CN107661540B - Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing - Google Patents

Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing Download PDF

Info

Publication number
CN107661540B
CN107661540B CN201711049313.6A CN201711049313A CN107661540B CN 107661540 B CN107661540 B CN 107661540B CN 201711049313 A CN201711049313 A CN 201711049313A CN 107661540 B CN107661540 B CN 107661540B
Authority
CN
China
Prior art keywords
solution
printing
chitosan
acid
calcium
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
CN201711049313.6A
Other languages
Chinese (zh)
Other versions
CN107661540A (en
Inventor
赵娜如
董怡帆
梁锦宁
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.)
South China University of Technology SCUT
Original Assignee
South China University of Technology SCUT
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 South China University of Technology SCUT filed Critical South China University of Technology SCUT
Priority to CN201711049313.6A priority Critical patent/CN107661540B/en
Publication of CN107661540A publication Critical patent/CN107661540A/en
Application granted granted Critical
Publication of CN107661540B publication Critical patent/CN107661540B/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
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/446—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with other specific inorganic fillers other than those covered by A61L27/443 or A61L27/46
    • 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
    • A61L27/56—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
    • B33Y70/00—Materials specially adapted for 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
    • B33Y80/00—Products made by additive manufacturing
    • 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

Abstract

The invention discloses a method for preparing a high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing. The method comprises the following steps: (1) dissolving chitosan in an acid solution to obtain solution A; (2) adding tetraethoxysilane into an acid solution for hydrolysis to obtain solution B; (3) adding a calcium source and a phosphorus source into the solution A to obtain solution C; (4) and adding the solution B into the solution C, uniformly stirring, removing bubbles in vacuum, then filling into a printing material cylinder, standing, and after the solution B is gelled, performing 3D printing molding by using a 3D printer to obtain the hybrid support. On the basis of a sol-gel method, nano-grade hydroxyapatite is introduced in situ; and the obtained support has the characteristics of high strength, controllable shape and size and the like by combining a 3D printing technology. The method has the advantages of simple operation, wide material source, mild preparation conditions, excellent biological and mechanical properties of the obtained material and wide application prospect in the fields of tissue engineering and regenerative medicine.

Description

Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing
Technical Field
The invention belongs to the technical field of biomedical materials, and particularly relates to a method for preparing a high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing.
Background
Large-area bone defects due to various causes such as trauma, infection, tumor, bone aging, congenital deformity, etc., are often treated by bone graft surgery. At present, the bone tissue engineering technology overcomes the defects of the traditional bone defect transplanting technology and provides a new choice for clinical bone repair treatment. And 3D printing is a new processing technology, can meet the requirements of micro and macro structures at the same time, is suitable for personalized customization, and has great advantages in bone tissue engineering.
The chitosan is a natural polysaccharide with positive charges, has a structure very similar to glycosaminoglycan, which is a main component of extracellular matrix, is one of main organic components in biological complexes in nature, and can provide a high molecular recognition point for the deposition of inorganic minerals as a macromolecular template. It has excellent biological compatibility, biodegradability, antibacterial property, easy film forming property and other excellent biochemical properties. In recent years, the method has been widely applied to various fields such as biology, medicine, food, cosmetics and the like, and the development and research of products thereof have attracted more and more attention from countries and research institutions. The application of pure chitosan as a material has certain limitation, and a great deal of research work has been done at home and abroad for modifying the chitosan to synthesize organic-inorganic hybrid or composite materials. The natural bone tissue is an organic/inorganic nano hybrid, wherein the inorganic matter is mainly nano hydroxyapatite. Therefore, from the viewpoint of bionics, constructing a bone repair material by simulating the composition, structural characteristics and mineralization process of human bone tissue has become a novel method for treating bone defects.
In recent years, organic-inorganic hybrid materials prepared by a sol-gel method have become a new research hotspot for bone repair materials. The hybrid material is a special composite material, organic components and inorganic components of the hybrid material can interact on a molecular level, no obvious phase interface exists between the organic components and the inorganic components in the hybrid material, the hybrid material realizes the full mixing of the molecular level, can combine the advantages of the inorganic materials in the aspects of mechanical strength and elastic modulus and the advantages of the high polymer materials in the aspect of toughness, makes up the defects of the two materials, realizes the temporary replacement of bones, and generates new bones along with the degradation induction of the materials.
Disclosure of Invention
The invention provides a method for preparing a high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by 3D printing, which is characterized in that an organic polymer and an inorganic substance are compounded on a molecular level by a sol-gel method, from the bionic angle, nano-grade hydroxyapatite is introduced in situ to prepare a hard tissue substitute material with a bionic structure, and the internal structure and the external form of the scaffold are accurately regulated and controlled by means of a 3D printing technology to prepare the scaffold with a specific shape. The preparation method is simple, the material source is wide, the preparation condition is mild, and the obtained three-dimensional scaffold has good mechanical property and biocompatibility and has wide application prospect in the fields of tissue engineering and regenerative medicine.
The invention is realized by the following technical scheme.
A method for preparing a high-strength hydroxyapatite-chitosan-silica hybrid scaffold by using 3D printing comprises the following steps:
(1) adding chitosan powder into an acid solution, and stirring to fully dissolve the chitosan powder to obtain a uniform and transparent chitosan solution; the acid solution is an aqueous solution of acetic acid, lactic acid and hydrochloric acid;
(2) adding tetraethoxysilane into an acid solution, stirring to fully hydrolyze the tetraethoxysilane to obtain clear and transparent hydrolysate, wherein the acid solution is an aqueous solution of hydrochloric acid, nitric acid, acetic acid and lactic acid;
(3) sequentially adding a calcium source and a phosphorus source into the chitosan solution obtained in the step (1), and stirring to fully dissolve the calcium source and the phosphorus source to obtain a mixed solution, wherein the calcium source is one of calcium acetate, calcium nitrate, calcium hydroxide, calcium hydrophosphate and calcium chloride; the phosphorus source is one of ammonium dihydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, phosphoric acid and triethyl phosphate;
(4) adding the hydrolysate obtained in the step (2) into the mixed solution obtained in the step (3), quickly and uniformly stirring, and removing bubbles in vacuum to obtain milky sol;
(5) putting the sol obtained in the step (4) into a 3D printing charging barrel, and standing to gelatinize the sol;
(6) using a 3D-BioplotterTMThe printing equipment extrudes the gel obtained in the step (5) into threads to be printed on a printing gasket under certain printing parameters to form a preset support structure, so that a three-dimensional porous support is obtained;
(7) fully gelatinizing the three-dimensional porous scaffold obtained in the step (6) to obtain a three-dimensional through porous scaffold;
(8) soaking the stent obtained in the step (7) in alkali liquor to remove the residual acid in the material;
(9) repeatedly washing the stent obtained in the step (8) with water until the pH value is neutral; and then drying to obtain a Hydroxyapatite (HA) -chitosan-silicon dioxide hybrid scaffold, and finally placing the dried scaffold in a constant temperature and humidity drying box for storage.
Preferably, the viscosity-average molecular weight of the chitosan in the step (1) is 10-84 ten thousand, and the deacetylation degree is 75-95%; the pH value of the acid solution is 2-6; the mass fraction of the chitosan solution is 4-12%.
Preferably, the molar concentration of the acid solution in the step (2) is 1-3.84 mol/L; the stirring time is 1-10 h.
Preferably, the amount of the calcium source and the phosphorus source in the step (3) is 0-30% of the mass of the chitosan, wherein the molar ratio of calcium to phosphorus is 1.67; the hybrid material comprises hydroxyapatite, chitosan and silicon dioxide.
Preferably, the sol in step (4) contains chitosan and SiO2The mass ratio of (A) to (B) is 1 to 2.
Preferably, the standing in the step (5) is to place the sol in a constant temperature and humidity box with the temperature of 4-40 ℃ and the humidity of 30-70% for 1-5 days.
Preferably, the parameter conditions of the printing in the step (6) are as follows: the extrusion pressure is 0-0.58 MPa, the printing speed is 10-50 mm/min, and the platform temperature is normal temperature.
Preferably, the diameter of the fiber of the three-dimensional porous support is set to be 0.2-1 mm during printing in the step (6), and the distance between the fibers is set to be 0.1-1 mm.
Preferably, the three-dimensional porous scaffold in the step (7) is fully gelatinized by placing the three-dimensional porous scaffold in a constant temperature and humidity box with the temperature of 20-30 ℃ and the humidity of 30-70% for curing for 3-15 days.
Preferably, the alkali solution in step (8) is an aqueous solution of sodium hydroxide, sodium carbonate, sodium bicarbonate, dipotassium hydrogen phosphate, disodium hydrogen phosphate or Tris.
Preferably, the temperature of the drying in step (9) is 40 ℃.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the invention adopts biodegradable polycation polysaccharide chitosan as a matrix material, and introduces inorganic reinforcement silicon dioxide and Hydroxyapatite (HA) by a sol-gel method and an in-situ precipitation method, thereby realizing hybridization of organic-inorganic materials at a molecular level, effectively improving the mechanical strength of a hybrid scaffold, and having good bioactivity.
(2) According to the invention, the organic-inorganic hybrid scaffold is prepared by means of a 3D printing technology, and the three-dimensional scaffold with different macro-morphologies and microstructures can be prepared by adjusting printing parameters while ensuring good mechanical properties of the scaffold.
Drawings
Fig. 1 is a schematic printing diagram of the preparation of a high-strength hydroxyapatite-chitosan-silica hybrid scaffold by 3D printing according to the present invention.
Fig. 2 is a scanning electron microscope image of a high-strength chitosan-silica hybrid scaffold prepared by 3D printing in example 1 of the present invention.
Fig. 3 is a scanning electron microscope image of a high-strength hydroxyapatite-chitosan-silica hybrid scaffold prepared by 3D printing in example 7 of the present invention.
Detailed Description
The invention is described in further detail below with reference to the figures and examples, which are only intended to illustrate the invention and are not to be construed as limiting the invention.
Example 1
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; according to CS/SiO2Adding the solution B into the solution A according to the mass ratio of 10/7, fully stirring to mix uniformly, removing bubbles in vacuum to obtain light yellow sol, putting the light yellow sol into a 3D printing material cylinder, and standing in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% for 56h to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model. The 3D printing process is shown in fig. 1.
Setting the printing parameters as follows: extrusion pressure: 0.35 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a constant temperature and humidity box with the temperature of 30 ℃ and the humidity of 50% for curing for 4D to ensure that the support is fully gelatinized; then the chitosan-silica hybrid scaffold is immersed into 1mol/L Tris aqueous solution (Tris), taken out after 24 hours, washed to be neutral by ultrapure water, and dried in an oven at 40 ℃ to obtain the three-dimensionally communicated chitosan-silica hybrid scaffold, as shown in figure 2.
The experiment shows that: the fiber diameter of the chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 200 μm, and the compressive strength and Young's modulus are shown in Table 1.
Example 2
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; respectively weighing Ca (NO) according to the mass ratio of CS to HA of 10/33)2.4H2O、H3PO4(ensuring that the molar ratio of Ca to P is 1.67) is sequentially added into the solution A, and the mixture is fully stirred until the mixture is uniformly mixed to obtain solution C; according to CS/SiO2Weighing the solution B according to a mass ratio of 10/7, adding the solution B into the solution C, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing for 48 hours in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.45 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a constant temperature and humidity box with the temperature of 30 ℃ and the humidity of 50% for curing for 4D to ensure that the support is fully gelatinized; and then immersing the hybrid scaffold into 1mol/L Tris aqueous solution, taking out after 24h, washing the hybrid scaffold to be neutral by using ultrapure water, and drying the hybrid scaffold in a drying oven at 40 ℃ to obtain the three-dimensionally communicated hydroxyapatite-chitosan-silicon dioxide hybrid scaffold.
The experiment shows that: the fiber diameter of the hydroxyapatite-chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 200 μm, and the compressive strength and young's modulus were as shown in table 1.
Example 3
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; respectively weighing Ca (NO) according to the mass ratio of CS to HA of 5/23)2.4H2O、H3PO4(ensuring that the molar ratio of Ca to P is 1.67) is sequentially added into the solution A, and the mixture is fully stirred until the mixture is uniformly mixed to obtain solution C; according to CS/SiO2Weighing the solution B according to the mass ratio of 10/7, adding the solution B into the solution C, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% for 46h to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.48 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a constant temperature and humidity box with the temperature of 30 ℃ and the humidity of 50% for curing for 4D to ensure that the support is fully gelatinized; and then immersing the hybrid scaffold into 1mol/L Tris aqueous solution, taking out after 24h, washing the hybrid scaffold to be neutral by using ultrapure water, and drying the hybrid scaffold in a drying oven at 40 ℃ to obtain the three-dimensionally communicated hydroxyapatite-chitosan-silicon dioxide hybrid scaffold.
The experiment shows that: the fiber diameter of the hydroxyapatite-chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 200 μm, and the compressive strength and young's modulus were as shown in table 1.
Example 4
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; respectively weighing Ca (NO) according to the mass ratio of CS to HA of 5/13)2.4H2O、H3PO4(ensuring that the molar ratio of Ca to P is 1.67) is sequentially added into the solution A, and the mixture is fully stirred until the mixture is uniformly mixed to obtain solution C; according to CS/SiO2Weighing the solution B according to a mass ratio of 10/7, adding the solution B into the solution C, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% for 50 hours to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.42 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a constant temperature and humidity box with the temperature of 30 ℃ and the humidity of 50% for curing for 4D to ensure that the support is fully gelatinized; and then immersing the hybrid scaffold into 1mol/L Tris aqueous solution, taking out after 24h, washing the hybrid scaffold to be neutral by using ultrapure water, and drying the hybrid scaffold in a drying oven at 40 ℃ to obtain the three-dimensionally communicated hydroxyapatite-chitosan-silicon dioxide hybrid scaffold.
The experiment shows that: the fiber diameter of the hydroxyapatite-chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 200 μm, and the compressive strength and young's modulus were as shown in table 1.
Example 5
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; respectively weighing Ca (NO) according to the mass ratio of CS to HA of 10/13)2.4H2O、H3PO4(ensuring that the molar ratio of Ca to P is 1.67) is sequentially added into the solution A, and the mixture is fully stirred until the mixture is uniformly mixed to obtain solution C; according to CS/SiO2Weighing the solution B according to a mass ratio of 10/7, adding the solution B into the solution C, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% for 52 hours to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.39 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a constant temperature and humidity box with the temperature of 30 ℃ and the humidity of 50% for curing for 4D to ensure that the support is fully gelatinized; and then immersing the hybrid scaffold into 1mol/L Tris aqueous solution, taking out after 24h, washing the hybrid scaffold to be neutral by using ultrapure water, and drying the hybrid scaffold in a drying oven at 40 ℃ to obtain the three-dimensionally communicated hydroxyapatite-chitosan-silicon dioxide hybrid scaffold.
The experiment shows that: the fiber diameter of the hydroxyapatite-chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 200 μm, and the compressive strength and young's modulus were as shown in table 1.
Example 6
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; according to CS/SiO2Weighing the solution B according to the mass ratio of 10/7, adding the solution B into the solution A, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% for 56h to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.35 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a refrigerator at the temperature of-20 ℃ for pre-freezing for 48h, and then freeze-drying in a vacuum drier for 2D; taking out the freeze-dried scaffold, immersing the freeze-dried scaffold in 1mol/L Tris aqueous solution, and taking out after 24 hours; repeatedly washing with ultrapure water to neutrality, and freeze-drying in a vacuum drier for 48h to obtain the three-dimensionally communicated chitosan-silicon dioxide hybrid scaffold.
The experiment shows that: the fiber diameter of the chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 450 μm, and the compressive strength and Young's modulus are shown in Table 1.
Example 7
(1) Adding Chitosan (CS) powder with viscosity-average molecular weight of 47 ten thousand and deacetylation degree of 85% into 4% (v/v) dilute acetic acid aqueous solution, stirring to dissolve completely, and making into chitosan stock solution with concentration of 8g/100ml, which is solution A; adding 20g of ethyl silicate into 13.85mL of acetic acid solution (4% (v/v)), and stirring at normal temperature for 6h to fully hydrolyze the ethyl silicate to obtain solution B; respectively weighing Ca (NO) according to the mass ratio of CS to HA of 10/33)2.4H2O、H3PO4(ensuring that the molar ratio of Ca to P is 1.67) is sequentially added into the solution A, and the mixture is fully stirred until the mixture is uniformly mixed to obtain solution C; according to CS/SiO2Weighing the solution B according to a mass ratio of 10/7, adding the solution B into the solution C, stirring to mix uniformly, removing bubbles in vacuum to obtain milky sol, putting the milky sol into a 3D printing material cylinder, and standing for 48 hours in a constant temperature and humidity box with the temperature of 40 ℃ and the humidity of 50% to fully gel.
(2) Pre-forming a bracket: using a 3D-BioplotterTMAnd the printing device is used for printing the three-dimensional support of the preset model.
Setting the printing parameters as follows: extrusion pressure: 0.45 MPa; printing speed: 35 mm/s; needle size inside diameter: 400 μm; platform temperature: and (5) normal temperature.
Parameters of the 3D printing support: fiber diameter: 400 μm; fiber spacing: 400 μm.
(3) And (3) post-treatment of the bracket: placing the support obtained by 3D printing in a refrigerator at the temperature of-20 ℃ for pre-freezing for 48h, and then freeze-drying in a vacuum drier for 2D; taking out the freeze-dried scaffold, immersing the freeze-dried scaffold in 1mol/L Tris aqueous solution, and taking out after 24 hours; repeatedly washing with ultrapure water to neutrality, and freeze-drying in a vacuum drier for 48h to obtain three-dimensionally connected hydroxyapatite-chitosan-silica hybrid scaffold, as shown in FIG. 3.
TABLE 1
Figure BDA0001452863440000081
The experiment shows that: the fiber diameter of the hydroxyapatite-chitosan-silica hybrid scaffold was about 200 μm, the fiber spacing was about 450 μm, and the compressive strength and young's modulus were as shown in table 1.

Claims (10)

1. A method for preparing a high-strength hydroxyapatite-chitosan-silica hybrid scaffold by using 3D printing is characterized by comprising the following steps:
(1) adding chitosan powder into an acid solution, and stirring to fully dissolve the chitosan powder to obtain a uniform and transparent chitosan solution; the acid solution is an aqueous solution of acetic acid, lactic acid or hydrochloric acid;
(2) adding tetraethoxysilane into an acid solution, stirring to fully hydrolyze the tetraethoxysilane to obtain clear and transparent hydrolysate, wherein the acid solution is an aqueous solution of hydrochloric acid, nitric acid, acetic acid or lactic acid;
(3) sequentially adding a calcium source and a phosphorus source into the chitosan solution obtained in the step (1), and stirring to fully dissolve the calcium source and the phosphorus source to obtain a mixed solution, wherein the calcium source is one of calcium acetate, calcium nitrate, calcium hydroxide, calcium hydrophosphate and calcium chloride; the phosphorus source is one of ammonium dihydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, phosphoric acid and triethyl phosphate;
(4) adding the hydrolysate obtained in the step (2) into the mixed solution obtained in the step (3), uniformly stirring, and removing bubbles in vacuum to obtain milky sol;
(5) putting the sol obtained in the step (4) into a 3D printing charging barrel, and standing to gelatinize the sol;
(6) extruding the gel obtained in the step (5) into threads by using 3D-Bioplotter ™ printing equipment, and printing the threads on a printing gasket to form a preset bracket structure to obtain a three-dimensional porous bracket;
(7) fully gelatinizing the three-dimensional porous scaffold obtained in the step (6) to obtain a three-dimensional through porous scaffold;
(8) soaking the stent obtained in the step (7) in alkali liquor to remove the residual acid in the material;
(9) repeatedly washing the bracket obtained in the step (8) with water until p H is neutral; and then drying to obtain the hydroxyapatite-chitosan-silicon dioxide hybrid scaffold.
2. The method according to claim 1, wherein the chitosan in step (1) has a viscosity average molecular weight of 10 to 84 ten thousand and a degree of deacetylation of 75 to 95%; the pH value of the acid solution is 2-6; the mass fraction of the chitosan solution is 4-12%.
3. The method according to claim 1, wherein the molar concentration of the acid solution in the step (2) is 1-3.84 mol/L; the stirring time is 1-10 h.
4. The method according to claim 1, wherein the amount of the calcium source and the phosphorus source used in step (3) is 0-30% of the total mass of the hybrid material, wherein the molar ratio of calcium to phosphorus is 1.67; the hybrid material comprises hydroxyapatite, chitosan and silicon dioxide.
5. The method according to claim 1, wherein the sol of step (4) comprises chitosan and SiO2The mass ratio of (1-2): 1.
6. the method of claim 1, wherein the standing in the step (5) is performed by placing the sol in a constant temperature and humidity chamber with a temperature of 4-40 ℃ and a humidity of 30-70% for 1-5 days.
7. The method according to claim 1, wherein the parameter conditions for printing in step (6) are: the extrusion pressure is 0-0.58 MPa, the printing speed is 10-50 mm/min, and the platform temperature is normal temperature.
8. The method according to claim 1, wherein the step (6) of printing sets the fiber diameter of the three-dimensional porous scaffold to be 0.2-1 mm and the fiber pitch to be 0.1-1 mm.
9. The method according to claim 1, wherein the three-dimensional porous scaffold in the step (7) is sufficiently gelled by placing the three-dimensional porous scaffold in a constant temperature and humidity chamber with a temperature of 20-30 ℃ and a humidity of 30-70% for curing for 3-15 days.
10. The method according to claim 1, wherein the alkali solution in step (8) is an aqueous solution of sodium hydroxide, sodium carbonate, sodium bicarbonate, dipotassium hydrogen phosphate, disodium hydrogen phosphate or tris (hydroxymethyl) aminomethane.
CN201711049313.6A 2017-10-31 2017-10-31 Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing Active CN107661540B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711049313.6A CN107661540B (en) 2017-10-31 2017-10-31 Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711049313.6A CN107661540B (en) 2017-10-31 2017-10-31 Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing

Publications (2)

Publication Number Publication Date
CN107661540A CN107661540A (en) 2018-02-06
CN107661540B true CN107661540B (en) 2020-12-22

Family

ID=61144377

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711049313.6A Active CN107661540B (en) 2017-10-31 2017-10-31 Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing

Country Status (1)

Country Link
CN (1) CN107661540B (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108525012A (en) * 2018-03-13 2018-09-14 华南理工大学 Gradient hybridization timbering material and preparation method thereof is repaired in a kind of bone cartilage integration
CN109251497A (en) * 2018-10-27 2019-01-22 林毅平 A kind of polylactic acid/hydroxy apatite composite material and preparation method thereof that acid fiber by polylactic is strengthened
CN110721346B (en) * 2019-10-30 2020-08-18 中国人民解放军总医院 Biological 3D printing ink and preparation method thereof
CN114524978B (en) * 2021-12-20 2023-04-21 华南理工大学 Chitosan/silicon dioxide nano hybrid material and biomimetic mineralization preparation method and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101601874A (en) * 2009-07-16 2009-12-16 浙江大学 The method for preparing high-strength hydroxylapatite and chitosan three-dimensional composite rod material
CN101848737A (en) * 2007-09-07 2010-09-29 皇家创新公司 Bioactive nanocomposite material
CN102008753A (en) * 2010-12-23 2011-04-13 福州大学 Organic-inorganic composite gel material for bone repair and preparation method thereof
CN102266590A (en) * 2011-07-15 2011-12-07 陕西师范大学 Preparation method of chitosan/hydroxyapatite three-dimensional porous composite bracket material
GB2528504A (en) * 2014-07-24 2016-01-27 Werner Ernst Ludwig Georg Muller Printable morphogenetic phase-specific chitosan-calcium-polyphosphate scaffold for bone repair
JP2017039253A (en) * 2015-08-19 2017-02-23 一般財団法人生産技術研究奨励会 Method for manufacturing carrier for tissue engineering, carrier for tissue engineering, method for manufacturing porous body, and porous body
CN106563169A (en) * 2016-11-04 2017-04-19 王淑芳 Preparation method for preparing nano silicon dioxide-contained collagen-hydroxyapatite bracket
CN106963980A (en) * 2017-04-17 2017-07-21 华南理工大学 A kind of preparation method of chitosan silicon dioxide nano-hybrid material and application

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101848737A (en) * 2007-09-07 2010-09-29 皇家创新公司 Bioactive nanocomposite material
CN101601874A (en) * 2009-07-16 2009-12-16 浙江大学 The method for preparing high-strength hydroxylapatite and chitosan three-dimensional composite rod material
CN102008753A (en) * 2010-12-23 2011-04-13 福州大学 Organic-inorganic composite gel material for bone repair and preparation method thereof
CN102266590A (en) * 2011-07-15 2011-12-07 陕西师范大学 Preparation method of chitosan/hydroxyapatite three-dimensional porous composite bracket material
GB2528504A (en) * 2014-07-24 2016-01-27 Werner Ernst Ludwig Georg Muller Printable morphogenetic phase-specific chitosan-calcium-polyphosphate scaffold for bone repair
JP2017039253A (en) * 2015-08-19 2017-02-23 一般財団法人生産技術研究奨励会 Method for manufacturing carrier for tissue engineering, carrier for tissue engineering, method for manufacturing porous body, and porous body
CN106563169A (en) * 2016-11-04 2017-04-19 王淑芳 Preparation method for preparing nano silicon dioxide-contained collagen-hydroxyapatite bracket
CN106963980A (en) * 2017-04-17 2017-07-21 华南理工大学 A kind of preparation method of chitosan silicon dioxide nano-hybrid material and application

Also Published As

Publication number Publication date
CN107661540A (en) 2018-02-06

Similar Documents

Publication Publication Date Title
CN107661540B (en) Method for preparing high-strength hydroxyapatite-chitosan-silicon dioxide hybrid scaffold by using 3D printing
Liu et al. In vitro evaluation of alginate/halloysite nanotube composite scaffolds for tissue engineering
CN106963980B (en) Preparation method and application of chitosan-silicon dioxide nano hybrid material
Ma et al. Synthesis and characterization of injectable self-healing hydrogels based on oxidized alginate-hybrid-hydroxyapatite nanoparticles and carboxymethyl chitosan
CN109453426A (en) A kind of Bone Defect Repari bioactive ceramics fibrous composite scaffold and preparation method thereof
Vueva et al. Silica/alginate hybrid biomaterials and assessment of their covalent coupling
Jiang et al. Preparation of cellulose nanofiber-reinforced gelatin hydrogel and optimization for 3D printing applications
CN111097068B (en) Bionic hydroxyapatite powder/gelatin/sodium alginate composite 3D printing support and preparation method thereof
CN107265426B (en) template-mediated synthesis of silicon-containing hydroxyapatite material and preparation method thereof
Abbasian et al. Biomimetic nylon 6-baghdadite nanocomposite scaffold for bone tissue engineering
CN107903336B (en) Creatine phosphate modified chitosan material and preparation method and application thereof
CN103386150B (en) Preparation method and application of glucomannan/chitosan composite membrane for conducting tissue regeneration
CN102604146A (en) Inorganic bioactive material and method for preparing polymer porous composite material
CN106421928A (en) Hydroxyapatite/polylactic acid composite material degradable in human bodies and method for preparing hydroxyapatite/polylactic acid composite material
Chen et al. Mineralized self-assembled silk fibroin/cellulose interpenetrating network aerogel for bone tissue engineering
CN106693050A (en) Preparation method for composite scaffold material based on collagen and collagen fibers
CN113234237A (en) Preparation method of high-strength nanocellulose/alginic acid composite hydrogel
Chen et al. In situ strategy for bone repair by facilitated endogenous tissue engineering
CN103007342B (en) Biodegradable and medical tricalcium phosphate/gamma-polyglutamic acid composite and preparation method thereof
CN104707179B (en) Oil-soluble/water-soluble organic-inorganic three-phase porous micro-nanometer composite bone repair material
Miao et al. Engineered bone tissues using biomineralized gelatin methacryloyl/sodium alginate hydrogels
CN1785443A (en) Method of improving anti collapsibility of calcium phosphate skeletal cement using denaturated starch
Abdal-hay et al. An in situ hydrothermal fabrication process of poly (vinyl alcohol)/apatite-like nanocomposites with improved thermal and mechanical properties
CN102875824A (en) Bioactive nano composite polyelectrolyte three-dimensional porous bone tissue engineering scaffold
WO2017028625A1 (en) Pleural/meningeal patch and preparation method therefor

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