CN113209368A - Animal-derived mineralized bone for treating bone diseases and cosmetic plastic and preparation method thereof - Google Patents
Animal-derived mineralized bone for treating bone diseases and cosmetic plastic and preparation method thereof Download PDFInfo
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Abstract
The invention discloses an animal-derived mineralized bone for treating bone diseases and cosmetic plastic and a preparation method thereof. The invention makes the prepared repair material have natural porous structure, bone conduction and bone induction functions by loading active factors on animal-derived mineralized bones, solves the problem that the artificially synthesized material cannot reach the pore structure, and simultaneously can replace allogeneic bones, thereby eliminating the problems of allogeneic bone antigenicity and ethical aspects.
Description
Technical Field
The invention belongs to the technical field of treating orthopedic diseases and beautifying and reshaping, and particularly relates to an animal-derived mineralized bone for treating orthopedic diseases and beautifying and reshaping and a preparation method thereof.
Background
When a human bone is damaged, the bone needs to be repaired. Natural autologous bone is the most ideal donor in terms of osteoconductivity, osteoinductivity and osteogenic capacity, and is called the "gold standard". However, due to its limited source, when the defect site is large, it is not possible to provide sufficient bone source. At present, aiming at bone defects in clinic, the used bone repair materials are various in types and mainly comprise artificially synthesized hydroxyapatite, organic bone cement, allogeneic bone, alpha-tricalcium phosphate, beta-tricalcium phosphate and other products.
Hydroxyapatite is the main component constituting human bones. At present, the artificially synthesized hydroxyapatite material has good biocompatibility, but has insufficient osteogenic activity. The organic bone cement mainly comprises polymethyl methacrylate (PMMA), the PMMA bone cement has good biocompatibility and mechanical property and is widely applied to clinical load-bearing bone filling, however, the PMMA bone cement has the characteristics of nondegradable property, large heat release amount in the curing process, easy inflammation reaction and the like, and the clinical application of the PMMA bone cement is limited. Allogeneic bone has the hidden danger of triggering immunological rejection and spreading diseases, and in addition, the risk of ethical problems also exists. Alpha-tricalcium phosphate and beta-tricalcium phosphate have the problems of over-high degradation speed, and unmatched material degradation speed and bone replacement speed.
At present, orthopedic products containing active factors such as bone morphogenetic protein (BMP-2) bring new eosin for bone repair due to specific induction performance of the orthopedic products, but the related products are clinically used, and the risk of ectopic osteogenesis, hyperosteogeny and even cancer is easily caused due to uncontrollable release dosage. Therefore, it is important to effectively control the loading and release of active factors and avoid the occurrence of adverse clinical side effects.
Disclosure of Invention
In view of the defects of the prior art, the invention provides the animal-derived mineralized bone for the orthopedic diseases and the cosmetic plastic and the preparation method thereof, the animal-derived mineralized bone and the preparation method thereof utilize hydroxyapatite particles to be processed by simulated body fluid solution, and then active factors are firmly adsorbed on the surface of the mineralized particles, so that on one hand, the problems that the artificially synthesized material cannot reach the pore structure and the allogeneic bone antigenicity and ethical aspects are eliminated; on the other hand, the release of calcium ions and loaded active factors is effectively controlled and regulated through mineralization treatment.
Another object of the present invention is to provide an animal-derived mineralized bone for bone diseases and cosmetic plastic, which has ultra-high crystallinity and crystal purity, and retains the trabeculae of natural bone, and the pores of which are interconnected to facilitate the adhesion and growth of osteoblasts, and a method for preparing the same.
The inventor researches and discovers that: the repair material for bones needs to be a component close to the constitution of human bones to be accepted by human bodies to the maximum extent, for example, hydroxyapatite materials, but the repair material needs to be combined with human bodies, not only the constitution of the material needs to be considered, but also the repair material has a structure capable of stably attaching active factors, so that the effect of the active factors can be fully exerted, and simultaneously allogeneic bones can be replaced, and the antigenicity and ethical problems of the allogeneic bones are eliminated.
Based on this, the invention is realized as follows:
an animal-derived mineralized bone for treating bone diseases and beautifying features that the hydroxyapatite particles are treated by the solution of simulative body fluid, and the active factor is adsorbed onto the surface of mineralized particles.
According to the invention, active factors are loaded on the mineralized hydroxyapatite, and the active factors are stably adsorbed on the surface of inorganic mineralized particles through electrostatic action. The mineralized bone of animal source is mainly composed of hydroxyapatite, and the surface of the mineralized bone has a microporous structure, so that on one hand, the mineralized bone of animal source solves the problem that the artificially synthesized material cannot reach a pore structure, and eliminates the problems of allogenic bone antigenicity and ethical aspect; on the other hand, the release of calcium ions and loaded active factors can be effectively controlled and regulated through mineralization treatment.
Further, the inorganic mineralized particles are animal bone-derived minerals obtained by processing the metaphyseal cancellous bone parts of the limbs and thighbones of mammals through the steps of cooking, hydrogen peroxide treatment, high-temperature primary calcination and the like.
Further, the animal-derived mineral is subjected to pulverization treatment to a particle size preferably between 100 μm and 500 μm, which is more suitable for cell growth.
The active factor is a biological factor which can be, but is not limited to, any active drug or functional protein, including any one of polypeptides or proteins derived from vascular endothelial growth factor, bone morphogenetic protein, serum protein, fibronectin, vitronectin, laminin, elastin, and collagen, and biologically active fragments thereof, and is selected according to the application scenario.
The simulated body fluid solution comprises: sodium chloride (120-140mM), potassium chloride (2-6mM), magnesium sulfate (0.2-0.6mM), magnesium chloride (0.5-1.2mM), sodium bicarbonate (4-100mM), calcium chloride (2-6mM), potassium dihydrogen phosphate (1-5mM), 4-hydroxyethyl piperazine ethanesulfonic acid (10-40mM), and the pH value is 6.8-7.0.
A process for preparing the mineralized bone from animal source for treating bone diseases and beautifying face and shaping includes such steps as dispersing the mineralized hydroxyapatite particles and active factor, and adsorbing.
Further, the ratio of the active factors to the mineralized hydroxyapatite particles is 20ng-500 mug/g, and the oscillation adsorption is carried out.
Further, the hydroxyapatite particles are animal-derived minerals which mainly pass through the metaphyseal cancellous bone portion of the femoral bones of the limbs of mammals, and cut the bone pieces into 2 to 3cm pieces3Cooking the small blocks, boiling water for 0.5-1h, cleaning with clear water, removing surface impurities, replacing clear water, and repeating the above steps for 3 times; then adding the cleaned bone blocks into the equal volume of mixed clear water and 30-50% hydrogen peroxide solution, and continuously boiling for 1-2h until all the bone blocks become white; cleaning the white bone blocks, adding clear water again, and boiling for 1 h; fishing out the bone blocks and washingAfter cleaning, drying for 1-2 hours; taking out the dried bone blocks, calcining at the temperature of 600-800 ℃ for 5-8h, and crushing to obtain the bone block.
Furthermore, the mineralized hydroxyapatite particles are prepared by carrying out mineralization treatment on the hydroxyapatite particles by a simulated body fluid solution, and are mainly prepared by soaking the crushed hydroxyapatite particles with the particle size of 100-500 mu m in the simulated body fluid solution, vibrating and mineralizing the crushed hydroxyapatite particles for 4-7 days by a shaking table at 37 ℃, replacing the solution every day and carrying out vacuum drying.
According to the invention, the mineralized hydroxyapatite particles are dispersed in the solution containing the active factors according to the proportion that the proportion of the active factors to the mineralized hydroxyapatite particles is 20ng-500 mug/g, and are subjected to oscillation adsorption, thus bringing positive effects to the loading and release of the active factors. Therefore, the prepared repair material has a natural porous structure, has bone conduction and bone induction functions by loading active factors on animal-derived mineralized bones, solves the problem that the artificially synthesized material cannot reach the pore structure, can replace allogeneic bones, and eliminates the problems of allogeneic bone antigenicity and ethical aspects. In particular to control the loading and release of active factors.
The repairing material has ultrahigh crystallinity and crystallization purity, keeps the trabecula of the natural bone, and has the mutually communicated pores which are beneficial to the adhesion and growth of osteoblasts.
The invention has the advantages of rich raw material sources, simple and convenient preparation process, low cost and convenient popularization and application.
Drawings
FIG. 1 is a schematic view of the microstructure of the mineralized coating morphology of animal-derived mineralized bone prepared by different concentrations of simulated body fluid solution under a scanning electron microscope.
FIG. 2 shows the ratio of bovine serum albumin loaded on mineralized animal bone derived from animal sources, which is obtained by using different concentrations of simulated body fluid solution.
FIG. 3 is a graph showing the release of calcium ions from animal-derived mineralized bone, which is obtained by simulating the concentration of body fluid solution according to the present invention.
FIG. 4 is a graph of bovine serum albumin released from mineralized animal bone derived from animal sources prepared by different concentrations of simulated body fluid solution realized by the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The animal-derived mineralized bone for orthopedic diseases and cosmetic plastic surgery is prepared by treating hydroxyapatite particles with a simulated body fluid solution and then stably adsorbing active factors on the surface of the mineralized particles. The preparation method comprises the steps of dispersing mineralized hydroxyapatite particles into a solution containing the active factors according to the proportion that the proportion of the active factors to the mineralized hydroxyapatite particles is 20ng-500 mug/g, and carrying out oscillation adsorption, so as to bring positive effects to the loading and releasing of the active factors. Specifically, the inorganic mineralized particles are dispersed in a solution containing the active factor (the concentration of the active factor solution is determined by the specific active factor, for example, the concentration of the Bovine Serum Albumin (BSA) can be 500. mu.g/ml) according to the ratio of the active factor to the particles being 20 ng-500. mu.g/g), and the shaking adsorption is carried out.
The animal-derived mineralized bone is obtained by loading an active factor on mineralized hydroxyapatite particles (dispersing the mineralized hydroxyapatite particles in a solution containing the active factor, and performing oscillation adsorption according to the proportion that the proportion of the active factor to the particles is 20ng-500 mu g/g), and stably adsorbing the active factor on the surfaces of the mineralized hydroxyapatite particles through electrostatic attraction. Wherein the animal-derived mineralized bone is animal-derived mineral, and is obtained by subjecting the metaphyseal cancellous bone part of the femoral bones of limbs of mammals to cooking, hydrogen peroxide treatment, high-temperature primary calcination and other steps (specifically, cutting the bone block into 2cm by a linear cutting machine)3And (3) putting the small blocks into a steamer for steaming, steaming for half an hour after water is boiled, cleaning with clear water, removing surface impurities, replacing the clear water, and repeating the steps for 3 times. Adding equal volume of clear water and 30-50% hydrogen peroxide solution into the cleaned bone pieces, and boiling for 1After 2 hours, all bone pieces turned white. The bone blocks are cleaned by clean water, and then the bone blocks are boiled for 1 hour by adding the clean water again. Taking out the bone blocks, cleaning, and drying in an oven at 70 deg.C for 1-2 hr. Taking out the dried bone blocks and putting the bone blocks into an incinerator to calcine for 5 to 8 hours at 800 ℃ to obtain the bone block composite material).
Furthermore, the animal bone-derived mineral is pulverized to a particle size suitable for cell growth when the particle size is 100 μm to 500 μm.
The hydroxyapatite particles are mineralized by a simulated body fluid solution, and are mainly prepared by soaking crushed hydroxyapatite particles with the particle size of 100-500 mu m in the simulated body fluid solution, vibrating and mineralizing the solution for 4-7 days at 37 ℃ in a shaking table, replacing the solution every day, so that a microporous structure is generated more favorably, and then drying the solution in vacuum.
The invention relates to mineralized hydroxyapatite particles loaded with active factors, which are firmly adsorbed on the surface of inorganic mineralized particles through electrostatic action. The mineralized hydroxyapatite particles mainly comprise inorganic substances, and the surfaces of the mineralized hydroxyapatite particles naturally have microporous structures, so that the mineralized hydroxyapatite particles not only solve the problem that artificial synthetic materials cannot reach pore structures, have good adsorption effect on active factors, but also eliminate the problems of allogenic bone antigenicity and ethical aspects. In particular to control the loading and release of active factors.
The biological factor may be, but is not limited to, any active drug or functional protein, including polypeptides or proteins derived from vascular endothelial growth factor, bone morphogenic proteins, serum proteins, fibronectin, vitronectin, laminin, elastin, and collagen, and biologically active fragments thereof. The biological factors are selected according to the scenario in which they are applied.
Example 1.
Preparing hydroxyapatite particles of animal bone origin: cutting the bone block into 2cm pieces by a linear cutting machine3And (3) putting the small blocks into a steamer for steaming, steaming for half an hour after water is boiled, cleaning with clear water, removing surface impurities, replacing the clear water, and repeating the steps for 3 times. Adding water and 30-50% hydrogen peroxide solution into cleaned bone, and boiling1-2 hours, until all bone pieces became white. The bone blocks are cleaned by clean water, and then the bone blocks are boiled for 1 hour by adding the clean water again. Taking out the bone blocks, cleaning, and drying in an oven at 70 deg.C for 1-2 hr. And taking out the dried bone blocks, and calcining the bone blocks in an incinerator at 800 ℃ for 5-8 hours to obtain the animal-derived hydroxyapatite. Pulverizing the bone blocks to a certain particle size, preferably 100-500 μm.
Mineralization: hydroxyapatite particles derived from animal bone were placed in a simulated body fluid solution comprising sodium chloride at a concentration of 120mM, potassium chloride at a concentration of 2mM, magnesium sulfate at a concentration of 0.2mM, magnesium chloride at a concentration of 0.5mM, sodium bicarbonate at a concentration of 4mM, calcium chloride at a concentration of 5mM, potassium dihydrogen phosphate at a concentration of 1mM, 4-hydroxyethylpiperazine ethanesulfonic acid at a concentration of 10mM, and a final pH of 6.8 (the amount was adjusted to cover all ranges). Soaking the hydroxyapatite particles in a simulated body fluid solution, performing shaking mineralization on the hydroxyapatite particles for 4 days at 37 ℃, replacing the solution every day, and performing vacuum drying to obtain the hydroxyapatite particles.
Loading: dispersing the hydroxyapatite particles derived from the animal bone into a Bovine Serum Albumin (BSA) solution according to the ratio of 20 mug/g of the active factors to the hydroxyapatite particles derived from the animal bone, and performing oscillation adsorption to enable BSA to be attached to the surfaces of the hydroxyapatite particles derived from the animal bone, thereby obtaining the mineralized bone derived from the animal after adsorption.
Example 2.
Preparing hydroxyapatite particles of animal bone origin: cutting the bone block into 2cm pieces by a linear cutting machine3And (3) putting the small blocks into a steamer for steaming, steaming for half an hour after water is boiled, cleaning with clear water, removing surface impurities, replacing the clear water, and repeating the steps for 4 times. Adding the cleaned bone blocks into equal volume of mixed clear water and 30-50% hydrogen peroxide solution, and boiling for 1-2 hr until all bone blocks become white. The bone blocks are cleaned by clean water, and then the bone blocks are boiled for 1 hour by adding the clean water again. Taking out the bone blocks, cleaning, and drying in an oven at 70 deg.C for 1-2 hr. And taking out the dried bone blocks, and calcining the bone blocks in an incinerator at 800 ℃ for 5-8 hours to obtain the animal-derived hydroxyapatite. Pulverizing the bone blocks to a certain particle size, preferably 100-500 μm.
Mineralization: hydroxyapatite particles derived from animal bones were placed in a simulated body fluid solution comprising sodium chloride at a concentration of 140mM, potassium chloride at a concentration of 6mM, magnesium sulfate at a concentration of 0.5mM, magnesium chloride at a concentration of 1.2mM, sodium bicarbonate at a concentration of 100mM, calcium chloride at a concentration of 6mM, potassium dihydrogen phosphate at a concentration of 5mM, 4-hydroxyethylpiperazine ethanesulfonic acid at a concentration of 40mM, and a final pH of 6.9. Soaking the hydroxyapatite particles in a simulated body fluid solution, performing shaking mineralization on the hydroxyapatite particles for 7 days at 37 ℃, replacing the solution every day, and performing vacuum drying to obtain the hydroxyapatite particles.
Loading: dispersing the hydroxyapatite particles derived from the animal bone into a Bovine Serum Albumin (BSA) solution according to the proportion of 500 mug/g of the active factors and the hydroxyapatite particles derived from the animal bone, and performing oscillation adsorption to enable BSA to be attached to the surfaces of the hydroxyapatite particles derived from the animal bone, thereby obtaining the mineralized bone derived from the animal after adsorption.
Example 3.
Preparing hydroxyapatite particles of animal bone origin: cutting the bone block into 3cm pieces by a linear cutter3And (3) putting the small blocks into a steamer for steaming, steaming for half an hour after water is boiled, cleaning with clear water, removing surface impurities, replacing the clear water, and repeating the steps for 3 times. Adding the cleaned bone blocks into equal volume of mixed clear water and 30-50% hydrogen peroxide solution, and boiling for 1-2 hr until all bone blocks become white. The bone blocks are cleaned by clean water, and then the bone blocks are boiled for 1 hour by adding the clean water again. Taking out the bone blocks, cleaning, and drying in an oven at 70 deg.C for 1-2 hr. And taking out the dried bone blocks, and calcining the bone blocks in an incinerator at 800 ℃ for 5-8 hours to obtain the animal-derived hydroxyapatite. Pulverizing the bone blocks to a certain particle size, preferably 100-500 μm.
Mineralization: hydroxyapatite particles derived from animal bones were placed in a simulated body fluid solution comprising the respective components at a concentration of 130mM sodium chloride, 4mM potassium chloride, 0.5mM magnesium sulfate, 1mM magnesium chloride, 60mM sodium bicarbonate, 5mM calcium chloride, 2mM potassium dihydrogen phosphate, 20mM 4-hydroxyethylpiperazine ethanesulfonic acid, and a final pH of 7.0. Soaking the hydroxyapatite particles in a simulated body fluid solution, performing shaking mineralization on the hydroxyapatite particles for 5 days at 37 ℃, replacing the solution every day, and performing vacuum drying to obtain the hydroxyapatite particles.
Loading: dispersing the hydroxyapatite particles derived from the animal bone into a Bovine Serum Albumin (BSA) solution according to the proportion of 300 mug/g of the active factors and the hydroxyapatite particles derived from the animal bone, and performing oscillation adsorption to enable BSA to be attached to the surfaces of the hydroxyapatite particles derived from the animal bone, thereby obtaining the mineralized bone derived from the animal after adsorption.
Therefore, the animal-derived mineralized bone prepared according to the invention has a microstructure of animal-derived hydroxyapatite particles mineralized by different mineralized solutions as shown in fig. 1. The loading ratio of BSA is shown in FIG. 2, and the loading ratio of BSA gradually decreases with increasing mineralization concentration. Meanwhile, calcium ions (as shown in figure 3) and BSA release (as shown in figure 4) are closely related to mineralization.
In a word, the surface of the animal-derived hydroxyapatite particles is loaded with active factors, so that the prepared repair material has a natural porous structure and has functions of bone conduction and bone induction, the problem that the artificially synthesized material cannot reach a pore structure is solved, simultaneously, allogeneic bone can be replaced, and the problems of allogeneic bone antigenicity and ethical aspects are solved. In particular to control the loading and release of active factors.
The invention has the advantages of rich raw material sources, simple and convenient preparation process, low cost and convenient popularization and application. Can be widely applied to various aspects such as bone repair, beauty treatment and the like.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An animal-derived mineralized bone for treating bone diseases and beautifying features that the particles of hydroxyapatite are treated by simulative body fluid solution, and the active factors are adsorbed onto the surface of mineralized particles.
2. The mineralized bone derived from animal for treating bone diseases and cosmetic plastic according to claim 1, wherein the hydroxyapatite particles are mineral derived from animal, and the mineral derived from animal is obtained by subjecting a metaphyseal cancellous bone portion of femoral bones of limbs of mammals to the steps of cooking, hydrogen peroxide treatment and high-temperature primary calcination.
3. The mineralized bone derived from animal for treating bone diseases and cosmetic plastic as claimed in claim 2, wherein the mineral derived from animal is pulverized to have a particle size of 100 μm to 2000 μm.
4. The mineralized bone derived from animal for treating bone diseases and cosmetic plastic as claimed in claim 1, wherein the simulated body fluid solution is a buffer solution composed of inorganic salts, and the concentrations of the components of the buffer solution are sodium chloride (120-140mM), potassium chloride (2-6mM), magnesium sulfate (0.2-0.6mM), magnesium chloride (0.5-1.2mM), sodium bicarbonate (4-100mM), calcium chloride (2-6mM), potassium dihydrogen phosphate (1-5mM), 4-hydroxyethylpiperazine ethanesulfonic acid (10-40mM), and the pH value is 6.8-7.0.
5. The mineralized bone derived from animal for treating bone diseases and cosmetic plastic diseases according to claim 1, wherein the active factor is a biological factor selected from the group consisting of vascular endothelial growth factor, bone morphogenetic protein, serum protein, fibronectin, vitronectin, laminin, elastin, collagen, and polypeptides or proteins derived from bioactive fragments thereof.
6. A process for preparing mineralized bone from animal source for treating bone diseases and beautifying features that the mineralized hydroxyapatite particles and active factor are dispersed and adsorbed.
7. The method for preparing mineralized bone derived from animal for treating bone diseases and cosmetic surgery as claimed in claim 6, wherein the active factor and the mineralized hydroxyapatite particles are in a ratio of 20ng-500 μ g/g, and the vibration adsorption is performed.
8. The method for producing an animal-derived mineralized bone for bone diseases and cosmetic surgery according to claim 7, wherein the hydroxyapatite particles are animal-derived minerals obtained by cutting a bone block into 2 to 3cm pieces through metaphyseal cancellous bone portions of femoral bones of limbs of mammals3Cooking the small blocks for 0.5-1h, cleaning and removing surface impurities; then adding the cleaned bone blocks into water and 30-50% hydrogen peroxide solution mixed in the same volume, and continuously boiling for 1-2h until all the bone blocks become white; cleaning the white bone blocks, adding water again and boiling for 1 h; fishing out the bone blocks, cleaning and drying for 1-2 hours; taking out the dried bone blocks, calcining at the temperature of 600-800 ℃ for 5-8h, and crushing to obtain the bone block.
9. The method according to claim 8, wherein the mineralized hydroxyapatite particles are prepared by mineralizing a simulated body fluid solution, soaking the mineralized hydroxyapatite particles with a particle size of 100 μm to 500 μm after being crushed in the simulated body fluid solution, vibrating and mineralizing the hydroxyapatite particles for 4 to 7 days by a shaker at 37 ℃, and vacuum drying.
10. The method of claim 9, wherein the active factor is a biological factor, and the biological factor can be any active drug or functional protein, including but not limited to vascular endothelial growth factor, bone morphogenetic protein, serum protein, fibronectin, vitronectin, laminin, elastin, collagen and any polypeptide or protein derived from bioactive fragments thereof.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113908341A (en) * | 2021-10-26 | 2022-01-11 | 深圳市迈捷生命科学有限公司 | Injectable bone induction repair material and preparation method thereof |
| CN113984853A (en) * | 2021-09-29 | 2022-01-28 | 广东省科学院健康医学研究所 | Equipment for in-vitro degradation and simulated mineralization and use method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1579562A (en) * | 2003-08-01 | 2005-02-16 | 中国人民解放军第四军医大学口腔医学院 | Preparation method for ceramic agg egate of same kind and different structure |
| DE102005016443A1 (en) * | 2004-11-23 | 2006-05-24 | Ossacur Ag | Bioabsorbable and mineralized material, useful for filling of bone defects, comprises a collagen matrix from aggregated collagen chains, where only the surface of the aggregated collagen chain is mineralized |
| US20070098756A1 (en) * | 2005-11-01 | 2007-05-03 | Keyvan Behnam | Bone Matrix Compositions and Methods |
| CN105561397A (en) * | 2015-12-31 | 2016-05-11 | 广西医科大学 | Production method of growth factor supported porous biologic ceramic artificial bone scaffold |
| CN108992708A (en) * | 2018-07-02 | 2018-12-14 | 西安巨子生物基因技术股份有限公司 | Modified bone meal in surface and preparation method thereof |
| CN109464397A (en) * | 2018-11-16 | 2019-03-15 | 中国药科大学 | A kind of graphene oxide multiple medicine based on biomineralization is total to loading system and its preparation method and application |
| KR20190053006A (en) * | 2017-11-09 | 2019-05-17 | 단국대학교 천안캠퍼스 산학협력단 | Method of preparation of biomimetic hydroxyapatite microspheres |
-
2021
- 2021-04-26 CN CN202110455615.3A patent/CN113209368A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1579562A (en) * | 2003-08-01 | 2005-02-16 | 中国人民解放军第四军医大学口腔医学院 | Preparation method for ceramic agg egate of same kind and different structure |
| DE102005016443A1 (en) * | 2004-11-23 | 2006-05-24 | Ossacur Ag | Bioabsorbable and mineralized material, useful for filling of bone defects, comprises a collagen matrix from aggregated collagen chains, where only the surface of the aggregated collagen chain is mineralized |
| US20070098756A1 (en) * | 2005-11-01 | 2007-05-03 | Keyvan Behnam | Bone Matrix Compositions and Methods |
| CN105561397A (en) * | 2015-12-31 | 2016-05-11 | 广西医科大学 | Production method of growth factor supported porous biologic ceramic artificial bone scaffold |
| KR20190053006A (en) * | 2017-11-09 | 2019-05-17 | 단국대학교 천안캠퍼스 산학협력단 | Method of preparation of biomimetic hydroxyapatite microspheres |
| CN108992708A (en) * | 2018-07-02 | 2018-12-14 | 西安巨子生物基因技术股份有限公司 | Modified bone meal in surface and preparation method thereof |
| CN109464397A (en) * | 2018-11-16 | 2019-03-15 | 中国药科大学 | A kind of graphene oxide multiple medicine based on biomineralization is total to loading system and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| ALEKSANDRA SZCZEŚ, ET AL: "Synthesis of hydroxyapatite for biomedical applications", 《ADVANCES IN COLLOID AND INTERFACE SCIENCE》 * |
| 曾戎等: "《生物医用仿生高分子材料》", 31 October 2010, 广州：华南理工大学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113984853A (en) * | 2021-09-29 | 2022-01-28 | 广东省科学院健康医学研究所 | Equipment for in-vitro degradation and simulated mineralization and use method thereof |
| CN113984853B (en) * | 2021-09-29 | 2024-03-01 | 广东省科学院健康医学研究所 | Equipment for in-vitro degradation and simulated mineralization and application method thereof |
| CN113908341A (en) * | 2021-10-26 | 2022-01-11 | 深圳市迈捷生命科学有限公司 | Injectable bone induction repair material and preparation method thereof |
| CN114949360A (en) * | 2021-10-26 | 2022-08-30 | 深圳市迈捷生命科学有限公司 | Injectable bone induction repair material and preparation method thereof |
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