CN101444646A - Gelatin/calcium phosphate composite porous microsphere for bone repair and preparation method and usage of the porous microsphere - Google Patents
Gelatin/calcium phosphate composite porous microsphere for bone repair and preparation method and usage of the porous microsphere Download PDFInfo
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Abstract
The invention belongs to the field of the preparation of gelatin derivative materials and in particular relates to a composite porous microsphere material formed by compounding gelatin, calcium phosphate and an antler polypeptide bioactive tissue or bone morphogenic protein (BMP). The composite porous microsphere is formed by uniformly dispersing, in a gelatin composite porous microsphere, the calcium phosphate and active-substance antler polypeptide or bone morphogenic protein which is capable of promoting bone growth; wherein, the weight of the active-substance antler polypeptide or the bone morphogenic protein accounts for millesimal to five percent of the weight of the gelatin; soluble calcium salt which forms the calcium phosphate ranges from 0.0003 to 0.03 mol in each gelatin; and the amount of soluble phosphate causes the ratio between Ca element and P element to range from 1:1 to 2:1. The composite porous microsphere material can also contain crosslinking agent. The composite porous microsphere material is a bone repair biological material which has good biocompatibility and good bone induction activity.
Description
Technical field
The invention belongs to the preparation field of gelatine derivative material, be particularly related to the composite porous microsphere material that gelatin and calcium phosphate and a kind of antler polypeptide biological activity tissue or bone morphogenetic protein (BMP) synthesize, this material is a kind of have excellent biological compatibility and active bone reparing biological material of induced osteogenesis.
Background technology
Gelatin is the product of collagen degradation, and it has not only kept collagen as the more peculiar advantages of biomaterial well, and compares with collagen, the gelatin no antigen, and no rejection implants; Gelatin is cheap and easy to get; Soluble in water; The detection means of its physicochemical properties developed comparative maturity, standard.Therefore, gelatin and calcium phosphate composite materials also are focuses in the present bone renovating material research.
Traditional gelatin and calcium phosphate composite materials mostly are granular pattern or block (B.S.Chang in irregular shape greatly, C.K.Lee, K.S.Hong, et al.Osteoconduction atporous hydroxyapatite with various pore configurations.Biomaterials, 2000,21:1291～1298.), this traditional material is when using as bone renovating material, can cause the intravital inflammatory reaction of patient, and new osteoplastic speed is lower, the skeletonization amount is less.In addition, traditional gelatin and calcium phosphate composite materials are all existing serious defective aspect mechanical property and the degradation rate, make this composite materials be applied to still to exist many problems when bone is repaired.
Studies show that: gelatin and hydroxyapatite (HAP) are combined into the microsphere form with certain form, the mechanical property and the degradation rate of bone renovating material have been improved, utilize the good pliability of this kind composite microsphere material, the various difform hole that filling bone defects causes.The most important thing is,, increased the contact area of implantation body and its surrounding bone tissue, promoted the compactness between interface and the interface greatly, can bring into play the skeletonization effect of material better with the bone renovating material of development spherical morphology.
Cornu Cervi Pantotrichum is that mammal is unique can regenerated bone class tissue, in the existing history in several thousand of pharmacopeia record.Exist multiple known somatomedin in the Cornu Cervi Pantotrichum, rebuild relevant many cells somatomedin TGF β 1 and TGF β 2 etc. as insulin-like growth factor I (IGF-1), growth hormone (GH) and with osteogenesis and bone, and think that the common existence and the comprehensive effect of these somatomedin are grown Cornu Cervi Pantotrichum fast just.The antler polypeptide that we extract from bright Cornu Cervi Pantotrichum is a main active, synthesizes the porous microsphere of one with gelatin and calcium phosphate, and this is still material itself on the technology of preparing of material no matter, on bone reparation and the therapeutic efficiency all being a raising greatly.
Application calcium microcosmic salt powder body such as Tsing-Hua University's face Yongnian and chitin/chitosan fiber are made complex (number of patent application: 200410034109.3), wherein also added a small amount of collagen (gelatin), and bone morphogenetic protein, confirm that through cell and zoopery this material has good biological activity and induced osteogenesis activity.(number of patent application: 01136246.4) in acid-sol stock solution, prepare the unformed powder body material of collagen/calcium phosphate by the sedimentation method, its composition and nature bone are similar for Cui Fuzhai etc.
University Of Tianjin's (number of patent application: 200410019979.3) propose a kind of preparation method that is rich in the complex microsphere of sodium alginate and hydroxyapatite, this method adopts hydroxy apatite powder and the common method that adds of sodium alginate, forms particle diameter at 200um～300um left and right sides complex microsphere by emulsification.
The present invention is by making the method for soluble calcium salt and microcosmic salt co-precipitation in the gelatin emulsion, be prepared into composite porous microspheres, add an amount of antler polypeptide or bone morphogenetic protein in the reaction, the preparation method that makes bone-repairing composite material and this composite yet there are no bibliographical information.
Summary of the invention
The purpose of this invention is to provide a kind of good biocompatibility, contain specific short bone active material (as: antler polypeptide or bone morphogenetic protein), easy prepared in batches, controlled, the cheap gelatin and the calcium phosphate composite porous microspheres that are used for the bone reparation of pattern.
A further object of the present invention provides and is used for the gelatin that bone repairs and the preparation method of calcium phosphate composite porous microspheres.
Another object of the present invention provides the gelatin that is used for the bone reparation and calcium phosphate composite porous microspheres in the purposes that is used for aspect the bone renovating material.
Gelatin that is used for the bone reparation of the present invention and calcium phosphate composite porous microspheres are a kind ofly calcium phosphate to be arranged and can impel the active substance antler polypeptide or the bone morphogenetic protein of osteogenesis at the gelatine microsphere homodisperse; Wherein, active substance antler polypeptide or bone morphogenetic protein are 1 ‰～5% of gelatin weight; The soluble calcium salt that forms calcium phosphate is 0.0003～0.03mol of every gram gelatin, and the amount of soluble phosphate makes that the ratio of Ca element and P element is 1:1～2:1, and the ratio of preferred Ca element and P element is 1.5:1.
Further homodisperse has cross-linking agent in the described gelatine microsphere, the amount of described cross-linking agent is the cross-linking agent that every gram gelatin adds 30ul～1000ul for the addition of cross-linking agent in reaction, preferably every gram gelatin adds the cross-linking agent of 500ul, and the mass concentration of cross-linking agent is 5 ‰～15%.
The described diameter of gelatin that bone repairs and calcium phosphate composite porous microspheres that is used for is at 500nm～500um.
The described diameter in gelatin that bone repairs and the hole on the calcium phosphate composite porous microspheres that is used for is at 10nm～20um.
Of the present inventionly be used for the gelatin of bone reparation and the preparation method of calcium phosphate composite porous microspheres may further comprise the steps:
1) at first takes by weighing a certain amount of gelatin powder body, add a certain amount of deionized water, be heated to 30～70 ℃, preparation quality concentration is 5%～30% gelatin solution, add soluble calcium salt then in gelatin solution, its addition is the soluble calcium salt that every gram gelatin powder body adds 0.0003～0.03mol; Subsequently, add the active substance antler polypeptide or the bone morphogenetic protein that can impel osteogenesis again, the addition of antler polypeptide or bone morphogenetic protein is 1 ‰～5% of gelatin weight;
2) mixed liquor that step 1) is made is poured in the oil that contains surfactant, stirs the water-in-oil emulsion that forms gelatin solution; The content of surfactant in oil is 1g/200ml oil～1g/20ml oil; The amount of used oil is every gram gelatin 5ml～250ml;
3) with step 2) the water-in-oil emulsion heating of the gelatin solution that makes, temperature is controlled at 30 ℃～70 ℃, and preferred temperature is 45 ℃, and continues to stir 0.5～3 hour, preferably stirs 1 hour, and mixing speed is 100 rev/mins～6000 rev/mins; Add the soluble phosphoric acid saline solution then, the mass concentration of soluble phosphoric acid saline solution is 5%～15%, and phosphatic addition makes the proportion control of Ca element in the mixed liquor and P element at 1:1～2:1, and the ratio of preferred Ca element and P element is 1.5:1;
4) mixed liquor that step 3) is made stirred 1～5 hour under 30 ℃～70 ℃ temperature, continued to stir down, with ice bath cooling 20～80 minutes; Form active substance antler polypeptide or bone morphogenetic protein that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis;
5) in the mixed liquor that step 4) makes, add cross-linking agent, the mass concentration of cross-linking agent is 5 ‰～15%, the addition of cross-linking agent is the cross-linking agent that every gram gelatin adds 30ul～1000ul, and preferably every gram gelatin adds the cross-linking agent of 500ul, continues to stir 20 minutes～100 minutes;
6) get step 4) or 5) mixed liquor that makes, add acetone, addition is 2～8 times of moisture content volume in the mixed liquor, preferably in the mixed liquor 3 of the moisture content volume times, continue to stir after 0.5～6 hour, preferably continue to stir after 1～2 hour, filter, obtain the crude product of composite porous microspheres;
7) after the crude product of the composite porous microspheres that step 6) is obtained is used the acetone cyclic washing, obtain being used for gelatin and the calcium phosphate composite porous microspheres that bone is repaired.
Described soluble calcium salt is selected from a kind of in calcium chloride, lime nitrate or the calcium acetate etc. or greater than a kind of mixture.
Described soluble phosphate is selected from a kind of in sodium dihydrogen phosphate, sodium hydrogen phosphate or the phosphoric acid hydrogen amine etc. or greater than a kind of mixture.
Described surfactant is the water-in-oil type surfactant, comprises a kind of among department class (SPAN) series (as SPAN80, SPAN60, SPAN20) or dioctyl sodium sulfosuccinate (AOT) surfactant.
A kind of in silicone oil, Oleum Glycines, olive oil, Oleum Camelliae, Oleum Brassicae campestris or mixed oil of described grease separation.
Described cross-linking agent is selected from a kind of in glutaraldehyde, formaldehyde, the dialdehyde glucose etc. or greater than a kind of mixture.
The gelatin and the calcium phosphate composite porous microspheres that are used for the bone reparation of the present invention's preparation, all show through cell experiment and zoopery: this composite porous microsphere material has excellent biological compatibility and induced osteogenesis activity, be a kind of tissue repair and treatment material of reliable degradable in vivo, material in can repairing as biological tissue's (as bone, cartilage, tooth) is with a wide range of applications.
The present invention makes gelatin solution with the conventional powder gelatin, then be dispersed in become gelatin solution in the oil bath water-in-oil emulsion as reaction medium, by in the water-in-oil emulsion of gelatin solution, adding the calcium salt and the phosphate of solubility, make soluble calcium salt and phosphate carry out precipitation in gelatine microsphere inside, the an amount of various short osteogenesis active substances (antler polypeptide or bone morphogenetic protein) that add in the course of reaction, finally obtain being used for gelatin and the calcium phosphate composite porous microspheres that bone is repaired, there is hole on this composite porous microspheres surface, is convenient to seeking connections with and growing of cell.
The present invention adds active hairy antler polypeptide or bone morphogenetic protein again on the basis of composite porous microspheres preferably preparing form, this is for promoting the bone marrow stromal cell differentiation, effects such as chondrocyte, epidermis cell and fibroblast proliferation have particularly effect such as ulcer skin ulcer face healing of regeneration of tangible accelerated bone tissue regeneration, peripheral nerve tissue and wound.This composite porous microsphere material can be used for bone, reparation that tooth is damaged; Can be used as the solid phase raw material of bone cement; Can be directly as the implant of the bone defect in the bone invasive procedures.
The specific embodiment
Embodiment 1.
1, accurately takes by weighing the beaker that 1 gram gelatin powder body is put into 100ml, add the 5ml deionized water, be placed on 50 ± 1 ℃ of stirred in water bath dissolvings, in beaker, add the required 0.0025mol lime nitrate solid of formation calcium phosphate subsequently, then add antler polypeptide 3mg, be made into mixed solution.
2, under rotating speed is 500 rev/mins stirring, above-mentioned solution is added drop-wise to 50ml lentamente to be dissolved with surfactant (SPAN 80, Oleum Glycines 1g/50ml) mutually in, form the water-in-oil emulsion of gelatin solution, reaction temperature remains on 45 ± 1 ℃, and continue to stir 2 hours, in emulsion system, drip mass concentration again and be 5% Na
2HPO
4Solution (the Ca/P mol ratio is 5:3).
3, after reaction finishes, 45 ± 1 ℃ reactant moves in the ice-water bath rapidly, and keeps stirring, and cools off 40 minutes, make the gelatine microsphere gelation contain calcium phosphate, form the active substance antler polypeptide that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis.
4, in cooled reaction system, add mass concentration and be 8% glutaraldehyde cross-linking agent 600ul and carry out crosslinkedly, stirred 30 minutes, adding acetone 30ml in system dewaters to the complex microsphere again.
5, reactant stops to stir, and is the composite porous microspheres crude product with serosity sucking filtration, filter cake, and it is used 30ml acetone repeated washing 3 times, and the end product that obtains is the powder of light yellow gelatin and calcium phosphoric acid composite porous microspheres.
This powder diameter is 25um ± 5um, and surface apertures is 1um ± 50nm.The composite porous microspheres powder by calcium phosphate, antler polypeptide and crosslinked after gelatin form, this composite porous microsphere material is used for bone, reparation that tooth is damaged; Solid phase raw material as bone cement; Be directly used in the implant of the bone defect in the bone invasive procedures.
Embodiment 2.
1, accurately takes by weighing the beaker that 1 gram gelatin powder body is put into 100ml, add the 5ml deionized water, be placed on 50 ± 1 ℃ of stirred in water bath dissolvings, in beaker, add subsequently and form the required 0.003mol calcium chloride solid of calcium phosphate, then, be made into mixed solution to wherein adding the 20mg antler polypeptide.
2, under rotating speed is 500 rev/mins stirring, above-mentioned solution is added drop-wise in the Fructus Canarii albi oil phase that 100ml is dissolved with surfactant (SPAN 60-1g/50ml) lentamente, form the water-in-oil emulsion of gelatin solution, reaction temperature remains on 45 ± 1 ℃, and continue to stir 2 hours, dripping mass concentration again in emulsion system is 10%Na
2HPO
4Solution (the Ca/P mol ratio is 5:3).
3, after reaction finishes, 45 ℃ ± 1 ℃ reactant moves in the ice-water bath rapidly, and keeps stirring, and cools off 40 minutes, make the gelatine microsphere gelation contain calcium phosphate, form the active substance antler polypeptide that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis.
4, in cooled reaction system, add acetone 30ml, continue to stir 40 minutes, the complex microsphere is dewatered,
5. reactant stops to stir, and is the composite porous microspheres crude product with serosity sucking filtration, filter cake, and it is used 30ml acetone repeated washing 4 times, the powder of the end product that obtains white gelatin/calcium phosphate composite porous microspheres.
This powder diameter is 25um ± 5um, and surface apertures is 1um ± 50nm.The composite porous microspheres powder by calcium phosphate, antler polypeptide and crosslinked after gelatin form, this composite porous microsphere material is used for bone, reparation that tooth is damaged; Solid phase raw material as bone cement; Be directly used in the implant of the bone defect in the bone invasive procedures.
Embodiment 3.
1, accurately takes by weighing the beaker that 1 gram gelatin powder body is put into 100ml, add the 5ml deionized water, be placed on 40 ± 1 ℃ of stirred in water bath dissolvings, in beaker, add subsequently and form the required 0.01mol calcium chloride solid of calcium phosphate, be made into the gelatin solution that contains certain calcium ion concentration, then to wherein adding the 20mg antler polypeptide, and it is uniformly dispersed.
2, under rotating speed is 200 rev/mins stirring, with above-mentioned solution be added drop-wise to lentamente 75ml be dissolved with surfactant (Oleum Camelliae of dioctyl sodium sulfosuccinate (AOT)-1g/50ml) mutually in, form the water-in-oil emulsion of gelatin solution, reaction temperature remains on 45 ± 1 ℃, and continue to stir 2 hours, dripping mass concentration again in emulsion system is 10%Na
2HPO
4Solution (the Ca/P mol ratio is 1.5:1).
3, after reaction finishes, 45 ± 1 ℃ reactant moves in the ice-water bath rapidly, and keeps stirring, and cools off 30 minutes, make the gelatine microsphere gelation contain calcium phosphate, form the active substance antler polypeptide that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis.
4, in cooled reaction system, adding mass concentration is that 4% formaldehyde 600ul is crosslinked, stirs 20 minutes, adds acetone 30ml again in system, and the complex microsphere is dewatered.
5, reactant stops to stir, and is the composite porous microspheres crude product with serosity sucking filtration, filter cake, and it is used 30ml acetone repeated washing 3 times, and the end product that obtains is the powder of light yellow gelatin/calcium phosphate composite porous microspheres.
This powder diameter is 45um ± 5um, and surface apertures is 5um ± 50nm.The composite porous microspheres powder by calcium phosphate, antler polypeptide and crosslinked after gelatin form, this composite porous microsphere material is used for bone, reparation that tooth is damaged; Solid phase raw material as bone cement; Be directly used in the implant of the bone defect in the bone invasive procedures.
Embodiment 4.
1, accurately takes by weighing the beaker that 1 gram gelatin powder body is put into 100ml, add the 5ml deionized water, be placed on 50 ± 1 ℃ of stirred in water bath dissolvings, in beaker, add subsequently and form the required 0.004mol calcium acetate solid of calcium phosphate, be made into the gelatin solution that contains certain calcium ion concentration, then to wherein adding bone morphogenetic protein 3mg, and stir.
2, under rotating speed is 200 rev/mins stirring, above-mentioned solution is added drop-wise in the mediation oil phase that 150ml is dissolved with surfactant (SPAN 80-1g/50ml) lentamente, form the water-in-oil emulsion of gelatin solution, reaction temperature remains on 45 ± 1 ℃, and continue to stir 2 hours, dripping mass concentration again in emulsion system is 5%NaH
2PO
4Solution (the Ca/P mol ratio is 2:1).
3, after reaction finishes, 45 ± 1 ℃ reactant moves in the ice-water bath rapidly, and keeps stirring, and cools off 50 minutes, make the gelatine microsphere gelation contain calcium phosphate, form the active substance bone morphogenetic protein that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis.
4, in cooled reaction system, 8% glutaraldehyde 600ul is crosslinked for the adding mass concentration, stirs 50 minutes, adds acetone 30ml again in system, and the complex microsphere is dewatered.
5, reactant stops to stir, and is the composite porous microspheres crude product with serosity sucking filtration, filter cake, and it is used 30ml acetone repeated washing 3 times, and the end product that obtains is the powder of light yellow gelatin/calcium phosphate composite porous microspheres.
This powder diameter is 45um ± 5um, and surface apertures is 5um ± 50nm.The composite porous microspheres powder by calcium phosphate, antler polypeptide and crosslinked after gelatin form, this composite porous microsphere material is used for bone, reparation that tooth is damaged; Solid phase raw material as bone cement; Be directly used in the implant of the bone defect in the bone invasive procedures.
Embodiment 5.
1, accurately takes by weighing the beaker that 1 gram gelatin powder body is put into 100ml, add the 5ml deionized water, be placed on 50 ± 1 ℃ of stirred in water bath dissolvings, in beaker, add subsequently and form the required 0.0008mol lime nitrate solid of calcium phosphate, be made into the gelatin solution that contains certain calcium ion concentration, then to wherein adding bone morphogenetic protein 20mg, and stir.
2, under rotating speed is 2000 rev/mins stirring, above-mentioned solution is added drop-wise in the Semen Allii Tuberosi oil phase that 200ml is dissolved with surfactant (SPAN 80-1g/50ml) lentamente, form the water-in-oil emulsion of gelatin solution, reaction temperature remains on 45 ± 1 ℃, and continue to stir 2 hours, dripping mass concentration again in emulsion system is 10% (NH
4)
2HPO
4Solution (the Ca/P mol ratio is 1.5:1).
3, after reaction finishes, 45 ± 1 ℃ reactant moves in the ice-water bath rapidly, and keeps stirring, and cools off 60 minutes, make the gelatine microsphere gelation contain calcium phosphate, form the active substance bone morphogenetic protein that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis.
4, in cooled reaction system, 8% formaldehyde 500ul is crosslinked for the adding mass concentration, stirs 60 minutes, adds acetone 25ml again in system, and the complex microsphere is dewatered.
5, reactant stops to stir, and is the composite porous microspheres crude product with serosity sucking filtration, filter cake, and it is used 30ml acetone repeated washing 3 times, and the end product that obtains is the powder of light yellow gelatin/calcium phosphate composite porous microspheres.
This powder diameter is 15um ± 5um, and surface apertures is 200nm ± 30nm.The composite porous microspheres powder by calcium phosphate, antler polypeptide and crosslinked after gelatin form, this composite porous microsphere material is used for bone, reparation that tooth is damaged; Solid phase raw material as bone cement; Be directly used in the implant of the bone defect in the bone invasive procedures.
Claims (13)
1. one kind is used for gelatin and the calcium phosphate composite porous microspheres that bone is repaired, and it is characterized in that: homodisperse has calcium phosphate and can impel the active substance antler polypeptide or the bone morphogenetic protein of osteogenesis in gelatine microsphere; Wherein, active substance antler polypeptide or bone morphogenetic protein are 1 ‰～5% of gelatin weight; The soluble calcium salt that forms calcium phosphate is 0.0003～0.03mol of every gram gelatin, and the amount of soluble phosphate makes that the ratio of Ca element and P element is 1:1～2:1.
2. composite porous microspheres according to claim 1 is characterized in that: homodisperse has cross-linking agent in the described gelatine microsphere.
3. composite porous microspheres according to claim 2 is characterized in that: the amount of described cross-linking agent is the cross-linking agent that every gram gelatin adds 30ul～1000ul for the addition of cross-linking agent in reaction, and the mass concentration of cross-linking agent is 5 ‰～15%.
4. according to claim 2 or 3 described composite porous microspheres, it is characterized in that: described cross-linking agent is selected from a kind of in glutaraldehyde, formaldehyde, the dialdehyde glucose or greater than a kind of mixture.
5. composite porous microspheres according to claim 1 is characterized in that: the amount of described soluble phosphate makes that the ratio of Ca element and P element is 1.5: 1.
6. composite porous microspheres according to claim 1 or 5 is characterized in that: described soluble phosphate is selected from a kind of in sodium dihydrogen phosphate, sodium hydrogen phosphate, the phosphoric acid hydrogen amine or greater than a kind of mixture.
7. composite porous microspheres according to claim 1 is characterized in that: described soluble calcium salt is selected from a kind of in calcium chloride, lime nitrate, the calcium acetate or greater than a kind of mixture.
8. composite porous microspheres according to claim 1 is characterized in that: the described diameter of gelatin that bone repairs and calcium phosphate composite porous microspheres that is used for is at 500nm～500um.
9. according to claim 1 or 8 described composite porous microspheres, it is characterized in that: the described diameter in gelatin that bone repairs and the hole on the calcium phosphate composite porous microspheres that is used for is at 10nm～20um.
10. the preparation method according to each described composite porous microspheres of claim 1～9 is characterized in that, this method may further comprise the steps:
1) get the gelatin powder body, add deionized water, the preparation gelatin solution adds soluble calcium salt then in gelatin solution, and its addition is the soluble calcium salt that every gram gelatin powder body adds 0.0003～0.03mol; Subsequently, add the active substance antler polypeptide or the bone morphogenetic protein that can impel osteogenesis again, the addition of antler polypeptide or bone morphogenetic protein is 1 ‰～5% of gelatin weight;
2) mixed liquor that step 1) is made is poured in the oil that contains surfactant, stirs the water-in-oil emulsion that forms gelatin solution; The content of surfactant in oil is 1g/200ml oil～1g/20ml oil; The amount of used oil is every gram gelatin 5ml～250ml;
3) with step 2) the water-in-oil emulsion heating of the gelatin solution that makes, temperature is controlled at 30 ℃～70 ℃, and continues to stir, and mixing speed is 100 rev/mins～6000 rev/mins; Add the soluble phosphoric acid saline solution then, phosphatic addition makes the proportion control of Ca element in the mixed liquor and P element at 1:1～2:1;
4) mixed liquor that step 3) is made stirs under 30 ℃～70 ℃ temperature, continues to stir down, cools off with ice bath; Form active substance antler polypeptide or bone morphogenetic protein that homodisperse in the gelatine microsphere has calcium phosphate and can impel osteogenesis;
5) add cross-linking agent in the mixed liquor that step 4) makes, the mass concentration of cross-linking agent is 5 ‰～15%, and the addition of cross-linking agent is the cross-linking agent that every gram gelatin adds 30ul～1000ul, continues to stir;
6) get step 4) or 5) mixed liquor that makes, add acetone, addition is 2～8 times of moisture content volume in the mixed liquor, continues to stir, and filters, and obtains the crude product of composite porous microspheres;
7) after the crude product of the composite porous microspheres that step 6) is obtained is used the acetone cyclic washing, obtain being used for gelatin and the calcium phosphate composite porous microspheres that bone is repaired.
11. method according to claim 10 is characterized in that: described surfactant is a kind of among SPAN series or the dioctyl sodium sulfosuccinate surfactant.
12. method according to claim 10 is characterized in that: a kind of in silicone oil, Oleum Glycines, olive oil, Oleum Camelliae, Oleum Brassicae campestris or mixed oil of described grease separation.
13. the purposes according to each described composite porous microspheres of claim 1～9 is characterized in that: described gelatin that bone repairs and the calcium phosphate composite porous microspheres of being used for can be as the material of the reparation of bone, tooth.
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| CN103041452A (en) * | 2011-10-13 | 2013-04-17 | 中国科学院理化技术研究所 | Porous bone repair framework material of gelatin/nano bone-like apatite, and preparation method and application thereof |
| CN103027740A (en) * | 2013-01-17 | 2013-04-10 | 北京中奥汇成生物材料科技有限公司 | Absorbable bone screw with slow-release biological factor and preparation method thereof |
| CN103467756A (en) * | 2013-09-18 | 2013-12-25 | 中国矿业大学 | Method for preparing chitosan/hydroxyapatite composite microspheres |
| CN104436311A (en) * | 2014-11-13 | 2015-03-25 | 吉林医药学院 | Pilose antler polypeptide-collagen-chitosan sustained-release material and preparation method thereof |
| CN104523341B (en) * | 2014-12-18 | 2017-06-20 | 中国人民解放军第四军医大学 | The manufacture method of the immediate implantation teeth with periodontal bioactivity |
| CN104523341A (en) * | 2014-12-18 | 2015-04-22 | 中国人民解放军第四军医大学 | Method for manufacturing immediately implanted tooth with periodontal bioactivity |
| CN104771785A (en) * | 2015-04-03 | 2015-07-15 | 周宏志 | Preparation method of bone repair material with neuropeptide inductive osteogenic activity |
| CN105858629A (en) * | 2016-03-30 | 2016-08-17 | 武汉亚洲生物材料有限公司 | Preparation method for inorganic microspheres, inorganic microspheres prepared thereby, and application of inorganic microspheres |
| CN105858629B (en) * | 2016-03-30 | 2018-07-17 | 武汉亚洲生物材料有限公司 | A kind of preparation method of inorganic microsphere and thus obtained inorganic microsphere and application thereof |
| CN106860913A (en) * | 2017-02-22 | 2017-06-20 | 昆明医科大学 | The application of antler polypeptide, tooth implant and its surface treatment method |
| CN112891622A (en) * | 2021-02-04 | 2021-06-04 | 武汉理工大学 | Porous calcium-magnesium dual-phase ceramic microsphere and preparation method thereof |
| CN113350573A (en) * | 2021-06-07 | 2021-09-07 | 王健 | Porous microsphere adhesive with osteoinductive capacity and preparation method thereof |
| CN113827767A (en) * | 2021-09-13 | 2021-12-24 | 西北大学 | Preparation method of novel microgel bone powder for repairing postoperative tissue of osteoma |
| CN113827767B (en) * | 2021-09-13 | 2022-05-17 | 西北大学 | Preparation method of novel microgel bone powder for repairing postoperative tissue of osteoma |
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