 minutes;
  • Step 5 Use the spinning solution obtained in step 4 to prepare nano-hydroxyapatite/fish collagen/polylactic acid-glycolic acid composite fiber membrane through an electrospinning machine.
  • the needle used is a flat-headed needle 23G, a flat receiver is used, and the spinning parameters are set It is: voltage 8kV, bolus injection speed 0.5mL/h, receiving distance 16cm;
  • Step 6 Use a grid-like receiver in step 5, take off the fiber membrane after continuous collection for 1 hour, and dry it in a vacuum drying oven for 3 days;
  • the morphology of the grid-like fiber membrane was observed by scanning electron microscopy. As shown in Fig. 5, the fiber membrane formed a uniform grid repeating unit with a grid size of about 500 ⁇ m.

Landscapes

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

Abstract

一种引导牙周硬软组织再生梯度材料及其制备方法。梯度材料包括3D打印支架层和静电纺丝纤维膜层。静电纺丝纤维膜的微孔结构可防止牙龈成纤维细胞向根面迁移从而起到机械屏障作用，同时利于营养和代谢物质的传输。3D打印工艺的高度可设计性能够使得支架层的结构和形状根据临床需要实现牙槽骨个性化修复。制备方法简单稳定，使牙周引导组织再生膜与牙槽骨修复支架材料一体化，具有实现临床定制化医疗的潜能。梯度材料具有良好的体内引导硬软组织修复效果，能够更好的应用于牙槽骨及牙周组织同步再生或修复。

Description

一种引导牙周硬软组织再生梯度材料及其制备方法 技术领域
本发明属于生物医用材料技术领域，具体涉及一种引导牙周硬软组织再生梯度材料及其制备方法。
背景技术
引导骨再生(Guided Bone Regeneration,GBR)技术是利用屏障膜阻止牙龈上皮向根表面迁移，从而为受损牙槽骨的重建和再生创造足够空间。引导组织再生技术为辅助治疗牙周疾病、促进骨组织再生提供了新策略。传统的不可降解GBR膜需要通过二次手术去除，会增加感染风险及患者痛苦，因而逐渐被淘汰；胶原基可降解膜由于其胶原成分大多来源于哺乳动物组织，因而存在疾病传播的风险，且存在降解过快的问题；生物合成高分子基可降解膜存在生物相容性不佳的问题。同时，临床上使用GBR膜时需在种植体周围结合使用骨粉以增强骨性结合及修复效果，但对于较大的牙槽骨缺损，植入的骨粉容易移位。基于此，本发明构建了一种集引导组织再生膜和骨充填双重作用一体化的梯度复合材料。
理想的组织修复材料既能从成分、结构上模拟天然组织的局部微环境且具有特定的生物学功能。静电纺丝技术作为一种操作简单、经济有效的制备超细纤维的方法，能制备微纳米级的纤维，纤维结构很好的模拟了细胞外基质结构，且所制备的纤维膜具有比表面积大、孔隙率高等特点，因而广泛应用于生物医学材料领域。3D打印作为一种极具发展前景的打印技术，可以产生具有良好互联性的高孔隙结构，可打印具有特定、复杂形状的组织工程支架材料，其工艺简单，制备的材料结构稳定、可控。
目前，常用于静电纺丝或生物3D打印的生物高分子材料包括左旋聚乳酸、聚己内酯、聚乳酸-羟基乙酸、壳聚糖、聚氨酯等。但是单一的高分子纤维往往由于力学性能较低或生物相容性较差而不能满足临床应用的需求。近年来，研究者通过对高分子纤维改性以期望获得性能优良的组织工程支架材料，并取得了一系列突破性进展。
现有技术中有一种壳聚糖与钙磷盐复合的引导组织再生膜的制备方法，该膜结构为致密层和疏松多孔层，具有良好的促成骨活性，但是壳聚糖基的生物材料存在力学性能较差的缺点。现有技术还有一种基于羟基磷灰石接枝聚丙交酯/聚乳酸共聚羟基乙酸电纺纤维膜的制备方法，构建了一种新的可生物降解的引导组织再生膜，复合膜展示出更好的力学性能，但该复合膜体系缺乏生物活性好的天然成分，如胶原蛋白、明胶等。
Brown等人通过静电纺丝制备了I型胶原改性聚乳酸-羟基乙酸的复合纤维膜，但胶原来源于鼠尾。目前临床使用商业化的胶原基引导组织再生膜(Bio-Gide，Bio-Mend等)也存在 缺陷：(1)由于其快速降解行为而丧失维持空间的能力；(2)胶原大多来源于哺乳动物器官或组织，如猪皮、牛跟腱等，存在潜在疾病传播的风险以及宗教限制等问题。
Qingqiang Yao等人以PCL为基体接枝氨基酸后通过3D打印制备了支架材料用于软硬骨一体化修复，但是用于牙周硬软组织再生修复材料也应具有一定的机械屏障作用，防止牙龈上皮细胞向根面迁移，因此支架材料应具有较致密的结构层发挥物理屏障作用。
发明内容
本发明的目的在于：基于上述研究现状和缺陷，本发明将传统的静电纺丝技术与新兴的生物3D打印技术相结合制备出能够同时引导牙周硬软组织再生的一体化修复梯度材料。
本发明采用的技术方案如下：
一种引导牙周硬软组织再生梯度材料，包括3D打印支架层和静电纺丝纤维膜层，所述3D打印支架层中羟基磷灰石含量高于静电纺丝纤维膜层，所述3D打印支架层的孔径大于静电纺丝纤维膜层，所述3D打印支架层的孔径大小为100-1000μm，所述静电纺丝纤维膜层的纤维直径大小为300-5000nm，所述静电纺丝纤维膜层的结构为无规分布或取向排列或网格状结构，静电纺丝纤维膜层厚度为0.08-1mm。
孔结构梯度变化，从纤维膜300-10000nm变化到支架的几百微米；成分的梯度变化，纤维膜中磷灰石含量较低，而3D支架中磷灰石含量较高：鱼胶原在纤维膜中含量较高，在多孔支架中较少，甚至可以不加。由此体现出，纤维膜与多孔支架在孔结构方面和成分含量方面的梯度变化。
鱼胶原可促进来源于软组织的细胞黏附、生长；高的磷灰石含量的支架可促进成骨性细胞的成骨分化。
梯度材料可为静电纺丝纤维膜与生物3D打印支架复合而得，具体复合方式可为纤维膜-3D打印支架形成的ABAB结构，该结构在三维方向上重复交替，孔结构及孔隙率呈重复交替式。
梯度材料可为静电纺丝纤维膜包裹在生物3D打印支架表面，形成硬-软梯度结构，促进支架材料与宿主组织的界面结合。
梯度材料可为静电纺丝纤维膜剪切成碎片后填充于3D打印支架孔结构中形成的复合支架材料。
梯度材料上层由孔隙率较小的静电纺丝纤维膜组成，能够有效阻止牙龈上皮细胞和牙龈结缔组织细胞向根面迁移，下层由具有较大孔隙率的3D打印支架组成。梯度材料具有优异的力学性能、可调控的降解速率、低免疫原性以及良好的体内生物活性，且在结构和成分上均可成梯度变化。
上述引导牙周硬软组织再生梯度材料的制备方法，包括以下步骤：
S1.将纳米羟基磷灰石分散于溶剂，超声分散1-2h，再加入鱼胶原和聚乳酸-羟基乙酸，震荡摇匀1.5-3h，再超声0.5-1h，得到纺丝液；其中超声分散方法为两步超声法；
S2.搅拌步骤S1中所得纺丝液使溶剂挥发，得到3D打印墨水；其中溶剂部分挥发，得到所需浓度的3D打印墨水；
S3.将步骤S1所得纺丝液通过静电纺丝法制备得到静电纺丝纤维膜层；
S4.将步骤S3得到的静电纺丝纤维膜层置于生物3D打印机平台上，使用步骤S2所得3D打印墨水通过生物3D打印机在静电纺丝纤维膜层上打印，构建静电纺丝纤维膜层与3D打印支架层复合的梯度材料。
上述制备方法中，也可先用3D打印墨水通过生物3D打印机打印3D打印支架层，然后在3D打印支架层上通过静电纺丝法制备静电纺丝纤维膜层，从而构建静电纺丝纤维膜层与3D打印支架层复合的梯度材料。
聚乳酸-羟基乙酸是经美国FDA批准应用于生物医药领域的生物高分子，鱼胶原作为蛋白分子引入到聚乳酸-羟基乙酸中能与其分子链发生相互作用形成分子网络结构，从而提高材料的力学性能。纳米羟基磷灰石作为人体骨主要的无机成分引入到聚乳酸-羟基乙酸中可赋予材料显著的生物活性和诱导成骨能力。
梯度材料可通过纳米羟基磷灰石负载生物因子或药物，如地塞米松、BMP、TGF、FGF、黄芩苷等，实现因子或药物的持续性释放，并赋予材料多功能性。
进一步地，静电纺丝纤维膜层中纳米羟基磷灰石含量为5-40wt％，鱼胶原的含量为1-30wt％；优选地，静电纺丝纤维膜层中纳米羟基磷灰石含量为10-25wt％，鱼胶原的含量为5-15wt％。
进一步地，3D打印支架层中纳米羟基磷灰石含量为10-70wt％。
进一步地，步骤S1中羟基磷灰石包括短棒状、针状、微球状及介孔羟基磷灰石，羟基磷灰石可由磷酸钙盐或硅酸钙盐替换。
进一步地，步骤S1中聚乳酸-羟基乙酸可由聚己内酯或聚乳酸或聚氨酯或壳聚糖替换。
进一步地，步骤S1中鱼胶原来源于鱼皮或鱼鳞，鱼为鳕鱼或罗非鱼或草鱼或鲢鱼。
进一步地，步骤S1中溶剂为三氟乙醇或六氟异丙醇或二氯甲烷或丙酮或N,N-二甲基甲酰胺或体积比7-9:1-3的三氟乙醇/N,N-二甲基甲酰胺混合溶液或体积比2-4:1的丙酮/N,N-二甲基甲酰胺混合溶液。
进一步地，步骤S3具体过程为：通过使用平板接收器、取向接收器、网格状接收器分别收集无规分布、取向排列、网格状的纤维膜，所述静电纺丝工艺参数设置为电压7-12kV、接收距离12-18cm、推注速度0.3-0.6mL/h；取向接收器辊筒转速为2000-4000r/min，网格接 收器的网格孔径大小为400-800μm；其中，优选的工艺参数为电压7-9kV、接收距离15cm、推注速度0.4-0.5mL/h，取向接收器辊筒优选转速为2500-3000r/min，网格接收器的优选网格孔径大小为500-600μm。
进一步地，步骤S4中通过生物3D打印技术制备3D打印支架层，挤出丝的直径为0.1-0.4mm，支架形状为正方体或圆柱状或菱柱或根据临床需要的其他个性化定制形状。
综上所述，由于采用了上述技术方案，本发明的有益效果是：
1、本发明中，梯度材料为静电纺丝纤维膜层与生物3D打印支架层复合构建的功能梯度材料，能实现牙周硬软组织同步修复，梯度材料的成分、孔结构、纤维膜与3D打印挤出丝的排列方式均可呈现梯度变化，相比于单一的纤维膜或者3D打印支架材料，该梯度材料具有更好的应用前景；
2、本发明梯度材料中纤维膜具有相对致密的微孔结构，能够作为机械屏障膜阻止牙龈成纤维细胞向根面迁移，同时引导牙周软组织修复；所述梯度材料中3D打印支架材料能引导牙槽骨再生，从而实现牙周硬软组织一体化修复；
3、本发明使用的鱼胶原为海洋生物来源，其氨基酸组成与哺乳动物相似，具有较高的生物相容性、较低的免疫原性、较高的细胞亲和力和生物降解性，其氨基酸序列与哺乳动物不同，具有不同的免疫表位，能避免疾病传播的风险，且没有宗教文化、伦理问题；
4、本发明通过在聚乳酸-羟基乙酸中引入少量鱼胶原，由于鱼胶原分子与聚乳酸-羟基乙酸分子链通过氢键作用形成分子网络结构，显著提高了材料的力学强度，且鱼胶原的引入改变了聚乳酸-羟基乙酸基体的主要降解行为，出现孔洞式降解，显著加快了梯度材料的降解速率，且通过调控鱼胶原的添加量调控材料的降解速率；
5、本发明通过静电纺丝制备纤维膜，模拟了细胞外基质结构，且操作简单、技术成熟、工艺稳定，通过生物3D打印技术制备支架具有极强的可设计性与重复性，能够根据缺损部位形状大小满足患者定制化需求，实现临床上个性化治疗。
附图说明
为了更清楚地说明本发明实施例的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，应当理解，以下附图仅示出了本发明的某些实施例，因此不应被看作是对范围的限定，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他相关的附图。
图1为纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜的形貌图；
图2为纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜的纤维直径分布图；
图3为纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜在磷酸盐缓冲液中降解8周后的形貌图；
图4为纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜(PFC5H15)在大鼠颅骨双侧缺损模型中植入4周后Micro-CT重建的缺损区骨修复效果图；
图5为网格状纤维膜形貌图；
图6为应力-应变曲线图；
图7为纤维膜与不同孔结构3D打印支架复合构建的梯度材料图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合附图及实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅用以解释本发明，并不用于限定本发明，即所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。通常在此处附图中描述和示出的本发明实施例的组件可以以各种不同的配置来布置和设计。
因此，以下对在附图中提供的本发明的实施例的详细描述并非旨在限制要求保护的本发明的范围，而是仅仅表示本发明的选定实施例。基于本发明的实施例，本领域技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例，都属于本发明保护的范围。
需要说明的是，术语“第一”和“第二”等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来，而不一定要求或者暗示这些实体或操作之间存在任何这种实际的关系或者顺序。而且，术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含，从而使得包括一系列要素的过程、方法、物品或者设备不仅包括那些要素，而且还包括没有明确列出的其他要素，或者是还包括为这种过程、方法、物品或者设备所固有的要素。在没有更多限制的情况下，由语句“包括一个……”限定的要素，并不排除在包括所述要素的过程、方法、物品或者设备中还存在另外的相同要素。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
实施例1
本发明较佳实施例提供的一种引导牙周硬软组织再生梯度材料的制备方法，原料包括：生工生物股份有限公司的鱼胶原，阿拉丁公司的六氟异丙醇，山东济南岱罡生物科技有限公司的聚乳酸-羟基乙酸，具体步骤如下：
步骤1：称取0.06g纳米羟基磷灰石分散于2mL六氟异丙醇中，用超声细胞粉碎机超声分 散1h；
步骤2：称取0.02g鱼胶原加入步骤1所得分散液，用恒温振荡器震荡摇匀10min；
步骤3：称取0.4g聚乳酸-羟基乙酸加入到步骤2所得混合溶液，在恒温振荡器中于25℃下震荡摇匀2h，得纺丝液；
步骤4：将步骤3所得纺丝液再次用超声细胞粉碎机超声分散30min；
步骤5：将步骤4所得纺丝液通过静电纺丝机制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜，所用针头为平头针23G，采用平板接收器，纺丝参数设置为：电压8kV，推注速度0.5mL/h，接收距离16cm；
步骤6：步骤5中使用网格状接收器，连续收集1h后取下纤维膜，置于真空干燥箱中干燥3天，即得静电纺丝纤维膜层；
步骤7：称取0.6g纳米羟基磷灰石分散于10mL二氯甲烷中，用超声波清洗机超声预分散20min；
步骤8：将步骤7所得分散液液用超声细胞粉碎机超声分散30min；
步骤9：称取0.2g鱼胶原加入步骤8所得分散液，用恒温振荡器震荡摇匀20min,得鱼胶原均匀分散在溶液里的混合悬浊液；
步骤10：称取4g聚乳酸-羟基乙酸加入到步骤9所得混合悬浊液，在恒温振荡器中于25℃下震荡摇匀2h；
步骤11：将步骤10所得混合溶液在通风橱中搅拌，测量其粘度为40±10mPa·s，得3D打印墨水；
步骤12：将步骤6所得静电纺丝纤维膜层置于生物3D打印机平台上，取步骤11所得墨水通过生物3D打印机在复合纤维膜上打印，制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合支架，所用针头为锥形针头，内径0.16-0.41mm，3D打印参数设置为：支架尺寸10×10×5mm 3，推出气压5bar，针头温度30℃，接收平台温度20℃，针头速度4mm/s，初始针头平台间距0.208mm，层间距0.208mm，初始针尖平台距离0.208mm，打印线间距0.3-0.8mm。
步骤13：将打印支架置于真空干燥箱中干燥3天，即得。
实施例2
本发明较佳实施例提供的一种引导牙周硬软组织再生梯度材料的制备方法，原料包括：生工生物股份有限公司的鱼胶原，阿拉丁公司的六氟异丙醇，山东济南岱罡生物科技有限公司的聚乳酸-羟基乙酸，具体步骤如下：
步骤1：称取0.06g纳米羟基磷灰石分散于2mL六氟异丙醇中，用超声细胞粉碎机超声分散1h；
步骤2：称取0.02g鱼胶原加入步骤1所得分散液，用恒温振荡器震荡摇匀10min；
步骤3：称取0.4g聚乳酸-羟基乙酸加入到步骤2所得混合溶液，在恒温振荡器中于25℃下震荡摇匀2h，得纺丝液；
步骤4：将步骤3所得纺丝液再次用超声细胞粉碎机超声分散30min；
步骤5：将步骤4所得纺丝液通过静电纺丝机制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜，所用针头为平头针23G，采用平板接收器，纺丝参数设置为：电压8kV，推注速度0.5mL/h，接收距离16cm；
步骤6：步骤5中使用网格状接收器，连续收集1h后取下纤维膜，置于真空干燥箱中干燥3天，即得静电纺丝纤维膜层；
步骤7：称取0.6g纳米羟基磷灰石分散于10mL二氯甲烷中，用超声波清洗机超声预分散20min；
步骤8：将步骤7所得分散液液用超声细胞粉碎机超声分散30min；
步骤9：称取0.2g鱼胶原加入步骤8所得分散液，用恒温振荡器震荡摇匀20min,得鱼胶原均匀分散在溶液里的混合悬浊液；
步骤10：称取4g聚乳酸-羟基乙酸加入到步骤9所得混合悬浊液，在恒温振荡器中于25℃下震荡摇匀2h；
步骤11：将步骤10所得混合溶液在通风橱中搅拌，测量其粘度为40±10mPa·s，得3D打印墨水；
步骤12：将步骤6所得静电纺丝纤维膜层置于生物3D打印机平台上，取步骤11所得墨水通过生物3D打印机在复合纤维膜上打印，打印一层后暂停，将步骤6所得静电纺丝纤维膜层置于3D打印第一层之上，重复以上操作十次，制备静电纺丝纤维膜与生物3D打印支架逐层交替形成的纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合梯度支架材料，所用针头为锥形针头，内径0.16-0.41mm，3D打印参数设置为：支架尺寸10×10×5mm 3，推出气压5bar，针头温度30℃，接收平台温度20℃，针头速度4mm/s，初始针头平台间距0.208mm，层间距0.2-0.3mm，初始针尖平台距离0.208mm，打印线间距0.3-0.8mm；
步骤13：将打印支架置于真空干燥箱中干燥3天，即得。
实验例1
实验采用静电纺丝方法制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸的复合纤维膜，具体步骤如下：
步骤1：称取0.06g纳米羟基磷灰石分散于2mL六氟异丙醇中，用超声细胞粉碎机超声分散1h；
步骤2：称取0.02g鱼胶原加入步骤1所得分散液，用恒温振荡器震荡摇匀10min；
步骤3：称取0.4g聚乳酸-羟基乙酸加入到步骤2所得混合溶液，在恒温振荡器中于25℃下震荡摇匀2h，得纺丝液；
步骤4：将步骤3所得纺丝液再次用超声细胞粉碎机超声分散30min；
步骤5：将步骤4所得纺丝液通过静电纺丝机制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜，所用针头为平头针23G，采用平板接收器，纺丝参数设置为：电压8kV，推注速度0.5mL/h，接收距离16cm；
步骤6：连续收集2h后，将纤维膜从接收器上取下，置于真空干燥箱中干燥3天，使溶剂充分挥发；
步骤7：静电纺丝制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸纤维膜；
使用扫描电镜观察纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸纤维膜的形貌，结果如图1所示，纤维光滑、无串珠形成；测量其直径分布，结果如图2所示，纤维平均直径为486±64nm，通过两步超声法实现了纳米羟基磷灰石在纤维中的均匀分布，纤维形貌良好。
通过扫描电镜观察体外降解实验8周后复合纤维膜的形貌，结果如图3所示，表明鱼胶原的引入改变了纤维的主要降解行为，纤维断裂、溶胀、腐蚀的同时，出现了孔洞式降解，显著加快了纤维膜的降解速率。
通过大鼠颅骨双侧缺损模型评价复合纤维膜体内骨修复效果，结果如图4所示，颅骨缺损区明显有新生骨组织形成，表明复合纤维膜在引导组织再生领域极具应用前景。
实验例2
实验采用静电纺丝方法制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸的复合纤维膜，具体步骤如下：
步骤1：称取0.06g纳米羟基磷灰石分散于2mL六氟异丙醇中，用超声细胞粉碎机超声分散1h；
步骤2：称取0.02g鱼胶原加入步骤1所得分散液，用恒温振荡器震荡摇匀10min；
步骤3：称取0.4g聚乳酸-羟基乙酸加入到步骤2所得混合溶液，在恒温振荡器中于25℃下震荡摇匀2h，得纺丝液；
步骤4：将步骤3所得纺丝液再次用超声细胞粉碎机超声分散30min；
步骤5：将步骤4所得纺丝液通过静电纺丝机制备纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜，所用针头为平头针23G，采用平板接收器，纺丝参数设置为：电压8kV，推注速度0.5mL/h，接收距离16cm；
步骤6：步骤5中使用网格状接收器，连续收集1h后取下纤维膜，置于真空干燥箱中干 燥3天；
通过扫描电镜观察网格状纤维膜形貌，结果如图5所示，纤维膜形成了均一的网格重复单元，网格尺寸大小为500μm左右。
实验例3
分别制备聚乳酸-羟基乙酸纤维膜(P)、鱼胶原/聚乳酸-羟基乙酸纤维膜(PFC5)以及纳米羟基磷灰石/鱼胶原/聚乳酸-羟基乙酸复合纤维膜(PFC5H15)，进行拉伸强度测试，结果如图6和下表1所示，可知鱼胶原的引入显著提高了纤维膜的拉伸强度。
表1纤维膜的力学性能表
Figure PCTCN2019095661-appb-000001
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (9)

  1. 一种引导牙周硬软组织再生梯度材料，其特征在于：包括3D打印支架层和静电纺丝纤维膜层，所述3D打印支架层中羟基磷灰石含量高于静电纺丝纤维膜层，所述3D打印支架层的孔径大于静电纺丝纤维膜层，所述3D打印支架层的孔径大小为100-1000μm，所述静电纺丝纤维膜层的纤维直径大小为300-5000nm，所述静电纺丝纤维膜层的结构为无规分布或取向排列或网格状结构，静电纺丝纤维膜层厚度为0.08-1mm。
  2. 权利要求1所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于，包括以下步骤：
    S1.将纳米羟基磷灰石分散于溶剂，超声分散1-2h，再加入鱼胶原和聚乳酸-羟基乙酸，震荡摇匀1.5-3h，再超声0.5-1h，得到纺丝液；
    S2.搅拌步骤S1中所得纺丝液使溶剂挥发，得到3D打印墨水；
    S3.将步骤S1所得纺丝液通过静电纺丝法制备得到静电纺丝纤维膜层；
    S4.将步骤S3得到的静电纺丝纤维膜层置于生物3D打印机平台上，使用步骤S2所得3D打印墨水通过生物3D打印机在静电纺丝纤维膜层上打印，构建静电纺丝纤维膜层与3D打印支架层复合的梯度材料。
  3. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述静电纺丝纤维膜层中纳米羟基磷灰石含量为5-40wt％，鱼胶原的含量为1-30wt％。
  4. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述3D打印支架层中纳米羟基磷灰石含量为10-70wt％。
  5. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述步骤S1中羟基磷灰石包括短棒状、针状、微球状及介孔羟基磷灰石，所述羟基磷灰石可由磷酸钙盐或硅酸钙盐替换。
  6. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述步骤S1中聚乳酸-羟基乙酸可由聚己内酯或聚乳酸或聚氨酯或壳聚糖替换。
  7. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述步骤S1中鱼胶原来源于鱼皮或鱼鳞，鱼为鳕鱼或罗非鱼或草鱼或鲢鱼。
  8. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述步骤S1中溶剂为三氟乙醇或六氟异丙醇或二氯甲烷或丙酮或N,N-二甲基甲酰胺或体积比7-9:1-3的三氟乙醇/N,N-二甲基甲酰胺混合溶液或体积比2-4:1的丙酮/N,N-二甲基甲酰胺混合溶液。
  9. 根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于，所述步骤S3具体过程为：通过使用平板接收器、取向接收器、网格状接收器分别收集无规分布、 取向排列、网格状的纤维膜，所述静电纺丝工艺参数设置为电压7-12kV、接收距离12-18cm、推注速度0.3-0.6mL/h；取向接收器辊筒转速为2000-4000r/min，网格接收器的网格孔径大小为400-800μm。
    根据权利要求2所述的引导牙周硬软组织再生梯度材料的制备方法，其特征在于：所述步骤S4中通过生物3D打印技术制备3D打印支架层，挤出丝的直径为0.1-0.4mm。
PCT/CN2019/095661 2019-05-30 2019-07-12 一种引导牙周硬软组织再生梯度材料及其制备方法 Ceased WO2020237785A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/771,221 US11696974B2 (en) 2019-05-30 2019-07-12 Method for preparing a functionally gradient material for guided periodontal hard and soft tissue regeneration

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910465240.1 2019-05-30
CN201910465240.1A CN110141687B (zh) 2019-05-30 2019-05-30 一种引导牙周硬软组织再生梯度材料及其制备方法

Publications (1)

Publication Number Publication Date
WO2020237785A1 true WO2020237785A1 (zh) 2020-12-03

Family

ID=67590033

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/095661 Ceased WO2020237785A1 (zh) 2019-05-30 2019-07-12 一种引导牙周硬软组织再生梯度材料及其制备方法

Country Status (3)

Country Link
US (1) US11696974B2 (zh)
CN (1) CN110141687B (zh)
WO (1) WO2020237785A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112604029A (zh) * 2020-12-30 2021-04-06 上海纳米技术及应用国家工程研究中心有限公司 用于牙槽骨修复的“三明治式”可降解支架的制备及产品和应用
CN115252906A (zh) * 2022-07-29 2022-11-01 佛山仙湖实验室 一种三明治结构支架及其制备方法和应用
CN116373357A (zh) * 2023-03-16 2023-07-04 北京市春立正达医疗器械股份有限公司 一种表面梯度化多孔结构的制备方法

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111110922B (zh) * 2019-12-25 2020-10-27 四川大学 一种用于3d生物打印的牙周生物模块及构建方法及应用
CN113117150B (zh) * 2019-12-31 2022-07-19 广州迈普再生医学科技股份有限公司 引导组织再生膜及其制备方法和应用
CN111214698B (zh) * 2020-01-22 2021-10-22 潍坊医学院附属医院 一种复合骨修复材料及其制备方法
CN111494720A (zh) * 2020-03-31 2020-08-07 东华大学 一种功能一体化可吸收引导组织再生膜及其制备方法
CN113332497B (zh) * 2021-04-30 2022-04-22 国家纳米科学中心 一种双面支架及其制备方法和应用
CN113230460A (zh) * 2021-05-07 2021-08-10 四川大学 一种引导口腔软硬组织一体化修复多孔支架及其制备方法
CN115094529A (zh) * 2022-07-11 2022-09-23 吉林大学第一医院 一种多孔取向性plga静电纺丝纤维及其制备方法
CN116120051A (zh) * 2023-02-13 2023-05-16 武汉理工大学 一种基于光固化的梯度生物复合陶瓷骨架的制备方法
CN117026517A (zh) * 2023-06-28 2023-11-10 广西桂科院华仁济瀛生物科技有限公司 一种静电纺丝纳米纤维膜、制备方法及应用
CN117298353A (zh) * 2023-09-28 2023-12-29 吉林大学 具有干细胞募集和免疫调控功能的引导组织再生纳米纤维膜的制备方法
CN118029056B (zh) * 2024-01-26 2026-04-24 中山大学附属口腔医院 一种多功能生物纤维膜及其制备方法和应用
CN118986551B (zh) * 2024-07-31 2026-01-09 中国科学院理化技术研究所 用于引导口腔软硬组织联合修复的梯度仿生支架
CN118814364B (zh) * 2024-09-19 2025-01-14 中国科学院大连化学物理研究所 一种耐高温阻燃纳米纤维膜及其制备方法和应用
CN119427741B (zh) * 2024-11-05 2025-09-30 吉林大学 一种多孔聚醚醚酮作为引导骨再生屏障膜的制备方法

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100853816B1 (ko) * 2007-02-23 2008-08-22 재단법인서울대학교산학협력재단 멤브레인형 생체활성 세라믹 부직포의 제조방법
WO2009054609A1 (en) * 2007-10-26 2009-04-30 Megagen Implant Co., Ltd. Bone regeneration membrane and method for manufacturing bone regeneration membrane
CN101584885A (zh) * 2009-06-25 2009-11-25 同济大学 具有梯度的三层引导组织再生膜的制备方法
CN104474589A (zh) * 2014-12-23 2015-04-01 山东国际生物科技园发展有限公司 一种引导组织再生膜及其制备方法与应用
CN106110407A (zh) * 2016-08-12 2016-11-16 上海交通大学医学院附属第九人民医院 一种引导性组织再生复合膜材料及其制备方法
CN107320787A (zh) * 2017-07-20 2017-11-07 南开大学 一种牙周修复用多孔纤维膜材料及其制备方法
WO2017203331A1 (en) * 2016-05-27 2017-11-30 Universität Bern Composition and methods for bone tissue engineering
CN107823704A (zh) * 2017-12-28 2018-03-23 广东泰宝医疗器械技术研究院有限公司 一种牙周组织再生修复膜及其制备方法
EP3311854A1 (en) * 2016-10-20 2018-04-25 Ústav Struktury A Mechaniky Hornin AV CR, V.V.I. A nanocomposite layer on the basis of collagen nanofibers, and a method of preparation thereof

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090286205A1 (en) * 2006-02-21 2009-11-19 Johnson Jason K Prefabricated Dental Crowns
CN100421736C (zh) * 2006-11-01 2008-10-01 华中科技大学 一种基于仿生结构的叠层梯度复合支架材料及其制备方法
CN100556468C (zh) * 2007-05-25 2009-11-04 浙江大学 一种膜状组织工程支架及其应用
CN101584886A (zh) * 2009-06-26 2009-11-25 安徽理工大学 层状仿生关节软骨修复与置换材料及其制备方法
CN101791431B (zh) * 2010-02-04 2012-11-07 同济大学 一种生物可降解促牙周组织再生膜片的制备方法
CN102166378B (zh) * 2011-01-13 2013-08-21 北京化工大学 引导组织再生膜及其制备方法
KR101328645B1 (ko) * 2011-02-28 2013-11-14 주식회사 원바이오젠 생분해성 고분자를 이용한 나노/마이크로 하이브리드 섬유 부직포 및 그 제조방법
CN103006359A (zh) * 2012-12-24 2013-04-03 汪泱 仿生三维立体组织工程支架及其制备方法
CN104414772A (zh) * 2013-09-06 2015-03-18 山东百多安医疗器械有限公司 体内可降解吸收的人工医用组织修复膜
CN104874027B (zh) * 2015-05-12 2018-04-27 上海大学 多药物可控负载梯度再生骨支架的制备方法
CN106880874A (zh) * 2017-01-16 2017-06-23 东华大学 一种人工复合疝气补片及其制备方法
CN107213529B (zh) * 2017-05-09 2021-04-20 苏州大学附属第二医院 一种用于提高成骨细胞粘附和成骨性能的可降解医用高分子三维材料的制备方法
AU2018266193C1 (en) * 2017-05-10 2025-05-29 Marquette University Medical and dental integrated multiphasic biomaterials for single or multi-tissue reconstruction/regeneration
CN108914368A (zh) * 2018-07-24 2018-11-30 高昕文 一种面膜用纤维无纺布的制备方法
CN109621001A (zh) * 2018-12-30 2019-04-16 山东百多安医疗器械有限公司 一种聚醚醚酮牙槽骨修复材料及个性化制作方法

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100853816B1 (ko) * 2007-02-23 2008-08-22 재단법인서울대학교산학협력재단 멤브레인형 생체활성 세라믹 부직포의 제조방법
WO2009054609A1 (en) * 2007-10-26 2009-04-30 Megagen Implant Co., Ltd. Bone regeneration membrane and method for manufacturing bone regeneration membrane
CN101584885A (zh) * 2009-06-25 2009-11-25 同济大学 具有梯度的三层引导组织再生膜的制备方法
CN104474589A (zh) * 2014-12-23 2015-04-01 山东国际生物科技园发展有限公司 一种引导组织再生膜及其制备方法与应用
WO2017203331A1 (en) * 2016-05-27 2017-11-30 Universität Bern Composition and methods for bone tissue engineering
CN106110407A (zh) * 2016-08-12 2016-11-16 上海交通大学医学院附属第九人民医院 一种引导性组织再生复合膜材料及其制备方法
EP3311854A1 (en) * 2016-10-20 2018-04-25 Ústav Struktury A Mechaniky Hornin AV CR, V.V.I. A nanocomposite layer on the basis of collagen nanofibers, and a method of preparation thereof
CN107320787A (zh) * 2017-07-20 2017-11-07 南开大学 一种牙周修复用多孔纤维膜材料及其制备方法
CN107823704A (zh) * 2017-12-28 2018-03-23 广东泰宝医疗器械技术研究院有限公司 一种牙周组织再生修复膜及其制备方法

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112604029A (zh) * 2020-12-30 2021-04-06 上海纳米技术及应用国家工程研究中心有限公司 用于牙槽骨修复的“三明治式”可降解支架的制备及产品和应用
CN115252906A (zh) * 2022-07-29 2022-11-01 佛山仙湖实验室 一种三明治结构支架及其制备方法和应用
CN116373357A (zh) * 2023-03-16 2023-07-04 北京市春立正达医疗器械股份有限公司 一种表面梯度化多孔结构的制备方法

Also Published As

Publication number Publication date
US20210402065A1 (en) 2021-12-30
US11696974B2 (en) 2023-07-11
CN110141687B (zh) 2021-10-08
CN110141687A (zh) 2019-08-20

Similar Documents

Publication Publication Date Title
WO2020237785A1 (zh) 一种引导牙周硬软组织再生梯度材料及其制备方法
Zhang et al. Electrospun PDLLA/PLGA composite membranes for potential application in guided tissue regeneration
Xu et al. 3D polycaprolactone/gelatin-oriented electrospun scaffolds promote periodontal regeneration
Xia et al. Emerging polymeric biomaterials and manufacturing techniques in regenerative medicine
Liu et al. Design and development of three-dimensional scaffolds for tissue engineering
Yao et al. In vitro angiogenesis of 3D tissue engineered adipose tissue
Chen et al. A three-dimensional dual-layer nano/microfibrous structure of electrospun chitosan/poly (d, l-lactide) membrane for the improvement of cytocompatibility
Bao et al. Effects of electrospun submicron fibers in calcium phosphate cement scaffold on mechanical properties and osteogenic differentiation of umbilical cord stem cells
CN113230460A (zh) 一种引导口腔软硬组织一体化修复多孔支架及其制备方法
Safi et al. Preparing polycaprolactone scaffolds using electrospinning technique for construction of artificial periodontal ligament tissue
CN106048744A (zh) 一种利用静电纺丝制备仿细胞外基质纳米纤维敷料的方法
Fu et al. Promoting bone regeneration via bioactive calcium silicate nanowires reinforced poly (ε-caprolactone) electrospun fibrous membranes
Wu et al. 3D-bioprinted alginate-based bioink scaffolds with β-tricalcium phosphate for bone regeneration applications
Ortega et al. Nanofibers and microfibers for osteochondral tissue engineering
Luo et al. Fabrication of a novel hierarchical fibrous scaffold for breast cancer cell culture
He et al. Osteogenic induction of bone marrow mesenchymal cells on electrospun polycaprolactone/chitosan nanofibrous membrane
Hu et al. Novel multi-functional microsphere scaffold with shape memory function for bone regeneration
CN113274553A (zh) 一种生物材料诱导的外泌体三维支架及其制备方法与应用
CN115364283A (zh) 一种仿生界面多层复合神经支架及其制备方法与应用
Mitropoulou et al. Development and evaluation of biodegradable core-shell microfibrous and nanofibrous scaffolds for tissue engineering applications
Tao et al. A novel fluffy PLGA/HA composite scaffold for bone defect repair
CN101249277A (zh) 三维多孔组织工程支架材料、其纤维粘结法制备及应用
Yuan et al. Electrospinning/3D printing-integrated porous scaffold guides oral tissue regeneration in beagles
CN1460526A (zh) 含有羟基磷灰石成分的多孔型骨修复体及其制备方法
CN106729972A (zh) 骨充填材料的组合物、预备品以及它们的制备方法和应用

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: 19931296

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19931296

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 19931296

Country of ref document: EP

Kind code of ref document: A1