CN108744060B - Bone repair material capable of being injected with multiple pore structures and preparation method thereof - Google Patents

Bone repair material capable of being injected with multiple pore structures and preparation method thereof Download PDF

Info

Publication number
CN108744060B
CN108744060B CN201810526290.1A CN201810526290A CN108744060B CN 108744060 B CN108744060 B CN 108744060B CN 201810526290 A CN201810526290 A CN 201810526290A CN 108744060 B CN108744060 B CN 108744060B
Authority
CN
China
Prior art keywords
dopamine
solution
acid copolymer
hyaluronic acid
repair material
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.)
Active
Application number
CN201810526290.1A
Other languages
Chinese (zh)
Other versions
CN108744060A (en
Inventor
孙勇
樊渝江
卢恭恭
张兴栋
王翔
陈铁军
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 CN201810526290.1A priority Critical patent/CN108744060B/en
Publication of CN108744060A publication Critical patent/CN108744060A/en
Application granted granted Critical
Publication of CN108744060B publication Critical patent/CN108744060B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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
    • A61L27/50—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
    • A61L27/02—Inorganic materials
    • A61L27/12—Phosphorus-containing materials, e.g. apatite
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/52—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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/602—Type of release, e.g. controlled, sustained, slow
    • 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
    • A61L2400/06—Flowable or injectable implant compositions
    • 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
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

Landscapes

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

Abstract

The invention provides a bone repair material capable of being injected with multiple pore structures and a preparation method thereof, the bone repair material comprises powder, a curing liquid A and a curing liquid B, the powder is hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, the curing liquid A is a collagen solution, the curing liquid B is a dopamine-hyaluronic acid aqueous solution, the powder is dispersed in the curing liquid A and then uniformly mixed with the curing liquid B, the pH value is adjusted to 7.5-8.5, phenolic hydroxyl groups of dopamine-hyaluronic acid can be oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, and the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres and the collagen are distributed in a three-dimensional network structure of the dopamine-hyaluronic acid copolymer hydrogel. The bone repair material has a multi-hole structure suitable for cell growth, can increase the tightness of combination between the bone repair material and autologous bones, and is expected to be applied to the fields of tissue repair such as skin filling and the like and cosmetology by adjusting the composition ratio.

Description

Bone repair material capable of being injected with multiple pore structures and preparation method thereof
Technical Field
The invention belongs to the field of bone repair materials, and relates to a bone repair material capable of being injected with a multi-pore structure and a preparation method thereof.
Background
The injectable bone repair material is a biomedical material for local bone defect and repair, can replace the traditional complex and time-consuming bone defect treatment modes such as autologous bone transplantation, allogeneic bone transplantation and the like, and has the advantages of small tissue damage, simplicity and convenience in operation, few surgical complications and the like. The ideal injectable bone repair material generally has the following conditions: good biocompatibility, good osteoconductivity and osteoinductivity, certain pore diameter and porosity, degradability and absorption in tissues and the like.
At present, the bone defect repair materials clinically used mainly comprise polymethyl methacrylate (PMMA) and alloy materials, and although the PMMA and the alloy materials have good mechanical properties, the PMMA is not degraded and has no biological activity, and the defect of toxic monomer residue exists; the metal has too high elastic modulus and is easy to wear and corrode in vivo, which provides requirements for developing a new generation of bone repair materials. In recent years, injectable bone repair materials have become a great hotspot for bone defect repair research. Injectable bone repair materials have been studied mainly in ceramics, polymers and composites of the two. The single ceramic material is not easy to form in vivo or loose after forming, and the single hydrogel material is easy to generate larger cavities due to degradation and can not meet the requirement of repairing larger bone defects. Accordingly, injectable ceramic/polymer composite scaffold materials have received great attention in recent years. Turner et al examined the effects of PMMA and CaP on spine repair using canines as models; weijie et al prepared a novel injectable nano apatite/polyurethane composite material; moreau et al, compound chitosan and Cap to prepare injectable bone repair materials. Although these materials have certain biocompatibility and can promote bone defect repair and reconstruction, the materials still have some defects, such as poor injection performance, no pore size and porosity suitable for cell growth, rapid degradation, unfavorable cell growth of acidic degradation products and the like. Moreover, the material is susceptible to temperature, strength and physiological environment factors, so that the cementation is not firm, and the bonding between the material and autologous bone is not tight, and the limit is well defined. This drawback greatly limits the use of ceramic/polymer composite stent materials in the medical field.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a bone repair material capable of being injected with a multi-pore structure and a preparation method thereof, so as to solve the problems that the existing bone repair material has no pore structure suitable for cell growth and is not tightly combined with autologous bone.
The bone repair material capable of being injected with the multiple pore structures comprises powder, curing liquid A and curing liquid B, wherein the powder is hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, the curing liquid A is a collagen solution, the curing liquid B is a dopamine-hyaluronic acid aqueous solution, the structural formula of the dopamine-hyaluronic acid is shown as a formula (I), the grafting rate of dopamine in the dopamine-hyaluronic acid is 1-50%,
Figure BDA0001676028850000021
dispersing the powder in the curing liquid A, then uniformly mixing the powder with the curing liquid B, adjusting the pH value to 7.5-8.5, oxidizing and crosslinking phenolic hydroxyl groups of dopamine-hyaluronic acid to form dopamine-hyaluronic acid copolymer hydrogel, and distributing hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres and collagen in a three-dimensional network structure of the dopamine-hyaluronic acid copolymer hydrogel.
In the technical scheme of the bone repair material capable of being injected with multiple pore structures, the grafting rate of dopamine in dopamine-hyaluronic acid is preferably 3% -8%.
In the above technical scheme of the bone repair material capable of injecting multiple pore structures, the content of hydroxyapatite in the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres is 1 wt.% to 90 wt.%, and preferably, the content of hydroxyapatite in the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres is 5 wt.% to 20 wt.%.
In the technical scheme of the bone repair material capable of being injected with the multiple pore structures, after the dopamine-hyaluronic acid hydrogel is formed by oxidative crosslinking of phenolic hydroxyl groups of dopamine-hyaluronic acid, the content of hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres in the bone repair material is 1-95 wt%, the content of collagen is 1-95 wt%, and the content of the dopamine-hyaluronic acid copolymer hydrogel is 1-95 wt%; preferably, the content of the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres in the bone repair material is 10-16 wt.%, the content of the collagen is 25-40 wt.%, and the content of the dopamine-hyaluronic acid copolymer hydrogel is 44-65 wt.%.
In the technical scheme of the bone repair material capable of being injected with the multiple pore structures, the concentration of collagen in the curing liquid A is preferably 10-30 mg/mL, and the concentration of dopamine-hyaluronic acid in the curing liquid B is preferably 20-30 mg/mL.
In the technical scheme of the bone repair material capable of being injected with the multiple pore structures, the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres are physical blends of hydroxyapatite and polylactic acid-glycolic acid copolymer porous microspheres, the hydroxyapatite is uniformly distributed in the polylactic acid-glycolic acid copolymer, and has mutually communicated pore channels, the pore diameter is 5-10 mu m, the particle diameter of the porous microspheres is 100-400 mu m, and the porosity is 60-90%. In the polylactic acid-glycolic acid copolymer component of the hydroxyapatite composite polylactic acid-glycolic acid porous microsphere, the mass ratio of polylactic acid to glycolic acid is (75-85) to (25-15).
In the technical scheme of the bone repair material capable of being injected with multiple pore structures, the dopamine-hyaluronic acid is obtained by grafting dopamine on the basis of sodium hyaluronate for modification, and the molecular weight of the sodium hyaluronate used as the modification basis is 0.1 w-400 w, preferably 8.9 w-200 w.
In the technical scheme of the bone repair material capable of being injected with the multiple pore structures, after the dopamine-hyaluronic acid copolymer hydrogel is formed by oxidative crosslinking of phenolic hydroxyl groups of dopamine-hyaluronic acid, the multiple pore structures consisting of a three-dimensional network structure in the hydrogel and a structure formed by communicating porous channels of porous microspheres are formed in the bone repair material.
The invention also provides a preparation method of the bone repair material capable of being injected with the multiple pore structures, which comprises the following steps:
(1) weighing 2-6 parts by weight of hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, 10-30 parts by weight of dopamine-hyaluronic acid and 5-15 parts by weight of collagen;
(2) dissolving collagen in 0.2-1 mol/L acetic acid to form a curing solution A, dispersing hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres in the curing solution A to form a dispersion solution, dissolving dopamine-hyaluronic acid in water to form a curing solution B, uniformly mixing the dispersion solution and the curing solution B, and adjusting the pH value to 7.5-8.5 before use to obtain the collagen-hyaluronic acid composite polylactic acid.
In the preparation method of the bone repair material capable of being injected with the multiple pore structures, the concentration of collagen in the curing liquid A is preferably 10-30 mg/mL, and the concentration of dopamine-hyaluronic acid in the curing liquid B is preferably 20-30 mg/mL.
In the preparation method of the bone repair material capable of being injected with multiple pore structures, the preparation method of the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microsphere comprises the following steps:
(1) dissolving a polylactic acid-glycolic acid copolymer in dichloromethane to form a polylactic acid-glycolic acid copolymer solution with the concentration of 0.03-0.06 g/mL, adding hydroxyapatite powder and an ammonium bicarbonate solution, performing ultrasonic emulsification, then adding an ammonium bicarbonate solution, and performing ultrasonic emulsification to obtain a W/O emulsion;
the mass ratio of the hydroxyapatite powder to the polylactic acid-glycolic acid copolymer is 3 (1-8), and the volume ratio of the ammonium bicarbonate solution to the polylactic acid-glycolic acid copolymer solution added each time is 0.5-1: 6; the concentration of the ammonium bicarbonate solution is 0.025-0.05 g/mL;
(2) injecting and transferring the W/O emulsion into a polyvinyl alcohol solution with the concentration of 10-25 g/L to form a W/O/W multiple emulsion, then adding water, stirring until dichloromethane is completely evaporated, washing the obtained product with water, and drying to obtain the water-soluble polyvinyl chloride/polyvinyl chloride composite emulsion; the volume ratio of the W/O emulsion to the polyvinyl alcohol solution to the water is (0.15-0.2): 1: 1.
When the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres are prepared, the ultrasonic emulsification time is preferably 5-20 min. And (3) washing the product in the step (2) with water, and filtering the product through a micron-sized filter sieve to obtain the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres with different particle sizes.
The preparation method of the injectable bone repair material with a multiple pore structure comprises the following steps:
(1) adding N-hydroxysuccinimide into a sodium hyaluronate aqueous solution, then adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, stirring and reacting for 1-3 hours, dropwise adding a dopamine hydrochloride aqueous solution, stirring and reacting for 10-14 hours, controlling the pH value to be within the range of 4.8-5.5 in the two stirring and reacting processes, and performing the operation of the step under the protection of nitrogen;
the molar ratio of the 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride to the N-hydroxysuccinimide to the dopamine hydrochloride to the carboxyl groups on the sodium hyaluronate is (1-6): 1-4): 1-3): 1, the concentration of the dopamine hydrochloride aqueous solution is 0.5-3 mmol/L, and the concentration of the sodium hyaluronate aqueous solution is 5-15 mg/mL;
(2) and (2) dialyzing the reaction solution obtained in the step (1) in water with the pH value of 3.0-3.5, and freeze-drying to obtain the compound.
Compared with the prior art, the invention has the following beneficial technical effects:
1. the invention provides a bone repair material capable of being injected with a multi-pore structure, which comprises powder, curing liquid A and curing liquid B, wherein the powder is hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, the curing liquid A is a collagen solution, the curing liquid B is a dopamine-hyaluronic acid aqueous solution, the powder is dispersed in the curing liquid A and then uniformly mixed with the curing liquid B, the pH value is adjusted to 7.5-8.5, then the dopamine-hyaluronic acid is crosslinked to form hydrogel, and the porous microspheres and the collagen are uniformly distributed in a three-dimensional network structure of the dopamine-hyaluronic acid copolymer hydrogel. The bone repair material is characterized in that a multi-pore structure formed by a three-dimensional network structure in hydrogel and a structure formed by communicating porous channels of porous microspheres is formed, the three-dimensional network structure is highly communicated, the porous microspheres are provided with the mutually communicated pore channels of 5-10 mu m, the porosity is up to more than 60%, and the existence of the multi-pore structure is beneficial to the slow controlled release of active ingredients such as protein preparations, micromolecular medicaments, active factors and the like, is beneficial to promoting the transmission of nutrient substances and the adhesion growth of cells, and provides an ideal three-dimensional space for cell proliferation. Overcomes the problems of the prior bone repair material caused by no pore structure suitable for cell growth.
2. In the bone repair material capable of being injected with the multiple pore structures, provided by the invention, the dopamine grafted and modified sodium hyaluronate is used as a basis for forming three-dimensional network hydrogel through crosslinking, phenolic hydroxyl on dopamine is converted into semiquinone free radicals and o-benzoquinone through redox reaction under an alkaline condition, and can generate covalent or non-covalent interaction with a substrate material, and the surface of bone tissue contains a large amount of-NH (NH)2Under alkaline conditions, phenolic hydroxyl groups in the polydopamine and the surface of bone tissues undergo Michael addition or Schiff-base reaction to generate covalent bonding with the surface of the bone tissues, and meanwhile, the polydopamine can also form an adhesion layer through non-covalent coupling effects such as metal coordination, hydrogen bonds, pi-pi stacking, hydroquinone charge transfer complexes and the like with the surface of a substrate, so that the introduction of the dopamine endows the bone repair material with certain bonding strength. Due to the fact that the grafting rate of dopamine in the dopamine-hyaluronic acid is proper, the dopamine-hyaluronic acid is beneficial to high mechanical strength and adhesion after phenolic hydroxyl of the dopamine-hyaluronic acid is oxidized and crosslinked, the improvement of the bonding tightness between the bone repair material and the autologous bone is facilitated, and the problems that the existing bone repair material is not firmly bonded with the autologous bone due to strength, physiological environment and environmental factors, and the bonding tightness and the well-defined limit are solved.
3. Due to the fact that the proportional relation among the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres and the collagen-dopamine-hyaluronic acid copolymer hydrogel is proper, the water content of the collagen and dopamine-hyaluronic acid copolymer hydrogel and the grafting rate of dopamine in dopamine-hyaluronic acid are proper, the bone repair material capable of being injected with the multiple pore structures has good injection performance, and meanwhile is endowed with good mechanical properties, and the bone repair material is beneficial to playing a better role in practical application.
4. The hyaluronic acid, the collagen and the polylactic acid-glycolic acid copolymer in the porous microspheres have good biocompatibility and biodegradability, so that the bone repair material capable of being injected with multiple pore structures has the characteristics of safety, degradability, good biocompatibility and the like. Meanwhile, the bone repair material provided by the invention also has ideal mechanical properties, cohesiveness, rheological property, injectability, proper curing time and material microstructure.
5. By adjusting the proportion of each component in the bone repair material provided by the invention, the formed material is expected to be applied to the fields of tissue repair and beauty treatment such as skin filling.
6. The invention also provides a method for preparing the injectable bone repair material with a multi-pore structure, which has the characteristics of simple operation, mild process conditions and low cost and is beneficial to realizing industrial production.
Drawings
Fig. 1 is an SEM photograph of PLGA porous microspheres prepared in example 1.
FIG. 2 is a partially enlarged SEM photograph of PLGA porous microspheres prepared in example 1.
FIG. 3 is PLGA prepared in example 13-HA2.5-250SEM photograph of (a).
FIG. 4 is PLGA prepared in example 13-HA1-250Partial SEM photograph of (a).
FIG. 5 is PLGA prepared in example 13-HA2.5-250Partial SEM photograph of (a).
FIG. 6 is PLGA prepared in example 13-HA2.5-250SEM photograph of (1).
FIG. 7 is an XRD pattern of HA-PLGA porous microspheres, and HA powder.
FIG. 8 is a thermogravimetric curve of a part of samples in example 1, wherein the samples numbered 1-5 are PLGA-250、PLGA3-HA1-250、PLGA3-HA2.5-250、PLGA3-HA5-250、PLGA3-HA8-250。
Fig. 9 is an SEM photograph of a cross section of the bone repair material prepared in example 5.
Fig. 10 is a partially enlarged SEM photograph of a cross section of the bone repair material prepared in example 5.
Fig. 11 is an SEM photograph of a cross section of the composite material prepared in comparative example 1.
Fig. 12 is a partially enlarged SEM photograph of a cross section of the composite material prepared in comparative example 1.
Fig. 13 is the injection thrust test result of the injectable bone repair material of example 6.
Fig. 14 is the results of mechanical property testing of the injectable bone repair material of example 7.
Detailed Description
The injectable bone repair material with multiple pore structure and the preparation method thereof provided by the invention are further illustrated by the following examples. It should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention, and those skilled in the art can make certain insubstantial modifications and adaptations of the present invention based on the above disclosure and still fall within the scope of the present invention.
Example 1
In this example, blank polylactic-co-glycolic acid (PLGA) porous microspheres and HA composite PLGA porous microspheres (HA-PLGA porous microspheres) having different Hydroxyapatite (HA) contents were prepared.
1. Preparation of PLGA porous microspheres
Weighing 1.8g of PLGA, adding the PLGA into 36mL of dichloromethane, stirring the mixture evenly at room temperature, wherein the molecular weight of the PLAG is 8.5W, the mass ratio of LA to GA is 85/15, then adding 6mL of 0.025g/mL ammonium bicarbonate solution, carrying out ultrasonic emulsification treatment for 5min, adding 6mL of 0.025g/mL ammonium bicarbonate solution, carrying out ultrasonic emulsification treatment for 15min to obtain evenly dispersed W/O emulsion, injecting and transferring the W/O emulsion into 300mL of 10g/L polyvinyl alcohol (PVA) solution to obtain W/O/W double emulsion, adding 300mL of deionized water, stirring the mixture overnight to ensure that the dichloromethane is completely evaporated, washing the mixture for multiple times by using the deionized water, filtering the mixture through a filter screen with the pore diameter of 250 mu m, centrifugally separating PLGA porous microspheres, freezing and drying the product which is marked as PLGA-250。
2. Preparation of HA-PLGA porous microspheres
Preparation of HA-PLAG porous microspheres with a particle size of about 250 μm: 1.8g PLGA was weighed out and added to 36mL dichloromethane, stirred at room temperature and homogenized, the molecular weight of PLAG was 8.5w, wherein the weight ratio of LA to GA was LA/GA 85/15, and thenAdding 600mg HA powder, adding 6mL of 0.025g/mL ammonium bicarbonate solution, carrying out ultrasonic emulsification treatment for 5min, adding 6mL of 0.025g/mL ammonium bicarbonate solution, carrying out ultrasonic emulsification treatment for 15min to obtain uniformly dispersed W/O emulsion, injecting and transferring the W/O emulsion into 300mL of 10g/L PVA solution to obtain W/O/W multiple emulsion, adding 300mL of deionized water, stirring overnight to ensure that dichloromethane is completely evaporated, washing with deionized water for multiple times, filtering through a filter screen with the pore diameter of 250 mu m, centrifuging to separate HA-PLGA porous microspheres, freeze-drying, and marking the product as PLGA3-HA1-250. According to preparation of PLGA3-HA1-250In the same manner, the amount of HA powder added was adjusted to 1.5g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA2.5-250。
According to preparation of PLGA3-HA1-250In the same way, the amount of HA powder added was adjusted to 3g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA5-250。
According to preparation of PLGA3-HA1-250In the same manner, the amount of HA powder added was adjusted to 4.8g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA8-250。
Preparing HA-PLAG porous microspheres with the particle size of about 400 mu m, weighing 1.8g PLGA, adding into 36mL dichloromethane, stirring uniformly at room temperature, the molecular weight of PLAG is 8.5W, wherein the mass ratio of LA to GA is LA/GA 75/25, adding 600mg HA powder, adding 3mL ammonium bicarbonate solution of 0.05g/mL, carrying out ultrasonic emulsification treatment for 10min, adding 3mL ammonium bicarbonate solution of 0.05g/mL, carrying out ultrasonic emulsification treatment for 20min to obtain uniformly dispersed W/O emulsion, injecting and transferring the W/O emulsion into 210mL PVA solution of 25g/L to obtain W/O/W multiple emulsion, adding 210mL deionized water, stirring overnight to ensure that dichloromethane is completely evaporated, washing with deionized water for multiple times, filtering through a filter screen with the pore size of 400 mu m, centrifuging to separate the HA-PLGA porous microspheres, freeze drying and marking the product as PLGA3-HA1-400。
According to preparation of PLGA3-HA1-400In the same manner, the amount of HA powder added was adjusted to 1.5g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA2.5-400。
According to preparation of PLGA3-HA1-400In the same way, the amount of HA powder added was adjusted to 3g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA5-400。
According to preparation of PLGA3-HA1-400In the same manner, the amount of HA powder added was adjusted to 4.8g to obtain HA-PLGA porous microspheres, denoted PLGA3-HA8-400。
Scanning electron microscope is adopted to observe the PLGA porous microspheres and the HA-PLGA porous microspheres prepared in the embodiment, FIG. 1 is an SEM (scanning electron microscope) picture of the PLGA porous microspheres, FIG. 2 is a local enlarged SEM picture of the PLGA porous microspheres, and FIG. 3 is a PLGA porous microsphere3-HA2.5-250SEM photograph of (1), FIG. 4 is PLGA3-HA1-250FIG. 5 is a partial SEM photograph of PLGA3-HA2.5-250FIG. 6 is a partial SEM photograph of PLGA3-HA2.5-250SEM photograph of (1). As can be seen from the scanning electron microscope image, the HA-PLGA porous microspheres have mutually communicated pore channels, the pore diameter is about 5-10 μm, and as can be seen from FIG. 6, the distribution uniformity of HA in PLGA is good.
Weighing HA-PLGA porous microspheres, PLGA porous microspheres and HA powder, pressing into tablets, and carrying out qualitative analysis on sample components by using an X-ray diffractometer, wherein the scanning range 2 theta is 10-80 degrees, the scanning step width is 0.05 degrees, the scanning step length is 0.2s, the working voltage is 40kV, the current is 30mA, and the XRD pattern of part of samples is shown in figure 7.
Weighing PLGA-HA porous microspheres and 3-6 mg of PLGA porous microspheres, performing thermogravimetric analysis, wherein the test temperature range is 25-1000 ℃, the heating rate is 20 ℃/min, observing and recording the mass change of the sample in the heating process, and the thermogravimetric curve of part of the sample is shown in figure 8.
The particle size, porosity and the like of the PLGA porous microspheres and the HA-PLGA porous microspheres which are finally prepared can be adjusted by adjusting the technological parameters for preparing the PLGA porous microspheres and the HA-PLGA porous microspheres, and the HA content in the HA-PLGA porous microspheres can also be adjusted. This example prepares PLGA porous microspheres having a particle size of about 250 μm, HA-PLAG porous microspheres having particle sizes of about 250 μm and 400 μm, and their particle sizes, porosities and hydroxyapatite contents are shown in Table 1.
TABLE 1
Figure BDA0001676028850000081
Example 2
In this example, dopamine-hyaluronic acid (HA-DOPA) was prepared by the following steps:
(1) dropwise adding an N-hydroxysuccinimide (NHS) solution with the concentration of 46mg/mL into a sodium hyaluronate (Mw ═ 340kDa) aqueous solution with the concentration of 11.5mg/mL, then dropwise adding a 1-ethyl- (3-dimethylaminopropyl) carbonyl diimine hydrochloride (EDCI) solution with the concentration of 150mg/mL, stirring and reacting for 2 hours, dropwise adding a dopamine hydrochloride aqueous solution with the concentration of 2mmol/L, stirring and reacting for 12 hours, controlling the pH value to be 5.0 in the two stirring and reacting processes, wherein the operation of the step is carried out under the protection of nitrogen, and the molar ratio of EDCI, NHS, dopamine hydrochloride to carboxyl on the sodium hyaluronate is 6:4:3: 1;
(2) and (2) dialyzing the reaction solution obtained in the step (1) in ultrapure water with the pH value of 3.5 for 48h by using a dialysis membrane (MW & lt 3.5-8kDa), carrying out vacuum freeze drying to obtain HA-DOPA with the dopamine grafting rate of 8%, and storing a sample in a dryer.
The grafting rate of dopamine in dopamine-hyaluronic acid can be changed by adjusting the molar ratio of EDCI, NHS, dopamine hydrochloride and carboxyl on sodium hyaluronate and the molecular weight of sodium hyaluronate, and the grafting rate of dopamine in dopamine-hyaluronic acid can be adjusted to be within the range of 3% -8% within the range of proportional relationship defined by the invention.
Example 3
In this example, HA-DOPA was prepared by the following steps:
(1) dropwise adding a 50mg/mL NHS solution into a15 mg/mL aqueous solution of sodium hyaluronate (Mw is 500kDa), then dropwise adding a150 mg/mL EDCI solution, stirring and reacting for 3h, dropwise adding a 0.5mmol/L dopamine hydrochloride aqueous solution, stirring and reacting for 14h, controlling the pH value to be 4.8 in the two stirring and reacting processes, wherein the operation of the step is carried out under the protection of nitrogen, and the molar ratio of EDCI, NHS, dopamine hydrochloride to carboxyl on the sodium hyaluronate is 1:2:2.5: 1;
(2) and (2) dialyzing the reaction solution obtained in the step (1) in ultrapure water with the pH value of 3.0 for 48h by using a dialysis membrane (MW & lt 3.5-8kDa), carrying out vacuum freeze drying to obtain HA-DOPA, and storing the sample in a dryer.
Example 4
In this example, HA-DOPA was prepared by the following steps:
(1) dropwise adding a 50mg/mL NHS solution into a 5mg/mL sodium hyaluronate (Mw is 90kDa), then dropwise adding a150 mg/mL EDCI solution, stirring and reacting for 1h, dropwise adding a 3mmol/L dopamine hydrochloride aqueous solution, stirring and reacting for 10h, controlling the pH value to be 5.5 in the two stirring and reacting processes, wherein the operation of the step is carried out under the protection of nitrogen, and the molar ratio of EDCI, NHS, dopamine hydrochloride to carboxyl on the sodium hyaluronate is 3:1:1: 1;
(2) and (2) dialyzing the reaction solution obtained in the step (1) in ultrapure water with the pH value of 3.5 for 48h by using a dialysis membrane (MW & lt 3.5-8kDa), carrying out vacuum freeze drying to obtain HA-DOPA, and storing the sample in a dryer.
Example 5
In this embodiment, the bone repair material with injectable multi-pore structure is prepared and tested by scanning electron microscope, and the steps are as follows:
(1) weighing PLGA3-HA1-25030mg of HA-DOPA150mg with dopamine grafting rate of 8 percent and 120mg of collagen (Col);
(2) adding Col into 0.5mol/L acetic acid, shaking on vortex oscillator to completely dissolve to form 24mg/mL Col concentration solidified solution A, and mixing PLGA3-HA1-250Adding the HA-DOPA into the curing liquid A, oscillating and mixing the HA-DOPA on a vortex oscillator uniformly to form a dispersion liquid, adding the HA-DOPA into ultrapure water, oscillating the ultrapure water on the vortex oscillator until the HA-DOPA is dissolved completely to form a curing liquid B with the HA-DOPA concentration of 30 mg/mL; uniformly mixing the dispersion liquid and the curing liquid B, taking 0.5mL of the uniformly mixed material, dropwise adding 0.2mol/L NaOH solution to adjust the pH value to 8.0, standing for 5min, and performing oxidative crosslinking on phenolic hydroxyl of HA-DOPA to form dopamine-hyaluronic acid copolymer hydrogel to obtain the bone collagenThe repair material, designated PLGA-HA-Col-HA-DOPA, was vacuum freeze dried to obtain a spongy solid. The cross section of the sample is observed by a scanning electron microscope, the SEM picture of the cross section and the local magnification SEM picture of the cross section are respectively shown in figures 9 and 10, and as can be seen from figures 9 and 10, the PLGA-HA-Col-HA-DOPA HAs a multi-pore structure after freeze drying, including a large network pore structure in the bone repair material, and PLGA distributed in the bone repair material3-HA1-250Also has a porous structure.
Comparative example 1
In order to compare with the bone repair material prepared in example 5, in this comparative example, without using PLGA microspheres, HA and collagen were directly dispersed in the hydrogel formed by HA-DOPA crosslinking to prepare a composite material and subjected to scanning electron microscopy testing, the steps were as follows:
(1) weighing HA powder 30mg, HA-DOPA150mg with dopamine grafting rate of 8% and collagen (Col)120 mg;
(2) adding Col into 0.5mol/L acetic acid, oscillating on a vortex oscillator until the Col is completely dissolved to form a solidified liquid A with the concentration of 24mg/mL, adding HA powder into the solidified liquid A, oscillating and uniformly mixing on the vortex oscillator to form a dispersion liquid, adding HA-DOPA into ultrapure water, oscillating on the vortex oscillator until the HA-DOPA is completely dissolved to form a solidified liquid B with the concentration of 30 mg/mL; and uniformly mixing the dispersion liquid and the curing liquid B, taking 0.5mL of the uniformly mixed material, dropwise adding 0.2mol/L NaOH solution to adjust the pH value to 8.0, standing until phenolic hydroxyl of HA-DOPA is oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, namely HA-Col-HA-DOPA, and carrying out vacuum freeze drying to obtain a spongy solid. The sample was frozen with liquid nitrogen and then cut vertically, and the cross section of the sample was observed with a scanning electron microscope, and the cross section and a partially enlarged SEM photograph of the cross section are shown in fig. 11 and 12, respectively.
Comparing fig. 9 and 10 with fig. 11 and 12, it can be seen that HA-Col-HA-DOPA does not have a dual pore structure, and HA is relatively poorly dispersed in the composite material, and agglomeration occurs.
Example 6
In this example, the injectable properties of the injectable bone repair material were examined, and in order to examine the injectable properties of the bone repair material, the following three samples were prepared and tested.
(1) The HA-PLGA porous microspheres prepared in example 1, the HA-DOPA and Col prepared in example 2 were weighed in the proportions shown in the following table:
PLGA3-HA2.5-250(mg) HA-DOPA(mg) Col(mg)
sample 1 2 10 5
Sample 2 4 20 10
Sample 3 6 30 15
(2) Adding Col into 0.5mL of 0.5mol/L acetic acid according to the raw material proportion of each sample, vibrating on a vortex oscillator until the mixture is clarified to form a solidified solution A, and mixingPLGA3-HA2.5-250Adding the HA-DOPA into the curing liquid A, vibrating and mixing the HA-DOPA uniformly on a vortex oscillator to form a dispersion liquid, adding the HA-DOPA into 0.5mL of ultrapure water, and vibrating the HA-DOPA on the vortex oscillator until the HA-DOPA is clear to form a curing liquid B; uniformly mixing the dispersion liquid and the curing liquid B, dropwise adding a NaOH solution with the concentration of 0.2mol/L to adjust the pH value to 7.5 to obtain samples 1-3, immediately sucking the samples 1-3 into a 1mL injector, and testing the injection thrust of the samples on an Shimadzu electronic universal mechanical testing machine, wherein the result is shown in figure 13, the curves from bottom to top in figure 13 are the test results of the samples 1, 2 and 3 in sequence, and as can be seen from figure 13, the injectable bone repair material has good injection performance.
Example 7
In this embodiment, the bone repair material capable of injecting a multi-pore structure is prepared and tested for mechanical properties, and the steps are as follows:
(1) weighing PLGA3-HA5-2502mg, 10mg of HA-DOPA prepared in example 2, Col 5 mg;
(2) adding Col into 1mol/L acetic acid, shaking on vortex oscillator to completely dissolve to form 30mg/mL Col concentration solidified solution A, and mixing PLGA3-HA5-250Adding the HA-DOPA into the curing liquid A, oscillating and mixing the HA-DOPA on a vortex oscillator uniformly to form a dispersion liquid, adding the HA-DOPA into ultrapure water, oscillating the ultrapure water on the vortex oscillator until the HA-DOPA is dissolved completely to form a curing liquid B with the HA-DOPA concentration of 20 mg/mL; and uniformly mixing the dispersion liquid and the curing liquid B, dropwise adding a NaOH solution with the concentration of 0.2mol/L to adjust the pH value to 8.5, and standing until the phenolic hydroxyl of HA-DOPA is oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel so as to obtain the injectable bone repair material. The hydrogel compression performance was tested using a universal mechanical tester DMA Q800, and the results are shown in fig. 14.
Example 8
In this example, the preparation of the injectable bone repair material with a multiple pore structure comprises the following steps:
(1) weighing PLGA3-HA8-25030mg, 150mg of HA-DOPA prepared in example 3, and 100mg of Col;
(2) adding Col into 0.2mol/L acetic acid, and vibrating with vortexOscillating on a shaker until the solution is completely dissolved to form solidified solution A with concentration of Col of 10mg/mL, and mixing PLGA3-HA8-250Adding the HA-DOPA into the curing liquid A, oscillating and mixing the HA-DOPA on a vortex oscillator uniformly to form a dispersion liquid, adding the HA-DOPA into ultrapure water, oscillating the ultrapure water on the vortex oscillator until the HA-DOPA is dissolved completely to form a curing liquid B with the HA-DOPA concentration of 20 mg/mL; and uniformly mixing the dispersion liquid and the curing liquid B, dropwise adding a NaOH solution with the concentration of 0.2mol/L to adjust the pH value to 8.0, and standing until the phenolic hydroxyl of HA-DOPA is oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, thus obtaining the bone repair material.
Example 9
In this example, the preparation of the injectable bone repair material with a multiple pore structure comprises the following steps:
(1) weighing PLGA3-HA1-40020mg, 120mg of HA-DOPA prepared in example 4, 80mg of Col;
(2) adding Col into 0.5mol/L acetic acid, shaking on vortex oscillator to completely dissolve to form solidified solution A with Col concentration of 15mg/mL, and mixing PLGA3-HA1-400Adding the HA-DOPA into the curing liquid A, oscillating and mixing the HA-DOPA on a vortex oscillator uniformly to form a dispersion liquid, adding the HA-DOPA into ultrapure water, oscillating the ultrapure water on the vortex oscillator until the HA-DOPA is dissolved completely to form a curing liquid B with the HA-DOPA concentration of 25 mg/mL; and uniformly mixing the dispersion liquid and the curing liquid B, dropwise adding a NaOH solution with the concentration of 0.2mol/L to adjust the pH value to 8.0, and standing until the phenolic hydroxyl of HA-DOPA is oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, thus obtaining the bone repair material.
Example 10
In this example, the preparation of the injectable bone repair material with a multiple pore structure comprises the following steps:
(1) weighing PLGA3-HA2.5-40050mg of HA-DOPA 260mg prepared in example 4 and Col 150 mg;
(2) adding Col into 0.5mol/L acetic acid, shaking on vortex oscillator to completely dissolve to form 20mg/mL Col concentration solidified solution A, and mixing PLGA3-HA2.5-400Adding into curing liquid A, mixing uniformly by oscillating on vortex oscillator to form dispersion, adding HA-DOPA into ultrapure waterIn the preparation process, the mixture is shaken on a vortex oscillator until the mixture is completely dissolved to form a solidification solution B with the HA-DOPA concentration of 30 mg/mL; and uniformly mixing the dispersion liquid and the curing liquid B, dropwise adding a NaOH solution with the concentration of 0.2mol/L to adjust the pH value to 8.0, and standing until the phenolic hydroxyl of HA-DOPA is oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, thus obtaining the bone repair material.

Claims (5)

1. The bone repair material capable of being injected with the multiple pore structures is characterized by comprising powder, curing liquid A and curing liquid B, wherein the powder is hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, the curing liquid A is a collagen solution, the curing liquid B is a dopamine-hyaluronic acid aqueous solution, the structural formula of the dopamine-hyaluronic acid is shown as a formula (I), the grafting rate of dopamine in the dopamine-hyaluronic acid is 3% -8%,
Figure FDA0002895510900000011
the concentration of collagen in the curing liquid A is 10-30 mg/mL, and the concentration of dopamine-hyaluronic acid in the curing liquid B is 20-30 mg/mL; the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microsphere has mutually communicated pore channels, the aperture is 5-10 mu m, the particle size of the porous microsphere is 100-400 mu m, and the porosity is 60-90%; in the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microsphere, the content of hydroxyapatite is 5-20 wt.%; in the polylactic acid-glycolic acid copolymer component of the hydroxyapatite composite polylactic acid-glycolic acid porous microsphere, the mass ratio of polylactic acid to glycolic acid is (75-85) to (25-15);
dispersing the powder in the curing liquid A, then uniformly mixing the powder with the curing liquid B, and adjusting the pH value to 7.5-8.5, wherein phenolic hydroxyl groups of dopamine-hyaluronic acid can be oxidized and crosslinked to form dopamine-hyaluronic acid copolymer hydrogel, and hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres and collagen are distributed in a three-dimensional network structure of the dopamine-hyaluronic acid copolymer hydrogel;
the preparation method of the bone repair material capable of injecting the multi-pore structure comprises the following steps:
(1) weighing 2-6 parts by weight of hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres, 10-30 parts by weight of dopamine-hyaluronic acid and 5-15 parts by weight of collagen;
(2) dissolving collagen in 0.2-1 mol/L acetic acid to form a curing solution A, dispersing hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres in the curing solution A to form a dispersion solution, dissolving dopamine-hyaluronic acid in water to form a curing solution B, uniformly mixing the dispersion solution and the curing solution B, and adjusting the pH value to 7.5-8.5 before use to obtain the collagen-hyaluronic acid composite polylactic acid.
2. The injectable bone repair material with a multiple pore structure according to claim 1, wherein after the phenolic hydroxyl group of the dopamine-hyaluronic acid is oxidized and crosslinked to form the dopamine-hyaluronic acid copolymer hydrogel, the content of the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microspheres in the bone repair material is 10-16 wt%, the content of the collagen is 25-40 wt%, and the content of the dopamine-hyaluronic acid copolymer hydrogel is 44-65 wt%.
3. The injectable bone repair material of multiple pore structure according to claim 1 or 2, wherein the dopamine-hyaluronic acid is modified by grafting dopamine on the basis of sodium hyaluronate, and the molecular weight of the sodium hyaluronate used as the modification base is 0.1 w-400 w.
4. The injectable bone repair material with multiple pore structures according to claim 1, wherein the preparation method of the hydroxyapatite composite polylactic acid-glycolic acid copolymer porous microsphere is as follows:
(1) dissolving a polylactic acid-glycolic acid copolymer in dichloromethane to form a polylactic acid-glycolic acid copolymer solution with the concentration of 0.03-0.06 g/mL, adding hydroxyapatite powder and an ammonium bicarbonate solution, performing ultrasonic emulsification, then adding an ammonium bicarbonate solution, and performing ultrasonic emulsification to obtain a W/O emulsion;
the mass ratio of the hydroxyapatite powder to the polylactic acid-glycolic acid copolymer is 3 (1-8), and the volume ratio of the ammonium bicarbonate solution to the polylactic acid-glycolic acid copolymer solution added each time is 0.5-1: 6; the concentration of the ammonium bicarbonate solution is 0.025-0.05 g/mL;
(2) injecting and transferring the W/O emulsion into a polyvinyl alcohol solution with the concentration of 10-25 g/L to form a W/O/W multiple emulsion, then adding water, stirring until dichloromethane is completely evaporated, washing the obtained product with water, and drying to obtain the water-soluble polyvinyl chloride/polyvinyl chloride composite emulsion; the volume ratio of the W/O emulsion to the polyvinyl alcohol solution to the water is (0.15-0.2): 1: 1.
5. The injectable multi-pore bone repair material according to claim 1 or 4, wherein the dopamine-hyaluronic acid is prepared by the following method:
(1) adding N-hydroxysuccinimide into a sodium hyaluronate aqueous solution, then adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, stirring and reacting for 1-3 hours, dropwise adding a dopamine hydrochloride aqueous solution, stirring and reacting for 10-14 hours, controlling the pH value to be within the range of 4.8-5.5 in the two stirring and reacting processes, and performing the operation of the step under the protection of nitrogen;
the molar ratio of the 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride to the N-hydroxysuccinimide to the dopamine hydrochloride to the carboxyl groups on the sodium hyaluronate is (1-6): 1-4): 1-3): 1, the concentration of the dopamine hydrochloride aqueous solution is 0.5-3 mmol/L, and the concentration of the sodium hyaluronate aqueous solution is 5-15 mg/mL;
(2) and (2) dialyzing the reaction solution obtained in the step (1) in water with the pH value of 3.0-3.5, and freeze-drying to obtain the compound.
CN201810526290.1A 2018-05-29 2018-05-29 Bone repair material capable of being injected with multiple pore structures and preparation method thereof Active CN108744060B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810526290.1A CN108744060B (en) 2018-05-29 2018-05-29 Bone repair material capable of being injected with multiple pore structures and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810526290.1A CN108744060B (en) 2018-05-29 2018-05-29 Bone repair material capable of being injected with multiple pore structures and preparation method thereof

Publications (2)

Publication Number Publication Date
CN108744060A CN108744060A (en) 2018-11-06
CN108744060B true CN108744060B (en) 2021-05-07

Family

ID=64003100

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810526290.1A Active CN108744060B (en) 2018-05-29 2018-05-29 Bone repair material capable of being injected with multiple pore structures and preparation method thereof

Country Status (1)

Country Link
CN (1) CN108744060B (en)

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109734937B (en) * 2019-01-08 2021-05-14 乐国平 Preparation method of VA @ PLGA-CS-HA/OHPPMC-HA antibacterial injectable hydrogel
CN109821061B (en) * 2019-01-18 2020-10-13 江南大学 Adhesive bionic gel with collagen spinning and hyaluronic acid as base materials
CN109880151A (en) * 2019-02-21 2019-06-14 上海市伤骨科研究所 A kind of preparation method and porous support materials of hydrogel porous microsphere
CN110128679B (en) * 2019-06-10 2021-10-08 西南交通大学 Preparation method of conductive double-layer hydrogel for electrical stimulation osteochondral integrated regeneration
CN110354078B (en) * 2019-08-12 2020-09-29 马晟利 Anti-candida albicans composition containing nisin and preparation method thereof
CN111068108B (en) * 2019-12-17 2022-03-15 联勤保障部队第九四〇医院 Bone grafting bracket for repairing bone defect of long tube of four limbs
CN111973804B (en) * 2020-09-17 2021-11-26 四川大学 Injectable bone repair adhesive loaded with stem cells and used for highly bionic active bone tissues and preparation method thereof
CN112138216B (en) * 2020-09-17 2021-10-26 四川大学 Hybrid cross-linked skull repairing porous membrane with highly bionic bone matrix and preparation method thereof
CN112076350B (en) * 2020-09-17 2022-01-07 四川大学 Biomimetic mineralized hydrogel with nano-micron composite structure and high mineral density as well as preparation method and application thereof
CN112279965A (en) * 2020-11-18 2021-01-29 四川大学 Preparation method of conductive adhesive hydrogel
CN113896906B (en) * 2021-09-03 2024-02-27 上海市伤骨科研究所 Charge-guided micro-nano adherable hydrogel and preparation method and application thereof
CN115926198A (en) * 2021-09-23 2023-04-07 四川大学 Injectable tissue regeneration type filler and preparation method thereof
WO2023093532A1 (en) * 2021-11-29 2023-06-01 爱美客技术发展股份有限公司 Composite gel, preparation method, and application
CN114259602B (en) * 2021-12-29 2022-09-30 江苏江山聚源生物技术有限公司 Recombinant collagen hydrogel for injection and preparation method thereof

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104174071A (en) * 2004-04-28 2014-12-03 安希奥设备国际有限责任公司 Compositions and systems for forming crosslinked biomaterials and associated methods of preparation and use
JP2009528080A (en) * 2006-02-27 2009-08-06 エイジェンシー・フォー・サイエンス，テクノロジー・アンド・リサーチ Hardenable bone cement
EP1891984A1 (en) * 2006-08-24 2008-02-27 Graftys Macroporous and highly resorbable apatitic calcium-phosphate cement
WO2009029049A1 (en) * 2007-08-30 2009-03-05 National University Of Singapore A bone and/or dental cement composition and uses thereof
CN101843918A (en) * 2010-05-27 2010-09-29 甘少磊 Composite bone repairing material based on nano-hydroxyapatite and hemihydrate calcium sulfate and preparation method thereof
CN103768658B (en) * 2012-10-18 2015-06-24 上海纳米技术及应用国家工程研究中心有限公司 Hydroxyapatite-loading polylactic acid porous microsphere and preparation method thereof
CN104056300B (en) * 2014-05-30 2016-03-30 浙江大学 A kind of polysaccharide-dopamine compound bio glue and application
CN106421929B (en) * 2016-09-22 2019-11-15 四川大学 A kind of injectable calcium phosphate/natural high molecular composite material and its preparation method and application

Also Published As

Publication number Publication date
CN108744060A (en) 2018-11-06

Similar Documents

Publication Publication Date Title
CN108744060B (en) Bone repair material capable of being injected with multiple pore structures and preparation method thereof
Yin et al. Preparation and characterization of macroporous chitosan–gelatin/β‐tricalcium phosphate composite scaffolds for bone tissue engineering
Janarthanan et al. Self-crosslinking hyaluronic acid–carboxymethylcellulose hydrogel enhances multilayered 3D-printed construct shape integrity and mechanical stability for soft tissue engineering
Levengood et al. Chitosan-based scaffolds for bone tissue engineering
Jin et al. In-situ formation of the hydroxyapatite/chitosan-alginate composite scaffolds
Zadegan et al. Synthesis and biocompatibility evaluation of cellulose/hydroxyapatite nanocomposite scaffold in 1-n-allyl-3-methylimidazolium chloride
CN110522948B (en) Injectable hydrogel and preparation method and application thereof
Rueda-Gensini et al. Graphene oxide-embedded extracellular matrix-derived hydrogel as a multiresponsive platform for 3D bioprinting applications
Wei et al. Nanocellulose based hydrogel or aerogel scaffolds for tissue engineering
CN110433331A (en) A kind of bioactive bracket and preparation method thereof
CN104004221B (en) Method for preparing polycaprolactone-keratin composite porous scaffolds
Zhang et al. Preparation of chitosan/hydroxyapatite guided membrane used for periodontal tissue regeneration
Jiang et al. Preparation of cellulose nanofiber-reinforced gelatin hydrogel and optimization for 3d printing applications.
CN101015712B (en) Process for preparation of polycaprolactone-chitosan network/hydroxyapatite porous compound support frame material
CN110396205B (en) Pickering high internal phase emulsion, 3D printing porous scaffold material and preparation method thereof
CN106421928A (en) Hydroxyapatite/polylactic acid composite material degradable in human bodies and method for preparing hydroxyapatite/polylactic acid composite material
Lin et al. Calcium phosphate cement reinforced by polypeptide copolymers
CN104910410A (en) Preparation method of RGD polypeptide grafted poly(maleic anhydride-hexamethylendiamine-DL-lactic acid)/modified hydroxyapatite porous composite material
CN105816918A (en) Aliphatic polyester-nano hydroxyapatite composite material and preparation method thereof
Cui et al. Freeze–thaw cycles for biocompatible, mechanically robust scaffolds of human hair keratins
CN113518804A (en) Ink material for 3D printing, preparation method and application
CN104001211A (en) Bone tissue engineering composite porous scaffold material and preparation method thereof
CN112138216A (en) Hybrid cross-linked skull repairing porous membrane with highly bionic bone matrix and preparation method thereof
CN101829355B (en) Hydrophilic extra-cellular matrix glycan/ hydrophobic aliphatic polyester composite material and preparation method thereof
CN116970304A (en) MnO (MnO) 2 -protein fiber composite particle gel ink and preparation and application thereof

Legal Events

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