CN103341206A - Calcium phosphate/collagen/bone-like apatite three-level bionic bone tissue engineering scaffold and preparation method thereof - Google Patents
Calcium phosphate/collagen/bone-like apatite three-level bionic bone tissue engineering scaffold and preparation method thereof Download PDFInfo
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Abstract
The invention provides a calcium phosphate/collagen/bone-like apatite three-level bionic bone tissue engineering scaffold and a preparation method thereof. The material of the scaffold possesses components similar to natural bone tissues, and an inorganic/organic/inorganic three-level bionic bone-like netted three-dimensional structure. The preparation method comprises: customizing a calcium phosphate ceramic which possesses a bionic porous structure similar to the natural bone micropore structure, filling collagen prepared by simulated body fluid (SBF) into the porous calcium phosphate ceramic base, low-temperature aging to simulate a biomineralization process, in-situ nucleating on the collagen organic macro-molecule matrix, and self-assembly crystallizing to form a third level bone-like apatite layer structure. The composite scaffold material has bionic material components and bionic microstructure both similar to the natural bone tissues, the mechanical properties and the biological activity of the material are substantially improved, and the composite scaffold material has a wide application prospect.
Description
Technical field
The present invention relates to a kind of three grades of biotic bone tissue engineering stent materials that are composited by calcium phosphate ceramic, collagen protein and osteoid apatite and preparation method thereof, belong to field of biomedical materials.
Technical background
Natural bone is a kind of complex biological composite with fine grading structure, it forms by common structure of organic and inorganic composition, its organic principle mainly is type i collagen albumen (Type I collagen), it gives osseous tissue elasticity and toughness, its inorganic constituents is mainly by calcium phosphate (Calcium phosphate), especially biological phosphorus lime stone (Biological aptite) constitutes, and it gives the enough rigidity of osseous tissue and intensity.The minimum osteon of natural bone starts from the deposition of nanocrystalline apatite on collagen fabric, collagen fiber provide the mineralising forming core site of apatite and control its orientation with regularly arranged array structure, orientated deposition forms length and is approximately 30-50 nm, width is 10-30 nm, thickness is the bone crystal unit of 2-10 nm, countless basic osteons forms osseous tissue basic structure by assembling in order, gives material structure and the mechanical property of natural bone uniqueness.
Desirable osseous tissue " reparation " or " replacement " strategy need satisfy the material composition of implant and damaged tissues quite or approach, the implant material structure meets the anatomical structure of damaged tissues as far as possible, have enough biological activitys simultaneously, integrate easily and repair surrounding bone tissue, and might have active and induce, repair, rebuild the biological function of osteanagenesis.For this reason, desirable bone tissue engineering stent material should have three-dimensional multistage network structure.Wherein the one-level underlying structure can be the composition degradable synthos similar to natural bone, requires perforation hole with specific about hundreds of micron size simultaneously, grows and the space of tissue reconstruction for support provides initial strength and cell; Secondary structure is the compound collagen stroma of internal stent, and support is played toughened and reinforced and raising biological activity effectively; Tertiary structure is mineralising nanometer hydroxyapatite network structure, and it is the important biomolecule active interface of support/tissue handing-over, for the growth of osseous tissue provides effective site and required calcium source.Therefore undertaken bionical syntheticly by composition, architectural feature and mineralization process to natural bone self, and then the bionic bone repair material that obtains to have better biological function is regarded as one of development priority of hard tissue repair.
Because natural bone mainly is to be composited by calcium phosphate and collagen protein in itself, therefore from bionical angle, calcium phosphate/collagen composite materials is the splendid selection of bone tissue restoration substitution material.Collagen protein is a kind of natural polymer, and it is the main organic principle of human body skin and bone, is the rich in protein of content in the bone matrix.Collagen protein has good biocompatibility and low immunity, is suitable as the substrate that new osteocyte attaches growth, is a kind of ideal material for bone tissue restoration.Existing research recently be devoted to develop the preparation collagen-base or with it as the reparation of the biomaterial that adds phase as osseous tissue, but the combination property of gained material and bionical degree still have big gap from application requirements.
Therefore, the invention provides three grades of bionical bone tissue engineering stent materials of a kind of calcium phosphate/collagen albumen/osteoid apatite and preparation method thereof.This timbering material is given mechanical property and the pore structure of timbering material excellence by customization one-level porous calcium phosphate ceramic, pour into compound secondary collagen fiber and mineralising form three grades of osteoid apatites can highly bionical natural bone composition and structure, further promote the support biological activity, be expected to promote the reconstruction of bearing position osseous tissue.
Summary of the invention
The invention provides a kind of three grades of bionical bone tissue engineering scaffold composites and preparation synthetic method thereof.
The present invention is achieved through the following technical solutions:
A kind of three grades of bionical bone tissue engineering stent materials, mainly by porous calcium phosphate ceramic, collagen protein, bone like apatite layer is composited, and has inorganic/three grades of biomimetic features of organic/inorganic.Described bionic bracket material is made of the three-level network structure, its primary structure is the porous calcium phosphate ceramic (as matrix material) of customization, secondary structure is the collagen fabric that is compound in the ceramic hole structure, the bone like apatite layer of tertiary structure for forming by bionical thing mineralising.Described tertiary structure height imitation natural bone tissue anatomical structure.
As optimal way, this bionical compound support frame material component can be adjusted flexibly, and the mass percent of compositions at different levels (Wt%) is: calcium phosphate ceramic 50%～90%, collagen protein 9.5～45%, osteoid apatite 0.1～5%.
As optimal way, described porous calcium phosphate ceramic material composition can be: hydroxyapatite, β phase tricalcium phosphate or biphasic calcium phosphate pottery.The bionical pore structure with multistage connection of described pottery can adopt hydrogen peroxide foaming or microsphere occupy-place method or slip casting method or additive method design customization.
As optimal way, described porous calcium phosphate ceramic pore structure characteristic is that ceramic overall porosity is 60%～95%, and macropore diameter is 100～500 microns, has micropore to connect mutually between the macropore, and the macropore inwall is covered with the capillary micropore.
As optimal way, described collagen protein is filled in and forms three-dimensional collagen fabric network structure in the ceramic hole.This structure is conducive to improve the mechanical property of material on the one hand, promotes toughened and reinforced effect, forms sealedly with the loose structure of ceramic matrix material on the other hand, makes both combinations more firm, and material is more stable, also is conducive to cell tactophily on it simultaneously.The more important thing is that described tridimensional network collagen can provide the forming core site for the mineralising deposition of calcium and phosphorus ions, oriented growth for bone like apatite layer provides organic formwork simultaneously, promote the formation of biomineralization and bone like apatite layer, and be that three grades of biomimetic features of described class evenly are dispersed throughout timbering material inside, rather than only appear at material surface.
As optimal way, described collagen protein is the I-collagen type.Collagen protein is a kind of GL-PP, contains a spot of gala acid and glucose, is the main component of extracellular matrix, also is one of most important composition in the human body simultaneously.It extensively is present among the tissues such as skin, skeleton, muscle, cartilage, joint of human body.Wherein mainly have the I-collagen type in human body os osseum tissue, it plays multi-efficiencies such as support, protection and reparation.Mainly by fibroblast or synthetic with the similar cell in its source such as osteoblast, it induces osteanagenesis reconstruction aspect that important biological effect is arranged promoting osteocyte propagation, differentiation to the I-collagen type.
As optimal way, described collagen protein adopts standard analog body fluid (simulated body fluid, SBF) dilution that has same ion concentration with serum before compound with calcium phosphate.
As optimal way, each ion concentration is Na among the described SBF
+=142 mM, K
+=5.0 mM, Ca
2+=2.5 mM, Mg
2+=1.5 mM, HCO
3 -=4.2 mM, Cl
-=148.0 mM, HPO
4 2-=1.0 mM, SO
4 2-=0.5 mM, pH=7.2.
As optimal way, described collagen protein is the aqueous solution of 20～50 mg/ml before dilution, and dilution back concentration is 5～20 mg/ml.The dilution of employing simulated body fluid for system provides abundant calcium and phosphorus ions, is conducive to the formation of bone like apatite layer in the mineralization process.
As optimal way, the bone like apatite layer of described tertiary structure forms three-dimensional needle-like, flake nano structure at collagen fabric, and this structure forms by the calcium and phosphorus ions mineralising precipitation that calcium phosphate ceramic substrate and SBF provide.Described nanostructured is more near the natural bone tissue structure, the forming process of altitude simulation natural bone tissue.
Another object of the present invention is to provide a kind of preparation method of above-mentioned three grades of bionical bone tissue engineering stent materials, its operating procedure comprises:
(a) the preparation IPN connects porous calcium phosphate ceramic, prepares ceramic porous structure idiosome, then through sintering porcelain into;
(b) configuration collagen protein diluent: prepare the collagen protein diluent with SBF solution dilution configuration collagen protein;
(c) composite collagen/ceramic secondary structure: the porous ceramics in the step (a) is immersed in the collagen protein diluent of preparation in the step (b), evacuation perfusion under the room temperature, thus the mode of collagen protein with the negative pressure of vacuum perfusion is pumped in the porous ceramics hole;
(d) leave standstill mineralising, with the lyophilization of step (c) products therefrom, left standstill again 1-3 days, will separate out third level nanometer bone like apatite layer for the template crystallographic orientation with the collagen fabric moisture evaporation or evaporation the time in the product;
(e) crosslinked encapsulation: step (c) products therefrom is carried out crosslinked, at last to the product encapsulation of sterilizing.
As preferably, in the described step (a), the pore structure of pottery adopts hydrogen peroxide foaming or microsphere occupy-place method or slip casting method to prepare ceramic porous structure idiosome, sinters porcelain into through Muffle furnace then, and the temperature of described sintering is 1000～1200 ℃.
As preferably, in the described step (b), by the pH value of described collagen protein diluent is finely tuned, make its PH 5.5～8.0, thereby make described bone like apatite layer form three-dimensional needle-like, flake nano network structure in the collagen fabric growth.
As preferably, in the described step (b), the collagen protein diluent concentration that disposes is 5～20 mg/ml.
As preferably, in the described step (c), described negative pressure filling process, vacuum requires 0～10 Pa, pressurize 1～3 hour.Thereby guarantee that the cancellated formation of three-dimensional collagen fabric guarantees composite quality.
As preferably, in the described step (c), described negative pressure filling process carries out down in that ultrasonic wave concussion is auxiliary, thus the compound uniformity of raising product.
As preferably, described step (c) repeats once at least.Increase the compound quantity of collagen protein, and further guarantee the cancellated formation of three-dimensional collagen fabric, improve the firm degree of material combination simultaneously, guarantee the stability of complex.
As preferably, crosslinked employing physical crosslinking method is carried out described in the step (e).As optimal way, described cross-linking method is UV-crosslinked (J Biomed Mater Res.1995,29 (11): 1373-1379), cross-linking radiation (Radiat Res.1964,22 (4): 606-621), xeothermic crosslinked (Chinese reconstruction surgical magazine .2005,19 (10): a kind of 826-830).Adopt physical method not introduce other chemical reagent, avoided poisonous, harmful substance to the pollution of bioceramic, guarantee biocompatibility and the safety of product.
The present invention also provides a kind of purposes of described three grades of bionical bone tissue engineering stent materials: used as human body hard tissue damaged reparation or bone tissue engineer timbering material.Particularly use it for bearing position osseous tissue repair in trauma.
As preferably, three grades of bionical bone tissue engineering stent materials of the present invention also can have the bone tissue engineering scaffold of specific drugs, biological function with compound preparations such as medicine, gene, cells.
Beneficial effect of the present invention:
1, biotic bone tissue engineering stent material of the present invention is raw material with calcium phosphate ceramic and collagen protein, matrix components from material component simulation natural bone, significantly promote biocompatibility and the biological activity of material, promoted its bone tissue restoration regeneration function.
2, timbering material of the present invention is by three grades of biomimetic features of calcium phosphate/collagen albumen/osteoid apatite, imitation natural bone tissue anatomical structure, this structure has improved the mechanical property of material on the one hand, on the other hand can be for cell provides the microenvironment similar with natural bone, help bone photo close cell adhesion, breed and induce new bone tissue to generate.
3, bionic bracket material of the present invention adopts grading system to make compound approach, be beneficial to control, custom materials characteristic in material preparation technologies at different levels, as optimizing the material hole structure at the porous ceramics preparatory phase, in composite collagen stage custom materials component, optimize material surface/interface biological characteristics at three grades of bone like apatite layer synthesis phases, thereby make the bionical natural bone of material height, possess good osteoinductive and osteogenesis ability, induce new bone tissue to generate rapidly, strengthen bone connection and healing function.
4, in the whole preparation process of bionic bracket material of the present invention, no chemical reaction and organic solvent participate in, and have avoided poisonous, harmful substance to the pollution of bioceramic.Product has good structural intergrity and mechanical property preferably, can be processed into different shape by the reparation requirement of damaged bone, satisfies multiple application demand.
Description of drawings
Fig. 1 is preparation technology's flow chart of the present invention;
The scanning electron microscope of sample segment (SEM) photo among Fig. 2 embodiment.Wherein (a) is the SEM shape appearance figure of pure porous calcium phosphate ceramic material among the embodiment; (b) be compound collagen fabric SEM shape appearance figure in the porous calcium phosphate ceramic material internal hole among the embodiment; (c) the lamellar osteoid apatite SEM shape appearance figure for generating on the collagen fabric template in the porous calcium phosphate ceramic material internal hole among the embodiment.
Fig. 3 is the cell in vitro cultivation results.Wherein (a) is the external 2 all laser co-focusing micrograies of cultivating altogether of mesenchymal stem cells MSCs (MSCs) of pure porous beta-TCP pottery (experiment contrast group); (b) be that β-TCP/Collagen binary compound rest and mesenchymal stem cells MSCs (MSCs) are in external laser co-focusing microgram of cultivating for 2 weeks altogether.(c) for three grades of biotic bone tissue engineering stents of the present invention exploitation and mesenchymal stem cells MSCs (MSCs) in external laser co-focusing microgram of cultivating for 2 weeks altogether.
Fig. 4 is experimental result in the animal body.Wherein (a) is for testing parallel control sample (pure porous bata-tricalcium phosphate pottery) through 2.5 months rabbit muscle of back implanting results; (b) be β-TCP/Collagen binary compound rest zoopery result; (c) be the new bone tissue pattern slice map of three grades of biotic bone tissue engineering stents of the present invention exploitation.
Specific implementation method:
For reaching above-mentioned purpose, the present invention prepares three grades of biotic bone tissue engineering stent materials of calcium phosphate/collagen albumen/osteoid apatite according to following method, below in conjunction with accompanying drawing technology of the present invention is described in further detail.
Preparation technology's flow process of the present invention as shown in Figure 1.Raw-material preparation is divided into three aspects, first design customization porous calcium phosphate ceramic body; It two is that the preparation of type i collagen protein solution is prepared, and it comprises the extraction, purification of collagen protein and to its immunogenic removing; It three is configuration standard simulated body fluids.The porous calcium phosphate ceramic that the present invention was suitable for mainly comprises following three classes: hydroxyapatite (Hydroxyapatite, HA), tricalcium phosphate pottery (beta-Tricalcium phosphate, β-TCP), and biphasic calcium phosphate pottery (Biphasic calcium phosphate, BCP), wherein the making of porous ceramics can be via hydrogen peroxide foaming (Chinese patent: CN1903384), microgranule occupy-place method (Chinese patent: CN 1268583C), slip casting method (United States Patent (USP): No.3090094) or the porous ceramics idiosome of additive method design customization with specific pore architectural feature, be sintered into porous calcium phosphate ceramic through Muffle furnace then.The pottery that the embodiment of the invention is used prepares idiosome by the hydrogen peroxide foaming, and ceramic overall porosity is 60%～95%, and macropore diameter is 100～500 microns, has micropore to connect mutually between the macropore.The collagen protein that the present invention is suitable for is all kinds collagen protein, the embodiment of the invention is used through sterilization, purification, is gone the present invention that demonstrates of the calf Corii Bovis seu Bubali type i collagen albumen of immune terminal amino acid, its original state is 20～50 mg/ml high purity concentrations, be configured to 5～20 mg/ml with perfusion composite porous calcium phosphate pottery through the dilution of standard analog body fluid, each constituent mass percentage ratio (Wt%) of synthetic after-poppet material is: calcium phosphate ceramic 50%～90%, collagen protein 9.5～45%, osteoid apatite 0.1～5%.
The following several optimum examples of the present invention of classifying as it should be understood that these embodiment only are used for the purpose of illustration, never limit protection scope of the present invention.
Embodiment 1
Be three grades of synthetic biotic bone tissue engineering stents of substrate with the hydroxyapatite porous ceramics.Its making concrete steps are as follows:
1) customization porous calcium phosphate ceramic product.The product hole requires: it is 80 ± 20 μ m that whole porosity 85% ± 5%, macropore diameter 400 ± 50 μ m, IPN connect micropore size, is cut into the thin discs of Φ 15 * 2 mm;
A) will be through the hydroxyapatite powder that Wet Method Reaction makes standard model sieve, filtering out diameter is the hydroxyapatite dry powder of 80～160 μ m, adopts the hydrogen peroxide foaming to prepare 20 * 20 * 50 mm porous ceramics idiosomes, low temperature＜100 ℃ oven dry subsequently;
B) ceramic idiosome is put into the Muffle furnace sintering, be warming up to 1100 ℃ from room temperature with the speed of 5 ℃/min and continue 2 hours sintering, furnace cooling then.Divide the thin round test piece that is cut into Φ 15 * 2 mm subsequently;
2) configuration collagen protein diluent.Will be through sterilization, purification, remove the calf Corii Bovis seu Bubali type i collagen albumen of immune terminal amino acid, be modulated to 5 mg/ml concentration with standard solution (SBF) dilution;
3) priming by vacuum composite porous ceramic, collagen protein.With the collagen protein diluent submergence porous ceramics for preparing, room temperature is evacuated to＜and 10 Pa pour into, and pressurize 3 hours was stirred 10 minutes through ultrasonic wave concussion again, repeated evacuation negative pressure filling process once.
4) leave standstill the synthetic bone like apatite layer of mineralising.The sample that has poured into collagen protein was left standstill 24-72 hour, the moisture in drying, the volatile material, inducing under forming core and the mediation of collagen fiber template, the inorganic mineral ion in the material system forms the osteoid apatite three-dimensional net structure with crystallization.
5) crosslinked.Adopted the crosslinked sample of UV-crosslinked method 8 hours, sterilization stores.
Embodiment 2:
(β-TCP) pottery is the synthetic three grades of biotic bone tissue engineering stents of matrix with the porous bata-tricalcium phosphate.The porous ceramic matrices suitable porosity is 85% ± 5%, macropore diameter is 400 ± 50 μ m, to connect micropore size be 80 ± 20 μ m to IPN, and all the other product preparation process and step are identical with embodiment 1.The scanning electron microscope picture of the porous bata-tricalcium phosphate pottery of gained is shown in (a) among Fig. 2, (b) is collagen fabric SEM shape appearance figure compound in the porous calcium phosphate ceramic material internal hole among Fig. 2, (c) lamellar osteoid apatite SEM shape appearance figure for generating on the collagen fabric template in the porous calcium phosphate ceramic material internal hole among Fig. 2.The mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels wherein is: calcium phosphate ceramic～85%, collagen protein～14.5%, osteoid apatite～0.5%.
Embodiment 3
Other condition and technology are with embodiment 2, and difference is to adopt the porous bata-tricalcium phosphate pottery of high porosity to be the synthetic three grades of biotic bone tissue engineering stents of matrix.Its ceramic porosity 95% ± 5%, macropore diameter are that 500 ± 100 μ m, IPN perforation micropore size are 100 ± 20 μ m.The mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels of gained is: calcium phosphate ceramic～80%, collagen protein～19%, osteoid apatite～1%.The lifting of pottery porosity is beneficial to the compound more collagen protein of perfusion, also is conducive to the generation of bone like apatite layer simultaneously.
Embodiment 4
Other condition and technology are with embodiment 2, difference is to adopt the collagen protein of variable concentrations to be prepared, its collagen protein is modulated to 15 mg/ml concentration through simulated body fluid (SBF), the mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels of gained is: calcium phosphate ceramic～85%, collagen protein～14.8%, osteoid apatite～0.2%.This presentation of results SBF solution is to the importance of synthesizing inorganic/organic/inorganic tertiary structure biologic bracket material, and we adopt deionized water dilution collagen protein to repeat step of the present invention in other experiments, can't obtain the bionic bracket material of tertiary structure.Reduced SBF content in the present embodiment and caused calcium, phosphate ion concentration in the material system to reduce, this is unfavorable for that the bone like apatite layer in the tertiary structure forms.
Embodiment 5
Three grades of biotic bone tissue engineering stents that embodiment 2 preparation is synthetic and mesenchymal stem cells MSCs (MSCs) were cultivated for 2 weeks altogether external, adopt diacetic acid fluorescein/propidium iodide (FDA/PI) that cell is dyeed, adopt laser confocal microscope to observe and take pictures then, the result shows shown in (c) among Fig. 3, MSCs has obtained tangible propagation in this biotic bone tissue engineering stent material, differentiation, compare the experiment contrast group: pure porous beta-TCP pottery (as among Fig. 3 a) and β-TCP/Collagen binary compound rest (as b among Fig. 3), observed MSCs cell quantity and density are bigger.This result shows that this three grades of biotic bone tissue engineering stent materials have better biocompatibility and biological activity, are beneficial to propagation and the differentiation of skeletonization relevant cell.
Embodiment 6
Three grades of biotic bone tissue engineering stent materials of embodiment 2 preparations are carried out experiment test in the animal body.The sample of Φ 3 * 6 mm is implanted the new zealand white rabbit muscle of back to be taken out in 2.5 months, carrying out decalcification handles, dyeing, sections observation freshman bone tissue of sclerous tissues pattern, the result is shown in (c) among Fig. 4, compare the experiment contrast group: pure porous beta-TCP pottery (as among Fig. 4 a) and result shown in β-TCP/Collagen two-stage compound rest (as b among Fig. 4), experiment shows through 2.5 months rabbit muscle of back implants, there is freshman bone tissue to form in the sample in the present embodiment, the formation situation of freshman bone tissue and bone formation rate obviously are better than testing the parallel control sample, this result shows that this three grades of biotic bone tissue engineering stent materials have induces osteanagenesis and speed the function that the bone defect repairing is rebuild.
With method of the present invention, the calcium phosphate/collagen albumen of preparing/osteoid apatite biotic bone tissue engineering stent material, highly bionical natural bone tissue on the space structure of material and the component, three grades of synthetic bionical complex timbering materials not only have good biological activity, the mechanical property of material has obtained remarkable lifting simultaneously, has broad application prospects clinically at orthopaedics.
Embodiment 7
Be the collagen protein diluent that 60% porous beta-TCP pottery is immersed in 5mg/ml with porosity, evacuation perfusion under the room temperature, vacuum is 10Pa, pressurize 1 hour after the lyophilization, was left standstill 2 days, product is carried out crosslinked, the sterilization encapsulation.The mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels of gained is: calcium phosphate ceramic～90%, collagen protein～9.5%, osteoid apatite～0.5%.
Embodiment 8
Be the collagen protein diluent that 95% porous beta-TCP pottery is immersed in 20mg/ml with porosity, evacuation perfusion under the room temperature, vacuum is 0.01Pa, pressurize 3 hours, described negative pressure filling process carries out under ultrasonic wave concussion is auxiliary, repeats above-mentioned submergence and filling process 5 times, after the lyophilization, left standstill 3 days, product is carried out crosslinked, sterilization encapsulation.The mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels of gained is: calcium phosphate ceramic～50%, collagen protein～45%, osteoid apatite～5%.
Embodiment 9
Be the collagen protein diluent that 82% porous beta-TCP pottery is immersed in 10mg/ml with porosity, evacuation perfusion under the room temperature, vacuum is 5Pa, pressurize 2 hours, described negative pressure filling process carries out under ultrasonic wave concussion is auxiliary, repeats above-mentioned submergence and filling process 2 times, after the lyophilization, left standstill 1 day, product is carried out crosslinked, sterilization encapsulation.The mass percent (Wt%) of calcium phosphate/collagen albumen/osteoid apatite compositions at different levels of gained is: calcium phosphate ceramic～84.9%, collagen protein～15%, osteoid apatite～0.1%.
The above only is the preferred embodiments of the present invention, only is illustrative for the purpose of the present invention, and nonrestrictive; Those of ordinary skills understand, and can carry out many changes to it in the spirit and scope that claim of the present invention limits, revise, even the equivalence change, but all will fall into protection scope of the present invention.
Claims (10)
1. biotic bone tissue engineering stent material is characterized in that it contains calcium phosphate, collagen protein and osteoid apatite, has inorganic/three grades of biomimetic features of organic/inorganic.
2. biotic bone tissue engineering stent material according to claim 1, it is characterized in that: described material has three grades of biomimetic features, and described calcium phosphate is porous ceramics, constitutes first order biomimetic features---porous calcium phosphate ceramic matrix; Described collagen protein is compounded in the described ceramic matrix, constitutes second level organic double compound biomimetic features; Described osteoid apatite is attached on described collagen protein and the pottery, constitutes third level inorganic composite biomimetic features.
3. biotic bone tissue engineering stent material according to claim 1 and 2 is characterized in that, the mass percent of each composition is: calcium phosphate 50%～90%, collagen protein 9.5～45%, osteoid apatite 0.1～5%.
4. biotic bone tissue engineering stent material according to claim 1 and 2 is characterized in that, described calcium phosphate is a kind of in hydroxyapatite, β phase tricalcium phosphate or the biphasic calcium phosphate pottery.
5. biotic bone tissue engineering stent material according to claim 2, it is characterized in that, the bionical pore structure of the multistage connection that described porous calcium phosphate ceramic matrix has, the pore structure characteristic of described pottery is porosity: 60%～95%, macropore diameter: 100～500 microns, have micropore to connect mutually between the macropore, the macropore inwall is covered with the capillary micropore.
6. biotic bone tissue engineering stent material according to claim 2 is characterized in that, described collagen protein is filled in and forms three-dimensional collagen fabric network structure in the ceramic hole.
7. biotic bone tissue engineering stent material according to claim 1 and 2 is characterized in that, described collagen protein disposes through standard analog body fluid (SBF) dilution before compound with calcium phosphate.
8. biotic bone tissue engineering stent material according to claim 1 and 2, it is characterized in that, described osteoid apatite forms bone like apatite layer in the collagen protein surface attachment, constitute the third level biomimetic features of described timbering material, described bone like apatite layer is three-dimensional needle-like or the flake nano structure of calcium and phosphorus ions mineralising formation of deposits on collagen fabric.
9. biotic bone tissue engineering stent preparation methods as claimed in claim 1 is characterized in that its making step comprises:
(a) calcium phosphate ceramic of bionical customization loose structure;
(b) preparation collagen protein diluent: with standard analog body fluid (SBF) dilution collagen protein;
(c) synthetic calcium phosphate/collagen albumen secondary structure is immersed in the porous ceramics in the step (a) in the collagen protein diluent of preparation in (b), evacuation perfusion under the room temperature;
(d) the synthetic bone like apatite layer of mineralising leaves standstill the material at low temperature that makes in the step (c) 1～3 day, and lyophilization again will be separated out the nanometer bone like apatite layer for the template crystallographic orientation with the collagen fabric moisture evaporation or evaporation the time in the product;
(e) crosslinked encapsulation: step (d) products therefrom is carried out crosslinked, at last to the product encapsulation of sterilizing.
10. the purposes of three grades of biotic bone tissue engineering stent materials as claimed in claim 1 or 2 is characterized in that, used as bone tissue engineering stent material.
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| CN110038158A (en) * | 2019-05-27 | 2019-07-23 | 中国人民解放军第四军医大学 | The formula and preparation method thereof of photocuring 3D printing Haversian system artificial bone scaffold |
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| CN103948959A (en) * | 2014-04-29 | 2014-07-30 | 东华大学 | Manufacturing method of polymer-base calcium phosphate/collagen three-dimensional composite bone scaffold |
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| JP2018529499A (en) * | 2015-12-07 | 2018-10-11 | ハンチョウ フアマイ メディカル デバイシズ カンパニー リミテッド | Composite material for bone repair based on decellularized biological tissue matrix material and method for preparing it |
| US10821207B2 (en) | 2015-12-07 | 2020-11-03 | Hangzhou Huamai Medical Devices Co., Ltd. | Composite materials for bone repair based on decellularized biological tissue matrix material and the preparation method thereof |
| CN107233619A (en) * | 2017-07-14 | 2017-10-10 | 中国人民解放军第四军医大学 | A kind of Porous titanium bone implant material of functionalization and preparation method thereof |
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| CN110038158A (en) * | 2019-05-27 | 2019-07-23 | 中国人民解放军第四军医大学 | The formula and preparation method thereof of photocuring 3D printing Haversian system artificial bone scaffold |
| CN114712557A (en) * | 2022-04-02 | 2022-07-08 | 中山大学附属口腔医院 | Hydrogel reinforced biological bone calcium apatite scaffold and manufacturing method thereof |
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