CN1891665A - Beta-tricalcium phosphate porous ceramic material for bone repair, and its preparing method and use - Google Patents
Beta-tricalcium phosphate porous ceramic material for bone repair, and its preparing method and use Download PDFInfo
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- CN1891665A CN1891665A CN 200510027618 CN200510027618A CN1891665A CN 1891665 A CN1891665 A CN 1891665A CN 200510027618 CN200510027618 CN 200510027618 CN 200510027618 A CN200510027618 A CN 200510027618A CN 1891665 A CN1891665 A CN 1891665A
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Abstract
This invention discloses a synthesizing method and its application about the porous ceramic material of beta-tricalcium phosphate which is used for bone renovation. The porosity ratio is 85~95 %, the average pore size of surgical sutures is 200-500 microns, and 90%-99.999% of the holes connect with each other. The material is an optimum option for synthesizing the support of the osseous graft for its high porosity ratio, degradation, and the good histocompatibility.
Description
Technical field
The present invention relates to field of tissue engineering technology and materialogy field, relate to a kind of porous ceramic film material and bone tissue engineering scaffold that bone is repaired that be used for particularly.
Background technology
Bone tissue engineer is to utilize the principle of engineering and life science and the new osseous tissue of method regeneration, repairing and alternative pathology or defective bone tissue, or promotes the technology of its function.
Repopulating cell on biomaterial scaffolds, the interior or vitro culture biological tissue through body, the position of subsequently their being implanted damaged or pathology is with repair deficiency or pathology osseous tissue.This is to adopt method of tissue engineering to make up new osseous tissue, or recovers one of approach of pathology osseous tissue function.
At present, on bone tissue engineer was used, the calcium phosphate stupalith was integrated performance with its good biocompatibility and skeletonization and is used to bone alternate material.Wherein, hydroxyapatite (HA) is the main component of vertebrates bone and tooth, good stability, and can induce new osteogenesis, be widely used in the reparation and the replacement material of sclerous tissues.But the HA absorbability is poor, and fragility is big, easily ruptures in the back that implants, and this has limited its application clinically.Therefore,, finally substituted, must adopt a kind of degradable material by osseous tissue as bone renovating material.In recent years, bata-tricalcium phosphate (β-TCP) since its biological degradability and biocompatibility get more and more people's extensive concerning preferably.After β-the TCP pottery implants, can degradation in vivo, the Ca that degrade, P enter the live body recycle system and form area of new bone, so are ideal engineering material of bone tissue comparatively, are subjected to various countries investigator's extensive concern at present.Studies show that β-TCP has good skeletonization transmissibility, clinical demonstration curative effect is better.Simultaneously, suitable vesicular structure will help quick vascularization and bone is grown into, and this is extremely important to obtaining to implant effect preferably.The porous beta-TCP support that preparation has the appropriate bore structure just becomes the focus that people study.
According to application target different with to material performance requirement, people have been developed multiple porous ceramics preparation technology, as porous plastics pickling process, foaming, interpolation pore-forming material method, sol-gel method etc.The porous plastics pickling process can be prepared high porosity (70-95%) porous ceramics with three-dimensional netted skeleton structure, and pore is to connect mutually.This manufacturing process proposes in U.S. Pat Pat.3090094, it utilizes the special construction of the three-dimensional netted skeleton of perforate that the organic foam body had, to expect evenly to be coated on the organic foam reticulate body with what prepare, and burn the organic foam body after the drying and obtain a kind of porous ceramics.Yet the shortcoming of this technology is that the porous ceramics physical strength of preparing is relatively poor, and ceramic aperture receives the aperture of machine foams and controls.
In sum, this area lack porosity height (＞85%), degradable, histocompatibility is good is used for the timbering material that bone is repaired, therefore, this area presses for the good timbering material that is used for the bone reparation of exploitation new high porosity (＞85%), histocompatibility.
Summary of the invention
The objective of the invention is to obtain to have high porosity, be fit to the aperture, and the timbering material of the bone degradable, that histocompatibility good (especially You Yi osteoconductive) reparation
In a first aspect of the present invention, a kind of beta-tricalcium phosphate porous ceramic material is provided, the porosity of described stupalith is 85～95%, mean pore size is the 200-500 micron, and the hole of 90%-99.999% interconnects.
In another preference, the resistance to compression hardness of described stupalith is 80-200Kpa, and mean pore size is the 300-500 micron.
In another preference, described stupalith is to be prepared by the method that contains following steps:
(a) with slurry dipping pore-forming template, slurry is fully entered in the hole of pore-forming template, the template that obtains flooding, wherein said slurry contains the β-TCP powder and 1～9 parts by weight of adhesive of 1～3 weight part, described pore-forming template can be calcined removal, and described slurry be the dipping template weight 4-1.5 doubly;
(b) the dipping template described in the step (a) is carried out drying, obtain the porous ceramics base substrate;
(c) the porous ceramics base substrate described in the step (b) is calcined at 900-1300 ℃, removed the pore-forming template, thereby obtain the agglomerating porous ceramics;
(d) porous ceramics in the cooling step (c) obtains described stupalith.
In a second aspect of the present invention, a kind of method for preparing beta-tricalcium phosphate porous ceramic material of the present invention also is provided, it contains following steps:
(a) with slurry dipping pore-forming template, slurry is fully entered in the hole of pore-forming template, the template that obtains flooding, wherein said slurry contains the β-TCP powder and 1～9 parts by weight of adhesive of 1～3 weight part, described pore-forming template can be calcined removal, and described slurry be the dipping template weight 4-1.5 doubly;
(b) the dipping template described in the step (a) is carried out drying, obtain the porous ceramics base substrate;
(c) the porous ceramics base substrate described in the step (b) is calcined at 900-1300 ℃, removed the pore-forming template, thereby obtain the agglomerating porous ceramics;
(d) porous ceramics in the cooling step (c) obtains described stupalith.
In another preference, described pore-forming template be the aperture be 23～40 holes/centimetre, the polyurethane sponge of porosity 95.0-99.9%, the polyurethane sponge of preferable porosity 98.0-99.9%.
In another preference, in described step (a) before, also comprise the pore-forming template is carried out pre-treatment: is alkali hydroxide soln immersion 12～24h of 1～10M with the pore-forming template in concentration, and/or the polyvinyl alcohol solution under 0.5～2wt% concentration soaks 12～24h.
In another preference, binding agent is selected from the described step (a): bio-vitric, methylcellulose gum, silicon sol or its combination.
In another preference, binding agent comprises methylcellulose gum, bio-vitric and silicon sol simultaneously described in the described step (a), wherein methylcellulose gum is 1～10wt% of powder quality, 3～20wt% of bio-vitric powder quality, silicon sol is 1～5 times of powder quality, Gu and the liquid/weight ratio of described slurry be 1～3.
In another preference, the drying conditions in the described step (b) is as follows: the at room temperature dry 24h-72h of porous ceramics base substrate, 40～100 ℃ of following dry 8-16h;
Perhaps, the calcination condition in the described step (c) is as follows:
Temperature rise rate with 0.1-1 ℃/min rises to 400-800 ℃ with the porous ceramics base substrate, preferred 550-650 ℃; Temperature rise rate with 2～5 ℃/min rises to 1100～1300 ℃ then, and is incubated 1-6 hour.
In a third aspect of the present invention, a kind of graft is provided, described graft contains beta-tricalcium phosphate porous ceramic material of the present invention and is inoculated in the stem cell of described stupalith, described stem cell is selected from bone marrow stroma stem cell or fat stem cell, and the inoculum size of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3Porous ceramic film material.
In a fourth aspect of the present invention, the purposes of beta-tricalcium phosphate porous ceramic material of the present invention is provided, it is used as the support for preparing bone graft.
Description of drawings
The optical microscope photograph of Fig. 1 porous beta-TCP pottery.
Scanning electron microscope (SEM) photo of Fig. 2 porous beta-TCP pottery.
After Fig. 3 (a) cultivated for 1 week, the stereoscan photograph of the bone marrow stroma stem cell of porous beta-TCP ceramic surface.Hole surface is almost covered by Polygons cell and extracellular matrix.Scale length is 300 μ m.
After Fig. 3 (b) cultivated for 1 week, the bone marrow stroma stem cell of porous beta-TCP ceramic surface is the stereoscan photograph of high-amplification-factor more.Scale length is 95 μ m.
After Fig. 4 (a) cultivated for 1 week, porous beta-TCP pottery section, the stereoscan photograph of bone marrow stroma stem cell.The hole inwall is coated with the Polygons cell, and extracellular matrix is also almost filled whole hole.Scale length is 95 μ m.
After Fig. 4 (b) cultivated for 1 week, porous beta-TCP pottery section, bone marrow stroma stem cell is the stereoscan photograph of high-amplification-factor more.Scale length is 37 μ m.
Fig. 5 (a) cultivated after 5 days, porous beta-TCP ceramic surface, the stereoscan photograph of fat stem cell.Hole surface is almost covered by Polygons cell and extracellular matrix.Scale length is 270 μ m.
Fig. 5 (b) cultivated after 5 days, and porous beta-TCP ceramic surface, fat stem cell be the stereoscan photograph of high-amplification-factor more.Scale length is 85 μ m.
Fig. 6 (a) cultivated after 5 days, porous beta-TCP pottery section, the stereoscan photograph of fat stem cell.Covered by cell and extracellular matrix in the hole.Scale length is 49 μ m.
Fig. 6 (b) cultivated after 5 days, porous beta-TCP pottery section, and fat stem cell is the stereoscan photograph of high-amplification-factor more.Scale length is 13.5 μ m.
Embodiment
The inventor is through extensive and deep research, by improving preparation technology, high porosity, suitable, the degradable beta-tricalcium phosphate porous ceramic material in aperture have been obtained, and the consistency that is surprised to find that this beta-tricalcium phosphate porous ceramic material and bone marrow stroma stem cell and fat stem cell is excellent especially, so be particularly suitable as the timbering material that bone is repaired.Finished the present invention on this basis.
Term
Term " purifying or isolating " refers to purifying or isolating material is substantially devoid of other cells, protein or polypeptide.
Term " xenotransplantation " refers to required biomaterial (as stem cell) is taken out and is applied to the method for another species object from a certain species.
Term " autotransplantation " refers to required biomaterial (as stem cell) is taken out and is applied to same patient's method from certain patient.
Term " heteroplastic transplantation " refers to required biomaterial (as stem cell) is taken out and is applied to another different patients' method from certain individuality of same species.
Porous ceramic film material
Porous bata-tricalcium phosphate stupalith of the present invention is that porosity is 85～95%, and mean pore size is 200-500 micron (a preferably 300-500 micron), and the hole of 90%-99% interconnects.
The raw material that is used to prepare stupalith of the present invention comprises: pore-forming template and slurry, described slurry contain β-TCP powder and binding agent.
Can be used for pore-forming template of the present invention and be not particularly limited, can be any porous sponge material, and needing only described pore-forming template can remove in calcination process, for example polyurethane sponge, EVC sponge and high-elastic sponge etc.Usually, the porosity of pore-forming template is 95～99.9%.
Be used for slurry of the present invention usually by the β-TCP powder and 1～9 parts by weight of adhesive of 1～3 weight part.
The particle diameter of β-TCP powder is generally 100nm～10 μ m, and β-TCP powder can prepare with chemical coprecipitation, also can buy from the market to obtain.
Can be used for tackiness agent of the present invention and be not particularly limited, representational tackiness agent example comprises (but being not limited to): methylcellulose gum, silicon sol or its combination.The inventor finds that a kind of excellent especially tackiness agent is a bio-vitric.Described bio-vitric composition comprises SiO
2, P
2O
5, CaO and CaF
2, by the common method preparation for preparing glass, its commodity are called Cerabone A-W.For example with SiO
234wt%, P
2O
516.2wt%, CaO 44.7wt% and CaF
24.6wt% prepares described bio-vitric with high-temperature melting method.By adding bio-vitric, can reduce calcining temperature, improve the mechanical property of porous ceramics.
The preparation method of stupalith of the present invention generally includes following steps:
(a) with slurry dipping pore-forming template, slurry is fully entered in the hole of pore-forming template, the template that obtains flooding; (b) the dipping template described in the step (a) is carried out drying, obtain the porous ceramics base substrate;
(c) the porous ceramics base substrate described in the step (b) is calcined at 900-1300 ℃, removed the pore-forming template, thereby obtain the agglomerating porous ceramics;
(d) porous ceramics in the cooling step (c) obtains described stupalith.
Above-mentioned dipping, drying, calcining and cooling step can carry out with the equipment and the method for this area routine.
Stem cell
The source of stem cell of the present invention is not particularly limited, and can be the stem cell in any source, and usually, stem cell of the present invention is from body or allochthonous stem cell.The position that obtains stem cell also is not particularly limited, and can be fat stem cell, bone marrow stroma stem cell or other stem cells.In addition, the also alternative stem cell of scleroblast is as the seed cell of bone tissue engineer structure.
Can be used for stem cell of the present invention can be preferably Mammals from any vertebrates, more preferably is primate, especially the people.
Although the stem cell from body is preferred, the source of the stem cell of allosome is more commonly used.Research shows, and different growths, the allogeneic stem cell of etap can and have in the allogeneic animal body of complete immunologic function in consistency difference in a organized way and form the stem cell tissue.
The method of separation and acquisition stem cell is as known in the art.A kind of preferable methods is density gradient centrifugation and enzyme digestion.
Cultural method of stem cell and nutrient solution also are to know in this area.A kind of preferable methods is at 37 ℃, saturated humidity, 5%CO with stem cell
2Cultivate in the incubator.Suitable nutrient solution comprises (but being not limited to): 1) DMEM substratum ((Gibco company)+5～20% foetal calf serums; 2) DMEM substratum+5～20% calf serums; 3) DMEM substratum+5～20% are from body (allosome) human serum.In addition, add various somatomedins (for example promoting the cytokine of stem cell growth etc.), various microbiotic, various inducible factor in the above-mentioned nutrient solution.
Be applicable to that stem cell of the present invention should be able to be in vivo or in-vitro multiplication.A kind of preferred stem cell is the bone marrow stroma stem cell of vitro culture.
Bone graft
Because the consistency of beta-tricalcium phosphate porous ceramic material of the present invention and bone marrow stroma stem cell and fat stem cell is very good, therefore be particularly suitable as the timbering material that bone is repaired.
Form stem cell-beta-tricalcium phosphate porous ceramic material mixture on that the bone marrow stroma stem cell of cultured and amplified in vitro and/or fat stem cell are inoculated into the biocompatibility excellence and the degradable multiporous stupalith, this " stem cell-porous ceramic film material " mixture is implanted to defect, degraded and absorbed gradually along with porous ceramic film material, new bone forming reaches the purpose of repairing bone defect.
The preparation method of organizational project bone graft of the present invention is easy, the bone marrow stroma stem cell of some amount and/or fat stem cell is inoculated in beta-tricalcium phosphate porous ceramic material gets final product.
The shape of engineered bone graft of the present invention is not particularly limited, can be moulding arbitrarily according to the shape of tissue defect.Usually, graft is a long strip shape.
Bone marrow stroma stem cell in the tissue-engineered bone of the present invention and/or fat stem cell concentration are about 0.5 * 10 usually
6/ cm
3(ceramics bracket volume) is to 5 * 10
8/ cm
3Or higher, preferably be 1 * 10
6/ cm
3To 1 * 10
8/ cm
3, more preferably be 5 * 10
6/ cm
3To 5 * 10
7/ cm
3Beta-tricalcium phosphate porous ceramic material.Usually, adjust bone marrow stroma stem cell and/or fat stem cell concentration, mix with degradation material then with nutrient solution.During mixing, the ratio of nutrient solution and degradation material is not particularly limited, but is advisable with the nutrient solution maximum that this material can adsorb.
In addition, in engineered bone graft of the present invention, also can add or compound other various cells, somatomedin, various microbiotic, thereby keep bone marrow stroma stem cell and/or fat stem cell phenotype, promote bone marrow stroma stem cell and/or fat stem cell growth, and promote tissue-engineered bone to grow in vivo.
Except engineered bone graft is implanted, also be placed in the external biological reactor and cultivate, thereby carry out the structure of tissue-engineered bone, have the tissue-engineered bone of certain histological structure, biochemical composition and biomechanical strength in external formation.
With the engineered bone graft that the inventive method forms, the damaged place of the bone that can directly implant.
Major advantage of the present invention is:
(a) high porosity.
(b) degradable.
(c) histocompatibility is good, and has excellent osteoconductive.
(d) Zhi Bei porous beta-TCP pottery can be compound with multiple stem cell, repairing bone defect.
(e) β of high porosity-TCP pottery helps sticking of a large amount of cells, repairing bone defect faster.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment 1A:
The preparation of porous beta-TCP pottery
Take by weighing 2g and cross the β-TCP powder of 100 order mesh screens, the methylcellulose gum that adds 3wt% in proportion is as binding agent, and 5wt% glass powder is as high-temperature agglomerant, after mixing, adds the SiO after the deionized water dilution
2Colloidal sol is as liquid phase, and liquid-solid ratio is 2.Fully stir, prepare rheological homogeneous β-TCP slurry preferably.The polyurethane sponge is used the polyvinyl alcohol solution immersion treatment of NaOH solution and the 1wt% of 8M respectively after washed with de-ionized water, blot standby then with filter paper.Pretreated sponge is immersed in the slurry, vacuumizes slurry is entered in the hole of sponge fully, and then unnecessary slurry is extruded.Dry 24h under the room temperature, dry 12h in 80 ℃ of baking ovens, the temperature rise rate with 0.5 ℃/min rises to 600 ℃ then, and the temperature rise rate with 2 ℃/min rises to 1100 ℃ again, insulation 3h, furnace cooling is promptly prepared the porous beta-TCP pottery.
β used in the present invention-TCP powder is by the preparation of wet-chemical coprecipitation method, and raw materials used is analytical pure Ca (NO
3)
24H
2O and (NH
4)
2HPO
4The diameter of particle that obtains is 200nm～500nm.
The porosity of beta-tricalcium phosphate porous ceramic is measured by conventional Archimedes's method, and porosity is 92.6 ± 1.1%.
The mensuration in aperture is measured 10 above through holes by image analysis software, and the mean pore size of each batch beta-tricalcium phosphate porous ceramic is 410 ± 123 μ m.The ultimate compression strength of porous ceramics is that (Shimadzu carries out on Japan), and intensity is 98.87 ± 22.72Kpa, is enough to satisfy the clinical application of bone graft support for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 1B
The preparation 2 of porous beta-TCP pottery
Take by weighing 2g and cross the β-TCP powder of 80～100 order mesh screens, the methylcellulose gum that adds 5wt% in proportion is as binding agent, and 10wt% glass powder is as high-temperature agglomerant, after mixing, adds the SiO after the deionized water dilution
2Colloidal sol is as liquid phase, and liquid-solid ratio is 1.5.Fully stir, prepare rheological homogeneous β-TCP slurry preferably.The polyurethane sponge is used the polyvinyl alcohol solution immersion treatment of NaOH solution and the 1wt% of 10M respectively after washed with de-ionized water, blot standby then with filter paper.Pretreated sponge is immersed in the slurry, vacuumizes slurry is entered in the hole of sponge fully, and then unnecessary slurry is extruded.Dry 24h under the room temperature, dry 12h in 80 ℃ of baking ovens, the temperature rise rate with 0.5 ℃/min rises to 600 ℃ then, and the temperature rise rate with 2 ℃/min rises to 1150 ℃ again, insulation 3h, furnace cooling is promptly prepared the porous beta-TCP pottery.
The porosity of the beta-tricalcium phosphate porous ceramic that embodiment 1A, 1B obtain is measured by conventional Archimedes's method, and porosity is 88.6 ± 3.1%.
The mensuration in aperture is measured 10 above through holes by image analysis software, and the mean pore size of the beta-tricalcium phosphate porous ceramic of each batch is 300 ± 133 μ m.。The ultimate compression strength of porous ceramics is that (Shimadzu carries out on Japan), and intensity is 134.8 ± 44.25Kpa, is enough to satisfy the clinical application of bone graft support for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 2
The preparation of organizational project bone graft
Cultivate and collect bone marrow stroma stem cell (BMSC), make 0.6 * 10 with the DMEM nutrient solution that contains 10% calf serum
6The cell suspension of/ml is added dropwise in the porous support materials.Hatch 2～5h in the incubator, make cell attachment after, slowly add 20ml and contain the serum nutrient solution, under 37 ℃, 5%CO
2Condition under cultivate 3～14 days after, a part of sample is fixed with 2.5% glutaraldehyde, CO
2Critical point drying is observed under the SEM Electronic Speculum behind the metal spraying; Another part returns to be implanted in the animal body.
Embodiment 3
The experimentation on animals of bone graft
Cultivate and collect fat stem cell, make 0.6 * 10 with the DMEM nutrient solution that contains 10% calf serum
6The cell suspension of/ml is added dropwise in the beta-tricalcium phosphate porous ceramic material of preparation among the embodiment 1.Press the beta-tricalcium phosphate porous ceramic material volumeter, the inoculum size of fat stem cell is 0.6 * 10
6Individual cell/cubic centimetre porous ceramic film material.
Hatch 2～5h in the incubator, make cell attachment after, slowly add 20ml and contain the serum nutrient solution, under 37 ℃, 5%CO
2Condition under cultivate 3～14 days after, a part of sample is fixed with 2.5% glutaraldehyde, CO
2Critical point drying is observed under the SEM Electronic Speculum behind the metal spraying; Another part returns to be implanted in the animal body.
The result:
(stupalith of β-TCP) has higher porosity to porous bata-tricalcium phosphate of the present invention, and the aperture is 300～500 μ m, and connects in the hole, is suitable as tissue engineering cell scaffold material (as Fig. 1-2).Because porous beta-TCP stupalith porosity of the present invention is higher, and is better with the consistency of bone marrow stroma stem cell and fat stem cell.In SEM photo (shown in Fig. 3-6), can find all to be covered in rack surface and the hole, and spread all over extracellular matrix by cell.Also demonstrate the extensive distribution of cell and extracellular matrix on the section structure of material.Show thus, bone marrow stroma stem cell and fat stem cell can be at porous beta-TCP ceramic surface well-growns, and after cultivating certain hour, because the good connectedness of porous ceramics, cell also can migrate to the material internal justacrine and go out a large amount of extracellular matrixs, and consistency is preferably arranged.For bone marrow stroma stem cell and fat stem cell, the porous beta-TCP pottery for preparing by the foam impregnation method is the good cell timbering material, can be as the used in tissue engineering bone renovating bracket material.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Claims (10)
1. a beta-tricalcium phosphate porous ceramic material is characterized in that, the porosity of described stupalith is 85～95%, and mean pore size is the 200-500 micron, and the hole of 90%-99.999% interconnects.
2. stupalith as claimed in claim 1 is characterized in that, the resistance to compression hardness of described stupalith is 80-200Kpa, and mean pore size is the 300-500 micron.
3. stupalith as claimed in claim 1 is characterized in that, described stupalith is to be prepared by the method that contains following steps:
(a) with slurry dipping pore-forming template, slurry is fully entered in the hole of pore-forming template, the template that obtains flooding, wherein said slurry contains the β-TCP powder and 1～9 parts by weight of adhesive of 1～3 weight part, described pore-forming template can be calcined removal, and described slurry be the dipping template weight 4-1.5 doubly;
(b) the dipping template described in the step (a) is carried out drying, obtain the porous ceramics base substrate;
(c) the porous ceramics base substrate described in the step (b) is calcined at 900-1300 ℃, removed the pore-forming template, thereby obtain the agglomerating porous ceramics;
(d) porous ceramics in the cooling step (c) obtains described stupalith.
4. a method for preparing the described beta-tricalcium phosphate porous ceramic material of claim 1 is characterized in that, contains following steps:
(a) with slurry dipping pore-forming template, slurry is fully entered in the hole of pore-forming template, the template that obtains flooding, wherein said slurry contains the β-TCP powder and 1～9 parts by weight of adhesive of 1～3 weight part, described pore-forming template can be calcined removal, and described slurry be the dipping template weight 4-1.5 doubly;
(b) the dipping template described in the step (a) is carried out drying, obtain the porous ceramics base substrate;
(c) the porous ceramics base substrate described in the step (b) is calcined at 900-1300 ℃, removed the pore-forming template, thereby obtain the agglomerating porous ceramics;
(d) porous ceramics in the cooling step (c) obtains described stupalith.
5. method as claimed in claim 4 is characterized in that, described pore-forming template be the aperture be 23～40 holes/centimetre, the polyurethane sponge of porosity 95.0-99.9%, the polyurethane sponge of preferable porosity 98.0-99.9%.
6. method as claimed in claim 4, it is characterized in that, in described step (a) before, also comprise the pore-forming template is carried out pre-treatment: is alkali hydroxide soln immersion 12～24h of 1～10M with the pore-forming template in concentration, and/or the polyvinyl alcohol solution under 0.5～2wt% concentration soaks 12～24h.
7. method as claimed in claim 4 is characterized in that, binding agent is selected from the described step (a): bio-vitric, methylcellulose gum, silicon sol or its combination;
Perhaps, binding agent comprises methylcellulose gum, bio-vitric and silicon sol simultaneously described in the described step (a), wherein methylcellulose gum is 1～10wt% of powder quality, 3～20wt% of bio-vitric powder quality, silicon sol is 1～5 times of powder quality, Gu and the liquid/weight ratio of described slurry be 1～3.
8. method as claimed in claim 4 is characterized in that, the drying conditions in the described step (b) is as follows: the at room temperature dry 24h-72h of porous ceramics base substrate, 40～100 ℃ of following dry 8-16h;
Perhaps, the calcination condition in the described step (c) is as follows:
Temperature rise rate with 0.1-1 ℃/min rises to 400-800 ℃ with the porous ceramics base substrate, preferred 550-650 ℃; Temperature rise rate with 2～5 ℃/min rises to 1100～1300 ℃ then, and is incubated 1-6 hour.
9. graft, it is characterized in that, described graft contains the described beta-tricalcium phosphate porous ceramic material of claim 1 and is inoculated in the stem cell of described stupalith, and described stem cell is selected from bone marrow stroma stem cell or fat stem cell, and the inoculum size of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3Porous ceramic film material.
10. the purposes of beta-tricalcium phosphate porous ceramic material as claimed in claim 1 is characterized in that, as the support of preparation bone graft.
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| RU2525211C1 (en) * | 2013-07-30 | 2014-08-10 | Михаил Васильевич Гилев | Surgical approach to lateral condyle of tibia for osteosynthesis in fractures |
| CN107185034A (en) * | 2017-05-04 | 2017-09-22 | 中国科学院上海硅酸盐研究所 | Bioceramic scaffold and its production and use is repaired in osteochondral defect integration |
| CN107638595A (en) * | 2016-07-21 | 2018-01-30 | 重庆润泽医药有限公司 | A kind of bone implant |
| CN110075357A (en) * | 2019-04-02 | 2019-08-02 | 浙江大学 | A kind of preparation method of fat stem cell and the compound bone renovating material of nanometer fibroin |
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| CN110092653A (en) * | 2019-05-08 | 2019-08-06 | 武汉理工大学 | A kind of degradable bata-tricalcium phosphate porous bioceramic scaffold of 3D printing and its preparation method and application |
| CN113101410A (en) * | 2021-03-22 | 2021-07-13 | 华南理工大学 | Tricalcium phosphate support with uniform mesopore and three-dimensional communicated hierarchical pore structure as well as preparation method and application of tricalcium phosphate support |
| CN113101410B (en) * | 2021-03-22 | 2022-05-24 | 华南理工大学 | Tricalcium phosphate support with uniform mesopore and three-dimensional communicated hierarchical pore structure as well as preparation method and application of tricalcium phosphate support |
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