CN101496909B - Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof - Google Patents
Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a polysaccharide/autosolidification calcium phosphate composite bone cement composition, which comprises the following components in percentage by weight: 10 to 95 weight percent of autosolidification calcium phosphate bone cement and 5 to 90 weight percent of polysaccharide. The bone cement composition can obtain bone cement with high strength, good toughness, strong plasticity, quick solidification, and good biocompatibility and degradability, thereby overcoming the defects of inadequate toughness and slow degradation in human bodies existing in the prior calcium phosphate bone cement materials and the defects existing in a plurality of enhancement methods in the prior art, and better meeting the requirement of operation use.
Description
Technical field
The invention belongs to the biomaterial for medical purpose field, relate to a kind of for filling or the damaged novel organic/inorganic composite material of injection reparation human body hard tissue.
Background technology
The repairing and treating that bone is damaged is the surgical thorny difficult problem of long-term puzzlement, and the various countries scientist is devoted to the research and development of desirable bone renovating material always.Last century, Brown and the Chow of the mid-80 U.S. invented a kind of self-curing type biological activity bone impairment renovation material---calcium phosphate bone cement (Calcium phosphate cement, CPC), it can be under the Human Physiology environment hydration and sclerosis voluntarily, finally be converted into to human bone and form similar hydroxyapatite (HA), and thermal discharge is few in the solidification process, can be according to any plastotype of bone defect shape, thereby caused that people pay close attention to greatly.Have good performance as the bone alternate material calcium phosphate bone cement, main manifestations is: good biocompatibility and biological safety, good biological degradability, guiding osteogenic activity.Calcium phosphate bone cement can change into natural bone and similarly form, can participate in metabolism behind the implant into body, the skeletonization by bone conduction effect, new osteogenesis (W.J.E.M.Habraken, J.G.C.Wolke, J.A.Jansen in absorbed bootable equivalent of while, ceramic composite is used for the research of organizational project medicine transmission timbering material, advanced drugs transmission 2007,59,234-248; Hockin H.K.Xu, Michael D.Weir, Elena F.Burguera and AlexisM.Fraser, injectable macropore calcium-phosphate cement support, biomaterial, 2006,27:4279-4287; Makoto Watanabe, Miyuki Tanaka, Makoto Sakurai and MiokoMaeda, Development of calcium phosphate cement (progress of calcium-phosphate cement) Journal of the European Ceramic Society (European ceramic journal), 2006,26:549-552).
Kind, performance and the theoretical research of calcium phosphate bone cement have all obtained considerable progress over nearly 20 years, have been widely used in clinical as bone renovating material.But intrinsic some shortcomings of bone cement self are partially long hardening time such as calcium phosphate bone cement, and adhesive property is relatively poor, and mechanical performance is not enough, and it is slower degrade, make its application be subject to a certain degree restriction, at present generally can only be for the reparation of non-heavy burden district bone.Many scholars carry out deep study on the modification to increase its mechanical strength to various calcium phosphate bone cements in recent years, adjust hardening time, improve its rheological characteristic and biological degradability (Lisa E.Carey, Hockin H.K.Xu, Jr., Carl G.Simon, Shozo Takagi, Laurence C.Chow, Premixed rapid-setting calciumphosphate composites for bone repair (premixing expediting setting type calcium phosphate composite bone repairing material), biomaterial (Biomaterials), 2005,26:5002-5014; Hockin H.K.Xu, Carl G.Simon, Fast setting calcium phosphate-chitosan scaffold:mechanical properties and biocompatibility. (solidify soon calcium-phosphate cement-chitosan stent: mechanical performance and Study on biocompatibility), Biomaterials (biomaterial), 2005,26:1337-1348).
Adding fiber molecule in calcium phosphate bone cement is the important method (Dai Honglian that improves anti-of bone cement, erosion-resisting characteristics, Li Shipu, Yan Yuhua, Wang Xinyu, Cao Xianying, Han Yingchao, Chen Xiaoming, Yuan Lin, Li Jianhua, a kind of preparation method of calcium phosphate composite bone cement, publication number CN1657483A, 2005; Li Yubao, Wei Jie, medical compound bio bone cement powder, bone cement liquid and medical compound bio bone cement, publication number CN1403168A, 2003).The fiber molecule complex method can be divided into that cross-linking agent is compound, plasticizer is compound, absorbable fibre is compound, natural fiber is compound etc.Composite principle according to material, in the fibre reinforced composites, fiber bears most of load, between matrix and fiber, play function served as bridge, and when matrix cracking expands to fiber and basal body interface, thereby in conjunction with suitable interface Anticrack or crackle is deflected reach adjustment interfacial stress, the effect that stops crackle to further expand.With regard to the degradation property of material, fiber reinforcement can be divided into non-degradation-type fiber reinforcement and degradable fiber reinforcement.Non-degradation-type fiber is the reinforcing material that early is used to improve the calcium phosphate bone cement mechanical property, its reinforced effects highly significant.Non-degradation-type fiber commonly used mainly comprises carbon fiber, glass fibre and high polymer fibre at present.Research finds that the absorption of polyoxy ion and serum albumin can suppress the growth of HAP crystal, may cause crystallite dimension to diminish, and form the staggered micro structure of crypto set more, polymer between multiple crystal bridge joint and be the reason that causes the calcium phosphate bone cement strength increase by the Plastic Flow absorbed energy.The self-curable calcium phosphate bone cement of compound two kinds of different reinforcing agents uses a kind of calcium phosphate bone cement of reinforcing agent to have higher chemiluminescence than tradition, and the cranium face and the plastic surgery that can be used for load-bearing repair.Studies show that, with polypropylene, nylon and carbon fiber add respectively in the calcium phosphate bone cement, although can cause because of the increase of porosity comprcssive strength to reduce, but toughness and the tensile strength of calcium phosphate bone cement have greatly been increased, this composite bone cement can be used for damaged (the Hockin H.K.Xu that restoration contour has certain flexibility, Janet B.Quinn, Shozo Takagi, Laurence C.Chow, Synergistic reinforcement of in situ hardening calcium phosphatecomposite scaffold for bone tissue engineering (the bone tissue engineer chemiluminescence of in-situ solidifying calcium phosphate bone cement), Biomaterials (biomaterial .), 2004:1029-1037).Sun Kangning etc. are dispersed in long-carbon-fiber the calcium phosphate bone cement Biocomposite material that makes imitative bone structure in the mould in a certain direction.This composite has good mechanical property and biocompatibility, can be applicable to (Sun Kangning, Zhao Ping, a kind of calcium phosphate bone cement Biocomposite materials and preparation method thereof such as the displacement of artificial bone in the medical operating and reparation, publication number CN1559887A, 2005).But above-mentioned weak point of adding fiber molecule in calcium phosphate bone cement is, because Presence of an interface problem between inorganic cement and the macromolecular fibre, therefore generally only in fiber addition situation seldom, just have reinforced effects, and toughening effect is often not obvious; Because these fibers that add are scarcely degraded, therefore can't take into account simultaneously mechanical performance and the degradation property of composite simultaneously.
Utilizing the degradable fibrous material to strengthen calcium phosphate bone cement is a kind of novel bone cement Enhancement Method that development in recent years is got up.The improvements of this method are that used degradable fibrous material can play effect stable and that strengthen at the initial stage of bone cement complex implant into body, and along with the progressively degraded of fiber, the cylindricality duct of its generation helps again growing into fast of the transmission of nutrient substance and blood vessel, cell, and the catabolite safety non-toxic.Therefore can say, biodegradable fiber is in the preparation of bone cement and played the dual function of pore and enhancing in using, avoided the negative effect (Lin that places for a long time human body to produce because of non-degradable material, et al, Process for producing fast-setting, bioresorbable calciumphosphate cements fast setting can absorb the preparation of calcium-phosphate cement. United States Patent (USP) 7,066,999,2006).The employing macromolecules such as Xu can absorb the long fibre element to be strengthened the TTCP/DCPA bone cement, the absorbability fiber of adding 25% in calcium phosphate bone cement, the diameter of fiber is 322 μ m, specimen after compound is placed 37 ℃ of saline, after 1,7,14,28,56 day, find 5 times of strength increases, toughness increases by 100 times, can keep 2-4 week with this intensity of difference and the toughness of fibrolysis speed.Research thinks that the adding of fiber makes calcium phosphate bone cement keep necessary intensity in the tissue regeneration process, and the hole that stays in calcium phosphate bone cement behind the fibrolysis is grown into newborn vascular tissue easily, provides good environment to osteanagenesis; Dissolving along with fiber, formed the cylindrical macropore of mutual perforation in the calcium phosphate bone cement, its bending strength is still high by 39% than the calcium phosphate bone cement of nothing macropore, high by 256% (the Hockin H.K.Xu of toughness, Janet B.Quinn, Calcium phosphate cement containing resorbablefibers for short-term reinforcement and macroporosity. (containing absorbable fibre is used for the short-term enhancing and forms macroporous calcium-phosphate cement), Biomaterials (biomaterial), 2002,23:193-202).In addition, can also strengthen calcium phosphate bone cement with the organic biological active material.Bigi etc. have attempted with gelatin the mechanical property of bone cement being improved, the result shows the comprcssive strength (A.Bigi that improves, B.Bracci, S.Panzavolta, Effect of added gelatin on theproperties of calcium phosphate cement. (gelatin is on the impact of calcium-phosphate cement performance), Biomaterials (biomaterial), 2004,25:2893-2899).Chitosan also is the bioactive substance that important being used for of a class improves the bone cement mechanical property.Xu and Takagi etc. have attempted chitosan is joined the method for preparing enhancement mode calcium phosphate cement/chitosan complex in the liquid phase of bone cement, the result shows the stability of bone cement and intensity (the Hockin H.K.Xu that is greatly improved, Carl G.Simon, Fast setting calcium phosphate-chitosan scaffold:mechanicalproperties and biocompatibility (premixing macropore calcium-phosphate cement support, the material science magazine), Biomaterials (biomaterial), 2007,18:1345-1353).Wang etc. have also investigated the reinforced effects of chitosan derivatives to bone cement, and they find the comprcssive strength after the chitosan of adding phosphorylation in calcium phosphate bone cement can obviously improve its curing, and think because Ca
2+So that newly-generated HA granule links together by polymer, comprcssive strength is improved with bonding action strong between chitosan.DosSantos etc. add Fypro when being in harmonious proportion material, intensity increases, finding simultaneously has radial crack in the fiber material around substrate, shown the break-through point (XiaohongWang that mechanical strength significantly improves once again, Jianbiao Ma, Yinong Wang, Binglin He, (phosphonized chitosan strengthens calcium-phosphate cement and is used for mouse tibia reparation research Bone repair in radii andtibias of rabbits with phosphorylated chitosan reinforced calciumphosphate cements, biomaterial) Biomaterials (biomaterial), 2002,23:4167-4176).
But the bone cement that above-mentioned prior art obtains remains further to be improved in strength character.
The micropore that is caused by water is a very important factor that affects the calcium phosphate bone cement mechanical strength.The researchs such as Barralet find that the adding of the 'alpha '-hydroxy acids sodium salt take sodium citrate as representative can cause Compressive Strength of Calcium Phosphate Cement significantly to improve, and think that the reason that causes comprcssive strength to improve is that the existence of sodium ion can reduce stable contact the between the calcium ion and hydroxy carboxylic acid ionic group in the solidify reaction process, thereby produce more free ion, and these free ions can be adsorbed on the particle surface of reactant and curing product, thereby increased their ζ-potential energy, improved the bone cement rheological property, when keeping the bone cement operability, reduced the content of water, and then cause the bone cement porosity to descend, intensity improves (Jake E.Barralet, Maryjane Tremayne, Kevin J.Lilley, Uwe Gbureck, Modificationof calcium phosphate cement with α-hydroxy acids and their salts (by 'alpha '-hydroxy acids and its esters modified phosphate calcium cement) .Chem Mater, (materials chemistry), 2005,17:1313-1319, Jake E Barralet, Mike Hofmann, Liam M.Grover, Uwe Gbureck, High-strength apatitic cement by modification with α-hydroxy acidsalts ('alpha '-hydroxy acids and its esters modification prepare high strength apatite cement), .Adv.Mater. (advanced material), 2003,15:2091-2094).
The collaborative enhancing of multiple material calcium phosphate bone cement is a completely new concept that proposes in recent years.The core part of this theory is to utilize between the multiple reinforcing material and mutually promotes, interactional relation reaches the purpose that improves the bone cement mechanical property.In the exploration in this field, the research work that Xu etc. carry out has certain representativeness.They are with degradable reticular fiber and chitosan forms the enhancing system and bone cement carries out compound, the result shows, the collaborative effect that strengthens of multicomponent will be apparently higher than one pack system reinforced effects (Hockin H.K.Xu, Elena F.Burguera, Lisa E.Carey, Strong, macroporous, and insitu-setting calcium phosphate cement-layered structures.Biomaterials, In Press, Corrected Proof, Available online 2007).In addition, also has other multiple Enhancement Method, such as Organic substance enhancing, whisker and the enhancing of inorganic ceramic granule etc.Matsuya etc. have reported a kind of copolymer calcium phosphate bone cement, this bone cement is that mixture with tetracalcium phosphate (TTCP) and calcium phosphate dibasic anhydrous (DCPA) is as solid phase, as liquid phase, both mix rear formation calcium phosphate bone cement compound system with the aqueous solution of poly-methyl ethyl ether-maleic acid (PMVE-Ma).PMVE-Ma can be by the hydrolysis of anhydride group by water dissolution, form corresponding maleic acid copolymer (PMVA-Ma), its existence makes calcium phosphate bone cement have higher mechanical strength, and can extend to above (the Yoko Matsuya of 30min hardening time, Shigeki Matsuya, Joseph M.Antonucci, Shozo Takagi, Laurence C.Chow, Akifumi Akamine, Effect of powder grinding onhydroxyapatite formation in a polymeric calcium phosphate cementprepared from tetracalcium phosphate and poly (methyl vinyl ether-maieicacid) (diameter of particle is on the impact by the polymeric calcium phosphate cement cement of tetracalcium phosphate and the preparation of poly-(methyl vinyl ether-maleic acid) system), Biomaterials (biomaterial,), 1999,20:691-697).Be used at present strengthening the whisker of calcium phosphate bone cement and inorganic particulate and mainly comprise silicon nitride crystal whisker, calcium carbonate crystal whisker, silicon carbide whisker and silanization material thereof and aluminium oxide, silicon dioxide inorganic particle etc.Studies show that and to make its original intensity obtain greatly to improve after above-mentioned substance and calcium phosphate bone cement is compound, even near intensity level (100-200MPa) (the Hockin H.K.Xu of cerebral cortex bone, Douglas T.Smith, C.G.Carl G.Simon, (the high strength bioactive composite bone repairing material that contains nanometer fused silica whisker), Biomaterials (biomaterial .), 2004,25:4615-4626; Ana I.Villacampa, Juan Ma.Garc í a-Ruiz, Synthesis of a new hydroxyapatite-silicacomposite material. (synthesizing of novel hydroxyapatite-Silicon stone composite), Journal ofCrystal Growth (crystal growth magazine,), 2000,211:111-115).The weak point of above-mentioned bone cement is: the component degradation that adds slowly or is not degraded, and is used for the generation that the bone reparation can affect new bone.
In sum, desirable bone alternate material should have good biocompatibility, can organize mutually with surrounding bone to merge, mechanical property should approach with natural bone, have suitable intensity and toughness, and can constantly degrade along with the material of growing into of tissue, finally replaced by freshman bone tissue.And calcium phosphate bone cement is a kind of important biological activity bone renovating material, although its Developments is very fast, still has at present some problem demanding prompt solutions, waits partially slowly such as undercapacity, curing rate.Existing Enhancement Method commonly used mostly introduces fiber or other adds composition, and strengthening body is the key of this type of composite research with the interface of the matrix problem of being combined; Most of fibers and some other add composition and can only just have reinforced effects seldom the time in addition, is difficult to make intensity and toughness to improve simultaneously.Simultaneously, because a lot of extraneous component itself is not degraded or degraded slowly, mechanical performance and the degradation property that therefore will take into account calcium phosphate bone cement are then more difficult.
Therefore, this area is demanded a kind of calcium phosphate composite bone cement and preparation method thereof urgently, described bone cement intensity height, good toughness, plasticity are strong, quick solidifying, have good biocompatibility and degradability, thereby overcome not enough, the slowly defective of in human body, degrading of toughness that existing calcium phosphate bone cement material exists, and the defective of several Enhancement Method existence, can satisfy better the requirement that operation is used.
Summary of the invention
The object of the invention is to obtain a kind of calcium phosphate composite bone cement compositions, described compositions can obtain that intensity height, good toughness, plasticity are strong, quick solidifying, have the bone cement of good biocompatibility and degradability.
Another object of the present invention is to obtain that a kind of intensity height, good toughness, plasticity are strong, quick solidifying, have the bone cement material of good biocompatibility and degradability.
A further object of the invention is to obtain a kind of method for preparing the bone cement material.
The present invention has a purpose to be to obtain a kind of tissue engineered implant that contains bone cement material of the present invention again.
A further object of the invention is to obtain a kind of purposes of bone cement material of the present invention, and it is as the support of preparation bone graft.
In a first aspect of the present invention, a kind of poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions is provided, its component and weight percent content comprise: 10%～95 % by weight self-curable calcium phosphate bone cement; And the poly-polysaccharide of 5%～90 % by weight.
In a specific embodiment of the present invention, described poly-polysaccharide is selected from natural polysaccharide, natural polysaccharide derivant or its combination,
Particularly, described natural polysaccharide is selected from glucosan, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannoglycan or its combination.
In a specific embodiment of the present invention, described natural polysaccharide derivant is selected from take natural polysaccharide as main chain, with the derivant of reactive functional groups modification;
Particularly, described reactive functional groups is selected from acrylate residue, acrylamide residue, acrylic acid residue, vinylpyrrolidone residue, crown ether residue, other reactive unsaturated group or its combination.
More specifically, described acrylate residue be with unsaturated double-bond the glycidyl methacrylate residue, with the methacrylic acid hydroxyl ethyl ester residue of unsaturated double-bond, with acrylic acid sulphonic acid ester residue or its combination of unsaturated double-bond; Described acrylamide residue is acrylamide residue, N-isopropyl acrylamide residue or its combination;
More specifically, the degree of modification of reactive functional groups described in the natural polysaccharide derivant on natural polysaccharide is to replace 1～30 reactive functional groups in per 100 natural polysaccharide unit.
In a specific embodiment of the present invention, the component of said self-curable calcium phosphate bone cement is selected from a kind of or its mixture in tricalcium phosphate, tetracalcium phosphate, OCP, calcium hydrogen phosphate, hydroxyapatite, fluor-apatite, the calcium pyrophosphate.
The bone cement material that further aspect of the present invention provides a kind of compositions of the present invention to make obtains described bone cement material by described self-curable calcium phosphate bone cement and described poly-polysaccharide composite.
In a specific embodiment of the present invention, described bone cement material is obtained by the complex method that may further comprise the steps:
The mixture of the poly-polysaccharide of 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight is provided;
In the presence of water soluble initiator, gather simultaneously the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture, form the bone cement material.
A further aspect of the invention provides a kind of method for preparing the bone cement material, and described method comprises the steps:
The mixture of the poly-polysaccharide of 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight is provided;
The poly-polysaccharide of described 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight carries out the compound described bone cement material that obtains;
Particularly, described complex method may further comprise the steps: in the presence of water soluble initiator, gather simultaneously the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture, form described bone cement material.
In a specific embodiment of the present invention,
Described mixture is obtained by following blend step: the poly-polysaccharide-disodium phosphate soln of preparation is as the consolidation liquid of self-curable calcium phosphate bone cement; Then the self-curable calcium phosphate bone cement is mixed with consolidation liquid, obtain described mixture;
Particularly, the solid-to-liquid ratio of described self-curable calcium phosphate bone cement and consolidation liquid is 0.1～5g/ml, and preferable range is 1～3g/ml.
In a specific embodiment of the present invention, the step of described cross-linking reaction and curing reaction comprises: add water soluble initiator in the described mixture, stir, be mixed into slurry; Described slurry is placed in 100 ± 10% humidity environments at 37 ± 5 ℃, until form the intercrossed network type bioactive composite structure, thus obtain described bone cement material;
Particularly, described water soluble initiator is persulfate-N, N, N ', N '-tetramethylethylenediamine, ammonium ceric nitrate or hydrogen peroxide-ferrous sulfate system;
Particularly, the concentration of described water soluble initiator is 1 * 10
-4～1 * 10
-1Mol/L, wherein preferred concentration is 1 * 10
-3～5 * 10
-1Mol/L;
Particularly, described slurry setting time is 0.5-60min, and be 4-48 hour hardening time, and wherein the preferred consolidation time is 15-30 hour.
The present invention provides a kind of tissue engineered implant on the other hand, and described graft contains bone cement material of the present invention and is inoculated in the stem cell of described bone cement material, and the inoculum concentration of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3The bone cement material.
A further aspect of the invention provides a kind of purposes of bone cement material, and it is as the support of preparation bone graft.
The specific embodiment
The invention discloses a kind of self-curable calcium phosphate bone-cement composite material that contains the natural polysaccharide derivant and preparation method thereof.The present invention adopts good biocompatibility, degradable natural polysaccharide derivant mutually compound with calcium phoshate bone cement powder, can realize simultaneously the crosslinking curing of calcium phosphate bone cement hydration and sclerosis and poly-polysaccharide derivates, shorten the presetting period of bone cement, the mechanical properties such as the comprcssive strength of raising calcium phosphate bone cement and toughness; The mastic that forms can random-shaping, hardens voluntarily under human body environment and humidity.The present invention can be used for that the bone that a variety of causes causes is damaged, filling reparation and the injectable repair materials of bone does not connect, bone delay in healing, is a kind of compound human body hard tissue repair materials of novel organic-inorganic with broad prospect of application.
In the term of the present invention " poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions ", the implication of described "/" refer to the two be " with " relation.
The implication of " natural polysaccharide derivant " of the present invention refers to, carries out the modification of reactive functionality and the chemical compound that obtains in the molecule of natural polysaccharide.Described method of modifying for example is graft reaction particularly.For instance, glucosan-acrylate derivative refers to be grafted with acrylate-functional groups in dextran molecule, thereby consists of a kind of poly-polysaccharide derivates.
Below dosage form describes in detail to various aspects of the present invention:
The self-curable calcium phosphate bone cement
Self-curable calcium phosphate bone cement of the present invention can adopt the traditional self-curable calcium phosphate bone cement in this area, for example is selected from particularly a kind of or its mixture in tricalcium phosphate, tetracalcium phosphate, OCP, calcium hydrogen phosphate, hydroxyapatite, fluor-apatite, the calcium pyrophosphate.Described self-curable calcium phosphate bone cement is not specifically limited, as long as can be in the presence of consolidation liquid bone cement compositions be solidified into the bone cement material.
The content of described self-curable calcium phosphate bone cement is generally 10%～95 % by weight, and preferred 25%～91 % by weight is by the total weight of described poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions.
The particle diameter of described bone cement is not specifically limited, only otherwise goal of the invention of the present invention is produced restriction to be got final product.Its particle diameter includes but not limited to 10-20 μ m.
Poly-polysaccharide
Poly-polysaccharide of the present invention is selected from natural polysaccharide, natural polysaccharide derivant or its combination usually.
Particularly, described natural polysaccharide is selected from glucosan, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannoglycan or its combination.
Particularly, described natural polysaccharide derivant is selected from take natural polysaccharide as main chain, with the derivant of reactive functional groups modification.
More specifically, described reactive functional groups is selected from acrylate residue, acrylamide residue, acrylic acid residue, vinylpyrrolidone residue, crown ether residue, other reactive unsaturated group or its combination.
More specifically, described acrylate residue be with unsaturated double-bond the glycidyl methacrylate residue, with the methacrylic acid hydroxyl ethyl ester residue of unsaturated double-bond, with acrylic acid sulphonic acid ester residue or its combination of unsaturated double-bond; Described acrylamide residue is acrylamide residue, N-isopropyl acrylamide residue or its combination.All with unsaturated double-bond, and higher reactivity is arranged on the poly-polysaccharide main chain of these several polymer, can form at short notice cross-linked structure by radical reaction, and can regulate the cross-linking reaction degree by the amount that accesses two keys.
More specifically, the degree of modification of reactive functional groups described in the natural polysaccharide derivant on natural polysaccharide is to replace 1～30 reactive functional groups in per 100 natural polysaccharide unit.
Natural polysaccharide is carried out modified with functional group and obtains the natural polysaccharide derivant belonging to the traditional reaction in this area.Described natural polysaccharide derivant also can be by commercially available acquisition.
The object lesson of producing the natural polysaccharide derivant is exemplified below (including but not limited to following example): take by weighing natural polysaccharide and be dissolved in the dimethyl sulfoxide (DMSO), being made into concentration is 1%-20% (g/ml) solution.Add an amount of acrylate or acrylamide, the mol ratio that makes Glucopyranose. ring in acrylate or acrylamide group and the poly-polysaccharide is 0.05～1.The control reaction temperature is 20-100 ℃, and reaction is 24-72 hour under argon shield.After reaction finishes, add isopropyl alcohol, sucking filtration also places deionization dialysis 12-72 hour, after lyophilization, obtains white fine hair shape product.The replacement degree that calculates acrylate-based in the product or acrylamide by proton nmr spectra is 5-60%.
The weight content of described poly-polysaccharide is 5%～90 % by weight, and preferred 9%～75 % by weight is by the total weight of described poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions.
The bone cement material
Bone cement material of the present invention obtains described bone cement material by described self-curable calcium phosphate bone cement and described poly-polysaccharide composite.
Usually, described bone cement is obtained by the complex method that may further comprise the steps:
The mixture of the poly-polysaccharide of 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight is provided;
In the presence of water soluble initiator, gather simultaneously the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture, form the bone cement material.
Particularly, the step of described cross-linking reaction and curing reaction comprises: add water soluble initiator in the described mixture, stir, be mixed into slurry; Described slurry is placed in 100 ± 10% humidity environments at 37 ± 5 ℃, until form the intercrossed network type bioactive composite structure, thus obtain described bone cement material;
Particularly, described water soluble initiator is persulfate-N, N, N ', N '-tetramethylethylenediamine, ammonium ceric nitrate or hydrogen peroxide-ferrous sulfate system;
Particularly, the concentration of described water soluble initiator is 1 * 10
-4～1 * 10
-1Mol/L, wherein preferred concentration is 1 * 10
-3～5 * 10
-1Mol/L;
Particularly, described slurry setting time is 0.5-60min, and be 4-48 hour hardening time, and wherein the preferred consolidation time is 15-30 hour.
Described slurry forms the intercrossed network type bioactive composite structure when solidifying.
The mixed method of described poly-polysaccharide and self-curable calcium phosphate bone cement can adopt the traditional mixed method in this area.Particularly for example, described mixture is obtained by following blend step: the poly-polysaccharide derivates-disodium phosphate soln (also can adopt other suitable solution) of preparation is as the consolidation liquid of self-curable calcium phosphate bone cement; Then the self-curable calcium phosphate bone cement is mixed with consolidation liquid, obtain described mixture.
Described consolidation liquid can adopt the traditional consolidation liquid in this area.And the concentration of consolidation liquid also is not specifically limited, only otherwise goal of the invention of the present invention is produced restriction to be got final product.
Usually, the 4%wt disodium phosphate soln is preparation calcium-phosphate cement consolidation liquid commonly used, and in the curing reaction of simple calcium phosphate bone cement, the consolidation liquid of this concentration is generally considered comparatively ideal curing system.In fact, and nonessential be sodium hydrogen phosphate and can not be other material, its concentration also can change.Therefore, the concentration of described disodium phosphate soln is not specifically limited, only otherwise goal of the invention of the present invention is produced restriction to be got final product.
The compound method of described consolidation liquid can adopt the traditional method in this area, for example will gather particularly polysaccharide derivates and be dissolved in the disodium phosphate soln, and being mixed with concentration is poly-polysaccharide/disodium phosphate soln of 0.1～3g/ml.
Can also add other biologically acceptable material in the consolidation liquid of the present invention.Particularly for example, the biotic factor that promotes osteogenic growth be can suitably add, the factor (SCF) or its combination connected such as bone morphogenetic protein (BMP), OP-1, bonegel, osteonectin (osteoconectin), bone calcification element (osteocalcin), skeleton.The biotic factor of described promotion osteogenic growth is different with affiliated animal classification, and the amount of interpolation is also different, and preferably the mass ratio with polymer 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 cell and Material cladding co-cultivation then can also add in culture fluid.
Particularly, the solid-to-liquid ratio of described self-curable calcium phosphate bone cement and consolidation liquid is 0.1～5g/ml, and preferable range is 1～3g/ml.
Design of the present invention is such:
The present invention selects suitable macromolecular material and calcium phosphate bone cement compound, form hybridization network, the HAP that aquation is generated is namely seeking connections with at the beginning of the nucleation on the high polymer network, carrying out along with hydration reaction, HAP crystal grain is grown at organic network, has finally formed take high polymer as " skeleton ", hydroxylapatite crystal is the composite construction of " muscle ".This structure has the presetting period of reduction, improves comprcssive strength, improves the advantages such as toughness and classification degraded with respect to the bone cement that is comprised of the hydrated product hydroxyapatite merely in the past, can satisfy human body heavy burden position to the requirement of intensity.With respect to present common Enhancement Method, the present invention seeks a kind of new compound thinking, namely adopt the method for interface IPN to make between the larger component of performance difference and produce firmly interface combination, thereby realize the complementation of different component performance or function, make it produce special synergism at macro property.And in the selection of enhancement component, considering that organism itself is constructed by biomacromolecules such as protein, saccharides forms, important component as Living organism, polysaccharide extensively exists in organism, playing an important role at control cell division and differentiation, adjusting Growth of Cells and the aspects such as old and feeble and the organic homergy that sustains life, is the important component that forms extracellular matrix.Simultaneously, polysaccharide also possesses the basic demand as bio-medical material, namely good biocompatibility and in vivo enzymolysis Cheng Yi be a class biodegradation absorption-type macromolecular material by the small-molecule substance that live body absorbs, has no side effect.Thereby select natural polysaccharide as wild phase, both overcome the undesirable shortcoming of the ubiquitous cellular affinity of synthesized polymer material, the problems such as immunoreation that the existing physiological characteristics of collagen stroma is unstable and may cause have also been avoided selecting, thereby so that have " hard and tough " between composite concurrently, can improve the degradability of calcium phosphate material simultaneously.
In a specific embodiment, the present invention forms inierpeneirating network structures by two kinds of cross-linked networks on the basis that the hydration reaction of the Raolical polymerizable of the two keys of poly-polysaccharide derivates and synthos carries out simultaneously.Characterize from infrared spectrum, X-ray diffraction spectrum etc., two reactions all occur, and the two keys in the poly-polysaccharide derivates disappear, and have simultaneously hydroxyapatite to generate, and these all illustrate the carrying out of two kinds of reactions.This two kinds of reactions are carried out simultaneously and finally to form the method for complex similar to the macromolecule interpenetrating networks, therefore are referred to as the intercrossed network type composite construction.
Owing to inorganic calcium microcosmic salt existence in the mixture and at 37 ℃, hydration reaction occurs in the environment of 100% humidity, carry out simultaneously cross-linking reaction separately in the system, therefore can form the network structure of inorganic/organic interface IPN, good two-phase interface is combined with to be beneficial to and transmits and dispersive stress, reduce the probability of two-phase interface defective and destruction, the IPN of network tangles and causes the increase of the system degree of cross linking simultaneously, can show significant synergism.
In a specific embodiment of the present invention, the present invention prepares the method for poly-polysaccharide/calcium phosphate bone cement hydridization interpenetrating networks, comprises following step:
(1) take by weighing natural polysaccharide and be dissolved in the dimethyl sulfoxide (DMSO), being made into concentration is 1%-20% (g/ml) solution.Add an amount of acrylate or acrylamide, the mol ratio that makes Glucopyranose. ring in acrylate or acrylamide group and the poly-polysaccharide is 0.05～1.The control reaction temperature is 20-100 ℃, and reaction is 24-72 hour under argon shield.After reaction finishes, add isopropyl alcohol, sucking filtration also places deionization dialysis 12-72 hour, after lyophilization, obtains white fine hair shape product.The replacement degree that calculates acrylate-based in the product or acrylamide by proton nmr spectra is 5-60%.
(2) preparation 1-10%wt disodium phosphate soln, 1-100mg/ml initiator solution.To gather polysaccharide derivates and be dissolved in the disodium phosphate soln, being mixed with concentration is poly-polysaccharide/disodium phosphate soln of 0.1～3g/ml, is added in the calcium phoshate bone cement powder by the certain mass ratio.Mix homogeneously, reconciling into pastel can implant, and also can place 37 ℃, 100% relative humidity environment to solidify 6-48 hour external, forms firming body, and then 50 ℃ of dryings implant.
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 cell.In addition, osteoblast also alternative stem cell as the seed cell of bone tissue engineer structure.
Can be used for stem cell of the present invention can be preferably mammal 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 stage of development can and have in the allogeneic animal body of complete immunologic function in a organized way compatibility difference and form the stem cell tissue.
The method of separation and acquisition stem cell is as known in the art.A kind of preferred method is density－gradient centrifuga－tion method and enzyme digestion.
The cultural method of stem cell and culture fluid also are well known in the art.A kind of preferred method is at 37 ℃, saturated humidity, 5%CO with stem cell
2Cultivate in the incubator.Suitable culture fluid comprises (but being not limited to): 1) DMEM culture medium ((Gibco company)+5～20% hyclones; 2) DMEM culture medium+5～20% calf serums; 3) DMEM culture medium+5～20% are from body (allosome) human serum.In addition, add various somatomedin (such as the cytokine that promotes stem cell growth etc.), various antibiotic, various inducible factor in the above-mentioned culture fluid.
Being applicable to 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 In vitro culture.
Bone graft
Because the compatibility of bone cement of the present invention 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-bone cement complex on the bone cement that bone marrow stroma stem cell and/or the fat stem cell of cultured and amplified in vitro is inoculated into the biocompatibility excellence, this " stem cell-bone cement " complex is implanted to defect, gradually degraded and absorbed along with the bone cement material, new bone formation reaches the purpose of repairing bone defect.
The preparation method of tissue engineering bone graft of the present invention is easy, the bone marrow stroma stem cell of some and/or fat stem cell is inoculated in the bone cement material gets final product.
The shape of engineered bone graft of the present invention is not particularly limited, can be according to the shape random-shaping of tissue defect.Usually, graft is strip.
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 with culture fluid, then mix with degradation material.During mixing, the ratio of culture fluid and degradation material is not particularly limited, but is advisable with the culture fluid 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 antibiotic, 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.
Beneficial effect of the present invention is:
(1) the invention provides the preparation method of a kind of novel natural polysaccharide/calcium phosphate bone cement hybrid material.The composite of selecting natural polysaccharide derivant with good biocompatibility and calcium microcosmic salt bone cement hydridization to form has good histocompatibility and degradability, can not cause in vivo the untoward reaction of tissue.
(2) among the present invention, macromolecule polysaccharide and inorganic calcium microcosmic salt are compared blending method commonly used by the hybrid systems of interpenetrating networks interface IPN, and composite material interface adhesive effect provided by the invention is stronger, the compatibility is better, is conducive to reach reinforced effects by the synergism between biphase.
(3) the inventive method preparation technology is simple, rate of polymerization is fast, the process of composite hybridization a step is finished when crosslinked and calcium microcosmic salt aquation is carried out simultaneously at macromolecule, need not to carry out again next step combination process, reaction condition is gentle, and whole reaction is all carried out at aqueous phase, has avoided with an organic solvent.
(4) simple firming body take inorganic matter as composition, whole mechanical performance is more crisp, lacking toughness.When bearing load, rigid collision can occur in the contact surface of material and bone, and the energy that the external force transmission comes can't absorb, and firming body collapses loose easily.The network that high polymer forms among the present invention can be brought into play the effect of elastic buffer, utilizes the deformation of hydrogel network to absorb external energy, keep the whole complete of firming body, thereby the resistance to compression of whole firming body, shock resistance are effectively promoted.
(5) the firming body degradation speed that is comprised of calcium phosphate bone cement merely is usually slow.The present invention selects the natural polysaccharide thing of biodegradable as the polymer moiety of composite, the degraded velocity ratio is very fast under the effect of enzyme in vivo, and because it is the skeleton of whole firming body, after high polymer is degraded, it is loose that firming body will become, more body fluid can be penetrated in the middle of the firming body, and complete firming body is divided into fritter so originally, thereby accelerates degradation speed.
(6) controllability.Biphase ratio in ratio of component by simple control reaction and the hybrid material can obtain the macromolecule polysaccharide of different two key substituent group numbers, thus the crosslink density of control macromolecule network, and intensity, degradability and the swellability of regulation and control hybrid material.
(7) applied widely.The hybrid material of the present invention's preparation can be used as pre-moulding bone renovating material, also can be used as curing materials in the art, and as the injectable bone renovating material, is used for the Minimally Invasive Surgerys such as vertebral body molding.
(8) the present invention selects to have good biocompatibility and degradability, has the natural polysaccharide derivant material of certain biomechanical property, react original position formation intercrossed network type Biocomposite material with calcium phosphate bone cement (CPC) under the water soluble initiator effect.The characteristic that this method utilizes interpenetrating networks to have is improved the comprcssive strength of calcium phosphate bone cement, and utilize substitution value variation (Degree of Substitution, DS) on the different and poly-polysaccharide derivates of initiator system ratio to realize the controlled of calcium phosphate bone cement setting time.
Chemical compound provided by the present invention can be synthetic by marketable material and traditional chemical transform mode.Can be by commercially available acquisition such as natural polysaccharide glucosan, chitosan, starch, cellulose, cyclodextrin etc., glucosan-acrylate, glucosan-N-isopropylacrylamide, chitosan-acrylate, Starch-acrylamide, cellulose-N-isopropylacrylamide, cyclodextrin-N-isopropylacrylamide, sodium alginate-N-isopropylacrylamide derivant all can obtain by the chemical conversion mode is synthetic.
Other aspects of the present invention are because the disclosure of this paper is apparent to those skilled in the art.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention 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, such as people such as Sambrook, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Unless otherwise defined or explanation, same meanings of being familiar with of all specialties used herein and scientific words and those skilled in the art.Any method similar or impartial to described content and material all can be applicable in the inventive method in addition.
Detailed description below by the specific embodiment of the inventive method is further set forth the present invention, but these examples are not used for limiting the present invention.
Embodiment 1
Take by weighing 0.3 gram glucosan-acrylate derivative (dextran molecule amount 40,000, DS=12, be to replace 12 acrylate in per 100 glucosan monomers, DS obtains by nuclear magnetic resonance hydrogen spectruming determining) be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid that 0.5g/ml contains glucosan; Taking by weighing the particle diameter that is comprised of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite is self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 1.5 grams of 10-20 μ m, adds above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state.
Then add persulfate (50mg/ml) 120 μ l, N, N, N ', N '-tetramethylethylenediamine (23mg/ml) 300 μ l insert behind the mix homogeneously in the tetrafluoroethene mould and make batten, place 37 ℃, solidify in 100% humidity environment, the batten presetting period is 2.48min.Comprcssive strength reaches 82.82MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.24 hours hardening times.
Embodiment 2
Take by weighing 0.3 gram glucosan-acrylate derivative (dextran molecule amount 40,000, DS=8.8 namely replace 8.8 acrylate in per 100 glucosan monomers) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid that 0.5g/ml contains glucosan; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 1.5 grams of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state.
Then add persulfate (50mg/ml) 120 μ l, N, N, N ', N '-tetramethylethylenediamine (23mg/ml) 300 μ l, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidify in 100% humidity environment, the batten presetting period is 2.48min.Comprcssive strength reaches 94.33MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 3
Take by weighing 0.3 gram glucosan-N-isopropylacrylamide derivant (DS=12 namely replaces 12 N-isopropylacrylamides in per 100 glucosan monomers) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid that 0.5g/ml contains glucosan; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 3 grams of the 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state.
Then add cerous nitrate ammonia salt initiators (2mg/ml) 200 μ l, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidifies in 100% humidity environment, and the batten presetting period is 2.48min.Comprcssive strength reaches 43.95MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 4
Take by weighing 1.5 gram chitosan-acrylate derivatives (DS=12 namely replaces 12 acrylate in per 100 chitosan monomers) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid of 0.5g/ml chitosan-containing; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 0.5 gram of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid its furnishing pasty state is obtained mixture.
Then add cerous nitrate ammonia salt initiators (2mg/ml) 200 μ l in described mixture, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidifies in 100% humidity environment, and the batten presetting period is 2.48min.Comprcssive strength reaches 80.24MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 5
Take by weighing 0.3 gram Starch-acrylamide derivant (DS=12 namely replaces 12 acrylamides in per 100 starch monomer molecules) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the amyloid consolidation liquid of 0.5g/ml; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 1.5 grams of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
Then add cerous nitrate ammonia salt initiators (2mg/ml) 120 μ l to mixture, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidifies in 100% humidity environment, and the batten presetting period is 1.33min.Comprcssive strength reaches 76.14MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 6
Take by weighing 0.3 gram cellulose-N-isopropylacrylamide derivant (DS=12 namely replaces 12 N-isopropylacrylamides in per 100 cellulose monomer molecules) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid of 0.5g/ml cellulose; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 3 grams of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
Then in described mixture, add persulfate (50mg/ml) 120 μ l, N, N, N ', N '-tetramethylethylenediamine (23mg/ml) 300 μ l, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidify in 100% humidity environment, the batten presetting period is 3.48min.Comprcssive strength reaches 50.68MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 7
Take by weighing 0.3 gram cyclodextrin-N-isopropylacrylamide derivant (DS=6 namely replaces 6 N-isopropylacrylamides in per 100 cyclodextrin monomer molecules) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid that 0.5g/ml contains cyclodextrin; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 3 grams of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
Then in described mixture, add persulfate (50mg/ml) 120 μ l, the slurry that obtains behind the sodium sulfite 50 μ l mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, place 37 ℃, solidify in 100% humidity environment, the batten presetting period is 5.68min.Comprcssive strength reaches 45.8MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
Embodiment 8
Take by weighing 0.3 gram sodium alginate-N-isopropylacrylamide derivant (DS=12 namely replaces 12 N-isopropylacrylamides in per 100 natural polysaccharide unit) and be dissolved in the 4%wt disodium phosphate soln, be mixed with the consolidation liquid that 0.5g/ml contains sodium alginate; Take by weighing self-curable calcium phosphate salt powder (Shanghai Ruibang Biological Material Co., Ltd.) 3 grams of the particle diameter 10-20 μ m that is formed by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
Then in described mixture, add persulfate (50mg/ml) 120 μ l, N, N, N ', N '-tetramethylethylenediamine (23mg/ml) 300 μ l, the slurry that obtains behind the mix homogeneously is inserted in the tetrafluoroethene mould and is made batten, places 37 ℃, solidify in 100% humidity environment, the batten presetting period is 3.25min.Comprcssive strength reaches 48.68MPa behind the bone dry, satisfies the part requirement of strength of human body heavy burden part.
All quote in this application as a reference at all documents that the present invention mentions, just as each piece document is quoted separately as a reference.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 limited range equally.
Claims (9)
1. poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions, its component and weight percent content comprise:
10%～95 % by weight self-curable calcium phosphate bone cement; And the poly-polysaccharide of 5%～90 % by weight, described poly-polysaccharide is selected from take natural polysaccharide as main chain, with the derivant of reactive functional groups modification;
Described reactive functional groups is selected from acrylate residue, acrylamide residue, acrylic acid residue;
Described acrylate residue be with unsaturated double-bond the glycidyl methacrylate residue, with the methacrylic acid hydroxyl ethyl ester residue of unsaturated double-bond, with acrylic acid sulphonic acid ester residue or its combination of unsaturated double-bond;
Described acrylamide residue is acrylamide residue, N-isopropyl acrylamide residue or its combination;
The degree of modification of described reactive functional groups on natural polysaccharide is to replace 1～30 reactive functional groups in per 100 natural polysaccharide unit.
2. compositions according to claim 1 is characterized in that, described natural polysaccharide is selected from glucosan, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannoglycan or its combination.
3. compositions according to claim 1, it is characterized in that the component of said self-curable calcium phosphate bone cement is selected from a kind of or its mixture in tricalcium phosphate, tetracalcium phosphate, OCP, calcium hydrogen phosphate, hydroxyapatite, fluor-apatite, the calcium pyrophosphate.
4. the bone cement material that makes of each described compositions according to claim 1～3 is characterized in that, obtains described bone cement material by described self-curable calcium phosphate bone cement and described poly-polysaccharide composite.
5. bone cement material as claimed in claim 4 is characterized in that, described bone cement material is obtained by the complex method that may further comprise the steps:
The mixture of the poly-polysaccharide of 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight is provided;
In the presence of water soluble initiator, gather simultaneously the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture, form the bone cement material.
6. method for preparing bone cement material as claimed in claim 4, described method comprises the steps:
The mixture of the poly-polysaccharide of 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight is provided;
The poly-polysaccharide of described 10%～95 % by weight self-curable calcium phosphate bone cement and 5%～90 % by weight carries out the compound described bone cement material that obtains;
Described compound may further comprise the steps: in the presence of water soluble initiator, gather simultaneously the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture, form described bone cement material.
7. method as claimed in claim 6 is characterized in that,
Described mixture is obtained by following blend step: the poly-polysaccharide-disodium phosphate soln of preparation is as the consolidation liquid of self-curable calcium phosphate bone cement; Then the self-curable calcium phosphate bone cement is mixed with consolidation liquid, obtain described mixture;
The solid-to-liquid ratio of described self-curable calcium phosphate bone cement and consolidation liquid is 0.1～5g/ml,
The step of described cross-linking reaction and curing reaction comprises: add water soluble initiator in the described mixture, stir, be mixed into slurry; Described slurry is placed in 100 ± 10% humidity environments at 37 ± 5 ℃, until form the intercrossed network type bioactive composite structure, thus obtain described bone cement material;
Described water soluble initiator is persulfate-N, N, N ', N '-tetramethylethylenediamine, ammonium ceric nitrate or hydrogen peroxide-ferrous sulfate system;
The concentration of described water soluble initiator is 1 * 10
-4～1 * 10
-1Mol/L;
Described slurry setting time is 0.5-60min, and be 4-48 hour hardening time.
8. a tissue engineered implant is characterized in that, described graft contains bone cement material claimed in claim 4 and is inoculated in the stem cell of described bone cement material, and the inoculum concentration of described stem cell is 2 * 10
6-5 * 10
7Individual cell/cm
3The bone cement material.
9. the purposes of a bone cement material as claimed in claim 4 is characterized in that, as the support of preparation bone graft.
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| CN115737911B (en) * | 2022-12-16 | 2024-06-21 | 华南理工大学 | High-strength and high-toughness bone repair composite material and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5180426A (en) * | 1987-12-28 | 1993-01-19 | Asahi Kogaku Kogyo K.K. | Composition for forming calcium phosphate type setting material and process for producing setting material |
| CN1785443A (en) * | 2005-10-20 | 2006-06-14 | 华南理工大学 | Method of improving anti collapsibility of calcium phosphate skeletal cement using denaturated starch |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5180426A (en) * | 1987-12-28 | 1993-01-19 | Asahi Kogaku Kogyo K.K. | Composition for forming calcium phosphate type setting material and process for producing setting material |
| CN1785443A (en) * | 2005-10-20 | 2006-06-14 | 华南理工大学 | Method of improving anti collapsibility of calcium phosphate skeletal cement using denaturated starch |
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