CN102107020A - Method for preparing artificial bone with three-dimensional bone-like structure and product prepared by method - Google Patents
Method for preparing artificial bone with three-dimensional bone-like structure and product prepared by method Download PDFInfo
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- CN102107020A CN102107020A CN2011100374152A CN201110037415A CN102107020A CN 102107020 A CN102107020 A CN 102107020A CN 2011100374152 A CN2011100374152 A CN 2011100374152A CN 201110037415 A CN201110037415 A CN 201110037415A CN 102107020 A CN102107020 A CN 102107020A
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- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 97
- 238000000034 method Methods 0.000 title claims abstract description 35
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 24
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- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims abstract description 15
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- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 13
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 13
- LQZZUXJYWNFBMV-UHFFFAOYSA-N dodecan-1-ol Chemical compound CCCCCCCCCCCCO LQZZUXJYWNFBMV-UHFFFAOYSA-N 0.000 claims description 13
- 235000010413 sodium alginate Nutrition 0.000 claims description 13
- 239000000661 sodium alginate Substances 0.000 claims description 13
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- 229910021641 deionized water Inorganic materials 0.000 claims description 12
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- 239000003795 chemical substances by application Substances 0.000 claims description 10
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Abstract
The invention discloses a method for preparing an artificial bone with a three-dimensional bone-like structure and a product prepared by the method. The method comprises the following steps of: preparing a green body with a three-dimensional bone-like structure by a three-dimensional laminated assembly molding process; drying and sintering the green body to obtain a high-strength hydroxyapatite (HA) scaffold with a three-dimensional structure or a high-strength beta-tricalcium phosphate (TCP) scaffold with a three-dimensional structure; and then compounding the high-strength HA scaffold or the high-strength beta-TCP scaffold with imitation bone solution with high bioactivity to obtain a HA or beta-TCP scaffold with a three-dimensional structure which has high bioactivity and high strength, wherein the HA or beta-TCP scaffold has high bioactivity, the compressive strength is more than 5Mpa, the porosity is 70 to 75 percent, and the aperture is 100 to 600mu m. The artificial bone with the three-dimensional bone-like structure which has high bioactivity and high strength can repair various bone defects and induce the bone defects into bones.
Description
Technical field
The present invention relates to artificial bone and preparation method thereof, relate in particular to the preparation method of the three-dimensional class bone structure of high bioactivity artificial bone and, belong to the artificial bone field by the product that this method prepares.
Background technology
The patent of research artificial bone and preparation method thereof is a lot of at present, and HA is not only arranged, and β-TCP material also has a large amount of composites.Compound patent of HA or Nano-HA and collagen such as US 7229971, US20170255422, US 200220018797, with JP 2003073304 or the like, though have the bone fusion faculty, initial strength is poor, do not have supporting role yet, some bone defect repair is had any problem.
Summary of the invention
Technical problem to be solved by this invention be overcome that the existing biological activity of existing imitative bone material is low, problem that intensity difference and implantation initial stage do not have the support effect, a kind of high bioactivity and high-intensity three-dimensional class bone structure artificial bone are provided.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of high bioactivity and high-intensity three-dimensional class bone structure artificial bone, its preparation method comprises:
(1) the ceramic mixed slurry of being formed with deionized water, sodium lauryl sulphate, lauryl alcohol and hydroxylapatite ceramic is a raw material, adds firming agent and initiator, adopts 3-D stacks assembled formation method to prepare the base substrate of three-dimensional class bone structure;
(2) base substrate with three-dimensional class bone structure obtains HA three dimensional structure support or β-TCP three dimensional structure support behind super-dry, sintering;
(3) HA three dimensional structure support or β-TCP three dimensional structure support are inserted in the imitative bone solution and soak, vacuum is drained, promptly.
In order to reach better preparation effect, in the step (1), the consisting of of described ceramic mixed slurry: contain the 20-40v% hydroxyapatite in the deionized water of every 100ml, 0.1-0.5 gram sodium lauryl sulphate and 0.3-0.8 gram lauryl alcohol; Above-mentioned each component is stirred, promptly.
Described firming agent comprises sodium alginate, is preferably the sodium alginate that concentration is 5wt%; Wherein the consumption of the sodium alginate that is added is the 5-10% that accounts for the mixed material cumulative volume;
Described initiator is the CaCl of 3M
2, its addition is identical with the consumption of sodium alginate, accounts for the 5-10% of mixed material cumulative volume;
In the step (2) with the base substrate of three-dimensional class bone structure after super-dry, obtained HA three dimensional structure support again through 900-1100 ℃ of sintering 2-6 hour; Or with the base substrate of three-dimensional class bone structure after super-dry, obtained β-TCP three dimensional structure support in sintering 2-4 hour through 1250 ℃-1350 ℃ again; Wherein, described drying mode is preferably under 37 ℃ the temperature dry 48 hours;
In order to reach better effect, with HA three dimensional structure support or β-TCP three dimensional structure support insert soak in the imitative bone solution before, preferably it was handled 80-120 hour with 10M NaOH aqueous solution soaking;
The preparation method of the imitative bone solution in the step (3) comprises: 0.1-0.5mg collagen-2 (rhBMP) is put into 1.5SBF (table 1) body fluid of 100ml, stir, promptly.
Soak time described in the step (4) is preferably 72-168 hour;
Prescription (the total amount 100ml of table 1 1.5SBF solution; 95.5ml deionized water wherein)
The present invention adopts 3-D stacks assembled formation method to prepare the base substrate of three-dimensional class bone structure; Behind super-dry, sintering, obtain high-intensity HA three dimensional structure support or through β-TCP three dimensional structure support; The imitative bone solution of itself and high bioactivity is carried out compoundly again, obtain the HA or the β-TCP support of high bioactivity and high-strength three-dimensional structure, its comprcssive strength is greater than 5MPa, porosity 70-75%, and the aperture is 100-600 μ m.High bioactivity of the present invention and high-intensity three-dimensional class bone structure artificial bone can be repaired various bones are damaged, but and induced osteogenesis.
Description of drawings
The process sketch map of Fig. 1 3-D stacks assembled formation.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Embodiment 1
(1) add 30vol%HA hydroxyapatite and 0.3 gram sodium lauryl sulphate and 0.3 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 8vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 8vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
The concrete preparation process of timbering material base substrate is as follows:
Exterior contour and inner aperture for the three dimensional structure complexity of reappearing natural bone, according to Granted publication is that the disclosed three dimensional gel laminated forming of CN 98124402.5C (patent No. ZL 98124402.5) system automatic moulding obtains the timbering material base substrate, and inner aperture can utilize the foaming preparation.
The concrete preparation process and the characteristics of three dimensional gel laminated forming system and timbering material base substrate are as follows:
Three dimensional gel laminated forming (3D Gel-lamination) is a kind of novel quick molding method, be rapid shaping (rapid prototyping, RP) product combining of thought and gel mold (Gel-casting) core technology adopts computer control three-dimensional fast shaping system of processing to prepare ceramic body.Utilize this technology can prepare the bone tissue engineering stent material with complex appearance and internal structure, thereby make the realization individuation be designed to possibility, promptly the practical situation according to patient customizes the artificial bone dummy.
The concrete preparation process of timbering material base substrate as shown in Figure 1.At first be full of ceramic size in slurry pool, after computer sent movement instruction, shaped platform dropped in the slurry pool, made and soaked full ceramic size on the shaped platform, rose to assigned address then, and scraper was scraped, and evenly spread on the shaped platform and had applied the skim slurry.At this moment delamination software " is cut " two dimensional image that goes out corresponding one deck from threedimensional model, be converted into the forming data signal and send to make-up machine, the initiator nozzle sprays the gel initiator according to signal at the correspondence position of thin layer, the slurry that receives initiator solidifies rapidly, form and the model consistent curing thin layer with definite shape of respective layer, i.e. a section in the Rapid Prototyping Process has been finished in preparation.Shaped platform continues motion afterwards, and new skim slurry is applied in the scraper shop, carries out the manufacturing of a new section, and connects as one with a last section.Circulation so repeatedly, the shaped platform position constantly descends, and base substrate is more and more higher, finishes up to whole blank forming.
Molding is taken off ceramic body after finishing, and rinses out uncured ceramic size residual on the base substrate with deionized water.This moment, ceramic body had certain intensity, was enough to keep the shape of its molding.Base substrate is put into climatic chamber, and slowly dry under the condition of 37 ℃ of temperature, humidity 60%, further sintering is removed the Organic substance in the base substrate, and obtains having definite shape and intensity timbering material.
The aperture of timbering material inside (aperture 100-600 μ m) can utilize the foaming preparation.Promptly by in ceramic size, adding suitable the surfactant sodium lauryl sulphate and the lauryl alcohol of specific consumption, produce uniform and stable in a large number, the foam of size appropriateness, it is reasonable to obtain pore-size distribution behind the gel, the porous support materials that the hole connectivity is good.
The three dimensional gel laminated process requires the stable foamed slurry of preparation, yet " foaming capacity " of slurry is distinguishing two notions with " foam stable ability ".Complexity that foam forms and growing amount what the former is meant; The latter refers to generate foamy persistency, i.e. the foamy life-span.In order to make the foam lasting stability of generation, in surfactant formula, add some cosurfactants, i.e. foam stabilizer usually.Surface tension is to influence the principal element that foam forms, but is not the determiner of foam stabilization.After adding lauryl alcohol in the sodium dodecyl sulfate solution, not only surface tension is lower, help bubbling, and surface viscosity is also bigger, can access more stable foam.So selected sodium lauryl sulphate as foaming agent in the experiment, lauryl alcohol is as foam stabilizer.
Further sintering is removed the Organic substance in the base substrate, and obtains having definite shape and intensity timbering material.
(2) step (1) is made the base substrate of three-dimensional class bone structure, after 37 ℃ of 48 hours dryings,, obtained the HA three dimensional structure support in 300-600 μ m aperture again through 1100 ℃ of sintering 4 hours; Make 10 in 10mm x 10mm x 2mm sample, wherein 5 are used to test comprcssive strength, and 5 are used to test alkaline phosphatase activity (ALP (U/g Protein (albumen)).
(3) HA three dimensional structure support sample was handled 96 hours with 10M NaOH;
(4) 1.5SBF (body fluid) that the collagen-2 (rhBMP-2) of 0.3mg is put into 100ml makes imitative bone solution;
(5) the HA three dimensional structure support that step (3) is handled is put into imitative bone solution immersion 72 hours; At room temperature vacuum is drained, and just makes high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 8.52 ± 1.21MPa;
The alkaline phosphatase activity (method of testing and the result of ALP (U/g Protein (albumen)) are as follows:
1.MC3T3-E1 osteoblast is cultivated:
The MC3T3-E1 osteoblast derives from the skull of newborn mice, and inducing culture is α-MEM culture medium, wherein contains 10% hyclone, 100U/ml green grass or young crops/streptomycin, 0.05g/L ascorbic acid and 10mmol/L sodium.
Prepared three-dimensional class bone structure artificial bone is put into 24 orifice plates, adopt the sterilization of 70% ethanol to spend the night, wash 3 times with phosphate buffer (PBS), and sterilized 2 hours with ultra-vioket radiation.Experiment is in every hole inoculation 10 of 24 orifice plates
5Individual cell finds that the cell growth and breeding is very fast.Keeping temperature is to contain 5%CO in 37 ℃, atmosphere
2, continue to cultivate, changed liquid every 3 days.
2. alkaline phosphatase activities detects:
Discard culture medium, wash 3 times, in every hole, add 250 μ l cell pyrolysis liquid (50mM Tris, 1mM MgCl then with PBS
2, 0.5%Triton X-100 pH7.6), smashes sample gently, freeze thawing 3 times.Then with sample transfer to the 1.5ml centrifuge tube, centrifugal 10 minutes of 4 ℃ of following 18000g, take out supernatant.
Measure alkali phosphatase (ALP) activity of osteoblast behind the certain natural law of inducing culture on the prepared three-dimensional class bone structure artificial bone, each sample is got 20 μ l lysis supernatants respectively, explanation according to test kit, the ALP activity of mensuration cell (Alkaline Phosphatase, Sigma).The mensuration number of times of every kind of sample is 3 times.
Osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone after 14 days and 21 days, is measured its ALP (U/g Protein) activity and is respectively: 28.11 ± 3.35U/g albumen; 52.23 ± 4.16U/g albumen; 68.15 ± 5.34U/g albumen.
Embodiment 2
(1) add 30vol% hydroxyapatite and 0.1 gram sodium lauryl sulphate and 0.5 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 5vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 5vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
The concrete preparation process of timbering material base substrate is seen embodiment 1.
(2) step (1) is made the base substrate of three-dimensional class bone structure, after 37 ℃ of 48 hours dryings,, obtained the HA three dimensional structure support in 300-600 μ m aperture again through 900 ℃ of sintering 6 hours; Make 10 in 10mm x 10mm x 2mm sample, wherein 5 are used to test comprcssive strength, and 5 are used to test alkaline phosphatase activity (ALP (U/g Protein (albumen)) (appendix 2 is seen in the ALP test).
(3) HA three dimensional structure support sample was handled 96 hours with 10M NaOH;
(4) 1.5SBF (body fluid) that the collagen-2 (rhBMP-2) of 0.3mg is put into 100ml makes imitative bone solution;
(5) the HA three dimensional structure support that step (3) is handled is put into imitative bone solution immersion 72 hours; At room temperature vacuum is drained, and just makes high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 8.26 ± 1.45MPa;
According to embodiment 1 described method,, after 14 days and 21 days, measure its ALP (U/g Protein (albumen)) activity and be respectively 29.67 ± 1.25U/g albumen osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone; 54.36 ± 2.05U/g albumen; 70.13 ± 4.72U/g albumen.
Embodiment 3
(1) add 30vol% hydroxyapatite and 0.5 gram sodium lauryl sulphate and 0.1 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 10vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 10vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
The concrete preparation process of timbering material base substrate is seen embodiment 1.
(2) step (1) is made the base substrate of three-dimensional class bone structure, after 37 ℃ of 48 hours dryings,, obtained the HA three dimensional structure support in 300-600 μ m aperture again through 1000 ℃ of sintering 5 hours; Make 10 in 10mm x 10mm x 2mm sample, wherein 5 are used to test comprcssive strength, and 5 are used to test alkaline phosphatase activity (ALP (U/g Protein (albumen)).
(3) HA three dimensional structure support sample was handled 96 hours with 10M NaOH;
(4) 1.5SBF (body fluid) that the collagen-2 (rhBMP-2) of 0.3mg is put into 100ml makes imitative bone solution;
(5) the HA three dimensional structure support that step (3) is handled is put into imitative bone solution immersion 72 hours; At room temperature vacuum is drained, and just makes high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 8.41 ± 1.34MPa;
According to embodiment 1 described method,, after 14 days and 21 days, measure its AL (U/g Protein) P activity and be respectively 28.65 ± 2.24U/g albumen osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone; 53.24 ± 4.07U/g albumen; 69.12 ± 4.78U/g albumen.
Embodiment 4
(1) add 40vol% hydroxyapatite and 0.2 gram sodium lauryl sulphate and 0.3 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 8vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 8vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
Adopt embodiment 1 described 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
(2) step (1) is made the base substrate of three dimensional structure, after 37 ℃ of 48 hours dryings,, obtained the β-TCP three dimensional structure support in 300-500 μ m aperture again through 1250 ℃ of sintering 4 hours;
(3) β-TCP three dimensional structure support was handled 96 hours with 10M NaOH.
(4) 1.5SBF (body fluid) that 0.2mg collagen-2 (rhBMP-2) is put into 100ml makes imitative bone solution;
(5) β-TCP three dimensional structure support of handling being put into imitative bone solution soaked 168 hours.At room temperature vacuum was drained 4 hours, just made degradable high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 6.53 ± 1.01MPa;
According to embodiment 1 described method,, after 14 days and 21 days, measure its AL (U/g Protein (albumen)) P activity and be respectively 19.13 ± 2.35U/g albumen osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone; 35.32 ± 4.15U/g albumen; 56.23 ± 5.12U/g albumen.
Embodiment 5
(1) add 40vol% hydroxyapatite and 0.5 gram sodium lauryl sulphate and 0.3 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 10vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 10vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
Adopt embodiment 1 described 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
(2) step (1) is made the base substrate of three dimensional structure, after 37 ℃ of 48 hours dryings,, obtained the β-TCP three dimensional structure support in 300-500 μ m aperture again through 1350 ℃ of sintering 3 hours;
(3) β-TCP three dimensional structure support was handled 96 hours with 10M NaOH.
(4) 1.5SBF (body fluid) that 0.2mg collagen-2 (rhBMP-2) is put into 100ml makes imitative bone solution;
(5) β-TCP three dimensional structure support of handling being put into imitative bone solution soaked 168 hours.At room temperature vacuum was drained 4 hours, just made degradable high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 6.34 ± 1.24MPa;
According to embodiment 1 described method,, after 14 days and 21 days, measure its ALP (U/g Protein) activity and be respectively 18.98 ± 2.56U/g albumen osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone; 33.78 ± 4.15U/g albumen; 54.93 ± 6.34U/g albumen.
Embodiment 6
(1) add 40vol% hydroxyapatite and 0.5 gram sodium lauryl sulphate and 0.1 gram lauryl alcohol in every 100ml deionized water, stirring obtains ceramic mixed slurry; The sodium alginate that adds the 5wt% that accounts for mixture cumulative volume 5vol% in above-mentioned ceramic mixed slurry adds the CaCl of the 3M that accounts for mixture cumulative volume 5vol% again as firming agent again
2As initiator, utilize the 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
Adopt embodiment 1 described 3-D stacks assemble method to make the base substrate of three-dimensional class bone structure;
(2) step (1) is made the base substrate of three dimensional structure, after 37 ℃ of 48 hours dryings,, obtained the β-TCP three dimensional structure support in 300-500 μ m aperture again through 1300 ℃ of sintering 3.5 hours;
(3) β-TCP three dimensional structure support was handled 96 hours with 10M NaOH.
(4) 1.5SBF (body fluid) that 0.2mg collagen-2 (rhBMP-2) is put into 100ml makes imitative bone solution;
(5) β-TCP three dimensional structure support of handling being put into imitative bone solution soaked 168 hours.At room temperature vacuum was drained 4 hours, just made degradable high bioactivity and high-intensity three-dimensional class bone structure artificial bone.
Test comprcssive strength on the mechanical test machine, comprcssive strength is 7.09 ± 1.21MPa;
According to embodiment 1 described method,, after 14 days and 21 days, measure its AL (U/g Protein) P activity and be respectively 19.03 ± 2.35U/g albumen osteoblast inducing culture 7 days on prepared three-dimensional class bone structure artificial bone; 34.21 ± 4.56U/g albumen; 55.23 ± 5.34U/g albumen.
Claims (10)
1. the preparation method of a high bioactivity and high-intensity three-dimensional class bone structure artificial bone may further comprise the steps:
(1) the ceramic mixed slurry of being formed with deionized water, sodium lauryl sulphate, lauryl alcohol and hydroxylapatite ceramic mixed slurry is a raw material, adds firming agent and initiator, adopts the 3-D stacks assemble method to prepare the base substrate of three-dimensional class bone structure;
(2) base substrate with three-dimensional class bone structure obtains HA three dimensional structure support or β-TCP three dimensional structure support behind super-dry, sintering;
(3) HA three dimensional structure support or β-TCP three dimensional structure support are inserted in the imitative bone solution and soak, vacuum is drained, promptly.
2. in accordance with the method for claim 1, it is characterized in that: the consisting of of the ceramic mixed slurry described in the step (1): contain the 20-40v% hydroxyapatite in the deionized water of every 100ml, 0.1-0.5 gram sodium lauryl sulphate and 0.3-0.8 gram lauryl alcohol.
3. in accordance with the method for claim 1, it is characterized in that: described firming agent comprises sodium alginate, is preferably the sodium alginate that concentration is 5wt%; Wherein the consumption of the sodium alginate that is added is the 5-10% that accounts for the mixture cumulative volume.
4. it is characterized in that in accordance with the method for claim 1: described initiator is the CaCl of 3M
2, the consumption of the initiator that is added is the 5-10% that accounts for the mixture cumulative volume.
5. in accordance with the method for claim 1, it is characterized in that: in the step (2) with the base substrate of three-dimensional class bone structure after super-dry, obtain HA three dimensional structure support through 900-1100 ℃ of sintering again; Or with the base substrate of three-dimensional class bone structure after super-dry, obtain β-TCP three dimensional structure support through 1250 ℃ of-1350 ℃ of sintering again.
6. in accordance with the method for claim 1, it is characterized in that: with HA three dimensional structure support or β-TCP three dimensional structure support insert soak in the imitative bone solution before, it was handled 80-120 hour with 10M NaOH aqueous solution soaking.
7. in accordance with the method for claim 1, it is characterized in that: the preparation method of the described imitative bone solution in the step (3) comprises: 0.1-0.5mg collagen-2 is dissolved in the 1.5SBF body fluid of 100ml, stirs, promptly.
8. it is characterized in that in accordance with the method for claim 1: the soak time described in the step (3) is 72-168 hour.
9. by the resulting product of any one preparation method of claim 1-8.
10. the described product of claim 9 is preparing the purposes of repairing in the bone injury material.
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