CN101496909A - Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof - Google Patents
Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof Download PDFInfo
- Publication number
- CN101496909A CN101496909A CNA2008100334204A CN200810033420A CN101496909A CN 101496909 A CN101496909 A CN 101496909A CN A2008100334204 A CNA2008100334204 A CN A2008100334204A CN 200810033420 A CN200810033420 A CN 200810033420A CN 101496909 A CN101496909 A CN 101496909A
- Authority
- CN
- China
- Prior art keywords
- bone cement
- polysaccharide
- calcium phosphate
- self
- residue
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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 damaged novel organic/inorganic composite material of reparation human body hard tissue that is used to fill or inject.
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 ideal bone renovating material always.Last century, the 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 voluntarily aquation solidify, finally be converted into to the human body 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, mainly show as: excellent biological compatibility and biological safety, favorable biological degradability, guiding osteogenic activity.Calcium phosphate bone cement can change into nature bone similarly to be formed, 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 the more inherent shortcomings of bone cement self are long partially hardening time such as calcium phosphate bone cement, and adhesive property is relatively poor, the mechanical performance deficiency, and it is slower to degrade, and makes its application be subjected to a certain degree restriction, generally can only be used for the reparation of non-heavy burden district bone at present.Many in recent years scholars carry out deep study on the modification to increase its mechanical strength to various calcium phosphate bone cements, 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 stop cracks can spread or crackle is deflected in conjunction with suitable interface and 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 the degradable fiber type strengthens.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 at present mainly comprises carbon fiber, glass fibre and high polymer fibre.The absorption of discovering polyoxy ion and serum albumin can suppress the crystalline growth of HAP, may cause crystallite dimension to diminish, and form the staggered micro structure of crypto set more, polymer between multiple crystal bridge joint and flow by plasticity that to absorb energy be the reason that causes calcium phosphate bone cement intensity to increase.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 collaborative potentiation 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 in the calcium phosphate bone cement respectively, though can cause comprcssive strength to reduce because of the increase of porosity, but the 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 collaborative potentiation 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 performance and biocompatibility, can be applicable to (Sun Kangning, Zhao Ping, a kind of calcium phosphate bone cement Biocomposite material and preparation method thereof, publication number CN1559887A, 2005) such as the displacement of artificial bone in the medical operating and reparations.But above-mentioned weak point of adding fiber molecule in calcium phosphate bone cement is, owing to have interface problem between inorganic cement and the macromolecular fibre, therefore generally only under 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 the mechanical performance and the degradation property of composite simultaneously simultaneously.
Utilizing degradable fiber type 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 fiber type material can play stable and enhanced effect 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 growing into fast of the transmission of nutrient substance and blood vessel, cell again, and the catabolite safety non-toxic.Therefore we can say, biodegradable fiber has played pore and enhanced dual function in the preparation of bone cement with in using, avoided the negative effect (Lin that places human body to produce for a long time 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).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, find that after 1,7,14,28,56 day intensity increases by 500, 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 organized easily neonatal blood vessels and grown into, and 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 no 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 to the calcium-phosphate cement Effect on Performance), Biomaterials (biomaterial), 2004,25:2893-2899).Chitosan also is the bioactive substance that more 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+Make newly-generated HA granule link together with intensive bonding action between chitosan, comprcssive strength is improved by polymer.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 on strength character.
The micropore that is caused by water is an important factors that influences the calcium phosphate bone cement mechanical strength.Barralet etc. discover with the sodium citrate to be that the adding of 'alpha '-hydroxy acids sodium salt of representative can cause calcium phosphate bone cement comprcssive strength 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 the more freedom 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 is formed the enhancing system and bone cement carries out compound, the result shows, the collaborative enhanced effect 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, 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 the back and form the 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-maleicacid) (diameter of particle is to the influence by the polymer phosphate calcium cement of tetracalcium phosphate and the preparation of poly-(methyl vinyl ether-maleic acid) system), Biomaterials (biomaterial,), 1999,20:691-697).Be used to strengthen the whisker of calcium phosphate bone cement and inorganic particulate at present 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 above-mentioned substance and calcium phosphate bone cement and can make its original intensity obtain greatly to improve after 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 influence new bone.
In sum, ideal bone alternate material should have good biocompatibility, can organize mutually with surrounding bone to merge, mechanical property should be approaching with nature bone, have suitable intensity and toughness, and can constantly degrade, finally replaced by freshman bone tissue along with the material of growing into of tissue.And calcium phosphate bone cement is a kind of important biological bone renovating material, though its correlational study makes much progress, still has some problem demanding prompt solutions at present, waits partially slowly as undercapacity, curing rate.Existing Enhancement Method commonly used introduces fiber mostly or other adds composition, strengthens body and combines the key that problem is this type of composite research with the interface of matrix; 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 toughness deficiency that existing calcium phosphate bone cement material exists, slow degradation defective in human body, and the defective of several Enhancement Method existence, can satisfy the requirement that operation is used better.
Summary of the invention
The objective 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 weight % self-curable calcium phosphate bone cement; And the poly-polysaccharide of 5%～90 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, it is main chain that described natural polysaccharide derivant is selected from the natural polysaccharide, has the derivant that reactive functional groups is modified;
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 is the glycidyl methacrylate residue that has unsaturated double-bond, the methacrylic acid hydroxyl ethyl ester residue that has unsaturated double-bond, the acrylic acid sulphonic acid ester residue that has unsaturated double-bond or its combination; Described acrylamide residue is acrylamide residue, N-isopropyl third rare amide residues 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 are compound.
In a specific embodiment of the present invention, described bone cement material is obtained by the complex method that may further comprise the steps:
10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 weight % are provided the mixture of poly-polysaccharide;
In the presence of water soluble initiator, gather the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture simultaneously, 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:
10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 weight % are provided the mixture of poly-polysaccharide;
The poly-polysaccharide of described 10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 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 the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture simultaneously, 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 formation intercrossed network type bioactive composite structure, thereby obtains 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 the calcium phosphate bone cement powder, can realize curing of calcium phosphate bone cement aquation and the crosslinking curing that gathers polysaccharide derivates simultaneously, shorten the presetting period of bone cement, mechanical properties such as the comprcssive strength of raising calcium phosphate bone cement and toughness; The mastic that forms can be moulding arbitrarily, sclerosis 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, the 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 "/" be meant the two be " with " relation.
The implication of " natural polysaccharide derivant " of the present invention is meant, 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 is meant and is grafted with acrylate-functional groups in dextran molecule, thereby constitutes 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 a kind of or its mixture in tricalcium phosphate, tetracalcium phosphate, OCP, calcium hydrogen phosphate, hydroxyapatite, fluor-apatite, the calcium pyrophosphate particularly.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 weight %, and preferred 25%～91 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 get 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, it is main chain that described natural polysaccharide derivant is selected from the natural polysaccharide, has the derivant that reactive functional groups is modified.
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 is the glycidyl methacrylate residue that has unsaturated double-bond, the methacrylic acid hydroxyl ethyl ester residue that has unsaturated double-bond, the acrylic acid sulphonic acid ester residue that has unsaturated double-bond or its combination; Described acrylamide residue is acrylamide residue, N-isopropyl third rare amide residues or its combination.All have unsaturated double-bond on the poly-polysaccharide main chain of these several polymer, and high reaction activity and high is arranged, can form cross-linked structure at short notice, and can regulate the cross-linking reaction degree by the amount that inserts two keys by radical reaction.
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.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.Calculate by proton nmr spectra that replacement degree acrylate-based or acrylamide is 5-60% in the product.
The weight content of described poly-polysaccharide is 5%～90 weight %, and preferred 9%～75 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 are compound.
Usually, described bone cement is obtained by the complex method that may further comprise the steps:
10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 weight % are provided the mixture of poly-polysaccharide;
In the presence of water soluble initiator, gather the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture simultaneously, 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 formation intercrossed network type bioactive composite structure, thereby obtains 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 get final product.
Usually, the 4%wt disodium phosphate soln is a 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 believed it is 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 get final product.
The compound method of described consolidation liquid can adopt the traditional method in this area, for example will gather polysaccharide derivates particularly 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 as bone morphogenetic protein (BMP), OP-1, bonegel, 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 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 the compound co-cultivation of cell and material 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 proper polymer material and calcium phosphate bone cement compound, form hybridization network, the HAP that aquation is generated is promptly 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 on organic network, and finally having formed with high polymer is the composite construction of " muscle " for " skeleton ", hydroxylapatite crystal.This structure has the presetting period of reduction, improves comprcssive strength, improves advantages such as toughness and classification degraded with respect to the bone cement of being made up of the hydrated product hydroxyapatite merely in the past, can satisfy the requirement of human body heavy burden position to intensity.With respect to present common Enhancement Method, the present invention seeks a kind of new compound thinking, promptly adopt the method for interface IPN to make the firm interface combination of generation between the bigger component of performance difference, thereby realize the complementation of different component performance or function, make it on macro property, produce special synergism.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, the growth of adjusting cell and aspects such as old and feeble and the organic homergy that earns a bare living, is the important component of forming extracellular matrix.Simultaneously, polysaccharide also possesses the basic demand as bio-medical material, promptly 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 unfavorable 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 for use, thereby make to 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 take place, and the two keys in the poly-polysaccharide derivates disappear, and have hydroxyapatite to generate simultaneously, 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 takes place in the environment of 100% humidity, carry out cross-linking reaction separately in the system simultaneously, 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 two-phase interface defective and destructive probability, 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.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.Calculate by proton nmr spectra that replacement degree acrylate-based or acrylamide is 5-60% in the product.
(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 phosphate 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 50 ℃ of dryings implant then.
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, the also alternative stem cell of osteoblast is 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 compatibility 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 and enzyme digestion.
Cultural method of stem cell and culture fluid 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 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 (for example promoting the cytokine of stem cell growth etc.), various antibiotic, various inducible factor in the above-mentioned culture fluid.
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 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 the 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, degraded and absorbed gradually along with the bone cement material, new bone formation reaches the purpose of repairing bone defect.
The preparation method of organizational project 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 moulding arbitrarily according to the shape of tissue defect.Usually, graft is a 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, mix with degradation material then with culture fluid.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 a kind of preparation method of novel natural polysaccharide/calcium phosphate bone cement hybrid material.The composite of selecting for use natural polysaccharide derivant with good biocompatibility and calcium microcosmic salt bone cement hydridization to form has the favorable tissue compatibility and degradability, can not cause the untoward reaction of tissue in vivo.
(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, helps reaching 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, the reaction condition gentleness, and entire reaction is all carried out at aqueous phase, has avoided with an organic solvent.
(4) simple is the firming body of composition with the inorganic matter, and whole mechanical performance is more crisp, lacking toughness.When bearing load, rigid collision can take place in the contact surface of material and bone, and the energy that the external force transmission comes can't absorb, and firming body collapses diffusing easily.The network that high polymer is formed 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 of being made up of calcium phosphate bone cement merely is slow usually.The present invention selects the polymer moiety of the natural polysaccharide thing of biodegradable as composite for use, the degraded velocity ratio is very fast down in 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 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 (Degree of Substitution DS) realizes the controlled of calcium phosphate bone cement setting time to utilize substitution value variation on the different and poly-polysaccharide derivates of initiator system ratio.
Chemical compound provided by the present invention can be synthetic by marketable material and traditional chemical transform mode.For example natural polysaccharide glucosan, chitosan, starch, cellulose, cyclodextrin etc. can be by commercially available acquisitions, and 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 conspicuous 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 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: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 in addition method similar or impartial to described content and material all can be applicable in the inventive method.
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 of being made up 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.
Add persulfate (50mg/ml) 120 μ l then, 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 replace 8.8 acrylate in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid its furnishing pasty state.
Add persulfate (50mg/ml) 120 μ l then, 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, place 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 replaces 12 N-isopropylacrylamides in promptly 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 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid its furnishing pasty state.
Add cerous nitrate ammonia salt initiators (2mg/ml) 200 μ l then, 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 replaces 12 acrylate in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid its furnishing pasty state is obtained mixture.
Add cerous nitrate ammonia salt initiators (2mg/ml) 200 μ l then 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 derivative (DS=12 replaces 12 acrylamides in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned glucosan/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
Add cerous nitrate ammonia salt initiators (2mg/ml) 120 μ l to mixture then, 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 replaces 12 N-isopropylacrylamides in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
In described mixture, add persulfate (50mg/ml) 120 μ l then, 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 replaces 6 N-isopropylacrylamides in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
In described mixture, add persulfate (50mg/ml) 120 μ l then, 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 replaces 12 N-isopropylacrylamides in promptly 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 particle diameter 10-20 μ m that form by calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned starch/sodium hydrogen phosphate consolidation liquid with its furnishing pasty state, obtain mixture.
In described mixture, add persulfate (50mg/ml) 120 μ l then, 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 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. poly-polysaccharide/self-curable calcium phosphate composite bone cement compositions, its component and weight divide ratio content to comprise in vain:
10%～95 weight % self-curable calcium phosphate bone cement; And the poly-polysaccharide of 5%～90 weight %.
2. compositions according to claim 1 is characterized in that, 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.
3. compositions according to claim 2 is characterized in that, it is main chain that described natural polysaccharide derivant is selected from the natural polysaccharide, has the derivant that reactive functional groups is modified;
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 is the glycidyl methacrylate residue that has unsaturated double-bond, the methacrylic acid hydroxyl ethyl ester residue that has unsaturated double-bond, the acrylic acid sulphonic acid ester residue that has unsaturated double-bond or its combination; Described acrylamide residue is acrylamide residue, N-isopropyl third rare amide residues 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.
4. 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.
5. a bone cement material that makes according to each described compositions of claim 1～4 is characterized in that, obtains described bone cement material by described self-curable calcium phosphate bone cement and described poly-polysaccharide are compound.
6, bone cement material as claimed in claim 5 is characterized in that, described bone cement material is obtained by the complex method that may further comprise the steps:
10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 weight % are provided the mixture of poly-polysaccharide;
In the presence of water soluble initiator, gather the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture simultaneously, form the bone cement material.
7, a kind of method for preparing the bone cement material, described method comprises the steps:
10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 weight % are provided the mixture of poly-polysaccharide;
The poly-polysaccharide of described 10%～95 weight % self-curable calcium phosphate bone cement and 5%～90 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 the cross-linking reaction of polysaccharide and the curing reaction of self-curable calcium phosphate bone cement in the described mixture simultaneously, form described bone cement material.
8, method as claimed in claim 7 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;
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;
Perhaps,
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 formation intercrossed network type bioactive composite structure, thereby obtains 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.
9. a tissue engineered implant is characterized in that, described graft contains the described bone cement material of claim 5 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.
10. the purposes of a bone cement material as claimed in claim 5 is characterized in that, as the support of preparation bone graft.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810033420 CN101496909B (en) | 2008-02-01 | 2008-02-01 | Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810033420 CN101496909B (en) | 2008-02-01 | 2008-02-01 | Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101496909A true CN101496909A (en) | 2009-08-05 |
| CN101496909B CN101496909B (en) | 2013-01-16 |
Family
ID=40944318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200810033420 Expired - Fee Related CN101496909B (en) | 2008-02-01 | 2008-02-01 | Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101496909B (en) |
Cited By (24)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101856509A (en) * | 2010-06-18 | 2010-10-13 | 华东理工大学 | Calcium magnesium injectable bone cement and preparation method and application thereof |
| CN102031690A (en) * | 2010-11-02 | 2011-04-27 | 西安交通大学 | Method for preparing organic silk-inorganic calcium hydrophosphate composite material by in-situ self-crystallization method |
| CN102698316A (en) * | 2012-05-16 | 2012-10-03 | 华东理工大学 | Rapid-curing adhesive bone repair material and preparation method thereof |
| CN102755668A (en) * | 2011-04-29 | 2012-10-31 | 天津市赛宁生物工程技术有限公司 | Medical plastic bone cement |
| CN103520774A (en) * | 2013-09-25 | 2014-01-22 | 胡建中 | Tissue-engineered artificial bone and building method thereof |
| CN103736149A (en) * | 2013-12-25 | 2014-04-23 | 暨南大学 | Injectable bioactive artificial bone material and preparation method and application thereof |
| CN103816566A (en) * | 2012-11-16 | 2014-05-28 | 赫罗伊斯医疗有限责任公司 | Antiseptic polymethylmethacrylate bone cement |
| CN103919632A (en) * | 2014-05-03 | 2014-07-16 | 王学建 | Artificial skull-repaired prosthesis manufacturing method |
| CN104368042A (en) * | 2014-11-19 | 2015-02-25 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of brushite bone cement |
| CN104587523A (en) * | 2014-06-10 | 2015-05-06 | 上海交通大学医学院附属第九人民医院 | Enriched bone marrow mesenchymal stem cell loaded self-curing calcium phosphate cement, and preparation method thereof |
| CN105461941A (en) * | 2015-12-23 | 2016-04-06 | 苏州大学 | Method for preparing self-curing high-strength macromolecular glue |
| CN106267365A (en) * | 2016-08-17 | 2017-01-04 | 林春梅 | A kind of enhancing activity injecting bone cement and preparation method thereof |
| CN106540317A (en) * | 2016-12-08 | 2017-03-29 | 苏州艾博迈尔新材料有限公司 | A kind of realizing controlled-release bone cement and preparation method thereof |
| CN106730002A (en) * | 2016-12-09 | 2017-05-31 | 苏州艾博迈尔新材料有限公司 | A kind of bone cement and preparation method thereof |
| CN107233627A (en) * | 2017-05-25 | 2017-10-10 | 华南理工大学 | A kind of calcium phosphate bone cement containing konjaku glucomannan and its preparation method and application |
| CN107648673A (en) * | 2017-09-18 | 2018-02-02 | 暨南大学 | A kind of compound calcium phosphate bone cement and preparation method and application |
| CN108219680A (en) * | 2017-12-15 | 2018-06-29 | 南京博物院 | For the adhesive composition of sclerotin historical relic reparation |
| CN108245704A (en) * | 2018-02-06 | 2018-07-06 | 重庆医科大学附属永川医院 | A kind of Orthopedic medical jointing material and preparation method thereof and application method |
| CN109276754A (en) * | 2018-09-25 | 2019-01-29 | 广州润虹医药科技股份有限公司 | A kind of somatotrophic injecting bone cement and preparation method thereof |
| CN109821058A (en) * | 2019-03-14 | 2019-05-31 | 广西大学 | A kind of antibacterial Orthopedic medical jointing material and preparation method thereof |
| CN111012946A (en) * | 2019-12-31 | 2020-04-17 | 广州润虹医药科技股份有限公司 | Thermo-sensitive calcium phosphate bone cement and preparation method thereof |
| CN112976222A (en) * | 2021-02-05 | 2021-06-18 | 金陵科技学院 | Bone cement 3D prints and stirs and extrusion device with double screw |
| CN113679892A (en) * | 2021-09-01 | 2021-11-23 | 山东明德生物医学工程有限公司 | Absorbable drug-loaded artificial bone and use method thereof |
| CN115737911A (en) * | 2022-12-16 | 2023-03-07 | 华南理工大学 | High-toughness bone repair composite material and preparation method thereof |
Family Cites Families (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 |
| CN100391550C (en) * | 2005-10-20 | 2008-06-04 | 华南理工大学 | Method of improving anti collapsibility of calcium phosphate skeletal cement using denaturated starch |
-
2008
- 2008-02-01 CN CN 200810033420 patent/CN101496909B/en not_active Expired - Fee Related
Cited By (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101856509A (en) * | 2010-06-18 | 2010-10-13 | 华东理工大学 | Calcium magnesium injectable bone cement and preparation method and application thereof |
| CN101856509B (en) * | 2010-06-18 | 2014-02-19 | 华东理工大学 | Calcium magnesium injectable bone cement and preparation method and application thereof |
| CN102031690A (en) * | 2010-11-02 | 2011-04-27 | 西安交通大学 | Method for preparing organic silk-inorganic calcium hydrophosphate composite material by in-situ self-crystallization method |
| CN102755668A (en) * | 2011-04-29 | 2012-10-31 | 天津市赛宁生物工程技术有限公司 | Medical plastic bone cement |
| CN102698316B (en) * | 2012-05-16 | 2014-08-13 | 华东理工大学 | Rapid-curing adhesive bone repair material and preparation method thereof |
| CN102698316A (en) * | 2012-05-16 | 2012-10-03 | 华东理工大学 | Rapid-curing adhesive bone repair material and preparation method thereof |
| CN103816566B (en) * | 2012-11-16 | 2016-08-24 | 赫罗伊斯医疗有限责任公司 | Antibiotic property polymethyl methacrylate bone cement |
| US9833472B2 (en) | 2012-11-16 | 2017-12-05 | Heraeus Medical Gmbh | Antiseptic polymethylmethacrylate bone cement |
| CN103816566A (en) * | 2012-11-16 | 2014-05-28 | 赫罗伊斯医疗有限责任公司 | Antiseptic polymethylmethacrylate bone cement |
| CN103520774A (en) * | 2013-09-25 | 2014-01-22 | 胡建中 | Tissue-engineered artificial bone and building method thereof |
| CN103736149A (en) * | 2013-12-25 | 2014-04-23 | 暨南大学 | Injectable bioactive artificial bone material and preparation method and application thereof |
| CN103736149B (en) * | 2013-12-25 | 2015-12-30 | 暨南大学 | A kind of injectable type biologically active artificial bone material and preparation method and application |
| CN103919632A (en) * | 2014-05-03 | 2014-07-16 | 王学建 | Artificial skull-repaired prosthesis manufacturing method |
| CN104587523A (en) * | 2014-06-10 | 2015-05-06 | 上海交通大学医学院附属第九人民医院 | Enriched bone marrow mesenchymal stem cell loaded self-curing calcium phosphate cement, and preparation method thereof |
| CN104368042A (en) * | 2014-11-19 | 2015-02-25 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of brushite bone cement |
| CN105461941A (en) * | 2015-12-23 | 2016-04-06 | 苏州大学 | Method for preparing self-curing high-strength macromolecular glue |
| CN105461941B (en) * | 2015-12-23 | 2019-02-19 | 苏州大学 | A kind of preparation method of self-curing high intensity macromolecular glue |
| CN106267365A (en) * | 2016-08-17 | 2017-01-04 | 林春梅 | A kind of enhancing activity injecting bone cement and preparation method thereof |
| CN106540317A (en) * | 2016-12-08 | 2017-03-29 | 苏州艾博迈尔新材料有限公司 | A kind of realizing controlled-release bone cement and preparation method thereof |
| CN106730002A (en) * | 2016-12-09 | 2017-05-31 | 苏州艾博迈尔新材料有限公司 | A kind of bone cement and preparation method thereof |
| CN107233627A (en) * | 2017-05-25 | 2017-10-10 | 华南理工大学 | A kind of calcium phosphate bone cement containing konjaku glucomannan and its preparation method and application |
| CN107233627B (en) * | 2017-05-25 | 2020-08-18 | 华南理工大学 | Calcium phosphate bone cement containing konjac glucomannan as well as preparation method and application thereof |
| CN107648673A (en) * | 2017-09-18 | 2018-02-02 | 暨南大学 | A kind of compound calcium phosphate bone cement and preparation method and application |
| CN108219680A (en) * | 2017-12-15 | 2018-06-29 | 南京博物院 | For the adhesive composition of sclerotin historical relic reparation |
| CN108245704A (en) * | 2018-02-06 | 2018-07-06 | 重庆医科大学附属永川医院 | A kind of Orthopedic medical jointing material and preparation method thereof and application method |
| CN109276754A (en) * | 2018-09-25 | 2019-01-29 | 广州润虹医药科技股份有限公司 | A kind of somatotrophic injecting bone cement and preparation method thereof |
| CN109821058A (en) * | 2019-03-14 | 2019-05-31 | 广西大学 | A kind of antibacterial Orthopedic medical jointing material and preparation method thereof |
| CN111012946A (en) * | 2019-12-31 | 2020-04-17 | 广州润虹医药科技股份有限公司 | Thermo-sensitive calcium phosphate bone cement and preparation method thereof |
| CN112976222A (en) * | 2021-02-05 | 2021-06-18 | 金陵科技学院 | Bone cement 3D prints and stirs and extrusion device with double screw |
| CN113679892A (en) * | 2021-09-01 | 2021-11-23 | 山东明德生物医学工程有限公司 | Absorbable drug-loaded artificial bone and use method thereof |
| CN115737911A (en) * | 2022-12-16 | 2023-03-07 | 华南理工大学 | High-toughness bone repair composite material and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101496909B (en) | 2013-01-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101496909B (en) | Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof | |
| US7713542B2 (en) | Three dimensional cell protector/pore architecture formation for bone and tissue constructs | |
| CN1302821C (en) | Preparation method of calcium orthophosphate bone cement degradable to pore in human body | |
| US6139578A (en) | Preparation of cell seeded ceramic compositions | |
| EP0613380B1 (en) | The use of monomeric/polymeric materials in the preparation of a curable composition for cartilage repair | |
| US20020055143A1 (en) | Bone precursor compositions | |
| Zhao et al. | Osteogenic media and rhBMP-2-induced differentiation of umbilical cord mesenchymal stem cells encapsulated in alginate microbeads and integrated in an injectable calcium phosphate-chitosan fibrous scaffold | |
| JP5406915B2 (en) | Biocompatible implant | |
| CN1972882A (en) | Macroporous, resorbable and injectible calcium phosphate-based cements (MCPC) for bone repair, augmentation, regeneration, and osteoporosis treatment | |
| CN101053673B (en) | High strength and tenacity degradable strontium calcium superphosphate composite bone cement and its preparation method | |
| Chen et al. | Injectable calcium sulfate/mineralized collagen‐based bone repair materials with regulable self‐setting properties | |
| CN110540404B (en) | Calcium phosphate bone cement with hollow through structure, preparation method and application thereof | |
| CN1226149A (en) | Injectable bio-active glass in dextran suspension | |
| CN100536935C (en) | Composite porous calcium phosphate bone cement and method for making same | |
| CN107050508B (en) | Injectable bone repair material and preparation method thereof | |
| CN101816808A (en) | Injectable porous high-strength bone repair material | |
| CN101530635B (en) | Bioactive bone-repairing cement material | |
| CN102552985B (en) | Silk fibroin/calcium phosphate bone cement-based porous composite material and preparation method thereof | |
| Anil et al. | Biomaterials for craniofacial tissue engineering and regenerative dentistry | |
| CN112190762B (en) | Injection type magnesium-based composite fiber reinforced calcium phosphate biological bone adhesive and preparation method and application thereof | |
| US8969430B2 (en) | Biocompatible ceramic-polymer hybrids | |
| KR102458881B1 (en) | A porous scaffold comprising a collagen and a polycarprolacton for regenerating the periodontal complex having improved healing characteristics, and method for preparing the same | |
| TheinHan et al. | Non‐rigid calcium phosphate cement containing hydrogel microbeads and absorbable fibres seeded with umbilical cord stem cells for bone engineering | |
| Uda et al. | Experimental studies on hydroxyapatite powder-carboxymethyl chitin composite: injectable material for bone augmentation | |
| CN113750289A (en) | Organic-inorganic composite bioactive material based on calcium phosphate and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130116 Termination date: 20180201 |