CN113171491A - Bone tissue repair material and preparation method of bone tissue engineering scaffold - Google Patents

Bone tissue repair material and preparation method of bone tissue engineering scaffold Download PDF

Info

Publication number
CN113171491A
CN113171491A CN202110456387.1A CN202110456387A CN113171491A CN 113171491 A CN113171491 A CN 113171491A CN 202110456387 A CN202110456387 A CN 202110456387A CN 113171491 A CN113171491 A CN 113171491A
Authority
CN
China
Prior art keywords
bone tissue
tissue engineering
engineering scaffold
soluble
water
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.)
Pending
Application number
CN202110456387.1A
Other languages
Chinese (zh)
Inventor
刘佳
唐毓金
郑川川
李凯
王翀
王立强
杨成亮
李野
黄可
蓝常贡
谢克恭
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.)
Shanghai Jiaotong University
Youjiang Medical University for Nationalities Affiliated Hospital
Original Assignee
Shanghai Jiaotong University
Youjiang Medical University for Nationalities Affiliated Hospital
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 Shanghai Jiaotong University, Youjiang Medical University for Nationalities Affiliated Hospital filed Critical Shanghai Jiaotong University
Priority to CN202110456387.1A priority Critical patent/CN113171491A/en
Publication of CN113171491A publication Critical patent/CN113171491A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/18—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
    • A61L27/02—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
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y10/00—Processes of additive manufacturing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y70/00—Materials specially adapted for additive manufacturing
    • B33Y70/10—Composites of different types of material, e.g. mixtures of ceramics and polymers or mixtures of metals and biomaterials
    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B26/00—Compositions of mortars, concrete or artificial stone, containing only organic binders, e.g. polymer or resin concrete
    • C04B26/02—Macromolecular compounds
    • C04B26/10—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • C04B26/18—Polyesters; Polycarbonates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—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
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B2111/00—Mortars, concrete or artificial stone or mixtures to prepare them, characterised by specific function, property or use
    • C04B2111/00034—Physico-chemical characteristics of the mixtures
    • C04B2111/00181—Mixtures specially adapted for three-dimensional printing (3DP), stereo-lithography or prototyping
    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B2111/00—Mortars, concrete or artificial stone or mixtures to prepare them, characterised by specific function, property or use
    • C04B2111/00474—Uses not provided for elsewhere in C04B2111/00
    • C04B2111/00836—Uses not provided for elsewhere in C04B2111/00 for medical or dental applications
    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B2201/00—Mortars, concrete or artificial stone characterised by specific physical values
    • C04B2201/50—Mortars, concrete or artificial stone characterised by specific physical values for the mechanical strength

Landscapes

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

Abstract

The invention relates to the technical field of regenerative medicine, and discloses a bone tissue repair material and a preparation method of a bone tissue engineering scaffold. The bone tissue repair material comprises the following components in parts by weight: 10-40 parts of oil-soluble high polymer material, 10-50 parts of biological ceramic powder, 50-100 parts of oily solvent, 0.1-1 part of water-soluble bone tissue repair drug, 2-30 parts of water, more than 0 part and not more than 10 parts of seed cells and 50-100 parts of culture medium. The bone tissue repair material forms a base material of the bone tissue engineering scaffold by the mutual matching of the oil-soluble polymer material and the biological ceramic powder, so that sufficient structure/mechanical properties are provided for the printed bone tissue engineering scaffold, and the personalized bone tissue engineering scaffold can be prepared by using 3D printing equipment; the osteoblast osteogenic differentiation is promoted through the action of the water-soluble bone tissue repair drug, so that the osteoblast is differentiated to form a complete bone tissue structure on the basis of the bone tissue engineering scaffold.

Description

Bone tissue repair material and preparation method of bone tissue engineering scaffold
Technical Field
The invention relates to the technical field of regenerative medicine, in particular to a bone tissue repair material and a preparation method of a bone tissue engineering scaffold.
Background
Bone atrophy, trauma, malignant tumor, periodontal disease and the like of different causes are common causes of bone defects. Generally, a bone defect having a defect range exceeding 50% of the bone circumference or a length exceeding 2cm or more is referred to as a large bone defect. Autologous bone grafting is the gold standard for clinical treatment of bone defects. However, bone grafting causes complications such as bleeding, hematoma, infection and chronic pain, and has limiting factors such as insufficient donor sources and excessive economic stress on patients. Therefore, the treatment strategy for large-area bone defects needs to be designed and developed to meet more and more clinical demands. Tissue engineering techniques have become one of the potential options available for repairing bone defects in recent years.
Many prosthetic materials (including single phase, multi-layered or having a gradient structure) have been used to guide bone tissue regeneration. Adult tissue cells and progenitor cells (including stem cells) are also seeded into scaffolds for in vivo and in vitro studies in bone tissue. Meanwhile, various growth factors are also used for inducing the directional differentiation of cells and accelerating the regeneration of bone tissues. Nevertheless, since bone tissue has considerable complexity, an effective bone tissue regeneration strategy has not yet been established. Therefore, the method has important theoretical significance for establishing a tissue engineering model capable of effectively repairing bone tissue injury and inducing bone tissue regeneration by adopting an advanced material preparation process, selecting a proper bone tissue repair drug, developing an effective growth factor delivery mode and further optimizing the interrelation among the scaffold structure/mechanical property, cell differentiation and bone repair drug slow release, can provide a new thought for explaining the biological essence of bone defect treatment, and has important significance for finally clinically transforming and treating bone defects.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the prior bone tissue repair material is difficult to obtain, and the prepared tissue engineering scaffold has poor structure/mechanical property and can not effectively regenerate bone tissues.
In order to solve the technical problems, the invention provides a bone tissue repair material which comprises the following components in parts by weight: 10-40 parts of oil-soluble high polymer material, 10-50 parts of biological ceramic powder, 50-100 parts of oily solvent, 0.1-1 part of water-soluble bone tissue repair drug and 2-30 parts of water.
Preferably, the oil-soluble polymer material is soluble in an organic solvent and insoluble in water, and the polymer material is one of racemic polylactic acid (PDLLA), polyglycolic acid (PGA), lactic acid-glycolic acid copolymer (PLGA), levorotatory polylactic acid (PLLA), copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV), and poly-beta-hydroxybutyrate (PHB).
Preferably, the oily solvent is Dichloromethane (DCM) and the water is deionized water.
Preferably, the water-soluble tissue repair agent is PR-171 (formula: C)40H57N5O7) FK506 (molecular formula: c44H69NO12) HA15 (molecular formula: c23H22N4O3S2) One or a mixture of several of them, so as to control the growth rate and growth amount of bone tissue by controlling the usage amount of each drug of the water-soluble tissue repair drug.
Preferably, the biological ceramic powder is one of nano calcium phosphate, beta-tricalcium phosphate and nano hydroxyapatite.
The invention also provides a preparation method of the bone tissue engineering scaffold based on the bone tissue repair material, which comprises the following steps:
s1, mixing the raw materials of the bracket: dissolving an oil-soluble polymer material in an oily solvent to prepare an oil-soluble polymer material/oily solvent solution, and dissolving the biological ceramic powder in the oil-soluble polymer material/oily solvent solution to prepare an oil-soluble polymer material/biological ceramic powder/oily solvent solution;
s2, preparing a water-soluble osteogenic solution: dissolving a water-soluble bone tissue repair drug in water to form a water-soluble osteogenic solution;
s3, preparation of forming ink: mixing the oil-soluble polymer material/biological ceramic powder/oily solvent solution prepared in the step S1 and the water-soluble osteogenic solution prepared in the step S2 uniformly to prepare the forming ink of the bone tissue engineering scaffold;
s4, printing the bone tissue engineering scaffold: obtaining the bone tissue engineering scaffold by the molding ink through printing equipment at a low temperature;
s5, freeze-drying the bone tissue engineering scaffold, and air-drying the bone tissue engineering scaffold to remove the solvent to obtain the finished product of the bone tissue engineering scaffold.
Preferably, the low temperature in the step S4 is-10 ℃ to-40 ℃.
Preferably, the porosity of the finished bone tissue engineering scaffold obtained in the step S5 is 40-60%, 60-85%, 85-95%, and the pore diameter is 100-500 μm, 500-1000 μm, 1000-2000 μm.
Compared with the prior art, the invention has the beneficial effects that:
1. the bone tissue repair material disclosed by the invention has the advantages that the manufacturing raw materials are common raw materials, the acquisition is easy, the resources are rich, and convenient conditions are provided for the popularization and the application of bone tissue engineering scaffolds; the bone tissue repair material of the invention forms the base material of the bone tissue engineering scaffold by the mutual matching of the oil-soluble polymer material and the biological ceramic powder, so as to provide enough structure/mechanical property for the printed bone tissue engineering scaffold; the material disclosed by the invention is convenient to prepare the bone tissue engineering scaffold, the personalized bone tissue engineering scaffold can be prepared by using 3D printing equipment, the bone tissue engineering scaffold with the needed personalized size and shape can be prepared according to a designed scheme, and the bone tissue engineering scaffold has good biomechanical strength, a bone-like tissue structure, anisotropic mechanical properties and a macro-microstructure; the osteoblast osteogenic differentiation is promoted through the action of the water-soluble bone tissue repairing medicine, so that the osteoblast is differentiated to form a complete bone tissue structure on the basis of the bone tissue engineering scaffold, the defective bone tissue obtains the strength and the shape equivalent to those of original bone tissue, and the bone tissue is effectively regenerated;
2. the preparation method of the bone tissue engineering scaffold can load water-soluble bone tissue repair drugs in situ, the preparation of the forming ink is simple, the printing and forming are carried out under the condition of-10 to-40 ℃, and then the bone tissue engineering scaffold can be shaped by freeze drying;
3. the preparation method of the bone tissue engineering scaffold adopts sequential low-temperature 3D printing to realize the load and controllable slow release of different water-soluble bone tissue repair drugs at different regions of the bone tissue engineering scaffold.
Drawings
FIG. 1 is an image of a tissue engineering scaffold prepared in example 1 of the present invention.
FIG. 2 is a display image of a tissue engineering scaffold prepared in example 2 of the present invention under a scanning electron microscope with a magnification of 200 μm.
FIG. 3 is a microscopic image of the tissue engineering scaffold prepared in example 2 of the present invention under a scanning electron microscope with a magnification of 100 μm.
FIG. 4 is a schematic diagram of the observation result of the inverted fluorescence microscope of the tissue engineering scaffold cultured for 3 days in example 2 of the present invention, wherein the white dots are the living cells.
Fig. 5 is a mechanical property test chart of the bone tissue engineering scaffold of example 2 of the present invention, in which the abscissa is the magnitude of displacement and the ordinate is the magnitude of force applied to the bone tissue engineering scaffold.
Detailed Description
The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
The 3D bioprinting apparatus used in the following examples is a multi-nozzle printer, and the printing ambient temperature can be reduced to 0 to-100 ℃.
Example 1: the embodiment provides a bone tissue repair material, which comprises the following components in parts by weight: 15 parts of oil-soluble high polymer material, 15 parts of biological ceramic powder, 50 parts of oily solvent, 1 part of water-soluble bone tissue repair medicine and 5 parts of water; the oil-soluble high polymer material is lactic acid-glycolic acid copolymer (PLGA); the oily solvent is dichloromethane; the biological ceramic is beta-tricalcium phosphate; the water is deionized water. The water-soluble bone tissue repair drug is PR 171.
The bone tissue engineering scaffold is prepared by using the bone tissue repair material according to the preparation method of the tissue engineering scaffold, and the preparation method comprises the following steps:
s1, preparing 10mL of 30% lactic acid-glycolic acid copolymer/dichloromethane solution, and then dissolving 3 g of beta-tricalcium phosphate powder in the lactic acid-glycolic acid copolymer/dichloromethane solution to be uniformly stirred to prepare the lactic acid-glycolic acid copolymer/beta-tricalcium phosphate powder/dichloromethane solution;
s2, dissolving 1 part of PR171 in deionized water to prepare a water-soluble osteogenic solution of PR 171;
s3, mixing the prepared lactic acid-glycolic acid copolymer/beta-tricalcium phosphate powder/dichloromethane solution with 1.5mL of water solution of the PR171 to obtain the paste-shaped ink for forming the bone tissue engineering scaffold;
s4, printing the molding ink on a low-temperature (-30 ℃) printing table through a nozzle according to the CAD model to mold the bone tissue engineering scaffold;
and S5, after the bone tissue engineering scaffold is printed, removing the solvent through low-temperature air drying, wherein the temperature at low temperature is 0-20 ℃, removing the solvent remaining in the bone cartilage structure, and completing the preparation of the bone tissue engineering scaffold to obtain a finished product of the bone tissue engineering scaffold, as shown in figure 1.
Example 2: the embodiment provides a bone tissue repair material, which comprises the following components in parts by weight: 20 parts of oil-soluble high polymer material, 20 parts of biological ceramic powder, 100 parts of oily solvent, 1 part of water-soluble bone tissue repair medicine and 30 parts of water; the oil-soluble high polymer material is lactic acid-glycolic acid copolymer (PLGA); the oily solvent is dichloromethane; the biological ceramic is beta-tricalcium phosphate; the water is deionized water. The water-soluble bone tissue repair drug is HA 15.
The preparation of the tissue engineering scaffold by using the printing material comprises the following steps:
s1, preparing 10mL of 30% lactic acid-glycolic acid copolymer/dichloromethane solution, and then dissolving 3 g of beta-tricalcium phosphate powder in the lactic acid-glycolic acid copolymer/dichloromethane solution to be uniformly stirred to prepare the lactic acid-glycolic acid copolymer/beta-tricalcium phosphate powder/dichloromethane solution;
s2, dissolving 1 part of HA15 in deionized water to prepare a water-soluble osteogenic solution of HA 15;
s3, mixing the prepared 10mL of lactic acid-glycolic acid copolymer/dichloromethane solution and 1.5mL of aqueous solution of LHA15 to obtain the cream-shaped forming ink of the bone tissue engineering scaffold;
s4, printing the molding ink on a low-temperature (-25 ℃) printing table through a nozzle according to the CAD model to mold the bone tissue engineering scaffold;
and S5, after the bone tissue engineering scaffold is printed, drying the residual solvent in the bone cartilage structure in air, and completing the preparation of the bone tissue engineering scaffold to obtain a finished bone tissue engineering scaffold, wherein the finished bone tissue engineering scaffold prepared in the embodiment 2 has a figure basically consistent with the appearance shown in the attached drawing 1, and the attached drawing is not provided herein, and reference is also made to the attached drawing 1. The prepared bone tissue engineering scaffold is developed by an electron microscope to obtain micrographs as shown in figures 2 and 3, and as can be seen from figures 2 and 3, the pores are approximately rectangular, the size of the larger pores is about 200 microns multiplied by 180 microns, the size of the smaller pores is 100 microns multiplied by 180 microns, the perfect pore structure reserves a large number of adhesion areas for cells, the porosity of the bone tissue engineering scaffold is 51%, and the pore diameter is 100-500 microns.
Biological activity assay
Culturing the prepared bone tissue engineering scaffold in a 5% carbon dioxide incubator at 37 ℃ for three days, and detecting the activity of cells after culturing. In the detection process, firstly, phosphate buffer solution is used for washing the surface of the cell-bone tissue engineering scaffold material composite body, then a dead and live cell detection kit is adopted, the cell-bone tissue engineering scaffold material composite body is placed in a culture medium containing dead and live cell fluorescent dye with working concentration (the storage concentration of the kit is 1000 times of moisture absorption in a DMEM culture medium), the incubation is carried out for half an hour in a cell culture box at 37 ℃, and then an inverted fluorescence microscope is used for observing the dead and live cells.
FIG. 4 is the observation result of an inverted fluorescence microscope of cells cultured for 3 days, in which white round dots are viable cells. The tissue engineering scaffold with the medicine for promoting the osteoblast activity and the biological ceramic powder can better provide help for cell adhesion, proliferation and expansion so as to promote the effective regeneration of bone tissues and form the bionic bone with similar structure and mechanical properties to the original bone.
Mechanical property testing
The bone tissue engineering scaffold prepared in example 2 was subjected to mechanical property test according to the test standards, and the experimental data was recorded in table 1.
Table 1: experimental data Table of bone tissue engineering scaffolds prepared in example 2
Figure BDA0003038962110000061
It can be seen from table 1 and fig. 5 that the bone tissue engineering scaffold prepared in example 1 has a maximum compressive strength of 0.682Mpa, a maximum displacement of 5.3372% at the maximum compressive strength, and a small change in displacement, has good mechanical properties, and is not easily deformed. It can be seen that, with the increase of the compressive strength, the displacement of the bone tissue engineering scaffold is firstly increased, and after the certain compressive strength is reached, the displacement of the bone tissue engineering scaffold is continuously increased, but the variation is gradually reduced.
In conclusion, the bone tissue repair material disclosed by the invention has the advantages that the preparation raw materials are common raw materials, the raw materials are easy to obtain and rich in resources, and convenience is provided for popularization and application of bone tissue engineering scaffolds; the bone tissue repair material of the invention forms the base material of the bone tissue scaffold by the mutual matching of the oil-soluble polymer material and the biological ceramic powder, so as to provide enough structure/mechanical property for the printed bone tissue engineering scaffold; the material disclosed by the invention is convenient to prepare the bone tissue engineering scaffold, the personalized bone tissue engineering scaffold can be prepared by using 3D printing equipment, the bone tissue engineering scaffold with the needed personalized size and shape can be prepared according to a designed scheme, and the bone tissue engineering scaffold has good biomechanical strength, a bone-like tissue structure, anisotropic mechanical properties and a macro-microstructure; the osteoblast osteogenic differentiation is promoted through the action of the water-soluble bone tissue repairing medicine, so that the osteoblast is differentiated to form a complete bone tissue structure on the basis of the bone tissue engineering scaffold, the defective bone tissue obtains the strength and the shape equivalent to those of original bone tissue, and the bone tissue is effectively regenerated;
the preparation method of the bone tissue engineering scaffold can load water-soluble bone tissue repair drugs in situ, the preparation of the forming ink is simple, the printing and forming are carried out under the condition of-10 to-40 ℃, and then the bone tissue engineering scaffold can be shaped by freeze drying; sequential low-temperature 3D printing is adopted, so that the load and controllable slow release of different water-soluble bone tissue repair drugs at different positions of the bone tissue engineering scaffold are realized.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and substitutions can be made without departing from the technical principle of the present invention, and these modifications and substitutions should also be regarded as the protection scope of the present invention.

Claims (8)

1. Bone tissue repair material, characterized in that: the paint comprises the following components in parts by weight: 10-40 parts of oil-soluble high polymer material, 10-50 parts of biological ceramic powder, 50-100 parts of oily solvent, 0.1-1 part of water-soluble bone tissue repair drug and 2-30 parts of water.
2. The bone tissue repair material according to claim 1, characterized in that: the oil-soluble high polymer material is dissolved in an organic solvent and is insoluble in water, and the high polymer material is one of racemic polylactic acid (PDLLA), polyglycolic acid (PGA), lactic acid-glycolic acid copolymer (PLGA), levorotatory polylactic acid (PLLA), copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV) and poly-beta-hydroxybutyrate (PHB).
3. The bone tissue repair material according to claim 1, characterized in that: the oily solvent is Dichloromethane (DCM), and the water is deionized water.
4. The bone tissue repair material according to claim 1, characterized in that: the water-soluble tissue repair drug is PR-171 (molecular formula: C)40H57N5O7) FK506 (molecular formula: c44H69NO12) HA15 (molecular formula: c23H22N4O3S2) One or a mixture of several of them.
5. The bone tissue repair material according to claim 1, characterized in that: the biological ceramic powder is one of nano calcium phosphate, beta-tricalcium phosphate and nano hydroxyapatite.
6. The preparation method of the bone tissue engineering scaffold is characterized by comprising the following steps of:
s1, mixing the raw materials of the bracket: dissolving an oil-soluble polymer material in an oily solvent to prepare an oil-soluble polymer material/oily solvent solution, and dissolving the biological ceramic powder in the oil-soluble polymer material/oily solvent solution to prepare an oil-soluble polymer material/biological ceramic powder/oily solvent solution;
s2, preparing a water-soluble osteogenic solution: dissolving a water-soluble bone tissue repair drug in water to form a water-soluble osteogenic solution;
s3, preparation of forming ink: mixing the oil-soluble polymer material/biological ceramic powder/oily solvent solution prepared in the step S1 and the water-soluble osteogenic solution prepared in the step S2 uniformly to prepare the forming ink of the bone tissue engineering scaffold;
s4, printing the bone tissue engineering scaffold: obtaining the bone tissue engineering scaffold by the molding ink through printing equipment at a low temperature;
s5, freeze-drying the bone tissue engineering scaffold, and air-drying the bone tissue engineering scaffold to remove the solvent to obtain the finished product of the bone tissue engineering scaffold.
7. The method for preparing a scaffold for bone tissue engineering according to claim 6, wherein: the low temperature in the step S4 is-10 ℃ to-40 ℃.
8. The method for preparing a scaffold for bone tissue engineering according to claim 6, wherein: the porosity of the finished bone tissue engineering scaffold obtained in the step S5 is 40-60%, or 60-85%, or 85-95%, and the pore diameter is 100-500 μm, or 500-1000 μm, or 1000-2000 μm.
CN202110456387.1A 2021-04-26 2021-04-26 Bone tissue repair material and preparation method of bone tissue engineering scaffold Pending CN113171491A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110456387.1A CN113171491A (en) 2021-04-26 2021-04-26 Bone tissue repair material and preparation method of bone tissue engineering scaffold

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110456387.1A CN113171491A (en) 2021-04-26 2021-04-26 Bone tissue repair material and preparation method of bone tissue engineering scaffold

Publications (1)

Publication Number Publication Date
CN113171491A true CN113171491A (en) 2021-07-27

Family

ID=76926506

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110456387.1A Pending CN113171491A (en) 2021-04-26 2021-04-26 Bone tissue repair material and preparation method of bone tissue engineering scaffold

Country Status (1)

Country Link
CN (1) CN113171491A (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105263438A (en) * 2013-03-14 2016-01-20 托莱多大学 Injectable biodegradable bone matrix for multiple myeloma lesion augmentation and osteoporosis
CN113198001A (en) * 2021-04-26 2021-08-03 右江民族医学院附属医院 Application of proteasome inhibitor PR171 in preparation of medicine for treating osteoporosis

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105263438A (en) * 2013-03-14 2016-01-20 托莱多大学 Injectable biodegradable bone matrix for multiple myeloma lesion augmentation and osteoporosis
CN113198001A (en) * 2021-04-26 2021-08-03 右江民族医学院附属医院 Application of proteasome inhibitor PR171 in preparation of medicine for treating osteoporosis

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
HURCHLA MA: "The epoxyketone-based proteasome inhibitors carfilzomib and orally bioavailable oprozomib have anti-resorptive and bone-anabolic activity in addition to anti-myeloma effects", 《LEUKEMIA》 *
LI YE: "3D Printing of Tricalcium Phosphate/Poly Lactic-co-glycolic Acid Scaffolds Loaded with Carfilzomib for Treating Critical-sized Rabbit Radial Bone Defects", 《INTERNATIONAL JOURNAL OF BIOPRINTING》 *
YOSHIKAWA T: "In vivo osteogenic capability of cultured allogeneic bone in porous hydroxyapatite: Immunosuppressive and osteogenic potential of FK506 in vivo", 《JOURNAL OF BONE AND MINERAL RESEARCH》 *
ZHENG CC: "3D-printed HA15-loaded beta-Tricalcium Phosphate/Poly (Lactic-co-glycolic acid) Bone Tissue Scaffold Promotes Bone Regeneration in Rabbit Radial Defects", 《INTERNATIONAL JOURNAL OF BIOPRINTING》 *
张帆: "PR171（Carfi Izomib）通过抑制PSME1、PSME2激活Wnt信号缓解骨质疏松发病的机制研究", 《中国优秀硕士学位论文全文数据库 (医药卫生科技辑)》 *

Similar Documents

Publication Publication Date Title
Litowczenko et al. Milestones and current achievements in development of multifunctional bioscaffolds for medical application
KR101131901B1 (en) Graphene oxide/biodegradable polymer nanofiber composites and preparation method thereof
Qian et al. Fabrication and in vitro biocompatibility of biomorphic PLGA/nHA composite scaffolds for bone tissue engineering
US20090117087A1 (en) Methods and compositions for printing biologically compatible nanotube composites of autologous tissue
CN101478934B (en) Bioengineered intervertebral discs and methods for their preparation
CA2619243C (en) Topographical templating of polymeric materials using cellular morphology
Li et al. Spatially featured porous chitosan conduits with micropatterned inner wall and seamless sidewall for bridging peripheral nerve regeneration
CN109364302A (en) A kind of preparation method of bone cartilage repair material and tissue engineering bracket
CN102319449B (en) Poly(lactic-co-glycolic acid)-based growth factor gradient release microsphere stent as well as preparation method and application thereof
CN111172100B (en) Biological 3D printing method for controlling cell orientation arrangement
CN104888277A (en) Cell-biological scaffold complex and 3D printing forming method thereof
CN1749319A (en) Supercritical CO 2Technology prepares the method for three-dimensional porous rack material for tissue engineering
CN111956864A (en) 3D printing composite support and preparation method and application thereof
KR101712862B1 (en) Porous polymer micro-beads, method for preparing thereof, and bio materials using the same
WO2007120840A2 (en) Methods and compositions for printing biologically compatible nanotube composites
CN109224132A (en) MULTILAYER COMPOSITE stomach wall repairs sticking patch bracket and preparation method thereof
CN113181429A (en) Method for preparing plastic long-section bone repair material and bone tissue engineering scaffold with controllable slow release of bioactive factors
Cai et al. Nanofiber composites in skeletal muscle tissue engineering
US11957815B2 (en) Ready to use biodegradable and biocompatible cell-based nerve conduit for nerve injury and a method of preparation thereof
EP2291556A2 (en) Nonwoven structure and method of fabricating the same
CN110859994B (en) Modified tussah silk fibroin 3D printing support and preparation method thereof
CN113171491A (en) Bone tissue repair material and preparation method of bone tissue engineering scaffold
Schäfer et al. Skeletal muscle tissue engineering
CN101837148A (en) Porous biodegradable stent and preparation method thereof
KR101667406B1 (en) Hydrophilic nanofiber scaffold comprising PVP-b-PCL block copolymers

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
RJ01 Rejection of invention patent application after publication

Application publication date: 20210727

RJ01 Rejection of invention patent application after publication