CN101596330B - Alpha-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones and preparation method thereof - Google Patents

Alpha-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones and preparation method thereof Download PDF

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CN101596330B
CN101596330B CN 200910088575 CN200910088575A CN101596330B CN 101596330 B CN101596330 B CN 101596330B CN 200910088575 CN200910088575 CN 200910088575 CN 200910088575 A CN200910088575 A CN 200910088575A CN 101596330 B CN101596330 B CN 101596330B
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CN101596330A (en
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毛克亚
王征
王岩
李江涛
杨云
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Beijing Zhongnuo Hengkang Biotechnology Co., Ltd.
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Abstract

The invention discloses a method for preparing alpha-calcium sulfate hemihydrates (alpha-CSH) /beta-tertiary calcium phosphate (beta-TCP) porous granular-type composite artificial bones, which is characterized in that the alpha-CSH is directly synthesized on the surface and/or in the pores of the beta-TCP granules synchronously by the hydrothermal synthesis process and finally the alpha-CSH/beta-TCP porous granular-type artificial bones is prepared. The invention has the advantages that the preparation process is simpler, the preparation period can be effectively reduced, and the preparation efficiency is improved; the proportion between alpha-CSH and beta-TCP phases of the granular-type composite artificial bones can be effectively controlled, thereby controlling the pore structure of the artificial bone granules; meanwhile, the invention guarantees the in-situ solidifying performance and degrading performance of the composite artificial bones, controls the evenness of the product and the porosity and pore diameter of the beta-TCP granules, improves the bone-formation performance and degrading performance of the composite artificial bones and further improves the quality of the product, thereby finally obtaining the composite artificial bones having self-solidifying performance and controllable degradation and having a certain biomechanical strength. The invention has good application and market prospect.

Description

α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones and preparation method thereof
Technical field
The present invention relates to a kind of preparation method of cmposite artificial bone and the product that is prepared from thereof, specifically utilize hydrothermal synthesis method directly the bata-tricalcium phosphate porous particle to be synthesized α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones, belong to field of medical materials.
Background technology
The inorganic compounding artificial bone belongs to the bioceramic material category, and in the past 50 years, bioceramic has experienced significant progress.Initial goal in research is not have biological activity, and has minimum interactive material with body.These bioceramices are known as first generation biomaterial.The research of second filial generation bioceramic material starts from twentieth century eighties, and it is having positive interaction as goal in research with biological tissue, and this class material has biological activity and resorbent characteristics.Arrived earlier 2000s, the medical science example has replaced tissue substitute by tissue regeneration.That is to say, bioceramic must be filled by the Cell and organism bioactive molecule, so the research work of third generation bioceramic begins at this point.The in recent years basic research of cmposite artificial bone and clinical use focus mainly concentrate on following a few class: hydroxyapatite, Corallium Japonicum Kishinouye heat conversion hydroxyapatite, bata-tricalcium phosphate (β-TCP), calcium-phosphate cement and calcium sulfate etc., these material sources are abundant, storage and easy to use, have good biocompatibility and avoid pathophorous advantage, but according to various disease is different to the performance requirement of bone-grafting material clinically, all there is certain shortcoming and defect.
For example: hydroox apatite artificial bone and calcium-phosphate cement material, although have good biocompatibility, because densification structure and the dissolubility of material are lower, so that substituting with new bone, the two degraded in body all can't realize.And the bata-tricalcium phosphate material of porous is as calcium phosphate biodegradability best material, though can realize growing into or substituting of new bone, but because the loose structure that material itself possesses, so that the mechanical property of this material reduces greatly, also there is simultaneously the poor problem of curing performance, plastotype and operating performance again.Therefore, the performance that more satisfactory polyethylene artificial bone composites should possess is: osteogenic activity and biocompatibility are good, and the new bone speed of growing into is consistent with material degradation speed, and has certain mechanical strength and clinical operability.
There is significant limitation in the patent of present published preparation polyethylene artificial bone composites.For example: application number is 200710063903.4 Chinese invention patent, discloses a kind of Preparation method and use of new-type of inorganic bone grafting material.This new-type of inorganic bone grafting material.The preparation method of this type of inorganic bone grafting material comprises: (1) preparation has the bata-tricalcium phosphate granule of microcellular structure; (2) preparation α-half-H 2 O calcium sulphate powder body; (3) the preparation mass percent is that bata-tricalcium phosphate granule and the mass percent of 20%-80% are 80%-20% α-half-H 2 O calcium sulphate powder body, forms α-half-H 2 O calcium sulphate powder layer in bata-tricalcium phosphate particle surface and/or hole.
The advantage of this invention is: based on α-half-H 2 O calcium sulphate and good biocompatibility and the vivo degradation performance of bata-tricalcium phosphate, the cmposite artificial bone with self-curing performance of preparation, the bone that can repair arbitrary shape is damaged, the mechanical strength of in-situ solidifying and fast quick-recovery skeleton.Along with the fast degradation of calcium sulfate, the bata-tricalcium phosphate granule with microcellular structure provides comparatively desirable support for new bone growth in vivo, and can be gradually by the area of new bone creeping substitution.This cmposite artificial bone wide material sources, storage and easy to use, function admirable, preparation technology is simple, practical, and manufacturing cycle is short and with low cost.But there is following defective in this invention preparation method: prepare at first respectively bata-tricalcium phosphate granule and α-half-H 2 O calcium sulphate powder body, then stirring with dehydrated alcohol and α-half-H 2 O calcium sulphate is pasty state, directly at bata-tricalcium phosphate particle surface spraying α-half-H 2 O calcium sulphate, the method can only form α-half-H 2 O calcium sulphate powder layer at the bata-tricalcium phosphate particle surface, and can not enter bata-tricalcium phosphate granule interior hole fully, and can not effectively control the spraying dosage of α-half-H 2 O calcium sulphate, porosity and the pore diameter of spraying uniformity and bata-tricalcium phosphate granule, thereby cause the bone formation performance of this cmposite artificial bone and degradation property to be difficult to control, and affect product quality.
Summary of the invention
The object of the invention is to for above-mentioned prior art shortcoming, preparation α-half-H 2 O calcium sulphate/bata-tricalcium phosphate (compound new technology of the hydro-thermal of porous particle type cmposite artificial bone of α-CSH/ β-TCP) is proposed, by on β-TCP porous particle surface and/or the synthetic α-CSH of hole inter-sync, carry out the modification of α-CSH/ β-TCP porous bone, the final pore structure that forms the granular pattern cmposite artificial bone of control, and play the purpose of regulating material degradation speed in body, realize finally by fully alternative from the body bone, obtain best bone defect repair effect.
For achieving the above object, the inventor's Research Thinking is: at first study the process kinetics for preparing porous beta-TCP with bovine cancellous bone, obtain the controlled near single-phase β of pore structure-TCP porous particle; And then research hydro-thermal synthesis process parameter on α-CSH at β-TCP porous particle surface out-phase forming core, grow up, distribution mode affect rule, realize the control to α-CSH/ β-TCP cmposite artificial bone mesopore structure.By curing performance, biocompatibility and the mechanism of degradation of studying this NEW TYPE OF COMPOSITE artificial bone, and to sign and the evaluation of its bone defect repair effect, feedback instructs the preparation technology of α-CSH/ β-TCP cmposite artificial bone, thereby obtain a kind of self-curing performance that has, and the degradation speed bone-grafting material consistent with the bone formation rate of bone defect repair in the implantable bone reaches desirable bone defect repair effect.
In order to realize purpose of the present invention, the concrete technical scheme of the present invention is:
A kind of preparation method of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones, it is directly to utilize synchronously synthetic α-half-H 2 O calcium sulphate of hydro-thermal synthesis process in bata-tricalcium phosphate porous particle surface and/or hole, finally is prepared into α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones.
Said method, its concrete steps are:
(1) preparation has the bata-tricalcium phosphate granule of natural pore structure;
(2) utilize hydro-thermal synthesis process to synthesize synchronously α-half-H 2 O calcium sulphate on the bata-tricalcium phosphate porous particle surface of step (1) preparation: to use CaSO 42H 2The O configuration concentration is 10～30% aaerosol solution, and then the bata-tricalcium phosphate granule with step (1) preparation is placed in one; The two is placed on temperature 110～140, under pressure 0.12～0.3MPa condition, behind reaction 6～12h, can separates out α-CSH at the porous particle surface crystallization of β-TCP.
After described step (2) reaction finishes, the solidliquid mixture of step (2) preparation in filtering more than 90 ℃, and is put into 120 ℃ baking oven and dried, namely finish the preparation of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones.In step (2), be 10～30% CaSO in concentration 42H 2Add crystal modifier in the O suspension, it is compound that described crystal modifier is that aluminum sulfate and citric acid are received, and accounts for CaSO 42H 2The 0.5wt% of O; Described aluminum sulfate is 50wt%～60wt%:40wt%～50wt% with the ratio that citric acid is received.The present invention tests and shows, the volume of crystal modifier has material impact to α-CSH performance.The voluntarily crystal modifier of preparation is used in this experiment, mixes according to a certain percentage with sodium citrate, aluminum sulfate.The crystal modifier that adds respectively different proportionings during steam pressure is tested, and inquires into the crystal modifier ratio to the impact of α-CSH pattern.Experimental result under the different proportioning crystal modifiers can be found out: crystal modifier aluminum sulfate and citric acid are received compositely proportional when being 60wt%:40wt% and 50wt%:50wt%, and synthetic α-CSH grain morphology is short cylinder, and distribution uniform.And when the crystal modifier compositely proportional was 60wt%:40wt%, the α of preparation-CSH crystallite dimension was larger, was about 30 μ m～50 μ m, and the crystal modifier compositely proportional is when being 50wt%:50wt%, and the α of preparation-CSH crystallite dimension is about 20 μ m～40 μ m, best results.
The concrete grammar that described step (1) preparation has the bata-tricalcium phosphate granule of natural pore structure is: adopt xenogenesis or homologous cancellus to slough behind the organic principle preroasting in the high-temperature calcination stove, 10 ℃/minutes of heating rates, 800 ℃ of calcinings 3 hours; The bone piece is soaked in (NH after taking out after will calcining 4) 2H PO 4In the solution 24 hours, remove unnecessary liquid, take out again 50 ℃ of oven dry 4 days of material; Material after the oven dry is put in the high-temperature calcination stove and calcines, 5～10 ℃/minutes of heating rates, 1100 ℃ of calcinings 1 hour slowly cooled to after the room temperature flowing water flushing 3 hours, rinsed with deionized water 2 times, 50 ℃ of oven dry 4 days.
Described (NH 4) 2H PO 4Concentration be that 0.6M～1.2M is best.
Xenogenesis or homologous cancellus are sloughed the concrete grammar of organic principle and are in the above-mentioned preparation method: get healthy bulls bone femoral head and distal femur spongy bone, after going cell, ungrease treatment, flowing water flushing, sodium hydroxide, hydrogen peroxide solution immersion use the distilled water rinsing clean, distilled water boiled 10 hours, the flowing water flushing is 3 hours again, rinsed with deionized water 2 times, then the 70%-100% gradient alcohol dehydration dried 4 days for 50 ℃.
The invention provides a kind of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones, it is that said method is prepared from.α of the present invention-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones structure is that bata-tricalcium phosphate porous particle surface and hole inside grow up to α-half-H 2 O calcium sulphate long column shape crystal.
α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones of the present invention serves many purposes, it is the good carrier of bone conduction and bone-inducting active material, therefore, the present invention can be used as the compound DBM of carrier and BMP repairs bone injury, or the open complicated war wound of composite antibiotic or chemotherapeutics treatment and a variety of causes osteomyelitis or the bone tumor that cause.
Concrete complex method is directly to add DBM, BMP, antibiotic or chemotherapeutics in cmposite artificial bone, has the self-curing performance, can directly inject use by special injecting apparatus, add other conventional medicine carrier and can improve its injection property, obtain better injection operation performance.
Advantage of the present invention and beneficial effect are:
1, raw material xenogenesis of the present invention or homologous cancellus are convenient can get, and can guarantee that synthetic α-CSH/ β-TCP Composite Bone granule has natural bone pore structure.
2, take β-TCP porous particle as the out-phase nucleating agent, in Hydrothermal Synthesis α-CSH, directly β-TCP porous particle surface void structure is carried out modification, so that preparation technology is simpler, can effectively shorten manufacturing cycle, improve preparation efficiency.
3, can be by regulating CaSO 42H 2The concentration of O solution, and the quantity of β-TCP porous particle, and the Hydrothermal Synthesis time, effectively control the biphase ratio of granular pattern cmposite artificial bone α-CSH and β-TCP, thereby the pore structure (hole dimension, pore-size distribution, pore communication) of control Composite Bone granule, guarantee simultaneously in-situ solidifying performance and the degradation property of cmposite artificial bone, final acquisition has the self-curing performance, controlled, as the to have certain biomechanical strength cmposite artificial bone of degrading.
4, the α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones of the inventive method preparation is when the bata-tricalcium phosphate particle surface forms α-half-H 2 O calcium sulphate powder layer, enter bata-tricalcium phosphate granule interior hole and form α-half-H 2 O calcium sulphate long column shape crystal, the uniformity of product and porosity and the pore diameter of bata-tricalcium phosphate granule have effectively been controlled, thereby improved bone formation performance and the degradation property of cmposite artificial bone, further improved product quality.
The invention will be further described below in conjunction with the specific embodiment, and embodiments of the present invention are not limited to this, every according to principle perhaps in disclosed by the invention, any this area of enforcement be equal to replacement, all belong to protection scope of the present invention.
Description of drawings
The XRD figure spectrum of 0 ℃ of calcination bovine cancellous bone of Figure 180;
000 ℃ of calcination of Figure 21 be soaked in 0.50molL-1NH4H2PO4 solution bovine cancellous bone XRD figure spectrum (HAP-◆, β-TCP-
Figure G2009100885752D00041
);
The pressure and temp profile of equilibrium figure of Fig. 3 calcium sulphate dihydrate and α-CSH (A), liquid phase water and gas phase water (B)
The XRD figure spectrum of Fig. 4 α-CSH powder body;
The NEW TYPE OF COMPOSITE artificial bone granule of Fig. 5 Hydrothermal Synthesis preparation;
Porous cmposite artificial bone after the porous Composite Bone structure of the compound front β-TCP of Fig. 6 hydro-thermal and hydro-thermal are compound;
The XRD figure spectrum of the compound artifact bone of Fig. 7 hydro-thermal;
As seen Fig. 8 porous beta-TCP particle detection result: A prepares the hole result of material; B, image after macropore amplifies; C, the microcellular structure of visible hole wall after amplifying; D, the XRD testing result;
Fig. 9 deposits the artificial bone hole of a large amount of α-CSH powder body artificial bone hole and deposition a small amount of α-CSH powder body;
Figure 10 α-CSH/ β-TCP cmposite artificial bone XRD figure spectrum;
Figure 11 cmposite artificial bone external degradation;
Figure 12 spinal fusion animal model;
Self-curing is implanted on Figure 13 spinal column right side can absorb cmposite artificial bone;
The outside 12 all mammographies were taken a picture after Figure 14 implanted spinal column;
The outside 12 all Micro-CT showed that embedded material and spinal fusion are integrated after Figure 15 implanted spinal column;
Figure 16 implants behind the spinal column outside 12 all histology and shows the area of new bone material internal of growing into;
The outside visible area of new bone of 12 all tetracycline markers formed after Figure 17 implanted spinal column;
Figure 18 implants at once outside drawing for the COB1 material in a left side; In be postoperative 4 all material degradation figure; The right side is postoperative 8 all material degradation figure;
Figure 19 left side is COB1 material 12 weeks after operation material degradation figure; The right side is postoperative 16 all material degradation figure;
Figure 20 implants at once outside drawing for the COB2 material in a left side; In be postoperative 4 all material degradation figure; The right side is postoperative 8 all material degradation figure;
Figure 21 left side is COB2 12 weeks after operation material degradation figure; The right side is postoperative 16 all material degradation figure;
Figure 22 implants at once outside drawing for the COB3 material in a left side; In be postoperative 4 all material degradation figure; The right side is postoperative 8 all material degradation figure;
Figure 23 left side is COB3 12 weeks after operation material degradation figure; The right side is postoperative 16 all material degradation figure.
The specific embodiment
The preparation of embodiment 1 α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones
(1) gets healthy bulls bone and reject its surperficial soft tissue, use internal diameter to drill through femoral head and distal femur spongy bone as the trepan of 8mm, approximately 8 * 12mm is cylindrical for gained bone block size, it is clean with the distilled water rinsing after cell, ungrease treatment are gone in flowing water flushing, sodium hydroxide, hydrogen peroxide solution immersion etc., distilled water boiled 10 hours, and the flowing water flushing is 3 hours again, rinsed with deionized water 2 times, then the 70%-100% gradient alcohol dehydration dried 4 days for 50 ℃.To be placed on through the material of above-mentioned processing preroasting in the high-temperature calcination stove, 10 ℃/minutes of heating rates were calcined 3 hours for 800 ℃.(the NH of 1M will be soaked in after the bone piece takes out after the calcining 4) 2HPO 4In the solution 24 hours, remove unnecessary liquid, take out again 50 ℃ of oven dry 4 days of material.Material after the oven dry is put in the high-temperature calcination stove and calcines, 5 ℃/minutes of heating rates, calcined 1 hour for 1100 ℃, flowing water washed 3 hours after slowly cooling to room temperature, rinsed with deionized water 2 times, 50 ℃ of oven dry can prepare nearly single-phase β-TCP porous particle in 4 days, its macropore diameter 200～600 μ m, and hole wall has the microcellular structure about diameter 1 μ m.
(2) use analytical pure CaSO 42H 2The O configuration concentration is the aaerosol solution of 20wt%, and adds crystal modifier, crystal modifier be aluminum sulfate (analytical pure, Beijing chemical reagents corporation) with citric acid receive (analytical pure, Beijing chemical reagents corporation) compound, account for CaSO 42H 2The 0.5wt% of O; Described aluminum sulfate is 50wt%～60wt%:40wt%～50wt% with the ratio that citric acid is received.Then β-TCP porous particle is placed in one.Then the two is placed in the airtight steam pressure still, the temperature in the autoclave pressure is controlled at 135 ℃, and pressure is controlled at 0.2MPa.Behind the reaction 7h, can separate out α-CSH at the porous particle surface crystallization of β-TCP.After reaction finishes, the solidliquid mixture in the autoclave in filtering more than 90 ℃, and is put into 120 ℃ baking oven and dried, the so far α after the modification-CSH/ β-TCP porous Composite Bone granule preparation is finished, and namely can be used as bone-grafting material after the package sterilization and uses.
Analysis for preparation method of the present invention:
(1) at first, preparation has the bata-tricalcium phosphate granule of natural pore structure;
With healthy xenogenesis or homologous cancellus through defat, processed after, be taken at dinectly bruning in the high temperature furnace, lower 3 hours of 800 ℃ of conditions, 5～10 ℃/minutes of heating rates, bovine cancellous bone after the calcination is that principal phase is the hydroxyapatite (HAP) by stoichiometric proportion, as shown in Figure 1.
By experiment as can be known, the technical parameter of calcination processing has a great impact to the transformation of β-TCP HAP.Take spongy bone as raw material, through variable concentrations NH 4H 2PO 4The immersion of solution, and the calcination processing of different temperatures can obtain the material of different phase compositions.At a certain temperature, the HPO that decomposites of soaking solution 4 2-Increase with soaking solution concentration, and HPO 4 2-The increase amount that causes HAP to change into β-TCP increase, (Fig. 1) can find out from XRD figure, HAP weakens by force at the peak gradually, β-TCP peak strengthens by force gradually, with the XRD figure of standard hydroxyapatite (HA) and β-TCP spectrum relatively, the characteristic diffraction peak that prepared near single-phase β-TCP granule XRD occurs at angle of diffraction 2 θ and characteristic diffraction maximum (main peak 2 θ=31.0 of standard β-TCP collection of illustrative plates; Inferior strong peak 2 θ=27.8,34.5,16.9,53.2 peaks weaken by force successively) coincide.Characteristic diffraction maximum (main peak 2 θs=31.8 close with HA also appear simultaneously, inferior strong peak 2 θ=31.2,31.9,25.9,49.5 the peak weakens by force successively), but its intensity obviously is weaker than the characteristic peak intensity of β-TCP, confirms that prepared material main component is β-TCP and contains a small amount of HA, does not find other composition characteristics diffraction maximums in the collection of illustrative plates.
Soaking solution concentration is increased to certain limit, with HPO 4 2-Ion consumes, HPO 4 2-Ion impels HAP to change into a little less than β-TCP effect more and more, simultaneously, and NH 4H 2PO 4The H that decomposites +Ion concentration improves, and impels HAP to change into Ca 2P 2O 7Act on more and more obviously, the dominant response that whole system occurs is that HAP changes into Ca 2P 2O 7Fig. 1 shows, 1100 ℃, and during 0.6M～1.2M, H +Impel HAP to change into Ca 2P 2O 7Play a major role, a large amount of HAP change into Ca 2P 2O 7, cause β-TCP content to reduce gradually Ca with the concentration increase 2P 2O 7Content increases gradually.According to above rule, prepared nearly single-phase β-TCP porous particle.The XRD testing result is seen Fig. 2.Collection of illustrative plates shows that principal phase is β-TCP, and contains a small amount of HAP.This is to generate HAP because β-TCP easily absorbs water to transform in wet environment, contains a small amount of HAP in the sample that causes preparing.Prepared near single-phase β-TCP is porous particle, and its macropore is 200～600 μ m, and hole wall has the microcellular structure about diameter 1 μ m.
(2) hydro-thermal synthesis process is at β-synchronously synthetic α-CSH in TCP porous particle surface.
Half-H 2 O calcium sulphate exists biphase, i.e. α phase and β phase, and their chemical components are identical, but the degree of crystallinity of α-CSH is far above β-CSH, so that the α type has the intensity of better plasticity and Geng Gao.The formation condition of the two also there are differences, and under 125～180 ℃ of dry air conditions, calcium sulphate dihydrate changes β-CSH into, and in 105～135 ℃ of saturated vapors and aqueous medium, calcium sulphate dihydrate changes α-CSH into, and this conclusion can be confirmed from Fig. 3.
As can be seen from Figure 3, the profile of equilibrium of calcium sulphate dihydrate and α-CSH (A) is in close proximity to the profile of equilibrium (B) of liquid phase water and gas phase water, and these two curves intersect near 100 ℃ the time.If calcium sulphate dihydrate has just formed α-CSH at liquid phase dissolved-recrystallize in greater than 1 atmospheric pressure cooker.1,2,3,4 spin cycle is finished in liquid phase fully on the curve, can form α-CSH of well-crystallized.On the contrary, be lower than under the atmospheric pressure, the water in the calcium sulphate dihydrate is just with the evaporation of dry saturated steam state, thereby generation microcosmic crystal is the microporous solid of loose gathering, Here it is beta-type semi-hydrated gypsum.
Therefore, the condition of Hydrothermal Synthesis just is defined as substantially: the sulfate dihydrate calcium powder is mixed slurry solid content: 15wt% with deionized water, place in the airtight pressure vessel, add simultaneously a certain amount of crystal modifier, it is compound that crystal modifier adopts aluminum sulfate and citric acid to receive, and accounts for the 0.5wt% of amount of gypsum; Under the pressure of 100～120 ℃ temperature and 0.1～0.15MPa, make calcium sulphate dihydrate dissolving recrystallize form α-CSH, then take out fast, after filtering with boiling water, put into the air dry oven freeze-day with constant temperature 4～6 hours, namely obtain α-CSH powder body.The present invention determines that hydro-thermal method synthesizes under the optimal processing parameter of α-half-H 2 O calcium sulphate, and the stable α-CSH powder body of preparing in the Hydrothermal Synthesis still is the α-CSH powder body of pure phase through X-ray diffraction analysis (XRD), as shown in Figure 4;
Because the bovine cancellous bone after the calcination is the material of loose structure, can play the effect of zeolite, therefore in the hydro-thermal recombination process of biphase material, reactive state is steady, the phenomenon of solution bumping do not occur.Take the porous particle of nearly single-phase β-TCP as the out-phase nucleating agent, directly β-TCP porous particle surface void structure is carried out modification when adopting Hydrothermal Synthesis α-CSH.The near single-phase β for preparing-TCP porous particle is placed the CaSO in the airtight steam pressure still 42H 2In the O solution, according to 70wt% α-CSH+30wt% β-TCP, 50wt% α-CSH+50wt% β-TCP, 30wt% α-different proportionings such as CSH+70wt% β-TCP, α-CSH at β-TCP porous particle surface forming core, grow up, directly prepare α-CSH/ β-TCP cmposite artificial bone.In the complex that obtains, after porous beta-TCP was coated, the NEW TYPE OF COMPOSITE artificial bone as shown in Figure 5 and Figure 6.Wherein can be by regulating CaSO at 10-30%wt% 42H 2The quantity of the concentration of O solution and β-TCP porous particle, the pore structure of control granular pattern cmposite artificial bone, obtain large bore dia 100～300 μ m, hole wall has the granular pattern cmposite artificial bone of microcellular structure, and make the α-CSH of pore surface generation grow up to the long column shape crystal, be beneficial to osteanagenesis after its degraded.Carry out X-ray diffraction analysis (XRD) after this artificial bone ground, the result shows that the multiporous biological bone material after compound is α-CSH, β-TCP and Ca 10(PO 4) 6(OH) 2The composite of (hydroxyapatite) three-phase, as shown in Figure 7.
Embodiment 2: physicochemical property and index detect
New product is detected, testing result and photo are provided, prove and realized goal of the invention.
The single-phase β of embodiment 1 preparation-TCP porous particle is carried out respectively scanning electron microscope, X-ray diffraction (XRD), porosity and biomechanics and detects.(* 40) visible resulting materials is cellular porous structure under the scanning electron microscope low power lens, the cellular space structure that keeps natural spongy bone, the aperture is similar to spongy bone with porosity, the macropore diameter size is between 50-600um, the densely covered microcellular structure of (* 3000) visible material macropore hole wall under high power lens, size is about 1um, and is connective good between hole, as shown in Figure 8.With the JCDS card contrast of standard β-TCP, the characteristic diffraction peak that the XRD of β-TCP porous particle occurs at angle of diffraction 2 θ (main peak 2 θ=31.0; Inferior strong peak 2 θ=27.8,34.5,16.9,53.2 the peak weakens by force successively), in the diffracting spectrum characteristic diffraction maximum close with standard HA diffracting spectrum also appearred simultaneously, but its characteristic peak intensity obviously is weaker than the intensity of the characteristic peak of β-TCP, does not find other composition characteristics diffraction maximums in the collection of illustrative plates, as shown in Figure 8.The porosity that mercury injection apparatus is measured the preparation material is 57.63%, show as bimodal curve, wherein previous peak is the hole situation of macropore in the material, the aperture is 403143.3nm-58433.5nm, the hole situation that the microcellular structure that second peak shows as material itself shows, pore size is 552.5-77.1nm.The comprcssive strength compressive strength that the biomechanics experiment machine is measured nearly single-phase β-TCP porous material test specimen is 4.47 ± 0.63MPa.
The α of embodiment 1 preparation-CSH/ β-TCP cmposite artificial bone is carried out respectively scanning electron microscope, XRD and biomechanics and detects.As seen grow up to a large amount of α-CSH long column shape crystal on β-TCP porous particle surface and hole inside under the scanning electron microscope, as shown in Figure 9, XRD can see the crest of α-CSH and β-TCP simultaneously, as shown in figure 10.Different according to α-CSH content, the compressive strength height that α-CSH/ β-TCP cmposite artificial bone adds behind the water cure is different, and the biomechanics experiment machine is measured the comprcssive strength compressive strength of α-CSH/ β-TCP cmposite artificial bone test specimen between 3MPa-10MPa.
Embodiment 3: external and vivo degradation experiment
The α of embodiment 1 preparation-CSH/ β-TCP cmposite artificial bone is prepared the standard that size is 12 * 8mm, be positioned over original ph and be in 7.4 the simulated body fluid, constant temperature is preserved in 37 ℃ of incubators, do not change liquid, measure once the 1st every day in week, each week is afterwards measured the pH value of a soak with the pH value analyzer, until pH value is constant, till no longer changing.The original ph that then above-mentioned test specimen is positioned over respectively be in 7.4 the 50ml SBF solution, to be positioned in 37 ℃ of incubators, constant temperature is preserved, and changes liquid once every 2 days, changes liquid 30ml at every turn; After being dried to constant weight, 4w, 8w, 12w, 16 all test specimens measure its residual heavy (Wr), Calculating material external degradation rate W%=[100 (Wo-Wr)/Wo].
In α-CSH/ β-TCP cmposite artificial bone degradation process, pH value prolongs gradually with degradation time and reduces, 7.4 5.52 when being reduced to gradually for 5 week when initial, and the last fortnight changes greatly, and only slightly reduction is basicly stable about 5.48 afterwards.In the degraded, the quality of cmposite artificial bone all increases constantly in time and reduces under simulated body fluid, and degraded is along with passage of time, and degradation speed slows down gradually.In the composite, along with α-CSH content increases, degradation speed also increases thereupon.Cmposite artificial bone has degraded approximately about 27.7%～42.4% in 16 whens week, as shown in figure 11.
48 of healthy new zealand white rabbits, male and female are not limit, body weight 2.5-3.0Kg, mix with sleep new II 1.5ml+ ketamine 2ml of speed, according to the intramuscular anesthesia of 0.2ml/kg body weight row, the anesthesia rear dorsal position of getting that comes into force, extremity fixing safe after, touch lateral femur condyle section, line makes marks, iodophor disinfection, spread aseptic hole-towel, do vertical stringer otch in outside femur condyle midpoint, be about 3cm, cut successively skin, subcutaneous, deep fascia and joint capsule and expose lateral side of femur and lateral meniscus outer rim.Apart from distal femur articular surface 4mm place, with solid brill (external diameter 2mm and 5mm) ecto-entad boring parallel with articular surface, the degree of depth is 12mm.Cause the damaged hole of bone of diameter 5mm * dark 12mm.Implant respectively than (7: 3,5: 5, be respectively COB1, COB2, COB3 at 3: 7) by experiment material β-TCP/ α-CSH different quality in the bone defect model of animals of different numberings.Each time point group specimen Micro CT scan, adopt Microview V2.1.2 three-dimensional reconstruction process software to carry out three-dimensional reconstruction, utilize system to carry software (the special-purpose bone analysis software of ABA), set the Target Segmentation value (COB1:2330Hu of material according to the average Hu value of measuring material behind the original material scan rebuilding, COB2:2039Hu, COB3:2419Hu, α-CSH:1395Hu, TCP:904Hu), calculate residual materials percentage by volume (Residual Materials Volume Fraction, RMVF) account for the percentage ratio of the damaged gross area of bone as residual materials, with the degradability of reflection material.
1 group of COB: Figure 18 implants at once outside drawing for the COB1 material in a left side.Relatively can find out, after surgery 4 whens week, visible embedded material edge begins degraded, and it is irregular that the embedded material periphery has become, and the degradation rate of embedded material is 17.80% (among Figure 18); After 8 weeks, reaching central authorities around the material has degraded, and the crack appears in material, and degradation rate is 31.60% (accompanying drawing 18 right sides); After 12 weeks, material is further degraded, and degradation rate is 44.81% (Figure 19 is left); In 16 weeks of postoperative, the material major part is degraded, and material is cracked, and degradation rate is 66.42% (Figure 19 is right).
2 groups of COB: Figure 20 implants at once outside drawing for the COB2 material in a left side.4 whens week after surgery, around the material and part central authorities begins degraded, degradation rate is 21.37% (among Figure 20); After 8 weeks, all there are degraded in material periphery and central authorities, and degradation rate is 36.49% (Figure 20 is right); After 12 weeks, material is further degraded, and degradation rate is 47.37% (Figure 21 is left); In 16 weeks of postoperative, the material major part is degraded, and material is cracked, and degradation rate is 73.75% (Figure 21 is right).
3 groups of COB: Figure 22 implants at once outside drawing for the COB3 material in a left side.After surgery 4 whens week, the material periphery begins degraded, and the embedded material periphery becomes irregular, and degradation rate is 22.71% (among Figure 22); After 8 weeks, material periphery and central authorities all begin degraded, and the crack appears in material, and degradation rate is 40.02% (Figure 22 is right); After 12 weeks, material is further degraded, and degradation rate is 59.48% (Figure 23 is left); In 16 weeks of postoperative, the material major part is degraded, and material is cracked, and degradation rate is 82.32% (Figure 23 is right).
Embodiment 4: zoopery
New product is used for zoopery, has the effect that invention will reach with proof.
36 of healthy new zealand white rabbits, male, body weight 2.5-3.0Kg sleeps new II 1.5ml+ ketamine 2ml mixing afterwards according to preserved skin after the intramuscular anesthesia of 0.2ml/kg body weight, iodophor disinfection, drape with speed, get the back median incision from breast 4 to breast 8 spinous process levels, successively cut skin, subcutaneous tissue, peel off along the spinous process both sides that to appear vertebral plate, transverse process terminal to transverse process, it is for subsequent use that rongeur bites away spinous process of thoracic vertebra, the worn vertebral plate of abrasive drilling and transverse process cortex, worn upper and lower zygapophysial joints.Can absorb cmposite artificial bone as experimental group to implant self-curing between right side vertebral plate and transverse process, the left side is got and is stung spinous process and the vertebral plate cortex removed and merge in contrast group as bone grafting between the capable vertebral plate of body bone and transverse process, such as Figure 12 and shown in Figure 13.
Observe the embedded material surrounding tissue behind the sacrifice of animal without the inflammatory reaction such as red and swollen, congested, confirm that this Biocompatibility is good.The visible material appearance profile is still complete during 4 week, and boundary is still clear between material and osseous tissue, and density of material is projecting normal bone tissues still, visible pore structure in the embedded material.New bone formation around visible material during 8 week, material is held by area of new bone and surface fiber tissue, and only at the visible bone-grafting material in surface, subregion, material is peeled off comparatively difficulty from implant bed; The density of material obviously lowers, and near the osseous tissue of surrounding normal, the appearance profile of material is difficult to distinguish, the visible newborn osseous tissue hole inside of growing into.The visible a large amount of new bone formation of material implant site are difficult to peel off from implant bed during 12 week, and Fusion levels is stiff such as one; The density of material further reduces, and is identical with the surrounding normal Bone density, and the profile of embedded material can't be differentiated, and only in the as seen a small amount of residual implant in the centre of material, the embedded material major part is substituted by freshman bone tissue.Such as Figure 14, Figure 15, Figure 16 and shown in Figure 17, can observe embedded material through molybdenum target photograph, Micro-CT, histology and tetracycline marker and degrade gradually, by from body bone creeping substitution, gradually with combine together the spinal bone fusion from the body bone.

Claims (9)

1. the preparation method of a α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones, it is directly to utilize synchronously synthetic α-half-H 2 O calcium sulphate of hydro-thermal synthesis process in bata-tricalcium phosphate porous particle surface and/or hole, finally be prepared into α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones, its concrete steps are:
(1) preparation has the bata-tricalcium phosphate granule of natural bone pore structure;
(2) utilize hydro-thermal synthesis process to synthesize synchronously α-half-H 2 O calcium sulphate on the bata-tricalcium phosphate porous particle surface of step (1) preparation: to use CaSO 42H 2The O compound concentration is the aaerosol solution of 10-30%, and then the bata-tricalcium phosphate granule with step (1) preparation is placed in one; The two is placed on temperature 110-140 ℃, under the pressure 0.12-0.3MPa condition, behind the reaction 6-12h, namely separates out α-CSH at the porous particle surface crystallization of β-TCP.
2. the preparation method of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 1, it is characterized in that: after step (2) reaction finishes, with the solidliquid mixture of step (2) preparation in filtering more than 90 ℃, and the baking oven of putting into 120 ℃ dries, and namely finishes the preparation of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones.
3. the preparation method of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 1 is characterized in that: in step (2), be the CaSO of 10-30% in concentration 42H 2Add crystal modifier in the O suspension, it is compound that described crystal modifier is that aluminum sulfate and citric acid are received, and accounts for CaSO 42H 2The 0.5wt% of O; Described aluminum sulfate is 50wt%～60wt%: 40wt%～50wt% with the ratio that citric acid is received.
4. the preparation method of a kind of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 1, it is characterized in that, the concrete grammar that described step (1) preparation has the bata-tricalcium phosphate granule of natural bone pore structure is: adopt xenogenesis or homologous cancellus to slough behind the organic principle preroasting in the high-temperature calcination stove, 10 ℃/minutes of heating rates were calcined 3 hours for 800 ℃; The bone piece is soaked in (NH after taking out after will calcining 4) 2HPO 4In the solution 24 hours, remove unnecessary liquid, take out again 50 ℃ of oven dry 4 days of material; Material after the oven dry is put in the high-temperature calcination stove and calcines, 5～10 ℃/minutes of heating rates, 1100 ℃ of calcinings 1 hour slowly cooled to after the room temperature flowing water flushing 3 hours, rinsed with deionized water 2 times, 50 ℃ of oven dry 4 days.
5. the preparation method of a kind of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 4 is characterized in that, described (NH 4) 2HPO 4Concentration be 0.6M～1.2M.
6. the preparation method of a kind of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 4, it is characterized in that, the concrete grammar that xenogenesis or homologous cancellus are sloughed organic principle is: get healthy bulls bone femoral head and distal femur spongy bone, after going cell, ungrease treatment, flowing water flushing, sodium hydroxide, hydrogen peroxide solution immersion use the distilled water rinsing clean, distilled water boiled 10 hours, the flowing water flushing is 3 hours again, rinsed with deionized water 2 times, then the 70%-100% gradient alcohol dehydration dried 4 days for 50 ℃.
7. a α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones is characterized in that, it is to be prepared from the described method of any one in the claim 1 to 6.
8. α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 7, it is characterized in that, described α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones structure is that bata-tricalcium phosphate porous particle surface and hole inside grow up to α-half-H 2 O calcium sulphate long column shape crystal.
9. the method for the compound decalcified bone matrix of α-calcium sulfate hemihydrates/beta-tertiary calcium phosphate porous granular-type composite artificial bones according to claim 8, bone morphogenetic protein, antibiotic or chemotherapeutics, it is characterized in that, directly in cmposite artificial bone, add decalcified bone matrix, bone morphogenetic protein, antibiotic or chemotherapeutics, have the self-curing performance, injection is used.
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