CN100546938C - Porous ceramic material for repairing bone and its production and application - Google Patents
Porous ceramic material for repairing bone and its production and application Download PDFInfo
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- CN100546938C CN100546938C CNB2005100305903A CN200510030590A CN100546938C CN 100546938 C CN100546938 C CN 100546938C CN B2005100305903 A CNB2005100305903 A CN B2005100305903A CN 200510030590 A CN200510030590 A CN 200510030590A CN 100546938 C CN100546938 C CN 100546938C
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Abstract
The invention provides a kind of porous ceramic material for repairing bone of a kind of porous ceramic material for repairing bone, the aperture of described porous ceramic film material is 200～500 μ m, porosity interconnects greater than 80%, 90%～99% hole, and ultimate compression strength is 300KPa-3MPa; Described stupalith contains the polymkeric substance in stupalith weight 0.2%～25%, and described polymkeric substance is the polymkeric substance of viscosity between 0.1～20PaS behind the wiring solution-forming.The invention also discloses the preparation method and the application of this porous ceramic film material.Porous ceramic film material intensity height of the present invention, the skeletonization transmissibility is strong, the preparation method is simple.
Description
Technical field
The present invention relates to technical field of biological material and field of tissue engineering technology, relate to porous ceramic material for repairing bone and its production and application particularly.
Background technology
Stupalith is as bone reparation and bone alternate material, and main drawback is that fragility is bigger, and its Young's modulus is difficult to be complementary with normal bone, has limited application clinically to a certain extent.Since the sixties, the correlative study that stupalith is used at orthopaedics is quite deep, but still limits to very much aspect clinical application, and major cause is the requirement that is difficult to reach clinical satisfaction.
The bioceramic material that is used for hard tissue repair comprises that hydroxyapatite (HAP), bata-tricalcium phosphate (TCP), calcium phosphate bone cement (CPC), bio-vitric (bioglass) etc. are multiple.Wherein β-TCP pottery molecular formula is Ca
3(PO
4)
2, ratio of calcium and phosphorus is 1.5, and is approaching with the ratio of calcium and phosphorus of normal bone tissues.The most outstanding characteristics of β-TCP are to degrade in vivo.As
Renooij W, wait (the bio-absorbable of the calcium phosphate ceramic of strontium 85 marks implantation dog-leg at Bioresorption of ceramicstrontium-85-labeled calcium phosphate implants in dog femora.A pilot study toquantitate bioresorption of ceramic implants of hydroxyapatite and tricalciumorthophosphate in vivo.; The forward position research of hydroxyapatite and the quantitative absorption in vivo of tricalcium orthophosphate pottery implant; Clinical plastic surgery correlative study) Clin Orthop Relat Res.1985Jul-Aug; (197): disclosed among the 272-85., experimentation on animals finds that β-TCP degraded in 24 weeks fully after implanting sucking pig palate mastoid process, with isotopic labeling HA and TCP, find that HA does not have degraded, and TCP is in 22 week degradeds 25%～30%.The biocompatibility of β-TCP is also very good, no fibrous tissue interval between bone and material, and bone directly contacts with material, and slight macrophage response is only arranged after implanting.There are some researches show that this degraded with β-TCP is relevant.The bone conduction ability of β-TCP is more intense, and β-TCP and HA are implanted the mandibular defect of sheep respectively, and the bone conduction effect of β-TCP is better than HA.The main drawback of β-TCP is that strength ratio is lower, so current research and application scale are not as good as HA.On the other hand, the essential coupling of the speed of material degradation and skeletonization, because the intensity of TCP is lower than other stupalith, if there is not skeletonization rapidly, the bone repair of TCP may be failed.
Different materials all respectively has relative merits in the damaged reparation of bone, the damaged requirement to repair materials of bone of different sites and size is also inequality, in addition, simple stupalith has only bone conduction effect and does not have inducing action, therefore, single material can expose inevitable defective when repairing bone defect.For addressing this problem, different materials and biotic factor are combined with the deficiency that may remedy various materials, play the damaged repair of bone better.As J.C.Brodie, Deng at Osteoblast interactions with calcium phosphateceramics modified by coating with type I collagen.J.Biomed.Mater.Res.73A:409-421, the 2005 (interactions between the calcium phosphate ceramic after scleroblast and type i collagen are protein modified; Bio-medical material research magazine) disclosed in, with HA, TCP and their mixture carry out finishing with collagen protein, can significantly improve propagation and the skeletonization integration (osteointegration) of scleroblast on the calcium orthophosphate base pottery in the early stage vitro culture.Also disclosed in Calvarial boneresponse to a tricalcium phosphate-genipin crosslinked gelatin composite. (skull is to the reaction of tricalcium phosphate/genipin cross-linked gelatin mixture) Biomaterials 26 (2005) 3065-3074 just like Chun-Hsu Yao etc., with TCP and gelatin-compounded reparation skull defeci, found that also has preferably the new bone situation of growing into.
As tissue engineering material, pottery is made the porous shape mostly, and the material that contains hole is grown into for area of new bone in bone the space is provided, and bone is contacted with material closely, and the osteoconductive that not only can increase material has also improved the stability of material in bone.For degradation material, the porous form can increase the total area of material, promotes the degraded of material.Porous material is also grown into for blood vessel and is provided the foundation.Blood vessel is grown into and has been brought the required various factors of skeletonization, mesenchymal cell, reaches the required nutrition of other osteogenesis.But, embedded material is made vesicular, can obviously weaken the intensity of material.The material that is used for bone tissue engineer should have fine intensity, so that can bear various loads in early days.On the other hand, the mechanics that too high intensity can change surrounding tissue distributes, and also is unfavorable for the formation of bone and moulds.
We utilize the foam impregnation method also to prepare the aperture at 200～500 μ m on the former study basis, and porosity has the various porous materials of intercommunicating pore structure greater than 80%, comprise porous beta-TCP pottery and Porous HAP stupalith.Confirm that through in vitro study this porous beta-TCP pottery helps the adhesion growth of bone with various kinds of cell such as stroma stem cell, fat stem cells, experimentation on animals shows also can well guide new osteogenesis, and good osteogenic activity is arranged.But because this porous beta-TCP pottery porosity height, maximum compressive strength only about 0.1Mpa, has limited its further application greatly.
In order to strengthen the intensity of porous ceramic film material, TCP pottery or other porous ceramicss have been carried out a lot of researchs with polymkeric substance is compound, but mostly other investigators be on the basis of polymkeric substance, add ceramic particle or fiber, thereby reach enhancing or compound purpose.As A.Bigi etc. at Bonelike apatite growth onhydroxyapatite-gelatin sponges from simulated body fluid.J Biomed Mater Res 59:709-714, the 2002 (growths of osteoid phosphatic rock on hydroxyapatite-gelfoam that simulated body fluid soaks; Bio-medical material research magazine). and Y Zhang etc. is at Cell growth and function oncalcium phosphate reinforced chitosan scaffolds.J Mater Sci Mater Med 15:255-260,2004. (growth and the functional expressions of cell in calcium phosphate enhanced chitosan stent; Materials science magazine-medical material fascicle) disclosed in.Mostly this method is the character based on polymkeric substance, and reinforced effects is not obvious.
The investigator is also arranged based on porous ceramics, polymkeric substance is introduced porous support, thereby realize compound by freeze-drying method.As Y Zhang, Deng at Three-dimensional macroporous calciumphosphate bioceramics with nested chitosan sponges for load-bearing bone implants.J Biomed Mater Res 61:1-8,2002 (the nested chitosan sponge of the three-dimensional macropore tricalcium phosphate biological ceramics bone implants that are used to bear a heavy burden; Bio-medical material is studied magazine) and .C Chang, Deng at Cartilage TissueEngineering on the Surface of a Novel Gelatin-Calcium-Phosphate BiphasicScaffold in a Double-Chamber Bioreactor.J Biomed Mater Res Part B:ApplBiomater 71B:313-321,2004 (in the double flute bio-reactor, a kind of new gelatin-calcium phosphate biphase ceramics rack surface cartilage tissue engineered; Bio-medical material research magazine B: disclosed the applying biological material).The matrix material that this method is prepared, the mass ratio of organic/inorganic thing is higher, and reinforced effects is also not too remarkable.
In sum, this area lacks a kind of intensity height, does not change porous ceramic film material of material character and preparation method thereof simultaneously again.Therefore, this area press for that exploitation a kind of intensity height, skeletonization transmissibility are strong, the simple porous ceramic film material of preparation method and preparation method thereof.
Summary of the invention
The object of the present invention is to provide that a kind of intensity height, skeletonization transmissibility are strong, the simple porous ceramic film material of preparation method and preparation method thereof.
In a first aspect of the present invention, a kind of porous ceramic material for repairing bone is provided, the aperture of described porous ceramic film material is 200～500 μ m, porosity interconnects greater than 80%, 90%～99% hole, and ultimate compression strength is 300KPa-3MPa; Described stupalith contains the polymkeric substance in stupalith weight 0.2%～25%, and described polymkeric substance is the polymkeric substance of viscosity between 0.1～20PaS behind the wiring solution-forming.
In the preference of the present invention, described stupalith is the bata-tricalcium phosphate stupalith.
In the preference of the present invention, described polymkeric substance comprises gelatin, collagen protein, chitosan, hyaluronic acid, alginate, fibrin gel, polyoxyethylene poly-oxygen propylene aether segmented copolymer, poly(lactic acid), polyglycolic acid or its combination.
The present invention provides the preparation method of porous ceramic material for repairing bone on the other hand, prepares with the method that may further comprise the steps:
(a) viscosity with described stupalith immersion 0.5～20% (w/v) is in the polymers soln of 0.1～20PaS, obtains the stupalith that the surface is aggregated the thing solution impregnation; The mass ratio of contained polymkeric substance and stupalith is 0.003～0.3 in the wherein said polymers soln;
(b) the infiltration stupalith that step (a) obtained takes out, and filter is done, and drying promptly obtains the stupalith of finishing.
In the preference of method of the present invention, may further comprise the steps:
(a) viscosity with described stupalith immersion 0.5～20% (w/v) is in the polymers soln of 0.1～20PaS, obtains the stupalith that the surface is aggregated the thing solution impregnation; The mass ratio of contained polymkeric substance and stupalith is 0.003～0.3 in the wherein said polymers soln;
(b) the infiltration stupalith that step (a) obtained takes out, and filter is done, and drying promptly obtains the stupalith of finishing.
In the preference of method of the present invention, the described polymkeric substance of step (a) comprises gelatin, collagen protein, chitosan, hyaluronic acid, alginate, fibrin gel, polyoxyethylene poly-oxygen propylene aether segmented copolymer, poly(lactic acid) or polyglycolic acid.
In the preference of the present invention, the polymers soln in the step (a) also adds a certain amount of (as 0.00625～0.1 μ g/mg) promotes the biotic factor of osteogenic growth.Preferably, described biotic factor is selected from Delicious peptide, osteonectin, bone calcification element, the skeleton connection factor or its combination.
In the preference of method of the present invention, the stupalith that step (b) obtains carries out step (a) and step (b) 1～5 time again, obtains the stupalith of finishing.
The present invention provides a kind of tissue engineered implant on the other hand, described graft contains porous ceramic material for repairing bone and is inoculated in the stem cell of described stupalith, the aperture of described porous ceramic film material is 200～500 μ m, porosity is greater than 80%, 90%～99% hole interconnects, and ultimate compression strength is 300KPa-3MPa; Described stupalith contains the polymkeric substance in stupalith weight 0.2%～25%, described polymkeric substance is the polymkeric substance of viscosity between 0.1～20PaS behind the wiring solution-forming, described stem cell is selected from bone marrow stroma stem cell or fat stem cell, and the inoculum size of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3Porous ceramic film material.
A further aspect of the present invention provides the purposes of porous ceramic film material, as the support of preparation bone graft.
Description of drawings
The stereoscan photograph of the porous beta-TCP pottery of Fig. 1 after 5% gelatin solution is modified;
The stereoscan photograph of the porous beta-TCP pottery of Fig. 2 after 2% chitosan solution is modified;
Fig. 3 modifies through 5% gelatin solution, the stereoscan photograph of β-TCP pottery that lyophilize obtains;
Before Fig. 4 finishing, the stereoscan photograph of porous beta-TCP pottery;
Fig. 5 bone marrow stroma stem cell is cultivated the stereoscan photograph after 7 days in the porous beta-TCP pottery.A left side: surface; Right: section;
Fig. 6 fat stem cell is cultivated the stereoscan photograph left side after 5 days in the porous beta-TCP pottery: the surface; Right: section.
Embodiment
The inventor is through extensive and deep research, porous ceramic film material is carried out finishing, unexpectedly with lower polymkeric substance/porous ceramics mass ratio, obtained to have the porous ceramics of higher-strength, high porosity, and adhesion, propagation, differentiation and the skeletonization transmissibility of the porous ceramics pair cell after find modifying do not influence even increase.Described porous ceramic film material is enhanced after by the polymers soln finishing, and behind the surface modification, does not change the porous ceramics pore structure, and ultimate compression strength is significantly improved.Finished the present invention on this basis.
The present invention also proposes a kind of method of finishing, makes the porous ceramics with high porosity that certain mechanical strength be arranged, and the porous ceramics after the finishing simultaneously can better guide the regeneration of bone.
Porous ceramic film material
Porous ceramic film material used in the present invention comprises bata-tricalcium phosphate (TCP), hydroxyapatite (HAP), bio-vitric (bioglass), TCP-HAP biphase ceramics or the mixture of various porous ceramicss.Can also adopt calcium phosphate bone cement (CPC) to substitute porous ceramic film material.
Preferably, porous ceramic film material is that bata-tricalcium phosphate (is the stupalith of β-TCP).It can make by general porous material preparation method, as porous plastics pickling process, foaming, interpolation pore-forming material method, sol-gel method, the preferably porous ceramic film material for preparing by the foam impregnation method.The aperture of porous beta-TCP pottery is 200～500 μ m (preferably, 300-500 μ m), and porosity has intercommunicating pore structure greater than 80%, and wherein the hole of 90%-99% interconnects.Preferably, the cellular type of 95-98% interconnects.The ultimate compression strength of porous beta-TCP pottery is 60Kpa-150kPa.After modifying, ultimate compression strength can reach 300KPa-3MPa.
By can prepare hydroxyapatite (HAP) porous ceramics with quadrat method, ultimate compression strength is 160.958 ± 19.63KPa, the HAP pottery after 1% gelatin solution is modified, and ultimate compression strength is 348.53 ± 17.98KPa; HAP pottery after 10% gelatin solution is modified, ultimate compression strength is 2.779 ± 0.483MPa.
Polymkeric substance
Polymkeric substance has no particular limits, so long as have certain viscosity behind the wiring solution-forming, and also easy film forming polymkeric substance all can be used as coating materials.The scope of the absolute viscosity η of the polymers soln that the present invention is suitable for is 0.1～20Pa, and this viscosity is measured by the rotational viscosimeter of routine.
The polymkeric substance that is suitable for comprises natural polymer, also comprises the synthetic polymkeric substance.Natural polymkeric substance such as gelatin, collagen protein, chitosan, hyaluronic acid, alginate, fibrin gel.Synthetic polymer such as Pluronic (polyoxyethylene poly-oxygen propylene aether segmented copolymer), poly(lactic acid), polyglycolic acid etc., the molecular weight of described polymkeric substance is 36KDa-100Kda, preferably, is 50KDa-80Kda.
During prepared polymer solution, the biotic factor that promotes osteogenic growth be can suitably add, the factor (SCF) or its combination connected as Delicious peptide (BMP), OP-1, bone gel, osteonectin (osteoconectin), bone calcification element (osteocalcin), skeleton.The biotic factor of described promotion osteogenic growth is with affiliated animal classification difference, and the amount of interpolation is also different, and preferably the mass ratio with polymkeric substance is 0.00625～0.1 μ g/mg, more preferably 0.01～0.08 μ g/mg.In the present invention, biotic factor can add when prepared polymer solution.If the compound co-cultivation of cell and material then can also add in nutrient solution.
The consumption of the polymkeric substance that finishing is used is: the mass ratio of polymkeric substance/TCP is 0.003～0.3, preferred 0.01-0.2; Wherein the quality of polymkeric substance is meant the quality of polymer solids part in the polymers soln.
The concentration of polymers soln is 0.5～20% (w/v), preferred 2～10% (w/v).Polymer concentration is low excessively, and then the film forming thickness deficiency a little less than the effect too of bracing frame, does not reach intensity of the present invention; Polymer concentration is too high, and then the plugging hole structure changes the structure in hole.
Surface modification method
The invention discloses the surface modification method of a kind of degradable multiporous β-TCP pottery.Described porous ceramics has high porosity, but intensity is low, can significantly improve the intensity of porous ceramics by surface modification method.
Described surface modification method is realized by following steps:
Be polymers soln at first, be used for porous ceramics is carried out surface treatment, carry out following step then the polymer formulation described in the above-mentioned embodiment;
(a) with in the polymers soln described in the immersion of porous beta-TCP pottery, make polymers soln partly or entirely soak into porous ceramic surface.Preferably, carry out vacuum stripping,, guarantee that polymers soln all soaks into porous ceramic surface until no bubble emersion;
(b) at last porous ceramics is taken out, filter is done, and carries out drying again.The drying of sample can adopt oven dry, also can adopt lyophilize.Preferably, dry 12～48h or 30～60 ℃ of down dry 12～48h under the room temperature, perhaps-20 ℃～-80 ℃ freeze overnight in back are done in filter, again or-50 ℃ of following lyophilizes, promptly obtain the stupalith after the finishing.
Stupalith after the above-mentioned finishing that obtains can carry out above-mentioned steps (a) and step (b) more repeatedly, for example carries out 1-5 time, preferably, carries out 2～3 times, obtains the stupalith of repeatedly finishing.
The intensity of the porous ceramic film material that obtains is 300KPa-3MPa, preferably is 800KPa-3MPa.
The mechanism that the TCP ceramics strength improves after the finishing is, the porous ceramics of foam impregnation method preparation, and the structure that has macropore and micropore and deposit, because porosity is higher, the ultimate compression strength of pottery is lower.After the polymers soln finishing, the micropore on the ceramic hole wall is aggregated the thing film and covers, and polymeric film has also played supporting role to whole porous ceramics simultaneously, thereby ceramics strength is significantly improved.Polymer concentration is high more, and film forming thickness is big more, and the effect of bracing frame is strong more, thereby intensity is also just high more.
Stem cell
The source of stem cell of the present invention is not particularly limited, and can be the stem cell in any source, and usually, stem cell of the present invention is from body or allochthonous stem cell.The position that obtains stem cell also is not particularly limited, and can be fat stem cell, bone marrow stroma stem cell or other stem cells.In addition, the also alternative stem cell of scleroblast is as the seed cell of bone tissue engineer structure.
Can be used for stem cell of the present invention can be preferably Mammals from any vertebrates, more preferably is primate, especially the people.
Although the stem cell from body is preferred, the source of the stem cell of allosome is more commonly used.Research shows, and different growths, the allogeneic stem cell of etap can and have in the allogeneic animal body of complete immunologic function in consistency difference in a organized way and form the stem cell tissue.
The method of separation and acquisition stem cell is as known in the art.A kind of preferable methods is density gradient centrifugation and enzyme digestion.
Cultural method of stem cell and nutrient solution also are to know in this area.A kind of preferable methods is at 37 ℃, saturated humidity, 5%CO with stem cell
2Cultivate in the incubator.Suitable nutrient solution comprises (but being not limited to): 1) DMEM substratum ((Gibco company)+5～20% foetal calf serums; 2) DMEM substratum+5～20% calf serums; 3) DMEM substratum+5～20% are from body (allosome) human serum.In addition, add various somatomedins (for example promoting the cytokine of stem cell growth etc.), various microbiotic, various inducible factor in the above-mentioned nutrient solution.
Be applicable to that stem cell of the present invention should be able to be in vivo or in-vitro multiplication.A kind of preferred stem cell is the bone marrow stroma stem cell of vitro culture.
Bone graft
Because the consistency of porous ceramic film material of the present invention, particularly beta-tricalcium phosphate porous ceramic material and bone marrow stroma stem cell and fat stem cell is very good, therefore be particularly suitable as the timbering material that bone is repaired.
Form stem cell-porous ceramic film material mixture on that the bone marrow stroma stem cell of cultured and amplified in vitro and/or fat stem cell are inoculated into the biocompatibility excellence and the degradable multiporous stupalith, this " stem cell-porous ceramic film material " mixture is implanted to defect, degraded and absorbed gradually along with porous ceramic film material, new bone forming reaches the purpose of repairing bone defect.
The preparation method of organizational project bone graft of the present invention is easy, with the bone marrow stroma stem cell of some amount and/or fat stem cell is inoculated in porous ceramic film material such as beta-tricalcium phosphate porous ceramic material gets final product.
The shape of engineered bone graft of the present invention is not particularly limited, can be moulding arbitrarily according to the shape of tissue defect.Usually, graft is a long strip shape.
Bone marrow stroma stem cell in the tissue-engineered bone of the present invention and/or fat stem cell concentration are about 0.5 * 10 usually
6/ cm
3(ceramics bracket volume) is to 5 * 10
8/ cm
3Or higher, preferably be 1 * 10
6/ cm
3To 1 * 10
8/ cm
3, more preferably be 5 * 10
6/ cm
3To 5 * 10
7/ cm
3Porous ceramic film material.Usually, adjust bone marrow stroma stem cell and/or fat stem cell concentration, mix with degradation material then with nutrient solution.During mixing, the ratio of nutrient solution and degradation material is not particularly limited, but is advisable with the nutrient solution maximum that this material can adsorb.
In addition, in engineered bone graft of the present invention, also can add or compound other various cells, somatomedin, various microbiotic, thereby keep bone marrow stroma stem cell and/or fat stem cell phenotype, promote bone marrow stroma stem cell and/or fat stem cell growth, and promote tissue-engineered bone to grow in vivo.
Except engineered bone graft is implanted, also be placed in the external biological reactor and cultivate, thereby carry out the structure of tissue-engineered bone, have the tissue-engineered bone of certain histological structure, biochemical composition and biomechanical strength in external formation.
With the engineered bone graft that the inventive method forms, the damaged place of the bone that can directly implant.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:ColdSpring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.Ratio and per-cent are based on weight, unless stated otherwise.
Embodiment 1
The gelatin solution of preparation 1% (w/v) concentration, porous beta-TCP pottery behind the sintering is put into gelatin solution, vacuumize degasification, until there not being the bubble emersion, then porous beta-TCP pottery is taken out, filter is done, drying 24h at least under the room temperature, 40 ℃ of dryings are 12h at least, promptly obtains enhanced porous beta-TCP pottery.
Porous ceramics after the finishing, pore structure are shown in accompanying drawing 1.The porous beta-TCP pottery still has intercommunicating pore structure, and the aperture does not change yet.By mensuration, the porous beta-TCP pottery after the finishing, porosity is 92.6 ± 1.07%.Ultimate compression strength is that (Shimadzu carries out on Japan), and the intensity of the porous beta-TCP pottery after 1% gelatin solution is modified is 387.83 ± 92Kpa for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 2
The gelatin solution of preparation 10% (w/v) concentration, the porous beta-TCP pottery is put into gelatin solution, vacuumize degasification, until there not being the bubble emersion, then porous beta-TCP pottery is taken out, filter is done, drying 24h at least under the room temperature, 40 ℃ of dryings are 12h at least, promptly obtains enhanced porous beta-TCP pottery.
Porous ceramics after the finishing, its porosity are 90.74 ± 1.99%.Ultimate compression strength is that (Shimadzu carries out on Japan), and the intensity of the porous beta-TCP pottery after 10% gelatin solution is modified is 2.05 ± 0.297Mpa for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 3
The chitosan solution of preparation 2% (w/v) concentration, the porous beta-TCP pottery is put into gelatin solution, vacuumize degasification, until there not being the bubble emersion, then porous beta-TCP pottery is taken out, filter is done, drying 24h at least under the room temperature, 60 ℃ of dryings are 12h at least, promptly obtains enhanced porous beta-TCP pottery.
Porous ceramics after the finishing, pore structure are shown in accompanying drawing 2.The porous beta-TCP pottery still has intercommunicating pore structure, and the aperture does not obviously change yet.By mensuration, the porous beta-TCP pottery after the finishing, porosity is 91.9 ± 1.1%.Ultimate compression strength is that (Shimadzu carries out on Japan), and the intensity of the porous beta-TCP pottery after 2% chitosan solution is modified is 484.85 ± 286.5Kpa for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 4
The gelatin solution of preparation 5% (w/v) concentration, porous beta-TCP pottery behind the sintering is put into gelatin solution, vacuumize degasification, until there not being the bubble emersion, then the porous beta-TCP pottery is taken out, filter is done ,-20 ℃ of freeze overnight, then-50 ℃ of following lyophilizes until the sample complete drying, promptly obtain enhanced porous beta-TCP pottery.
By the porous ceramics that freeze-drying method obtains, pore structure is shown in accompanying drawing 3.The porous beta-TCP pottery still has intercommunicating pore structure, and the aperture does not change yet.The visible film sample of hole wall surface gelatin since porous ceramic surface to adhere to gelatin less, the common netted gelatin structure of freeze-drying is not obvious.Ultimate compression strength is that (Shimadzu carries out on Japan), modifies through 5% gelatin solution, and the intensity of the porous beta-TCP pottery after the lyophilize is 201.37 ± 17.06Kpa for the omnipotent testing experiment machine of AG-1 in model.
Embodiment 5
Being equipped with the HAP porous ceramics by the foam impregnation legal system, after 1100 ℃ of calcinings, is that (Shimadzu carries out intensity test on Japan), and its ultimate compression strength is 160.958 ± 19.63KPa for the omnipotent testing experiment machine of AG-1 in model.The gelatin solution of preparation 10% (w/v) concentration is put into gelatin solution with the Porous HAP pottery, vacuumizes degasification, until there not being the bubble emersion, then the Porous HAP pottery to be taken out, filter is done, dry 24h under the room temperature, drying 12h at least in 37 ℃ of baking ovens, surveying its ultimate compression strength is 2.779 ± 0.483MPa.
Porous ceramic film material of the present invention for example the porous bata-tricalcium phosphate (stupalith of β-TCP) has higher porosity, and the aperture is 300～500 μ m, and connects in the hole, is suitable as tissue engineering cell scaffold material (as Fig. 4).Because porous beta-TCP stupalith porosity of the present invention is higher, and is better with the consistency of bone marrow stroma stem cell and fat stem cell.In SEM photo (shown in Fig. 5-6), can find all to be covered in rack surface and the hole, and spread all over extracellular matrix by cell.Also demonstrate the extensive distribution of cell and extracellular matrix on the section structure of material.Show thus, bone marrow stroma stem cell and fat stem cell can be at porous beta-TCP ceramic surface well-growns, and after cultivating certain hour, because the good connectedness of porous ceramics, cell also can migrate to the material internal justacrine and go out a large amount of extracellular matrixs, and consistency is preferably arranged.For bone marrow stroma stem cell and fat stem cell, the porous beta-TCP pottery for preparing by the foam impregnation method is the good cell timbering material, can be as the used in tissue engineering bone renovating bracket material.
Major advantage of the present invention is:
(1) simple to operate, be convenient to promote;
(2) do not change the pore structure of original porous ceramics;
(3) effectively improve the intensity of high porosity porous ceramics material;
(4) can add multiple biologically active factors in the operating process, thereby further improve clinical efficacy as the bone material for repairing.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Claims (10)
1. a bone reparation stupalith is characterized in that, the aperture of described stupalith is 200～500 μ m, and porosity interconnects greater than 80%, 90%～99% hole, and ultimate compression strength is 300KPa-3MPa; Described stupalith contains the polymkeric substance in stupalith weight 0.2%～25%, and described polymkeric substance is the polymkeric substance of viscosity between 0.1～20PaS behind the wiring solution-forming;
Described polymkeric substance is selected from gelatin, collagen protein, chitosan, hyaluronic acid, alginate, fibrin gel, polyoxyethylene poly-oxygen propylene aether segmented copolymer, poly(lactic acid), polyglycolic acid or its combination.
2. stupalith as claimed in claim 1 is characterized in that, described stupalith is the bata-tricalcium phosphate stupalith.
3. stupalith as claimed in claim 1 is characterized in that, the concentration behind the described polymkeric substance wiring solution-forming counts 2～10% with w/v.
4. as any described stupalith of claim 1 to 3, it is characterized in that, prepare with the method that may further comprise the steps:
(a) viscosity with stupalith immersion 0.5～20% (w/v) is in the polymers soln of 0.1～20PaS, obtains the stupalith that the surface is aggregated the thing solution impregnation; The mass ratio of contained polymkeric substance and stupalith is 0.003～0.3 in the wherein said polymers soln;
(b) the infiltration stupalith that step (a) obtained takes out, and filter is done, and drying promptly obtains the stupalith of finishing.
5. the preparation method of a stupalith as claimed in claim 1 is characterized in that, may further comprise the steps:
(a) viscosity with stupalith immersion 0.5～20% is in the polymers soln of 0.1～20PaS, obtains the stupalith that the surface is aggregated the thing solution impregnation; The mass ratio of contained polymkeric substance and stupalith is 0.003～0.3 in the wherein said polymers soln; Described polymkeric substance is selected from gelatin, collagen protein, chitosan, hyaluronic acid, alginate, fibrin gel, polyoxyethylene poly-oxygen propylene aether segmented copolymer, poly(lactic acid) or polyglycolic acid;
(b) the infiltration stupalith that step (a) obtained takes out, and filter is done, and drying promptly obtains the stupalith of finishing.
6. method as claimed in claim 5 is characterized in that, the concentration of the described polymers soln of step (a) counts 2～10% with w/v.
7. method as claimed in claim 5, it is characterized in that, polymers soln in the step (a) also adds the biotic factor that promotes osteogenic growth, and described biotic factor is selected from Delicious peptide, and the mass ratio of described biotic factor and polymkeric substance is 0.00625～0.1 μ g/mg.
8. method as claimed in claim 5 is characterized in that, the stupalith that step (b) obtains carries out step (a) and step (b) 1～5 time again, obtains the stupalith of finishing.
9. tissue engineered implant, it is characterized in that, described graft contains the described stupalith of claim 1 and is inoculated in the stem cell of described stupalith, and described stem cell is selected from bone marrow stroma stem cell or fat stem cell, and the inoculum size of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3Stupalith.
10. the purposes of stupalith as claimed in claim 1 is characterized in that, as the support of preparation bone graft.
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| GB2511825A (en) | 2013-03-14 | 2014-09-17 | Ostomycure As | Implant |
| WO2016096939A1 (en) * | 2014-12-16 | 2016-06-23 | Ceramtec Gmbh | Spinal cages and instruments for inserting same |
| CN107308503A (en) * | 2017-06-16 | 2017-11-03 | 无锡贝迪生物工程股份有限公司 | A kind of plasticity artificial bone and preparation method thereof |
| CN107812236A (en) * | 2017-10-31 | 2018-03-20 | 桂林市漓江机电制造有限公司 | A kind of high-strength and high ductility bone repair material |
| CN108310454B (en) * | 2018-03-20 | 2020-04-17 | 山东大学 | Gradient biological ceramic material coated with gelatin/chitosan composite porous membrane and preparation method thereof |
| CN108273127A (en) * | 2018-04-23 | 2018-07-13 | 周荣 | A kind of preparation method of enzymolysis type bio-vitric dental pulp repair materials |
| CN108793845A (en) * | 2018-07-05 | 2018-11-13 | 叶露穗 | A kind of calcium phosphate bioactive ceramics and preparation method thereof |
| US11433159B2 (en) * | 2019-07-26 | 2022-09-06 | Warsaw Orthopedic, Inc. | Hydratable and flowable implantable compositions and methods of making and using them |
| US11498880B2 (en) | 2019-07-26 | 2022-11-15 | Warsaw Orthopedic, Inc. | Calcium phosphate granules and methods of making them |
| CN112220964A (en) * | 2020-10-19 | 2021-01-15 | 西安点云生物科技有限公司 | Composite biological ceramic powder, composite biological ceramic artificial bone prepared from composite biological ceramic powder and preparation method of composite biological ceramic artificial bone |
| CN116553907A (en) * | 2023-05-10 | 2023-08-08 | 中国科学技术大学 | Porous ceramic material of cross-scale half-tube-like structure, preparation method thereof and metal mold |
| CN116617455A (en) * | 2023-05-29 | 2023-08-22 | 重庆生物智能制造研究院 | Preparation method of porous biological ceramic artificial bone scaffold with bioactivity |
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