CN101496909B - Polysaccharide/calcium orthophosphate composite bone cement and preparation method thereof - Google Patents

Abstract

The invention discloses a polysaccharide/self-curing calcium phosphate composite bone cement composition. The composition and weight percentage content include: 10%-95% by weight of self-curing calcium phosphate bone cement; and 5%-90% by weight of polysaccharide . The bone cement composition of the present invention can obtain a bone cement with high strength, good toughness, strong plasticity, fast curing, good biocompatibility and degradability, thereby overcoming the lack of toughness existing in existing calcium phosphate bone cement materials. The defects of slow degradation in the human body and the defects of several enhancement methods in the prior art can better meet the requirements of surgical use.

Description

Polysaccharide/calcium phosphate composite bone cement and its preparation method

technical field

The invention belongs to the field of medical biomaterials and relates to a novel organic/inorganic composite material used for filling or injecting to repair human hard tissue defects.

Background technique

The repair and treatment of bone defects has long been a thorny problem for surgeons. Scientists from all over the world have been devoting themselves to the research and development of ideal bone repair materials. In the mid-1980s, Brown and Chow in the United States invented a self-curing bioactive bone defect repair material—calcium phosphate cement (Calcium phosphate cement, CPC), which can be hydrated and solidified by itself in the physiological environment of the human body. It is finally transformed into hydroxyapatite (HA) which is similar to the composition of human bone, and the heat release is less during the curing process, and it can be shaped arbitrarily according to the shape of the bone defect, which has attracted great attention. As a bone substitute material, calcium phosphate bone cement has excellent properties, mainly as follows: good biocompatibility and biosafety, good biodegradability, and osteogenic activity. Calcium phosphate bone cement can be converted into a composition similar to natural bone. After being implanted in the human body, it can participate in metabolism, form bone through osteoconduction, and guide the same amount of new bone formation while being absorbed (W.J.E.M.Habraken, J.G.C.Wolke, J.A.Jansen, Research on Ceramic Composite Materials as Scaffold Materials for Drug Delivery in Tissue Engineering, Advanced Drug Delivery 2007, 59, 234-248; Hockin H.K.Xu, Michael D.Weir, Elena F.Burguera and AlexisM.Fraser, Injectable Macroporous Phosphoric Acid Calcium cement scaffold, Biomaterials, 2006, 27: 4279-4287; Makoto Watanabe, Miyuki Tanaka, Makoto Sakurai and MiokoMaeda, Development of calcium phosphate cement (research progress of calcium phosphate cement) Journal of the European Ceramic Society (European Ceramic Society) , 2006, 26:549-552).

In the past 20 years, the types, properties and theoretical research of calcium phosphate bone cement have made great progress, and it has been widely used in clinical practice as a bone repair material. However, some inherent shortcomings of bone cement itself, such as calcium phosphate bone cement has a long curing time, poor bonding performance, insufficient mechanical properties, and slow degradation, which limits its application to a certain extent. Currently, it can only be used in non-load-bearing areas. Bone repair. In recent years, many scholars have carried out in-depth modification research on various calcium phosphate bone cements to increase their mechanical strength, adjust the curing time, and improve their rheology and biodegradability (Lisa E. Carey, Hockin H.K. Xu, Jr., Carl G .Simon, Shozo Takagi, Laurence C.Chow, Premixed rapid-setting calcium phosphate composites for bone repair (premixed rapid-setting calcium phosphate composite bone repair material), Biomaterials (Biomaterials), 2005, 26: 5002-5014; Hockin H.K. Xu, Carl G. Simon, Fast setting calcium phosphate-chitosan scaffold: mechanical properties and biocompatibility. 26:1337-1348).

Adding fiber molecules to calcium phosphate bone cement is an important method to improve the tensile and impact properties of bone cement (Dai Honglian, Li Shipu, Yan Yuhua, Wang Xinyu, Cao Xianying, Han Yingchao, Chen Xiaoming, Yuan Lin, Li Jianhua, Preparation of a calcium phosphate composite bone cement Methods, Publication No. CN1657483A, 2005; Li Yubao, Wei Jie, Medical composite bio-bone cement powder, bone cement liquid and medical composite bio-bone cement, Publication No. CN1403168A, 2003). Fiber molecular compounding methods can be divided into crosslinking agent compounding, plasticizer compounding, absorbable fiber compounding, natural fiber compounding, etc. According to the composite principle of materials, in fiber-reinforced composite materials, fibers bear most of the load and act as a bridge between the matrix and fibers, and when the matrix cracks extend to the fiber-matrix interface, combined with an appropriate interface to prevent cracks from expanding or make The crack is deflected to adjust the interfacial stress and prevent the crack from further expanding. In terms of the degradation performance of materials, fiber reinforcement can be divided into non-degradable fiber reinforcement and degradable fiber reinforcement. Non-degradable fiber is an early reinforcing material used to improve the mechanical properties of calcium phosphate cement, and its reinforcing effect is very significant. Currently commonly used non-degradable fibers mainly include carbon fibers, glass fibers and polymer fibers. The study found that the adsorption of polyoxygen ions and serum proteins can inhibit the growth of HAP crystals, which may lead to smaller grain sizes and the formation of denser interlaced microstructures, polymer bridging between various crystals and energy absorption through plastic flow It is responsible for the increased strength of calcium phosphate bone cement. Self-curing calcium phosphate cement combined with two different enhancers has higher synergistic reinforcement than conventional calcium phosphate cement with one enhancer, and can be used in load-bearing craniofacial and orthopedic repairs. Studies have shown that adding polypropylene, nylon and carbon fiber to calcium phosphate bone cement, although the compressive strength can be reduced due to the increase in porosity, it greatly increases the toughness and tensile strength of calcium phosphate bone cement, which This kind of composite bone cement can be used to repair defects with a certain degree of curvature (Hockin H.K.Xu, Janet B.Quinn, Shozo Takagi, Laurence C.Chow, Synergistic reinforcement of in situ hardening calcium phosphate composite scaffold for bone tissue engineering Synergistic reinforcement of in situ solidified calcium phosphate bone cement), Biomaterials (biomaterials.), 2004: 1029-1037). Sun Kangning and others dispersed long carbon fibers in a mold along a certain direction to obtain a calcium phosphate bone cement biocomposite material with a bone-like structure. The composite material has good mechanical properties and biocompatibility, and can be applied to the replacement and repair of artificial bone in medical operations, etc. (Sun Kangning, Zhao Ping, A calcium phosphate bone cement biocomposite material and its preparation method, Publication No. CN1559887A, 2005). However, the disadvantage of adding fiber molecules to the calcium phosphate bone cement mentioned above is that because there are interface problems between the inorganic cement and the polymer fibers, it generally only has a reinforcing effect when the amount of fibers added is very small, while toughening The effect is often not obvious; at the same time, since most of these added fibers do not degrade, it is impossible to take into account the mechanical properties and degradation properties of the composite material at the same time.

The use of degradable fiber materials to enhance calcium phosphate bone cement is a new bone cement reinforcement method developed in recent years. The improvement of this method is that the degradable fiber material used can stabilize and strengthen the bone cement composite in the initial stage of implantation into the human body, and as the fiber gradually degrades, the columnar channels it produces can help For the delivery of nutrients and the rapid growth of blood vessels and cells, and the degradation products are safe and non-toxic. Therefore, it can be said that the degradable fiber has played a dual role of pore formation and reinforcement in the preparation and application of bone cement, avoiding the negative effects caused by long-term placement of non-degradable materials in the human body (Lin, et al, Process for producing preparation of fast-setting, bioresorbable calcium phosphate cements fast-setting absorbable calcium phosphate cement. US Patent 7,066,999, 2006). Xu et al. used polymer absorbable long cellulose to strengthen TTCP/DCPA bone cement, added 25% absorbable fibers to the calcium phosphate bone cement, and the diameter of the fibers was 322 μm. The composite specimens were placed in 37°C saline Among them, after 1, 7, 14, 28, and 56 days, it was found that the strength increased by 5 times, and the toughness increased by 100 times. The strength and toughness can be maintained for 2-4 weeks depending on the fiber dissolution rate. Studies have shown that the addition of fibers keeps the calcium phosphate bone cement necessary for the regeneration process, and the pores left in the calcium phosphate bone cement after the fibers dissolve make it easy for new blood vessels to grow in, providing bone tissue regeneration. Good environment; with the dissolution of fibers, interpenetrating cylindrical macropores are formed in the calcium phosphate bone cement, and its flexural strength is still 39% higher than that of calcium phosphate bone cement without macropores, and its toughness is 256% higher (Hockin HKXu , Janet B. Quinn, Calcium phosphate cement containing resorbable fibers for short-term reinforcement and macroporosity. 202). In addition, calcium phosphate cements can be enhanced with organic bioactive substances. Bigi et al. tried to improve the mechanical properties of bone cement with gelatin, and the results showed that the compressive strength had been improved to a certain extent (A.Bigi, B.Bracci, S.Panzavolta, Effect of added gelatin on the properties of calcium phosphate cement.( Effect of Gelatin on the Properties of Calcium Phosphate Cement), Biomaterials, 2004, 25: 2893-2899). Chitosan is also an important bioactive substance used to improve the mechanical properties of bone cement. Xu and Takagi tried adding chitosan to the liquid phase of bone cement to prepare enhanced calcium phosphate bone cement/chitosan composites, and the results showed that the stability and strength of bone cement were greatly improved. (Hockin HKXu, Carl G. Simon, Fast setting calcium phosphate-chitosan scaffold: mechanical properties and biocompatibility (premixed macroporous calcium phosphate cement scaffold, Journal of Materials Science), Biomaterials (biomaterials), 2007, 18: 1345-1353). Wang et al. also investigated the enhancement effect of chitosan derivatives on bone cement. They found that adding phosphorylated chitosan to calcium phosphate bone cement can significantly improve the compressive strength after ^curing . The strong bonding between chitosan makes the newly generated HA particles linked together through the polymer, which improves the compressive strength. DosSantos et al. added polyamide fiber when blending the material, and the strength was improved. At the same time, it was found that there were radial cracks in the matrix of the material around the fiber, which showed the breakthrough point of a significant increase in mechanical strength again (XiaohongWang, Jianbiao Ma, Yinong Wang, Binglin He, Bone repair in radii and tibias of rabbits with phosphorylated chitosan reinforced calcium phosphate cements (phosphorylated chitosan reinforced calcium phosphate cement for mouse tibia repair research, Biomaterials (biomaterials), 2002, 23: 4167-4176).

However, the strength performance of the bone cement obtained by the above-mentioned prior art needs to be further improved.

Micropores caused by water are a very important factor affecting the mechanical strength of calcium phosphate cement. Barralet et al found that the addition of α-hydroxy acid sodium salts represented by sodium citrate can lead to a significant increase in the compressive strength of calcium phosphate bone cement, and believe that the reason for the increase in compressive strength is that the presence of sodium ions will reduce the curing reaction process The stable connection between the calcium ion and the hydroxycarboxylic acid ion group generates more free ions, and these free ions will be adsorbed on the particle surface of the reactant and the solidified product, thereby increasing their ζ- Potential energy, improves the rheological properties of the bone cement, reduces the water content while maintaining the operability of the bone cement, which in turn leads to a decrease in the porosity of the bone cement and an increase in strength (Jake E. Barralet, Maryjane Tremayne, Kevin J. Lilley, Uwe Gbureck, Modification of calcium phosphate cement with α-hydroxy acids and their salts (modified calcium phosphate cement through α-hydroxy acids and their salts). Chem Mater, (Material Chemistry), 2005, 17: 1313-1319, Jake E Barralet , Mike Hofmann, Liam M.Grover, Uwe Gbureck, High-strength apatitic cement by modification with α-hydroxy acid salts (alpha-hydroxy acid and its salt modified to prepare high-strength apatite cement), .Adv.Mater.( Advanced Materials), 2003, 15:2091-2094).

Multi-material synergistic enhancement of calcium phosphate bone cement is a new concept proposed in recent years. The core of this concept is to use the mutual promotion and interaction relationship between various reinforcing materials to improve the mechanical properties of bone cement. In the exploration of this field, the research work carried out by Xu et al. is representative to a certain extent. They combined degradable reticular fiber and chitosan composition reinforcement system with bone cement, and the results showed that the effect of multi-component synergistic reinforcement was significantly higher than that of single-component reinforcement (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, there are many other reinforcement methods, such as organic reinforcement, whisker and inorganic ceramic particle reinforcement, etc. Matsuya et al reported a copolymer calcium phosphate bone cement, the bone cement is a mixture of tetracalcium phosphate (TTCP) and anhydrous calcium hydrogen phosphate (DCPA) as the solid phase, polymethyl ethyl ether-maleic acid The aqueous solution of the copolymer (PMVE-Ma) is used as the liquid phase, and the two are mixed to form a calcium phosphate bone cement composite system. PMVE-Ma can be dissolved by water through the hydrolysis of acid anhydride groups to form the corresponding condensed malic acid copolymer (PMVA-Ma). Its existence makes calcium phosphate bone cement have higher mechanical strength, and the curing time can be extended To more than 30min (Yoko Matsuya, Shigeki Matsuya, Joseph M.Antonucci, Shozo Takagi, Laurence C.Chow, Akifumi Akamine, Effect of powder grinding onhydroxyapatite formation in a polymeric calcium phosphate cement prepared from etracalcium phosphate (tetracalcium phosphate) (Effect of powder particle size on polymeric calcium phosphate cements prepared from tetracalcium phosphate and poly(methyl vinyl ether-maleic acid) systems), Biomaterials, 1999, 20:691-697). Whiskers and inorganic particles currently used to strengthen calcium phosphate bone cement mainly include silicon nitride whiskers, calcium carbonate whiskers, silicon carbide whiskers and their silanized substances, and inorganic particles of alumina and silica. Studies have shown that combining the above substances with calcium phosphate cement can greatly improve the original strength, even close to the strength level of cerebral cortex bone (100-200MPa) (Hockin H.K.Xu, Douglas T.Smith, C.G.Carl G. Simon, (High-strength bioactive composite bone repair material containing nano-fused silica whiskers), Biomaterials (biomaterials.), 2004, 25: 4615-4626; Ana I. Villacampa, Juan Ma. García-Ruiz, Synthesis of a new hydroxyapatite-silicacomposite material. (New hydroxyapatite-silica composite material synthesis), Journal ofCrystal Growth (Crystal Growth Journal,), 2000, 211: 111-115). The disadvantage of the above-mentioned bone cement is that the added components degrade slowly or not, and the use for bone repair will affect the formation of new bone.

In summary, an ideal bone substitute material should have good biocompatibility, be able to integrate with the surrounding bone tissue, have mechanical properties close to those of natural bone, have appropriate strength and toughness, and be able to grow with tissue growth. The input material is continuously degraded and eventually replaced by new bone tissue. Calcium phosphate bone cement is an important bioactive bone repair material. Although its related research progresses rapidly, there are still some problems to be solved, such as insufficient strength and slow curing speed. Most of the existing commonly used reinforcement methods introduce fibers or other external components. The interfacial bonding between the reinforcement and the matrix is the key to the research of this type of composite material; most fibers and other external components can only be reinforced when the amount of addition is small. As a result, it is difficult to simultaneously improve strength and toughness. At the same time, because many external components do not degrade or degrade slowly, it is more difficult to balance the mechanical properties and degradation properties of calcium phosphate bone cement.

Therefore, there is an urgent need for a calcium phosphate composite bone cement and a preparation method thereof in this area. The bone cement has high strength, good toughness, strong plasticity, fast curing, good biocompatibility and degradability, thereby overcoming the existing calcium phosphate The lack of toughness of bone cement materials, the defects of slow degradation in the human body, and the defects of several reinforcement methods can better meet the requirements of surgical use.

Contents of the invention

The purpose of the present invention is to obtain a calcium phosphate composite bone cement composition, which can obtain bone cement with high strength, good toughness, strong plasticity, fast curing, good biocompatibility and degradability.

Another object of the present invention is to obtain a bone cement material with high strength, good toughness, strong plasticity, fast curing, good biocompatibility and degradability.

Yet another object of the invention is to obtain a method for producing bone cement material.

Another object of the present invention is to obtain a tissue engineered graft containing the bone cement material of the present invention.

Yet another object of the present invention is to obtain the use of the bone cement material according to the invention as a scaffold for the preparation of bone grafts.

In the first aspect of the present invention, a polysaccharide/self-curing calcium phosphate composite bone cement composition is provided, and its components and weight percent content include: 10% to 95% by weight of self-curing calcium phosphate bone cement; and 5% ~90% polysaccharide by weight.

In a specific embodiment of the present invention, the polypolysaccharide is selected from natural polysaccharides, natural polysaccharide derivatives or combinations thereof,

Specifically, the natural polysaccharide is selected from dextran, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannan or combinations thereof.

In a specific embodiment of the present invention, the natural polysaccharide derivatives are selected from natural polysaccharide-based derivatives modified with reactive functional groups;

Specifically, the reactive functional group is selected from acrylate residues, acrylamide residues, acrylic acid residues, vinylpyrrolidone residues, crown ether residues, other reactive unsaturated groups or combinations thereof.

More specifically, the acrylate residues are glycidyl methacrylate residues with unsaturated double bonds, hydroxyethyl methacrylate residues with unsaturated double bonds, Acrylic sulfonate residues or combinations thereof; the acrylamide residues are acrylamide residues, N-isopropylacrylamide residues or combinations thereof;

More specifically, the degree of modification of the reactive functional groups on the natural polysaccharide in the natural polysaccharide derivatives is 1-30 reactive functional groups per 100 natural polysaccharide units.

In a specific embodiment of the present invention, the components of the self-curing calcium phosphate bone cement are selected from the group consisting of tricalcium phosphate, tetracalcium phosphate, octacalcium phosphate, calcium hydrogen phosphate, hydroxyapatite, fluorapatite, One or a mixture of calcium pyrophosphate.

Another aspect of the present invention provides a bone cement material prepared from the composition of the present invention, which is obtained by compounding the self-curing calcium phosphate bone cement and the polysaccharide.

In a specific embodiment of the present invention, the bone cement material is obtained by a composite method comprising the following steps:

Provide a mixture of 10% to 95% by weight of self-curing calcium phosphate bone cement and 5% to 90% by weight of polysaccharide;

In the presence of the water-soluble initiation system, the cross-linking reaction of the polysaccharide and the curing reaction of the self-curing calcium phosphate bone cement proceed simultaneously in the mixture to form a bone cement material.

Still another aspect of the present invention provides a method for preparing bone cement material, said method comprising the steps of:

Provide a mixture of 10% to 95% by weight of self-curing calcium phosphate bone cement and 5% to 90% by weight of polysaccharide;

The bone cement material is obtained by compounding the 10%-95% by weight self-curing calcium phosphate bone cement and 5%-90% polysaccharide by weight;

Specifically, the composite method includes the following steps: in the presence of a water-soluble initiator system, the cross-linking reaction of polysaccharide and the curing reaction of self-curing calcium phosphate bone cement are simultaneously carried out in the mixture to form the bone cement material.

In a specific embodiment of the present invention,

The mixture is obtained through the following mixing steps: preparing a polysaccharide-disodium hydrogen phosphate solution as a curing solution of self-curing calcium phosphate bone cement; then mixing the self-curing calcium phosphate bone cement with the curing solution to obtain the mixture;

Specifically, the solid-to-liquid ratio of the self-curing calcium phosphate bone cement to the solidifying liquid is 0.1-5 g/ml, preferably in the range of 1-3 g/ml.

In a specific embodiment of the present invention, the steps of crosslinking reaction and curing reaction include: adding a water-soluble initiator system to the mixture, stirring evenly, and mixing into a slurry; placed in a 100±10% humidity environment until an interpenetrating network biocomposite structure is formed, thereby obtaining the bone cement material;

Specifically, the water-soluble initiation system is persulfate-N, N, N', N'-tetramethylethylenediamine, ammonium cerium nitrate, or hydrogen peroxide-ferrous sulfate system;

Specifically, the concentration of the water-soluble initiator system is 1×10 ^-4 to 1×10 ^-1 mol/L, and the preferred concentration is 1×10 ^-3 to 5×10 ^-1 mol/L;

Specifically, the setting time of the slurry is 0.5-60 minutes, and the curing time is 4-48 hours, wherein the preferred curing time is 15-30 hours.

Another aspect of the present invention provides a tissue engineered graft, the graft contains the bone cement material of the present invention and stem cells inoculated on the bone cement material, and the seeding amount of the stem cells is 2×10 ⁶ -5×10 ⁷ cells/cm ³ bone cement material.

Still another aspect of the present invention provides a use of a bone cement material as a scaffold for preparing a bone graft.

Detailed ways

The invention discloses a self-curing calcium phosphate bone cement composite material containing natural polysaccharide derivatives and a preparation method thereof. The present invention uses biocompatibility, degradable natural polysaccharide derivatives and calcium phosphate bone cement powder to compound, can simultaneously realize calcium phosphate bone cement hydration and solidification and polysaccharide derivative cross-linking and solidification, and shorten the life span of bone cement. The initial setting time improves the mechanical properties of calcium phosphate bone cement such as compressive strength and toughness; the formed paste can be shaped arbitrarily and hardens itself under the human environment and humidity. The invention can be used for filling and repairing and injectable repairing material of bone defect, nonunion and delayed bone union caused by various reasons, and is a novel organic-inorganic composite human body hard tissue repairing material with broad application prospects.

In the term "polysaccharide/self-curing calcium phosphate composite bone cement composition" of the present invention, the meaning of "/" refers to the relationship between the two is "and".

The "natural polysaccharide derivative" in the present invention means a compound obtained by modifying a reactive functional group in a molecule of a natural polysaccharide. Specifically, the modification method is, for example, a grafting reaction. For example, the dextran-acrylate derivative refers to the acrylate functional group grafted into the dextran molecule, thereby forming a polysaccharide derivative.

The dosage forms of various aspects of the present invention are described in detail below:

self-curing calcium phosphate bone cement

The self-curing calcium phosphate bone cement of the present invention can adopt the traditional self-curing calcium phosphate bone cement in this field, specifically for example selected from tricalcium phosphate, tetracalcium phosphate, octacalcium phosphate, calcium hydrogen phosphate, hydroxyapatite, fluorophosphorus One of limestone, calcium pyrophosphate or a mixture thereof. The self-curing calcium phosphate bone cement is not particularly limited, as long as the bone cement composition can be cured into a bone cement material in the presence of a curing liquid.

The content of the self-setting calcium phosphate bone cement is usually 10%-95% by weight, preferably 25%-91% by weight, calculated from the total weight of the polysaccharide/self-setting calcium phosphate composite bone cement composition.

The particle size of the bone cement is not specifically limited, as long as it does not limit the purpose of the invention. Its particle size includes but is not limited to 10-20 μm.

polysaccharide

Polysaccharides of the present invention are generally selected from natural polysaccharides, natural polysaccharide derivatives or combinations thereof.

Specifically, the natural polysaccharide is selected from dextran, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannan or combinations thereof.

Specifically, the natural polysaccharide derivatives are selected from derivatives with natural polysaccharides as the main chain and modified with reactive functional groups.

More specifically, the reactive functional group is selected from acrylate residues, acrylamide residues, acrylic acid residues, vinylpyrrolidone residues, crown ether residues, other reactive unsaturated groups or combinations thereof.

More specifically, the acrylate residues are glycidyl methacrylate residues with unsaturated double bonds, hydroxyethyl methacrylate residues with unsaturated double bonds, Acrylic sulfonate residue or a combination thereof; the acrylamide residue is acrylamide residue, N-isopropylacrylamide residue or a combination thereof. The polysaccharide main chains of these polymers all have unsaturated double bonds and have high reactivity. They can form cross-linked structures in a short time through free radical reactions, and can be modified by the amount of double bonds. To adjust the degree of cross-linking reaction.

More specifically, the degree of modification of the reactive functional groups on the natural polysaccharide in the natural polysaccharide derivatives is 1-30 reactive functional groups per 100 natural polysaccharide units.

Modification of functional groups on natural polysaccharides to obtain natural polysaccharide derivatives is a traditional reaction in this field. The natural polysaccharide derivatives can also be obtained commercially.

Specific examples of preparing natural polysaccharide derivatives are as follows (including but not limited to the following examples): Weigh natural polysaccharides and dissolve them in dimethyl sulfoxide (DMSO) to form a concentration of 1%-20% (g/ ml) solution. An appropriate amount of acrylate or acrylamide is added, so that the molar ratio of the acrylate or acrylamide group to the glucopyranose ring in the polysaccharide is 0.05-1. The reaction temperature is controlled at 20-100° C., and the reaction is carried out under the protection of argon for 24-72 hours. After the reaction, isopropanol was added, suction filtered and dialyzed in a deionized environment for 12-72 hours, and a white fluffy product was obtained after freeze-drying. The substitution degree of acrylate group or acrylamide in the product is calculated by hydrogen nuclear magnetic resonance spectrum to be 5-60%.

The weight content of the polysaccharide is 5%-90% by weight, preferably 9%-75% by weight, calculated from the total weight of the polypolysaccharide/self-curing calcium phosphate composite bone cement composition.

bone cement material

The bone cement material of the present invention is obtained by compounding the self-curing calcium phosphate bone cement and the polysaccharide.

Typically, the bone cement is obtained by a composite method comprising the following steps:

Provide a mixture of 10% to 95% by weight of self-curing calcium phosphate bone cement and 5% to 90% by weight of polysaccharide;

In the presence of the water-soluble initiation system, the cross-linking reaction of the polysaccharide and the curing reaction of the self-curing calcium phosphate bone cement proceed simultaneously in the mixture to form a bone cement material.

Specifically, the steps of cross-linking reaction and curing reaction include: adding a water-soluble initiator system to the mixture, stirring evenly, and mixing into a slurry; placed until an interpenetrating network biocomposite structure is formed, thereby obtaining the bone cement material;

Specifically, the water-soluble initiation system is persulfate-N, N, N', N'-tetramethylethylenediamine, ammonium cerium nitrate, or hydrogen peroxide-ferrous sulfate system;

Specifically, the concentration of the water-soluble initiator system is 1×10 ^-4 to 1×10 ^-1 mol/L, and the preferred concentration is 1×10 ^-3 to 5×10 ^-1 mol/L;

Specifically, the setting time of the slurry is 0.5-60 minutes, and the curing time is 4-48 hours, wherein the preferred curing time is 15-30 hours.

The interpenetrating network biocomposite structure is formed when the slurry solidifies.

The mixing method of the polysaccharide and the self-curing calcium phosphate bone cement can adopt a traditional mixing method in the art. Specifically, for example, the mixture is obtained by the following mixing steps: preparing polysaccharide derivative-disodium hydrogen phosphate solution (other suitable solutions can also be used) as the solidification solution of self-curing calcium phosphate bone cement; The bone cement is mixed with the curing fluid to obtain said mixture.

The solidifying liquid can be a conventional solidifying liquid in the art. And the concentration of the solidified liquid is not specifically limited, as long as it does not limit the purpose of the invention.

Generally, 4%wt disodium hydrogen phosphate solution is commonly used in the preparation of calcium phosphate cement. In the curing reaction of simple calcium phosphate cement, the curing solution of this concentration is generally considered to be an ideal curing system. In fact, it is not necessarily disodium hydrogen phosphate but other substances, and the concentration can also vary. Therefore, the concentration of the disodium hydrogen phosphate solution is not specifically limited, as long as it does not limit the purpose of the present invention.

The preparation method of the solidification liquid can adopt the traditional method in this field, specifically, for example, polysaccharide derivatives are dissolved in a disodium hydrogen phosphate solution to prepare a polysaccharide/disodium hydrogen phosphate solution with a concentration of 0.1-3 g/ml .

Other biologically acceptable substances can also be added to the solidification solution of the present invention. Specifically, for example, biological factors that promote osteogenic growth can be added appropriately, such as bone morphogenetic protein (BMP), OP-1, bonegel, osteonectin (osteoconectin), osteocalcin (osteocalcin), bone connection factor (SCF ) or a combination thereof. The amount of the biological factor that promotes osteogenic growth varies with the type of animal it belongs to. The mass ratio of the biological factor to the polymer is preferably 0.00625-0.1 μg/mg, more preferably 0.01-0.08 μg/mg. In the present invention, biological factors can be added when preparing the polymer solution. If cells and materials are co-cultured together, they can also be added to the culture medium.

Specifically, the solid-to-liquid ratio of the self-curing calcium phosphate bone cement to the solidifying liquid is 0.1-5 g/ml, preferably in the range of 1-3 g/ml.

Design of the present invention is such:

The present invention selects suitable polymer materials and calcium phosphate bone cement to form a hybrid network, so that the HAP generated by hydration is attached to the polymer network at the beginning of nucleation, and as the hydration reaction proceeds, the HAP crystals The particles grow on the organic network, and finally form a composite structure with high polymer as the "skeleton" and hydroxyapatite crystals as the "muscle". Compared with the previous bone cement composed solely of hydration product hydroxyapatite, this structure has the advantages of reducing initial setting time, increasing compressive strength, improving toughness and hierarchical degradation, and can meet the strength requirements of the weight-bearing parts of the human body. Compared with the current common enhancement methods, the present invention seeks a new composite idea, that is, adopts the method of interfacial interpenetration to generate a firm interfacial bond between components with large performance differences, so as to realize the complementary properties or functions of different components , so that it produces a special synergistic effect on macroscopic properties. In the selection of enhanced components, considering that the organism itself is formed by the construction of biological macromolecules such as proteins and sugars, as an important component of living organisms, polysaccharides widely exist in organisms, and play an important role in controlling cell division and differentiation. It plays an important role in regulating cell growth and aging and maintaining the normal metabolism of living organisms. It is an important component of the extracellular matrix. At the same time, polysaccharides also have the basic requirements as biomedical materials, that is, excellent biocompatibility and can be enzymatically decomposed into small molecular substances that are easily absorbed by the living body and have no toxic side effects. They are a type of biodegradable and absorbable polymer materials. Therefore, choosing natural polysaccharides as the reinforcing phase not only overcomes the disadvantages of unsatisfactory cell affinity commonly found in synthetic polymer materials, but also avoids the problems of unstable physiological properties and possible immune reactions caused by the selection of collagen matrix. Therefore, the composite materials are both "hard and tough", and at the same time, the degradability of calcium phosphate materials can be improved.

In a specific embodiment, the present invention forms an interpenetrating network structure through two kinds of cross-linked networks on the basis of simultaneous free radical polymerization of double bonds of polysaccharide derivatives and hydration of calcium phosphate salt. According to infrared spectrum, X-ray diffraction spectrum and other characterizations, both reactions occur, the double bond in the polysaccharide derivative disappears, and hydroxyapatite is formed at the same time, which all indicate that the two reactions proceed. The method that the two reactions proceed simultaneously and finally form a complex is similar to the polymer interpenetrating network, so it is called an interpenetrating network type composite structure.

Due to the existence of inorganic calcium and phosphate salts in the mixture and the hydration reaction occurs in an environment of 37°C and 100% humidity, the respective crosslinking reactions are carried out in the system at the same time, so a network structure with interpenetrating inorganic/organic interfaces can be formed, and a good two The combination of phase interface is conducive to the transmission and dispersion of stress, reducing the possibility of defects and damage at the two-phase interface. At the same time, the interpenetrating entanglement of the network leads to the increase of the crosslinking degree of the system, which can show a significant synergy.

In a specific embodiment of the present invention, the method for preparing polysaccharide/calcium phosphate bone cement hybrid interpenetrating network of the present invention includes the following steps:

(1) Weigh the natural polysaccharide and dissolve it in dimethyl sulfoxide (DMSO) to prepare a solution with a concentration of 1%-20% (g/ml). An appropriate amount of acrylate or acrylamide is added, so that the molar ratio of the acrylate or acrylamide group to the glucopyranose ring in the polysaccharide is 0.05-1. The reaction temperature is controlled at 20-100° C., and the reaction is carried out under the protection of argon for 24-72 hours. After the reaction, isopropanol was added, suction filtered and dialyzed in a deionized environment for 12-72 hours, and a white fluffy product was obtained after freeze-drying. The substitution degree of acrylate group or acrylamide in the product is calculated by hydrogen nuclear magnetic resonance spectrum to be 5-60%.

(2) Prepare 1-10%wt disodium hydrogen phosphate solution and 1-100mg/ml initiator solution. Dissolving polysaccharide derivatives in disodium hydrogen phosphate solution, preparing a polysaccharide/disodium hydrogen phosphate solution with a concentration of 0.1-3 g/ml, and adding it to calcium phosphate bone cement powder according to a certain mass ratio. It can be mixed evenly and reconciled into a paste, which can be implanted in the body. It can also be cured in vitro at 37°C and 100% relative humidity for 6-48 hours to form a cured body, dried at 50°C, and then implanted in the body.

stem cell

The source of the stem cells of the present invention is not particularly limited, and may be stem cells from any source. Generally, the stem cells of the present invention are autologous or allogeneic stem cells. There is no particular limitation on where the stem cells are obtained, and they may be fat stem cells, bone marrow stromal stem cells or other stem cells. In addition, osteoblasts can also replace stem cells as seed cells for bone tissue engineering.

Stem cells that can be used in the present invention can be from any vertebrate, preferably a mammal, more preferably a primate, especially a human.

Although autologous stem cells are preferred, allogeneic sources of stem cells are more commonly used. Studies have shown that allogeneic stem cells of different growth and developmental stages can form stem cell tissues in allogeneic animals with differences in histocompatibility and complete immune function.

Methods of isolating and obtaining stem cells are known in the art. A preferred method is density gradient centrifugation and enzymatic digestion.

Stem cell culture methods and culture medium are also well known in the art. A preferred method is to culture the stem cells in an incubator at 37° C., saturated humidity, and 5% CO ₂ . Suitable culture medium includes (but is not limited to): 1) DMEM medium ((Gibco company)+5～20% fetal calf serum; 2) DMEM medium+5～20% calf serum; 3) DMEM medium +5-20% autologous (allogeneic) human serum. In addition, various growth factors (such as cytokines that promote the growth of stem cells, etc.), various antibiotics, and various induction factors are added to the above-mentioned culture solution.

Stem cells suitable for use in the present invention should be able to proliferate in vivo or in vitro. A preferred stem cell is bone marrow stromal stem cell cultured in vitro.

bone graft

Because the bone cement of the invention has very good compatibility with bone marrow stromal stem cells and fat stem cells, it is particularly suitable as a scaffold material for bone repair.

Bone marrow stromal stem cells and/or adipose stem cells cultured and expanded in vitro are seeded on bone cement with excellent biocompatibility to form a stem cell-bone cement complex, and this "stem cell-bone cement" complex is implanted into the defect site, With the gradual degradation and absorption of bone cement materials, new bone is formed to achieve the purpose of repairing bone defects.

The preparation method of the tissue engineering bone graft of the present invention is simple and convenient, and only a certain amount of bone marrow stromal stem cells and/or fat stem cells are inoculated on the bone cement material.

The shape of the tissue-engineered bone graft of the present invention is not particularly limited, and can be shaped arbitrarily according to the shape of tissue defect. Typically, grafts are long strips.

The concentration of bone marrow stromal stem cells and/or adipose stem cells in the tissue engineered bone of the present invention is usually about 0.5×10 ⁶ /cm ³ (ceramic scaffold volume) to 5×10 ⁸ /cm ³ or higher, preferably 1× 10 ⁶ /cm ³ to 1×10 ⁸ /cm ³ , more preferably 5×10 ⁶ /cm ³ to 5×10 ⁷ /cm ³ porous ceramic material. Usually, the concentration of bone marrow stromal stem cells and/or adipose stem cells is adjusted with a culture medium, and then mixed with a degradable material. When mixing, the ratio of the culture solution to the degradable material is not particularly limited, but it is appropriate that the material can absorb the maximum amount of the culture solution.

In addition, in the tissue engineered bone graft of the present invention, various other cells, growth factors, and various antibiotics can also be added or compounded, so as to maintain the phenotype of bone marrow stromal stem cells and/or adipose stem cells, promote bone marrow stromal stem cells and/or Or adipose stem cell growth, and promote tissue engineered bone growth in vivo.

In addition to implanting tissue-engineered bone grafts into the body, they are also cultured in an in vitro bioreactor to construct tissue-engineered bones, which have a certain histological structure, biochemical composition, and biomechanical strength in vitro. tissue engineered bone.

The tissue engineered bone graft formed by the method of the invention can be directly implanted into the bone defect in the body.

The beneficial effects of the present invention are:

(1) The present invention provides a preparation method of a novel natural polysaccharide/calcium phosphate bone cement hybrid material. The composite material formed by hybridizing natural polysaccharide derivatives with good biocompatibility and calcium phosphate bone cement has good tissue compatibility and degradability, and will not cause adverse tissue reactions in vivo.

(2) In the present invention, the hybrid system in which polymer polysaccharides and inorganic calcium phosphate salts are interpenetrated through the interpenetrating network interface, compared with the commonly used blending method, the composite material interface provided by the present invention has stronger interfacial adhesion and relatively Capacitance is better, which is beneficial to achieve the enhancement effect through the synergy between the two phases.

(3) The preparation process of the inventive method is simple, the polymerization rate is fast, and the process of composite hybridization is completed in one step when the polymer crosslinking and calcium phosphate salinization are carried out simultaneously, without the need to carry out the next step of the composite process, the reaction conditions are mild, and the whole reaction All carried out in the aqueous phase, avoiding the use of organic solvents.

(4) The cured body composed solely of inorganic substances has relatively brittle overall mechanical properties and lacks toughness. When the load is applied, the contact surface between the material and the bone will collide rigidly, the energy transmitted by the external force cannot be absorbed, and the solidified body will easily collapse. In the present invention, the network composed of polymers can play the role of elastic buffer, and the deformation of the hydrogel network can be used to absorb external energy and keep the overall integrity of the solidified body, so that the compression and impact resistance of the entire solidified body can be effectively improved. .

(5) The degradation rate of the solidified body composed solely of calcium phosphate bone cement is usually relatively slow. The present invention selects the biodegradable natural polysaccharide as the macromolecular part of the composite material, and the degradation speed is relatively fast under the action of enzymes in the body, and because it is the skeleton of the whole cured body, when the polymer is degraded, it will be cured The body will become loose, so that more body fluid can penetrate into the middle of the solidified body, so that the originally complete solidified body is divided into small pieces, thereby accelerating the degradation rate.

(6) Controllability. By simply controlling the reaction component ratio and the ratio of the two phases in the hybrid material, polymer polysaccharides with different numbers of double bond substituents can be obtained, thereby controlling the cross-linking density of the polymer network, and regulating the strength and degradability of the hybrid material and swelling.

(7) The scope of application is wide. The hybrid material prepared by the invention can be used as a pre-shaped bone repair material, as an intraoperative solidified material, and as an injectable bone repair material for minimally invasive operations such as vertebral body shaping.

(8) The present invention selects a natural polysaccharide derivative material with good biocompatibility and degradability and certain biomechanical properties, and reacts with calcium phosphate bone cement (CPC) under the action of a water-soluble trigger system to form an interaction in situ. Wear network biocomposites. This method utilizes the characteristics of the interpenetrating network to improve the compressive strength of calcium phosphate bone cement, and utilizes the different proportions of the initiating system and the degree of substitution (Degree of Substitution, DS) on polysaccharide derivatives to realize the setting time of calcium phosphate bone cement. controllable.

The compounds provided by the present invention can be synthesized by commercially available raw materials and traditional chemical transformation methods. For example, natural polysaccharide dextran, chitosan, starch, cellulose, cyclodextrin, etc. can be obtained commercially, dextran-acrylate, dextran-isopropylacrylamide, chitosan-acrylic acid Esters, starch-acrylamide, cellulose-isopropylacrylamide, cyclodextrin-isopropylacrylamide, sodium alginate-isopropylacrylamide derivatives can all be synthesized by chemical transformation.

Other aspects of the invention will be apparent to those skilled in the art from the disclosure herein.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples is usually according to conventional conditions, such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions described in the manufacture conditions recommended by the manufacturer.

Unless otherwise defined or stated, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the method of the present invention.

The present invention is further illustrated below through the detailed description of specific embodiments of the method of the present invention, but these examples are not used to limit the present invention.

Example 1

Weigh 0.3 g of dextran-acrylate derivatives (dextran molecular weight 40,000, DS=12, that is, 12 acrylates are replaced in every 100 dextran monomers, DS is obtained by H NMR spectroscopy) and dissolved In 4%wt disodium hydrogen phosphate solution, prepare 0.5g/ml dextran-containing solidification solution; weigh the self-curing solidification solution consisting of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite with a particle size of 10-20 μm Calcium phosphate salt powder (Shanghai Ruibang Biomaterials Co., Ltd.) 1.5 grams, add the above-mentioned dextran/disodium hydrogen phosphate solidified solution to adjust it into a paste.

Then add persulfate (50mg/ml) 120μl, N,N,N',N'-tetramethylethylenediamine (23mg/ml) 300μl, mix well and fill it into a tetrafluoroethylene mold to make a sample, Put it in a 37°C, 100% humidity environment for curing, and the initial setting time of the sample is 2.48min. After complete drying, the compressive strength reaches 82.82MPa, which meets the partial strength requirements of the weight-bearing part of the human body. Curing time 24 hours.

Example 2

Weigh 0.3 grams of dextran-acrylate derivatives (dextran molecular weight 40,000, DS=8.8, that is, 8.8 acrylates are replaced in every 100 dextran monomers) and dissolved in 4%wt disodium hydrogen phosphate solution , be prepared into 0.5g/ml solidified solution containing dextran; Weigh the self-curing calcium phosphate salt powder (Shanghai Ruibang Biomaterials Co., Ltd.) Co., Ltd.) 1.5 g, add the above-mentioned dextran/disodium hydrogen phosphate solidification solution to make it into a paste.

Then add persulfate (50mg/ml) 120μl, N,N,N',N'-tetramethylethylenediamine (23mg/ml) 300μl, mix the slurry obtained after filling into tetrafluoroethylene mold Formed into a spline, placed in a 37°C, 100% humidity environment for curing, and the initial setting time of the spline was 2.48min. After complete drying, the compressive strength reaches 94.33MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 3

Weigh 0.3 g of dextran-isopropylacrylamide derivatives (DS=12, that is, 12 isopropylacrylamides are replaced per 100 dextran monomers) and dissolve in 4%wt disodium hydrogen phosphate solution, Prepare 0.5g/ml dextran-containing solidification solution; weigh 10-20 μm self-curing calcium phosphate powder composed of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite (Shanghai Ruibang Biomaterials Co., Ltd.) 3 grams, add the above-mentioned dextran/disodium hydrogen phosphate solidification solution to adjust it into a paste.

Then add 200 μl of cerium nitrate ammonium salt initiator (2mg/ml), mix the obtained slurry and fill it into a tetrafluoroethylene mold to make a sample, place it at 37°C, solidify in a 100% humidity environment, and the sample will initially set The time is 2.48min. After completely drying, the compressive strength reaches 43.95MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 4

Take by weighing 1.5 grams of chitosan-acrylate derivatives (DS=12, that is, replace 12 acrylates in every 100 chitosan monomers) and dissolve in 4%wt disodium hydrogen phosphate solution to prepare 0.5g/ml Chitosan-containing solidification solution; 0.5 grams of self-curing calcium phosphate salt powder (Shanghai Ruibang Biomaterials Co., Ltd.) with a particle size of 10-20 μm composed of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite were weighed, and added The above-mentioned dextran/disodium hydrogen phosphate solidified solution was adjusted into a paste to obtain a mixture.

Then, 200 μl of cerium nitrate ammonium salt initiator (2 mg/ml) was added to the mixture, and the slurry obtained after mixing was filled into a tetrafluoroethylene mold to make a sample, and placed at 37° C. and cured in a 100% humidity environment. , the initial setting time of the spline is 2.48min. After complete drying, the compressive strength reaches 80.24MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 5

Weigh 0.3 grams of starch-acrylamide derivatives (DS=12, that is, 12 acrylamides are replaced in every 100 starch monomer molecules) and dissolve in 4%wt disodium hydrogen phosphate solution to prepare 0.5g/ml starch-containing Solidifying liquid; Take by weighing 1.5 grams of self-curing calcium phosphate salt powder (Shanghai Ruibang Biomaterials Co., Ltd.) of particle diameter 10-20 μm that is made up of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite, add above-mentioned dextran/ Disodium hydrogen phosphate solidified liquid was adjusted into a paste to obtain a mixture.

Then add cerium nitrate ammonium salt initiator (2mg/ml) 120 μ l to the mixture, and the slurry obtained after mixing is filled into a tetrafluoroethylene mold to make a sample, placed at 37 ° C, solidified in a 100% humidity environment, and the sample The initial setting time is 1.33min. After complete drying, the compressive strength reaches 76.14MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 6

Weigh 0.3 g of cellulose-isopropylacrylamide derivatives (DS=12, that is, 12 isopropylacrylamides are replaced per 100 cellulose monomer molecules) and dissolve in 4%wt disodium hydrogen phosphate solution to prepare into 0.5g/ml cellulose-containing solidified solution; weigh the self-curing calcium phosphate salt powder with a particle size of 10-20 μm consisting of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite (Shanghai Ruibang Biomaterials Co., Ltd.) 3 grams, adding the above-mentioned starch/disodium hydrogen phosphate solidification solution to adjust it into a paste to obtain a mixture.

Then, 120 μl of persulfate (50 mg/ml) and 300 μl of N, N, N’, N’-tetramethylethylenediamine (23 mg/ml) were added to the mixture, and the slurry obtained after mixing evenly was filled into four The sample was made in a vinyl fluoride mold, and placed in an environment of 37°C and 100% humidity for curing. The initial setting time of the sample was 3.48 minutes. After complete drying, the compressive strength reaches 50.68MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 7

Weigh 0.3 g of cyclodextrin-isopropylacrylamide derivatives (DS=6, that is, 6 isopropylacrylamides are replaced per 100 cyclodextrin monomer molecules) and dissolve in 4%wt disodium hydrogen phosphate solution , prepared into a 0.5g/ml solidified solution containing cyclodextrin; weigh the self-curing calcium phosphate salt powder (Shanghai Ruibang Biomaterials Co., Ltd. Co., Ltd.) 3 grams, add the above-mentioned starch/disodium hydrogen phosphate solidification solution and adjust it into a paste to obtain a mixture.

Then add 120 μl of persulfate (50mg/ml) to the mixture, and mix the slurry obtained after 50 μl of sodium bisulfite is filled into a tetrafluoroethylene mold to make a sample, and place it at 37° C. in a 100% humidity environment Medium curing, the initial setting time of the spline is 5.68min. After complete drying, the compressive strength reaches 45.8MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

Example 8

Weigh 0.3 g of sodium alginate-isopropylacrylamide derivative (DS=12, that is, per 100 natural polysaccharide units replace 12 isopropylacrylamide) and dissolve in 4%wt disodium hydrogen phosphate solution to prepare 0.5g/ml solidified solution containing sodium alginate; weigh self-curing calcium phosphate salt powder with a particle size of 10-20 μm consisting of calcium hydrogen phosphate, tetracalcium phosphate and hydroxyapatite (Shanghai Ruibang Biomaterials Co., Ltd.) 3 grams, adding the above-mentioned starch/disodium hydrogen phosphate solidification solution to adjust it into a paste to obtain a mixture.

Then, 120 μl of persulfate (50 mg/ml) and 300 μl of N, N, N’, N’-tetramethylethylenediamine (23 mg/ml) were added to the mixture, and the slurry obtained after mixing evenly was filled into four The sample was made in a vinyl fluoride mold, and placed in an environment of 37° C. and 100% humidity for curing. The initial setting time of the sample was 3.25 minutes. After completely drying, the compressive strength reaches 48.68MPa, which meets the partial strength requirements of the weight-bearing part of the human body.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (9)

1. A polysaccharide/self-curing calcium phosphate composite bone cement composition, its components and weight percent content comprising: 10% to 95% by weight of self-curing calcium phosphate bone cement; and 5% to 90% by weight of polysaccharides, which are selected from natural polysaccharides as the main chain and derivatives modified with reactive functional groups; The reactive functional group is selected from acrylate residues, acrylamide residues, acrylic acid residues; The acrylate residues are glycidyl methacrylate residues with unsaturated double bonds, hydroxyethyl methacrylate residues with unsaturated double bonds, sulfonic acid esters with unsaturated double bonds residues or combinations thereof; The acrylamide residue is an acrylamide residue, an N-isopropylacrylamide residue or a combination thereof; The degree of modification of the reactive functional groups on the natural polysaccharide is to replace 1-30 reactive functional groups per 100 natural polysaccharide units. 2. composition according to claim 1, is characterized in that, described natural polysaccharide is selected from dextran, chitosan, cellulose, alginic acid, starch, cyclodextrin, xanthan gum, glucomannan or a combination thereof. 3. composition according to claim 1 is characterized in that, the component of said self-curing calcium phosphate bone cement is selected from tricalcium phosphate, tetracalcium phosphate, octacalcium phosphate, calcium hydrogen phosphate, hydroxyapatite , fluorapatite, calcium pyrophosphate or a mixture thereof. 4. A bone cement material obtained from the composition according to any one of claims 1 to 3, wherein the bone cement material is obtained by compounding the self-curing calcium phosphate bone cement and the polysaccharide . 5. bone cement material as claimed in claim 4, is characterized in that, described bone cement material is obtained by the composite method that comprises the following steps: Provide a mixture of 10% to 95% by weight of self-curing calcium phosphate bone cement and 5% to 90% by weight of polysaccharide; In the presence of the water-soluble initiation system, the cross-linking reaction of the polysaccharide and the curing reaction of the self-curing calcium phosphate bone cement proceed simultaneously in the mixture to form a bone cement material. 6. A method for preparing bone cement material as claimed in claim 4, said method comprising the steps of: Provide a mixture of 10% to 95% by weight of self-curing calcium phosphate bone cement and 5% to 90% by weight of polysaccharide; The bone cement material is obtained by compounding the 10%-95% by weight self-curing calcium phosphate bone cement and 5%-90% polysaccharide by weight; The composite comprises the following steps: in the presence of a water-soluble trigger system, the cross-linking reaction of the polysaccharide and the curing reaction of the self-curing calcium phosphate bone cement are carried out simultaneously in the mixture to form the bone cement material. 7. The method of claim 6, wherein, The mixture is obtained through the following mixing steps: preparing a polysaccharide-disodium hydrogen phosphate solution as a curing solution of self-curing calcium phosphate bone cement; then mixing the self-curing calcium phosphate bone cement with the curing solution to obtain the mixture; The solid-to-liquid ratio of the self-curing calcium phosphate bone cement to the curing liquid is 0.1 to 5 g/ml, The steps of the crosslinking reaction and curing reaction include: adding a water-soluble initiator system to the mixture, stirring evenly, and mixing into a slurry; the slurry is placed in an environment of 37±5°C and 100±10% humidity until forming an interpenetrating network biocomposite structure, thereby obtaining the bone cement material; The water-soluble initiation system is persulfate-N, N, N', N'-tetramethylethylenediamine, ammonium cerium nitrate, or hydrogen peroxide-ferrous sulfate system; The concentration of the water-soluble initiator system is 1×10 ^-4 ~ 1×10 ^-1 mol/L; The setting time of the slurry is 0.5-60 minutes, and the curing time is 4-48 hours. 8. A tissue engineered graft, characterized in that, the graft contains the bone cement material according to claim 4 and stem cells inoculated on the bone cement material, and the seeding amount of the stem cells is 2× 10 ⁶ -5×10 ⁷ cells/cm ³ bone cement material. 9. A use of the bone cement material as claimed in claim 4, characterized in that it is used as a scaffold for preparing bone grafts. the