CN109134915B - Rare earth phosphate/bioactive polymer three-dimensional porous composite material, and preparation method and application thereof - Google Patents

Rare earth phosphate/bioactive polymer three-dimensional porous composite material, and preparation method and application thereof Download PDF

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CN109134915B
CN109134915B CN201810831941.8A CN201810831941A CN109134915B CN 109134915 B CN109134915 B CN 109134915B CN 201810831941 A CN201810831941 A CN 201810831941A CN 109134915 B CN109134915 B CN 109134915B
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phosphate
rare earth
dimensional porous
chitosan
trivalent
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CN109134915A (en
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郭亚平
赵培培
柯勤飞
郭翠香
张川建
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Shanghai Normal University
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    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/28—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a liquid phase from a macromolecular composition or article, e.g. drying of coagulum
    • 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
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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    • 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
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    • 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
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/58—Materials at least partially resorbable by the body
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    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
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    • C08J2367/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
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    • C08K3/00—Use of inorganic substances as compounding ingredients
    • C08K3/32—Phosphorus-containing compounds
    • C08K2003/321—Phosphates
    • C08K2003/328—Phosphates of heavy metals

Abstract

The invention belongs to the field of inorganic non-metallic materials, organic bioactive high polymer materials and biomedical materials, and particularly relates to a rare earth phosphate/bioactive high polymer three-dimensional porous composite material, a preparation method and application thereof. The preparation method comprises the following steps: uniformly mixing rare earth phosphate and acid-containing bioactive polymer material solution, and freeze-drying; soaking in alkali solution, washing to neutrality, and freeze drying. The structure of the bone conduction membrane has higher porosity and connectivity, good bioactivity, biocompatibility, biodegradability, mechanical property and mechanical property, promotes cell adhesion and growth, promotes bone and osteoinduction, and has wide clinical application prospect.

Description

Rare earth phosphate/bioactive polymer three-dimensional porous composite material, and preparation method and application thereof
Technical Field
The invention belongs to the field of inorganic non-metallic materials, organic bioactive polymer materials and biomedical materials, and particularly relates to a rare earth phosphate/bioactive polymer three-dimensional porous composite material, and a preparation method and application thereof.
Background
Repair and reconstruction of bone defects remains a significant challenge to clinicians, particularly large bone defects caused by trauma, infection, injury, or genetic deformity. To overcome this problem, many bone repair materials having bone forming activity and osteoinductive ability have been clinically used. The most common bone repair materials are tricalcium phosphate (beta-TCP), Hydroxyapatite (HA), Bioactive Glass (BG) and Chitosan (CS), which all have good bioactivity and biocompatibility, but their limited osteoinductive properties do not meet the therapeutic needs of patients for osteoporosis and metabolic disorders. Therefore, the development and design of new materials for bone defects or bone injuries remains a research focus of great interest.
Osteoinductivity can be improved in several ways. First, drug release can improve the osteogenic inductive properties of the material. For example, the osteogenic ratio of HM-ZSM-5/CS/DOX spheroids can be increased by the cumulative release ratio of the DOX drug. Second, growth factors exert osteogenic effects by activating the corresponding signal transduction and regulating osteoblast gene transcription. Third, the reactive metal ions can improve biological responses, including cell proliferation, differentiation, and bone regeneration. For example, the release of Mg and Si ions from the m-MS/PBSu composite scaffold can improve the biological response, improve biocompatibility and osteogenesis.
The prior article reports that an environment favorable for cell growth can be created through the synergistic stimulation of Sr ions and a porous nano-grid structure. In addition, the release rate of Ag ions and the addition of ZnO both reduce cytotoxicity, and provide inherent bacterial resistance and good osteogenic capacity, as well as rapid bone fusion. However, active metal ions such as Zn, Mg, Ag, etc. do not meet the clinical requirements of bone defects at present. Therefore, there is an urgent need to develop a novel bone material having not only good biocompatibility but also good osteoinductive properties.
In recent years, rare earth elements have been widely turned offAnd (6) note. Trace rare earth elements are also found in human bodies and play an important role in cell differentiation, metabolism and tissue regeneration. Luminescent rare earth nanoparticles are increasingly used in nanomedicine due to their excellent physicochemical properties, such as biomedical imaging agents, drug carriers, biomarkers, and the like. Cerium oxide nanoparticles, as catalysts, have surprising pharmacological potential since their antioxidant properties derive from CeO2Middle Ce3+Ions. The study also showed that the cerium oxide nanoparticles were non-toxic to the cells.
Medical therapeutic applications of rare earth-based composites have been extensively studied, including kidney disease and diabetes, as well as the toxicity of rare earths to humans. When used to control hyperphosphatemia, LaCO3Mild metabolic acidosis can be helped to be corrected by increasing serum baking soda concentration. The study showed that La (NO)3)3，La(dpp)3And la (xt) no significant liver or kidney toxicity was found. La (CO)3) The duration of vascular calcification can be delayed. There are also a few studies showing that rare earth composite materials have an effect of treating tumors, and lanthanum-conjugated nanoparticles have been used as effective X-ray radiotherapy drugs for treating cancers. Lanthanum chloride can inhibit the proliferation of tumor cells and induce apoptosis by up-regulating apoptosis-related genes, and at the same time, can regulate the expression of apoptosis proteins in vivo and in vitro by changing cell cycles. To date, rare earth metal-based scaffolds such as La have been rarely used in biomaterials and their role in osteogenesis has not been clear.
The invention provides a rare earth-based bioactive polymer composite material, which not only has good biocompatibility, but also can activate related passages through released rare earth ions to quickly promote osteogenesis, and also has the effects of accelerating cell proliferation and differentiation and the like.
Disclosure of Invention
The invention aims to provide a rare earth phosphate/bioactive polymer three-dimensional porous composite material which has a three-dimensional through porous channel structure, has high porosity and connectivity, good bioactivity, biocompatibility, biodegradability, mechanical properties and mechanical properties, promotes cell adhesion and growth, promotes bone and osteoinduction and has a wide clinical application prospect.
The invention also provides a preparation method of the rare earth phosphate/bioactive polymer three-dimensional porous composite material, which is simple and easy to operate, low in production cost, short in production period, strong in operability, almost free of waste or pollutants and environment-friendly.
The invention has the technical scheme that the rare earth phosphate/bioactive polymer three-dimensional porous composite material is formed by mutually adhering sheet structures of bioactive polymer material doped with rare earth phosphate particles to form a three-dimensional through porous channel structure, and the rare earth phosphate particles are uniformly distributed on the surface and inside the sheet structures. The bioactive high polymer material is doped with a flaky structure of rare earth phosphate particles to form a pore wall of a porous channel structure, and the bioactive high polymer material is coated on the surfaces of the rare earth phosphate particles attached to the surfaces of the flaky structure. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The aperture of the three-dimensional through multi-pore channel structure is 10-200 mu m, preferably 50-100 mu m; the porosity is 85-95%.
The bioactive high molecular material is selected from any one or combination of chitosan, collagen, polyvinylpyrrolidone, polyhydroxybutyrate and polycaprolactone.
The rare earth phosphate comprises lanthanum phosphate, cerium phosphate, gadolinium phosphate, ytterbium phosphate, europium phosphate or samarium phosphate, and the particle size of the rare earth phosphate is 50 nm-5 mu m.
The mass ratio of the rare earth phosphate to the bioactive high polymer material is 1: 0.5-5, preferably 1: 0.5 to 2, more preferably 1: 1.
the preparation method of the rare earth phosphate/bioactive polymer three-dimensional porous composite material comprises the following steps:
(1) and (3) uniformly mixing the rare earth phosphate and the acid-containing bioactive polymer material solution, and freeze-drying to obtain the rare earth phosphate precursor/bioactive polymer composite material.
(2) And (3) soaking the rare earth phosphate precursor/bioactive polymer material in alkali liquor, washing to be neutral, and freeze-drying to obtain the rare earth phosphate/bioactive polymer three-dimensional porous composite material. The alkali liquor soaking can neutralize the acid in the rare earth phosphate precursor/bioactive polymer material and promote the deposition of bioactive polymer material.
In order to prepare a certain shape, the step (1) is specifically that the rare earth phosphate and the acid-containing bioactive polymer material solution are uniformly mixed, placed in a mould, and then freeze-dried.
In the step (1), the dosage ratio of the rare earth phosphate to the bioactive polymer material is 1: 0.5-5, preferably 1: 0.5 to 2, more preferably 1: 1; the concentration of the bioactive polymer material is 0.005 g/L-saturated solution, preferably 10 g/L-saturated solution, and more preferably 20 g/L-40 g/L; preferably, the concentration is 40 g/L; contains 0.2-5% of acid by volume fraction, specifically, 0.5-5% of organic acid by volume fraction, preferably 2%; the volume fraction of the inorganic acid is 0.2% to 1%, preferably 0.5%.
The acid comprises organic acid and inorganic acid, the organic acid comprises acetic acid, formic acid, propionic acid, oxalic acid or citric acid, etc., preferably acetic acid; the inorganic acid includes hydrochloric acid, sulfuric acid, phosphoric acid, etc., and hydrochloric acid is preferred.
Step (1), freeze-drying for 30 min-700 h at-85-0 ℃ and 1-50 pa. The temperature of freeze drying is preferably-85 ℃ to-60 ℃, and more preferably-80 ℃; the vacuum degree of freeze drying is preferably 1-10 pa, more preferably 1-2 pa; the freeze drying time is preferably 24-120 h, and more preferably 48 h.
The preparation method of the acid-containing bioactive polymer material solution comprises the following steps: adding bioactive polymer material into acid-containing solvent, and mixing.
In the acid-containing bioactive polymer material solution, the volume fraction of acid is 0.2-5%, specifically, the volume fraction of organic acid is 0.5-5%, preferably 2%; the volume fraction of the inorganic acid is 0.2% to 1%, preferably 0.5%.
The acid comprises organic acid and inorganic acid, the organic acid comprises acetic acid, formic acid, propionic acid, oxalic acid or citric acid, etc., preferably acetic acid; the inorganic acid includes hydrochloric acid, sulfuric acid, phosphoric acid, etc., and hydrochloric acid is preferred.
In the acid-containing bioactive polymer material solution, the concentration of the bioactive polymer material is 0.005 g/L-saturated solution, preferably 10 g/L-saturated solution, and further preferably 20 g/L-40 g/L; preferably, it is 40 g/L.
The solvent is selected from any one or combination of water, alcohols or esters, the alcohols comprise ethanol, methanol or propanol, and ethanol is preferred; esters include glycerol or ethyl acetate, preferably ethyl acetate.
The preparation method of the rare earth phosphate comprises the following steps:
a. adding soluble trivalent rare earth salt or soluble trivalent rare earth salt solution into soluble phosphate solution with the pH value of 7-12 at the temperature of 10-60 ℃ under the condition of stirring and uniformly mixing;
b. after mixing, stirring for 0.5-2 h at 70-90 ℃, then stirring for 24-48 h at 10-60 ℃, filtering and washing to obtain a neutral precipitate;
c. drying the neutral precipitate for 3-30 h at 50-90 ℃, and calcining for 2-6 h at 500-1000 ℃ to prepare the rare earth phosphate.
Step a, the preparation method of the soluble phosphate solution with the pH value of 7-12 comprises the following steps: dissolving soluble phosphate in water, and adding ammonia water to adjust the pH value to 7-12.
Step a, the preparation method of the soluble trivalent rare earth salt solution comprises the following steps: dissolving the soluble trivalent rare earth salt in water.
Step a, the soluble phosphate comprises a dihydrogen phosphate, a hydrogen phosphate or an orthophosphate, preferably a hydrogen phosphate. The soluble trivalent rare earth salt comprises trivalent lanthanum salt, trivalent cerium salt, trivalent gadolinium salt, trivalent ytterbium salt, trivalent europium salt or trivalent samarium salt.
In the step a, the pH value of the soluble phosphate solution is preferably 11-11.5.
And a preferable scheme of the step a is that under the conditions of stirring and 20-40 ℃, soluble trivalent rare earth salt or soluble trivalent rare earth salt solution is added into the soluble phosphate solution with the pH value of 7-12 and is uniformly mixed.
Step a, slowly adding soluble trivalent rare earth salt or a soluble trivalent rare earth salt solution into a soluble phosphate solution, specifically, dropwise adding the soluble trivalent rare earth salt solution into the soluble phosphate solution.
In the uniformly mixing system of the step a, the molar ratio of the trivalent rare earth ions to the phosphate radicals is 1: 0.5-5, preferably 1: 1; the concentration of the trivalent rare earth ions is 0.05-0.1 mol/L, preferably 0.06-0.07 mol/L.
B, the first stirring temperature is preferably 90 ℃, and the first stirring time is preferably 1 h; the post-stirring temperature is preferably 20-40 ℃, and the post-stirring time is preferably 24-36 h. Preferably, the mixture is stirred for 1 hour at 90 ℃ and then for 24 hours at 40 ℃.
C, the first drying temperature is preferably 80-90 ℃, and the first drying time is 5-6 hours; the post-calcining temperature is preferably 800-1000 ℃, and the post-calcining time is preferably 3-4 h. Preferably, the drying is performed for 5-6 h at 90 ℃, and then the calcination is performed for 3h at 1000 ℃.
The particle size of the rare earth phosphate prepared in the step c is 50 nm-5 mu m.
Step (2), the concentration of the alkali liquor is 0.05-0.2 mol/L, preferably 0.05-0.1 mol/L; the alkali contained in the alkali liquor is selected from any one or combination of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, ammonium carbonate and potassium bicarbonate, and preferably the combination of sodium hydroxide or potassium hydroxide and any one of sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, ammonium carbonate and potassium bicarbonate.
Soaking in alkali liquor for 4-48 h at the temperature of 20-50 ℃, wherein the soaking temperature of the alkali liquor is preferably 30-40 ℃, and more preferably 30 ℃; the time of soaking in alkali liquor is preferably 24 h; the pressure of the alkali liquor soaking is normal pressure-2 MPa.
And (2) washing with deionized water until the pH value is approximately equal to 7.0.
And (2) freeze-drying for 30 min-700 h at-85-0 ℃ and 1-50 pa. The temperature of freeze drying is preferably-85 ℃ to-60 ℃, and more preferably-80 ℃; the vacuum degree of freeze drying is preferably 1-10 pa, more preferably 1-2 pa; the freeze drying time is preferably 24-120 h, and more preferably 48 h.
The rare earth phosphate/bioactive polymer three-dimensional porous composite material prepared by the method has a three-dimensional through porous channel structure with uniform micropore size and distribution, high porosity and connectivity, good biocompatibility, mechanical property and mechanical property, and improved contact performance with bone defect tissues; good biodegradability, release rare earth ions along with biodegradation, improve cell activity and promote cell growth and adhesion; the bone regeneration promoting agent has good bone promoting and bone inducing effects, improves the bone regeneration capability, has certain reinforcing and promoting effects on the repair of bone defect parts, and is suitable for preparing bone repair materials.
Compared with the prior art, the invention has the beneficial effects that:
(1) the rare earth phosphate/bioactive polymer three-dimensional porous composite material prepared by the invention has a three-dimensional through porous channel structure, high porosity and large pore diameter, is beneficial to cell adhesion, spreading and growth, and transformation to bone tissues, and improves bone tissue repair activity.
(2) The rare earth phosphate/bioactive polymer three-dimensional porous composite material prepared by the invention has good mechanical and mechanical properties, good bioactivity, biocompatibility, biodegradability and low immunogenicity, and is an excellent bone repair material.
(3) The rare earth phosphate/bioactive polymer three-dimensional porous composite material takes soluble rare earth salt, soluble phosphate and bioactive polymer material as raw materials, adopts a freeze-drying method, has cheap and easily-obtained raw materials, simple preparation process, low cost investment, strong operability, short period and almost no waste, and is an economic and environment-friendly synthesis method.
Drawings
FIG. 1 is an SEM image of a lanthanum phosphate/chitosan three-dimensional porous scaffold prepared in example 1.
Fig. 2 is an XRD pattern of chitosan (a), lanthanum phosphate particles (b), lanthanum phosphate/chitosan three-dimensional porous scaffold (c) prepared in example 1.
Fig. 3 is a diagram of chitosan (a), lanthanum phosphate particles (b), lanthanum phosphate/chitosan three-dimensional porous scaffold prepared in example 1 (c) FIRT.
FIG. 4 shows beta-phase calcium phosphate/chitosan scaffolds (beta-TCP/CS, A and C), lanthanum phosphate/chitosan three-dimensional porous scaffold (LaPO) prepared in example 14SEM images of cell adhesion of/CS, B and D), wherein C is an enlarged view of a portion E in A, and D is an enlarged view of a portion F in B.
Fig. 5 is a lanthanum ion release curve diagram of the lanthanum phosphate/chitosan three-dimensional porous scaffold prepared in example 1 in simulated body fluid.
FIG. 6 shows beta-phase calcium phosphate/chitosan scaffolds (beta-TCP/CS, A and C), gadolinium phosphate/chitosan three-dimensional porous scaffold prepared in example 5 (GdPO)4Micro-CT images of skull defect repair of/CS, B and D).
FIG. 7 shows a beta-phase calcium phosphate/chitosan scaffold (beta-TCP/CS, A and C), gadolinium phosphate/chitosan three-dimensional porous scaffold prepared in example 5 (GdPO)4The morphological staining graph of bone tissue of/CS, B and D), wherein C is the magnified image of the E part in A, and D is the magnified image of the F part in B.
Detailed Description
The present invention is further illustratively described in detail below with reference to specific examples. The description of the embodiments is provided to assist understanding of the present invention, but the present invention is not limited thereto.
Example 1
(1) 2.9877g La (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Separately dissolved in 100mL of deionized water to prepare La (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11.5 with O, and then adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 20 ℃ for mechanical stirring and La (NO) is added dropwise3)3Adding dropwise the aqueous solution, stirring at 90 deg.C for 1 hr, stirring at 20 deg.C for 24 hr, and filteringWashing to obtain neutral precipitate, drying at 50 deg.C for 12 hr, calcining at 500 deg.C for 6 hr to obtain lanthanum phosphate (LaPO)4) Powder with a particle size of 50nm to 5 μm.
(3) And adding 4.0g of chitosan into 100mL of acetic acid solution with the volume fraction of 1%, and stirring to completely dissolve the chitosan until the chitosan is clear and transparent.
(4) And (4.0 g of lanthanum phosphate powder is added into the chitosan solution prepared in the step (3), and the mixture is magnetically stirred uniformly to obtain a lanthanum phosphate precursor/chitosan slurry.
(5) And (3) transferring the lanthanum phosphate precursor/chitosan slurry prepared in the step (4) to a mold with the diameter of 12mm multiplied by 18mm multiplied by the height, and freeze-drying the lanthanum phosphate precursor/chitosan slurry for 48 hours at the temperature of minus 80 ℃ and under the pressure of 1-2 Pa to obtain the preliminarily molded lanthanum phosphate precursor/chitosan three-dimensional porous scaffold.
(6) And (3) transferring the preliminarily formed lanthanum phosphate precursor/chitosan three-dimensional porous scaffold into a NaOH solution with the concentration of 0.2mol/L, taking out after reacting for 1 day at 20 ℃, washing with deionized water until the pH value is approximately equal to 7.0, freeze-drying (the same freeze-drying condition as the step (5)), and converting the lanthanum phosphate precursor/chitosan three-dimensional porous scaffold into a lanthanum phosphate/chitosan three-dimensional porous scaffold with the aperture of 10-200 mu m and the porosity of 85-95%.
The scanning electron micrograph of the lanthanum phosphate/chitosan three-dimensional porous scaffold prepared in this example is shown in fig. 1, and lanthanum phosphate (LaPO) is doped with chitosan4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely lanthanum phosphate (LaPO)4) Particles are uniformly distributed on the surface and inside of the sheet structure, and lanthanum phosphate (LaPO) attached to the surface of the sheet structure4) The surface of the particles is coated with chitosan. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD patterns of the chitosan (a), the lanthanum phosphate particles (b) and the lanthanum phosphate/chitosan three-dimensional porous scaffold (c) prepared in this example are shown in fig. 2, and the lanthanum phosphate/chitosan three-dimensional porous scaffold has no impurity peak and has characteristic peaks of both chitosan and lanthanum phosphate.
FTIR patterns of chitosan (a), lanthanum phosphate particles (b) and lanthanum phosphate/chitosan three-dimensional porous scaffold (c) prepared in this example are shown in FIG. 3, and lanthanum phosphate/chitosan three-dimensional multi-porous scaffoldThe pore scaffold has the characteristic peaks of both chitosan and lanthanum phosphate and is 993cm-1The characteristic peak of absorption of phosphate radical appears.
As can be seen from fig. 2 and 3, the method of the present invention has prepared a three-dimensional porous scaffold having lanthanum phosphate and chitosan as components.
Beta-phase calcium phosphate/Chitosan (beta-TCP/CS) scaffolds and lanthanum phosphate/Chitosan (LaPO) prepared in this example4CS) cell adhesion diagram of the three-dimensional porous scaffold is shown in figure 4, A and C are cell adhesion diagrams of a beta-phase calcium phosphate/chitosan scaffold, and C is an enlarged view of a part E in A; b and D are cell adhesion figures of the lanthanum phosphate/chitosan three-dimensional porous scaffold, and D is an enlarged view of a part F in the B. As shown in fig. 4, compared to the β -phase calcium phosphate/chitosan scaffold, the cells spread well on the surface of the lanthanum phosphate/chitosan three-dimensional porous scaffold, the bioactivity remains intact, and the biocompatibility is good.
Lanthanum phosphate/chitosan (LaPO) prepared in this example was assayed at 37 ℃ for 6h, 12h, 24h, 48h, 72h, 96h, and 120h, respectively4The concentration of lanthanum ions contained in the CS three-dimensional porous scaffold in simulated body fluid is obtained to obtain lanthanum phosphate/chitosan (LaPO)4CS) lanthanum ion release profile of three-dimensional porous scaffold, lanthanum ion was slowly released in the simulated body fluid due to slow degradation of chitosan attached to the surface of lanthanum ion, which was released at an effective concentration of 0.6 μmoL in the simulated body fluid, as shown in fig. 5.
Example 2
(1) 2.9877g La (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Separately dissolved in 100mL of deionized water to prepare La (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 20 ℃ for mechanical stirring and La (NO) is added dropwise3)3Adding the aqueous solution, stirring at 70 deg.C for 1 hr, stirring at 20 deg.C for 24 hr, filtering, and mixingWashing to obtain neutral precipitate, drying at 90 deg.C for 5 hr, calcining at 1000 deg.C for 6 hr to obtain lanthanum phosphate (LaPO)4) Powder with a particle size of 50nm to 5 μm.
(3) And (3) adding 2.0g of collagen into 100mL of acetic acid solution with the volume fraction of 4%, and stirring to completely dissolve the collagen until the collagen is clear and transparent.
(4) And (4.0 g of lanthanum phosphate powder is added into the collagen solution prepared in the step (3), and the mixture is magnetically stirred uniformly to obtain a lanthanum phosphate precursor/collagen slurry.
(5) And transferring the lanthanum phosphate precursor/collagen slurry into a mold with the diameter of 12mm multiplied by 18mm multiplied by the height, and freeze-drying for 72 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily formed lanthanum phosphate precursor/collagen three-dimensional porous scaffold.
(6) Transferring the preliminarily formed lanthanum phosphate precursor/collagen three-dimensional porous scaffold to Na with the concentration of 0.2mol/L2CO3And (3) reacting in the solution at 30 ℃ for 1 day, taking out, washing with deionized water until the pH value is approximately equal to 7.0, and freeze-drying (the freeze-drying condition is the same as the step (5)), so that the lanthanum phosphate precursor/collagen three-dimensional porous scaffold is converted into a lanthanum phosphate/collagen three-dimensional porous scaffold, the aperture is 10-200 mu m, and the porosity is 85-95%.
The structure of the lanthanum phosphate/collagen three-dimensional porous scaffold prepared in this example shown in the scanning electron microscope is similar to that of example 1, and lanthanum phosphate (LaPO) is doped with collagen4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely lanthanum phosphate (LaPO)4) Particles are uniformly distributed on the surface and inside of the sheet structure, and lanthanum phosphate (LaPO) attached to the surface of the sheet structure4) The surface of the particles is coated with collagen. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the lanthanum phosphate/collagen three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold comprising lanthanum phosphate and collagen as components was prepared.
The cell adhesion result of the lanthanum phosphate/collagen three-dimensional porous scaffold prepared in the embodiment is similar to that of the embodiment 1, the cell spreading state is good, and the biocompatibility is good.
Example 3
(1) 2.9961g Ce (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolved in 100mL of deionized water to prepare Ce (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 40 ℃ for mechanical stirring and simultaneously dripping Ce (NO)3)3Adding the aqueous solution dropwise, stirring at 90 deg.C for 2 hr, stirring at 40 deg.C for 24 hr, filtering, washing to obtain neutral precipitate, drying at 90 deg.C for 5 hr, calcining at 1000 deg.C for 3 hr to obtain cerous phosphate (CePO)4) Powder with a particle size of 50nm to 5 μm.
(3) And adding 4.0g of collagen into 100mL of hydrochloric acid solution with the volume fraction of 0.5%, and continuously stirring to completely dissolve the collagen until the collagen is clear and transparent.
(4) And adding 2.0g of cerium phosphate powder into the collagen solution, and uniformly stirring by magnetic force to obtain a cerium phosphate precursor/collagen slurry.
(5) And transferring the cerium phosphate precursor/collagen slurry into a mold with the diameter of 12mm multiplied by 18mm multiplied by the height, and freeze-drying for 24 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily molded cerium phosphate precursor/collagen three-dimensional porous scaffold.
(6) And (3) transferring the preliminarily formed cerium phosphate precursor/collagen three-dimensional porous scaffold to a NaOH solution with the concentration of 0.05mol/L, reacting for 1 day at 30 ℃, taking out, washing with deionized water until the pH value is approximately equal to 7.0, freeze-drying (the same freeze-drying condition as the step (5)), and converting the cerium phosphate precursor/collagen three-dimensional porous scaffold into the cerium phosphate/collagen three-dimensional porous scaffold with the aperture of 10-200 mu m and the porosity of 85-95%.
The structure of the cerium phosphate/collagen three-dimensional porous scaffold prepared in this example, which is shown in the scanning electron micrograph, is similar to that of example 1, and cerium phosphate (CePO) is doped with collagen4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely cerium phosphate (CePO)4) Cerium phosphate (CePO) with particles uniformly distributed on and in the surface of the sheet structure and attached on the surface of the sheet structure4) The surface of the particles is coated with collagen. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the cerium phosphate/collagen three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold comprising cerium phosphate and collagen as components was prepared.
The cell adhesion result of the cerium phosphate/collagen three-dimensional porous scaffold prepared in the example is similar to that of the scaffold prepared in the example 1, the cell spreading state is good, and the biocompatibility is good.
Example 4
(1) 2.9961g Ce (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolved in 100mL of deionized water to prepare Ce (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11.5 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 20 ℃ for mechanical stirring and simultaneously dripping Ce (NO)3)3Adding the aqueous solution dropwise, stirring at 90 deg.C for 1 hr, stirring at 20 deg.C for 24 hr, filtering, washing to obtain neutral precipitate, drying at 60 deg.C for 24 hr, calcining at 500 deg.C for 6 hr to obtain cerous phosphate (CePO)4) Powder with a particle size of 50nm to 5 μm.
(3) 100mL of 2% acetic acid solution is added with 4.0g of polyhydroxybutyrate, and the solution is stirred continuously to dissolve the polyhydroxybutyrate until the solution is clear and transparent.
(4) Adding 4.0g of cerium phosphate powder into the polyhydroxybutyrate solution, and uniformly stirring by magnetic force to obtain a cerium phosphate precursor/polyhydroxybutyrate slurry.
(5) And transferring the cerium phosphate precursor/polyhydroxybutyrate slurry to a mold with the diameter of 12mm multiplied by 18mm multiplied by the height, and freeze-drying for 48 hours at the temperature of-85 ℃ and under the condition of 1-2 Pa to obtain the cerium phosphate precursor/polyhydroxybutyrate three-dimensional porous scaffold.
(6) Transferring the cerium phosphate precursor/polyhydroxybutyrate three-dimensional porous scaffold into a NaOH solution with the concentration of 0.1mol/L, reacting for 1 day at 30 ℃, taking out, washing with deionized water until the pH value is approximately equal to 7.0, freeze-drying (the same freeze-drying condition as the step (5)), and converting the cerium phosphate precursor/polyhydroxybutyrate three-dimensional porous scaffold into a cerium phosphate/polyhydroxybutyrate three-dimensional porous scaffold with the aperture of 10-200 mu m and the porosity of 85-95%.
The scanning electron micrograph of the cerium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in this example shows a structure similar to that of example 1, and the polyhydroxybutyrate-doped cerium phosphate (CePO) is used4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely cerium phosphate (CePO)4) Cerium phosphate (CePO) with particles uniformly distributed on and in the surface of the sheet structure and attached on the surface of the sheet structure4) The surface of the granule is coated with polyhydroxybutyrate. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the cerium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold composed of cerium phosphate and polyhydroxybutyrate as constituent components was prepared.
The cell adhesion result of the cerium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in the embodiment is similar to that of the scaffold prepared in the embodiment 1, the cell spreading state is good, and the biocompatibility is good.
Example 5
(1) 3.11gGd (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolving in 100mL deionized water to prepare Gd (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 25 ℃ for mechanical stirring and Gd (NO) is added dropwise3)3Stirring the aqueous solution at 90 deg.C for 1 hr after dropwise addition, stirring at 25 deg.C for 24 hr, filtering and washing to obtain neutral precipitate, drying at 90 deg.C for 10 hr, calcining at 1000 deg.C for 3 hr to obtain gadolinium phosphate (GdPO)4) Powder with a particle size of 50nm to 5 μm.
(3) 100mL of acetic acid solution with volume fraction of 1 percent is taken, 4.0g of chitosan is added, and stirring is continuously carried out to dissolve the chitosan until the chitosan is clear and transparent.
(4) And adding 4.0g of gadolinium phosphate powder into the chitosan solution, and uniformly stirring by magnetic force to obtain gadolinium phosphate precursor/chitosan slurry.
(5) And transferring the gadolinium phosphate precursor/chitosan slurry into a mold with the diameter being multiplied by 18mm (the diameter being multiplied by the height), and freeze-drying for 48 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily molded gadolinium phosphate precursor/chitosan three-dimensional porous scaffold.
(6) Transferring the preliminarily formed gadolinium phosphate precursor/chitosan three-dimensional porous scaffold to Na with the concentration of 0.05mol/L2CO3And (3) reacting in the solution at 30 ℃ for 1 day, taking out, washing with deionized water until the pH value is approximately equal to 7.0, and freeze-drying (the freeze-drying condition is the same as the step (5)), so that a gadolinium phosphate precursor/chitosan three-dimensional porous scaffold is converted into a gadolinium phosphate/chitosan three-dimensional porous scaffold, the aperture is 10-200 mu m, and the porosity is 85-95%.
The scanning electron micrograph of the gadolinium phosphate/chitosan three-dimensional porous scaffold prepared in this example shows a structure similar to that of example 1, and the gadolinium phosphate (GdPO) is doped with chitosan4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely gadolinium phosphate (GdPO)4) The particles are uniformly distributed on the surface and inside of the sheet structure, and gadolinium phosphate (GdPO) attached to the surface of the sheet structure4) The surface of the particles is coated with chitosan. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the gadolinium phosphate/chitosan three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold comprising gadolinium phosphate and chitosan as constituent components was prepared.
The cell adhesion result of the gadolinium phosphate/chitosan three-dimensional porous scaffold prepared in this example is similar to that of example 1, the cell spreading state is good, and the biocompatibility is good.
Beta-phase calcium phosphate/chitosan (beta-TCP/CS) scaffolds and gadolinium phosphate/chitosan (GdPO) prepared in this example4CS) skull defect repair X-ray Micro tomography imaging (Micro-CT) of three-dimensional porous scaffolds is shown in fig. 6. In fig. 6, a is skull defect imaging at 0 weeks after the implantation of the beta-phase calcium phosphate/chitosan scaffold, C is skull defect imaging at 12 weeks after the implantation of the beta-phase calcium phosphate/chitosan scaffold, and B is implanted gadolinium phosphate/chitosan (GdPO)4Per CS) skull defect imaging at 0 week after three-dimensional porous scaffold, D is implanted gadolinium phosphate/chitosan (GdPO)4/CS) skull defect imaging at 12 weeks post three-dimensional porous scaffold.
As can be seen from FIG. 6, gadolinium phosphate/chitosan (GdPO) prepared in this example was implanted compared to a beta-phase calcium phosphate/chitosan scaffold4After the/CS) three-dimensional porous scaffold, the repair area of skull defect is obviously increased, and the gadolinium phosphate/chitosan (GdPO) prepared in the embodiment4the/CS) three-dimensional porous scaffold has obvious bone-promoting effect.
For the respective implants of beta-phase calcium phosphate/chitosan (beta-TCP/CS) scaffold and gadolinium phosphate/chitosan (GdPO) prepared in this example4CS) three-dimensional porous scaffolds bone tissue repaired for 12 weeks was morphologically and histologically stained, and the staining results are shown in fig. 7. In fig. 7, A is a histological staining pattern of the implanted beta-phase calcium phosphate/chitosan scaffold, and C is an enlarged view of the portion E in A; b is implanted gadolinium phosphate/chitosan (GdPO)4CS) histological staining pattern of three-dimensional porous scaffold, D is the enlarged view of F part in B.
As can be seen from FIG. 7, gadolinium phosphate/chitosan (GdPO) prepared in this example was implanted compared to a beta-phase calcium phosphate/chitosan scaffold4After the/CS) three-dimensional porous scaffold, the repair area of the bone defect is obviously increased, and the gadolinium phosphate chitosan (GdPO) prepared in the embodiment4the/CS) three-dimensional porous scaffold has obvious bone-promoting effect.
The bone-promoting effect of the rare earth phosphate/bioactive polymer three-dimensional porous scaffolds prepared in examples 1 to 4 is similar to that of example 5, and shows a significant bone-promoting effect.
Example 6
(1) 3.223gYb (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolved in 100mL of deionized water to prepare Yb (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 25 ℃ for mechanical stirring and Yb (NO) is dripped into the oil bath kettle3)3Adding dropwise the aqueous solution, stirring at 90 deg.C for 1 hr, stirring at 25 deg.C for 24 hr, filtering, washing to obtain neutral precipitate, drying at 90 deg.C for 6 hr, calcining at 1000 deg.C for 3 hr to obtain YbPO (YbPO)4) Powder with a particle size of 50nm to 5 μm.
(3) 100mL of 0.5 volume percent hydrochloric acid solution is added with 4.0g of polyhydroxybutyrate, and the mixture is continuously stirred to be dissolved until the mixture is clear and transparent.
(4) Adding 4.0g of ytterbium phosphate powder into the polyhydroxybutyrate solution, and uniformly stirring by magnetic force to obtain the ytterbium phosphate precursor/polyhydroxybutyrate slurry.
(5) And transferring the ytterbium phosphate precursor/polyhydroxybutyrate slurry into a mold with the diameter being multiplied by 18mm (the height being multiplied by the diameter), and freeze-drying for 96 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily formed ytterbium phosphate precursor/polyhydroxybutyrate three-dimensional porous support.
(6) Transferring the preliminarily formed ytterbium phosphate precursor/polyhydroxybutyrate three-dimensional porous scaffold into a NaOH solution with the concentration of 0.2mol/L, reacting for 1 day at 30 ℃, taking out the scaffold, washing the scaffold with deionized water until the pH value is approximately 7.0, freeze-drying (the same freeze-drying condition as the step (5)), converting the ytterbium phosphate precursor/polyhydroxybutyrate three-dimensional porous scaffold into a ytterbium phosphate/polyhydroxybutyrate three-dimensional porous scaffold with the aperture of 10-200 mu m and the porosity of 85-95%.
Scanning electron microscope image of the ytterbium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in the embodiment shows a structureYtterbium phosphate (YbPO) was doped with polyhydroxybutyrate in a similar manner to example 14) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely ytterbium phosphate (YbPO)4) Ytterbium phosphate (YbPO) with particles uniformly distributed on and in the surface of the sheet structure and attached on the surface of the sheet structure4) The surface of the granule is coated with polyhydroxybutyrate. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the ytterbium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold composed of ytterbium phosphate and polyhydroxybutyrate as constituent components was prepared.
The cell adhesion result of the ytterbium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in the embodiment is similar to that of the scaffold prepared in the embodiment 1, the cell spreading state is good, and the biocompatibility is good.
The bone-promoting effect of the ytterbium phosphate/polyhydroxybutyrate three-dimensional porous scaffold prepared in the embodiment is similar to that of the embodiment 5, and the bone-promoting effect is obvious.
Example 7
(1) 3.063g of Sm (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolved in 100mL of deionized water to prepare Sm (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4The aqueous solution is put into an oil bath kettle at 40 ℃ for mechanical stirring and Sm (NO) is dripped into the oil bath kettle3)3Adding dropwise water solution, stirring at 90 deg.C for 1 hr, stirring at 40 deg.C for 24 hr, filtering, washing to obtain neutral precipitate, drying at 60 deg.C for 30 hr, calcining at 1000 deg.C for 3 hr to obtain samarium phosphate (SmPO)4) Powder with a particle size of 50nm to 5 μm.
(3) And (3) adding 2.0g of polycaprolactone into 100mL of acetic acid solution with the volume fraction of 4%, and continuously stirring to dissolve the polycaprolactone until the polycaprolactone is clear and transparent.
(4) And adding 4.0g of samarium phosphate powder into the polycaprolactone solution, and magnetically stirring uniformly to obtain the samarium phosphate precursor/polycaprolactone slurry.
(5) And transferring the samarium phosphate precursor/polycaprolactone slurry to a mold with the diameter of 12mm multiplied by 18mm (the diameter is multiplied by the height), and freeze-drying for 48 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily molded samarium phosphate precursor/polycaprolactone three-dimensional porous scaffold.
(6) Transferring the preliminarily formed samarium phosphate precursor/polycaprolactone three-dimensional porous scaffold to Na with the concentration of 0.05mol/L2CO3And (3) reacting in the solution at 30 ℃ for 1 day, taking out the scaffold, washing the scaffold with deionized water until the pH value is approximately 7.0, and freeze-drying (the freeze-drying condition is the same as the step (5)), so that the samarium phosphate precursor/polycaprolactone three-dimensional porous scaffold is converted into the samarium phosphate/polycaprolactone three-dimensional porous scaffold, the aperture is 10-200 mu m, and the porosity is 85-95%.
The scanning electron micrograph of the samarium phosphate/polycaprolactone three-dimensional porous scaffold prepared in this example shows a structure similar to that of example 1, and the polycaprolactone-doped samarium phosphate (SmPO) is used4) The flaky structure of the particles is a pore wall, and the particles are mutually adhered to form a three-dimensional through porous channel structure, namely samarium phosphate (SmPO)4) Particles are uniformly distributed on the surface and inside of the sheet structure, and samarium phosphate (SmPO) is attached to the surface of the sheet structure4) The particle surface is wrapped with polycaprolactone. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the samarium phosphate/polycaprolactone three-dimensional porous scaffold prepared in this example showed the results similar to those of example 1, and a three-dimensional porous scaffold composed of samarium phosphate and polycaprolactone as a constituent was prepared.
The cell adhesion result of the samarium phosphate/polycaprolactone three-dimensional porous scaffold prepared by the embodiment is similar to that of the embodiment 1, the cell spreading state is good, and the biocompatibility is good.
The bone-promoting effect of the samarium phosphate/polycaprolactone three-dimensional porous scaffold prepared in the embodiment is similar to that of the embodiment 5, and the samarium phosphate/polycaprolactone three-dimensional porous scaffold shows an obvious bone-promoting effect.
Example 8
(1) 3.077gEu (NO)3)3·6H2O and 0.9112g (NH)4)2HPO4Respectively dissolved in 100mL of deionized water to prepare Eu (NO)3)3Aqueous solution and (NH)4)2HPO4An aqueous solution.
(2) To (NH)4)2HPO4Adding NH to the aqueous solution3·H2Adjusting pH to 11 with O, and adding (NH)4)2HPO4Putting the aqueous solution into a 40 ℃ oil bath kettle, mechanically stirring and simultaneously dropwise adding Eu (NO)3)3Adding the aqueous solution dropwise, stirring at 90 deg.C for 1 hr, stirring at 40 deg.C for 24 hr, filtering, washing to obtain neutral precipitate, drying at 90 deg.C for 5 hr, calcining at 1000 deg.C for 3 hr to obtain europium phosphate (EuPO)4) Powder with a particle size of 50nm to 5 μm.
(3) 100mL of 2% acetic acid solution is added with 4.0g of chitosan, and the mixture is continuously stirred to be dissolved until the mixture is clear and transparent.
(4) 4.0g of europium phosphate powder is added into the chitosan solution, and the mixture is magnetically stirred uniformly to obtain europium phosphate precursor/chitosan slurry.
(5) And transferring the europium phosphate precursor/chitosan slurry into a mold with the diameter of 12mm multiplied by 18mm multiplied by the height, and freeze-drying for 48 hours at the temperature of-60 ℃ and under the condition of 1-2 Pa to obtain the preliminarily molded europium phosphate precursor/chitosan three-dimensional porous scaffold.
(6) And (3) transferring the preliminarily formed europium phosphate precursor/chitosan three-dimensional porous scaffold into a NaOH solution with the concentration of 0.1mol/L, reacting for 1 day at 30 ℃, taking out, washing with deionized water until the pH value is approximately equal to 7.0, freeze-drying (the same freeze-drying condition as the step (5)), and converting the europium phosphate precursor/chitosan three-dimensional porous scaffold into the europium phosphate/chitosan three-dimensional porous scaffold, wherein the aperture is 10-200 mu m, and the porosity is 85-95%.
The scanning electron micrograph of the europium phosphate/chitosan three-dimensional porous scaffold prepared in this example shows a structure similar to that of example 1, and europium phosphate (EuPO) is doped with chitosan4) The flaky structure of the particles is pore walls which are mutually adhered to form a three-dimensional through porous channel structure, namely europium phosphate (EuPO)4) The particles are uniformly distributed on the surface and inside of the sheet structure, and the phosphorus is attached to the surface of the sheet structureEuropium acid (EuPO)4) The surface of the particles is coated with chitosan. The size and the distribution of micropores of the three-dimensional through porous channel structure are uniform.
The XRD pattern and FTIR pattern of the europium phosphate/chitosan three-dimensional porous scaffold prepared in this example showed results similar to those of example 1, and a three-dimensional porous scaffold comprising europium phosphate and chitosan as constituent elements was prepared.
The cell adhesion result of the europium phosphate/chitosan three-dimensional porous scaffold prepared in the embodiment is similar to that of the europium phosphate/chitosan three-dimensional porous scaffold prepared in the embodiment 1, the cell spreading state is good, and the biocompatibility is good.
The bone-promoting effect of the europium phosphate/chitosan three-dimensional porous scaffold prepared in the example is similar to that of the scaffold prepared in the example 5, and the scaffold shows a remarkable bone-promoting effect.
The embodiments described above are intended to facilitate the understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims (9)

1. A rare earth phosphate/bioactive polymer three-dimensional porous composite material is characterized in that sheet structures of bioactive polymer material doped with rare earth phosphate particles are adhered to each other to form a three-dimensional through porous channel structure, and the rare earth phosphate particles are uniformly distributed on the surface and inside of the sheet structures;
the bioactive high polymer material is selected from any one or a combination of chitosan, collagen, polyvinylpyrrolidone, polyhydroxybutyrate and polycaprolactone, the rare earth phosphate is lanthanum phosphate, cerium phosphate, gadolinium phosphate, ytterbium phosphate, europium phosphate or samarium phosphate, and the particle size of the rare earth phosphate is 50 nm-5 mu m; the pore diameter of the multi-pore channel structure is 10-200 mu m, and the porosity is 85-95%.
2. The preparation method of the rare earth phosphate/bioactive polymer three-dimensional porous composite material as claimed in claim 1, which is characterized by comprising the following steps:
(1) uniformly mixing rare earth phosphate and acid-containing bioactive polymer material solution, and freeze-drying to obtain a rare earth phosphate precursor/bioactive polymer composite material;
(2) and (3) soaking the rare earth phosphate precursor/bioactive polymer material in alkali liquor, washing to be neutral, and freeze-drying to obtain the rare earth phosphate/bioactive polymer three-dimensional porous composite material.
3. The preparation method according to claim 2, wherein in the step (1), the ratio of the rare earth phosphate to the bioactive polymer material is 1: 0.5-5, the concentration of the bioactive polymer material is 0.005 g/L-saturated solution, and the bioactive polymer material contains 0.2-5% of acid by volume fraction, wherein the acid comprises organic acid and inorganic acid.
4. The preparation method according to claim 2, wherein the step (1) is freeze-dried at-85 ℃ to 0 ℃ for 30min to 700h at 1 to 50 pa.
5. The method for preparing a rare earth phosphate according to claim 2, wherein the method for preparing a rare earth phosphate comprises the steps of:
a. adding soluble trivalent rare earth salt or soluble trivalent rare earth salt solution into soluble phosphate solution with the pH = 7-12 at the temperature of 10-60 ℃ under the stirring condition, and uniformly mixing;
b. after mixing, stirring for 0.5-2 h at 70-90 ℃, then stirring for 24-48 h at 10-60 ℃, filtering and washing to obtain a neutral precipitate;
c. drying the neutral precipitate for 3-30 h at 50-90 ℃, and calcining for 2-6 h at 500-1000 ℃ to prepare the rare earth phosphate.
6. The preparation method according to claim 5, wherein in step a, the soluble phosphate comprises dihydrogen phosphate, hydrogen phosphate or orthophosphate, and the soluble trivalent rare earth salt comprises trivalent lanthanum salt, trivalent cerium salt, trivalent gadolinium salt, trivalent ytterbium salt, trivalent europium salt or trivalent samarium salt; in the uniformly mixing system of the step a, the molar ratio of the trivalent rare earth ions to the phosphate radicals is 1: 0.5 to 5; the concentration of the trivalent rare earth ions is 0.05-0.1 mol/L.
7. The preparation method according to claim 2, characterized in that in the step (2), the alkali liquor is soaked for 4-48 h at 20-50 ℃, the concentration of the alkali liquor is 0.05-0.2 mol/L, and the alkali contained in the alkali liquor is selected from any one or combination of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, ammonium carbonate and potassium bicarbonate.
8. The preparation method according to claim 2, wherein the step (2) is freeze-dried at-85 ℃ to 0 ℃ for 30min to 700h at 1 to 50 pa.
9. The use of the rare earth phosphate/bioactive polymer three-dimensional porous composite material of claim 1 in the preparation of bone repair materials.
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