WO2018000793A1 - Degradable magnesium/zinc-containing calcium phosphate-calcium sulfate porous composite biological scaffold - Google Patents

Abstract

A degradable magnesium/zinc-containing calcium phosphate-calcium sulfate porous composite biological scaffold, which is prepared by: processing a bovine calcined cancellous bone mineral porous scaffold by using a quaternary system containing magnesium source/zinc source/sulfur source/phosphorus source, taking out the product, drying, and carrying out high-temperature calcination. The magnesium/zinc-containing calcium phosphate-calcium sulfate porous composite biological scaffold possesses a favorable three-dimensional intercommunication mesh structure, and possesses good osteoconductivity, degradability, mechanical strength, and biocompatibility. Calcium sulfate whiskers with a high length/diameter ratio grow in the meshes, thereby increasing the specific surface area of the material, and thus improving the cell adhesion.

Description

Calcium phosphate-calcium sulfate porous composite biological scaffold capable of degrading magnesium and zinc Technical field

The invention relates to the field of medical materials, in particular to a calcium phosphate-calcium sulfate porous composite biological scaffold which can degrade magnesium and zinc.

Background technique

Bone transplantation is a tissue transplant that is only less than blood transfusion. The research and development of bone substitute materials is one of the focuses of medical research. There are researches on the mechanism of bone substitute materials and the body in China's 12th Five-Year National Project.

The human bones and bones have elements such as Ca, P, C, O, H, S, Fe, Mg, Cu, Si, Zn, Mn, Na, K, etc., and there are extensive homogenous replacement behaviors in human bone mineralization, human bones. Has a complex composition and structure. In the design process of bone tissue engineering scaffolds or artificial bones, it is important to consider the complex composition and structure of human bones such as severe mineralized tissues; human bones cannot be completely replaced by the limited properties provided by a single material, and more importantly, the scaffolds It is also necessary to provide a three-dimensional porous microstructure for the regeneration of bone tissue to guide the differentiation and proliferation of cells, and to maintain or quickly obtain sufficient mechanical strength to meet the mechanical requirements of the material to be replaced. The ideal bone graft replacement material or bone tissue engineering scaffold material should have the following conditions: 1) good bone conductivity, material with ideal three-dimensional interpenetrating mesh structure, as high porosity and specific surface area as possible; 2) Osteoinductive; 3) good biocompatibility and surface chemistry and microstructure supporting bone cell growth and functional differentiation; 4) good biodegradability; 5) part of the material that bears bone conduction must have Sufficient mechanical strength and load carrying capacity; 6) easy to process.

In a review in 2010, Johan et al. classified New Zealand's clinically available bone graft replacement materials into four categories: 1. Single-phase calcium-phosphorus materials, including hydroxyapatite-converted bioceramics, and synthetic hydroxyl groups. A kind of apatite cement, β-tricalcium phosphate artificial ceramics; 2, composite materials include tetracalcium phosphate / calcium hydrogen phosphate, 62.5% α-tricalcium phosphate / 26.8% anhydrous calcium hydrogen phosphate / 8.9% calcium carbonate /1.8% hydroxyapatite, 60% hydroxyapatite/40% b-tricalcium phosphate, 73% b-tricalcium phosphate/21% calcium dihydrogen phosphate/5% magnesium hydrogen phosphate, tetracalcium phosphate/hydrogen phosphate 6 kinds of synthetic cements such as calcium/amorphous calcium phosphate, α-tricalcium phosphate/calcium carbonate/calcium dihydrogen phosphate, and the formula is 80% tricalcium phosphate/20% calcium hydrogen phosphate artificial ceramic; 3, single There are four kinds of mud or particles prepared by calcium sulphate; 4, one type of silicon-containing bioglass. Apatite and calcium sulfate are currently the most common alternative materials or components for bone transplantation in clinical practice. At present, there is a lack of ideal bone graft replacement materials in clinical practice, mainly in the ideal three-dimensional interconnected mesh structure, great porosity and specific surface area, degradability, osteoconductivity, osteoinductivity and mechanical strength. It.

The artificial bones with ideal three-dimensional interconnected mesh microstructures have been transformed from animal materials: two of them are porous hydroxyapatite ceramic bones prepared from high temperature sintering process of bovine cancellous bone. The three-dimensional interpenetrating micro-structure of bovine cancellous bone natural bone ore is close to human bone mineral composition, with good biocompatibility, good bone conductivity and good compressive strength, free of high temperature sintering procedures. Xenogeneic bone immune rejection and pathogen introduction are possible and easy to process. The porous hydroxyapatite from bovine bone has a good porosity of 60-90% and is rich in bovine bone resources, and has a pore diameter of 390-1360 μm, a bone graft replacement material slightly larger than 150-400 μm and an ideal pore diameter of the bone tissue engineering scaffold; It has good compressive strength of 1-20MPa; implanted in the body is beneficial to bone repair cell recruitment, blood vessel entry, oxygen and tissue fluid exchange, providing good physiological activity space and adhesion support for bone repair cells; its huge disadvantages The bone mineral-hydroxyapatite obtained by high-temperature sintering of bovine cancellous bone is too stable, and the degradation in the body is too slow and long. The solubility in calcium-phosphorus bone grafting material is the lowest, and the degradation rate is far from matching with the formation speed of new bone. , can not continue to release Higher concentrations of calcium and other osteogenesis beneficial ions, and therefore lack of good osteogenic activity, is not conducive to bone repair and transformation.

The ideal degradation rate is another important requirement for artificial bone or bone tissue engineering scaffolds. The ideal artificial bone degradation rate should match the formation speed of new bones, and further provide space for guiding new bone formation while gradually degrading into new bone substitutes. The osteogenesis beneficial ions such as calcium ions continuously released during the degradation process provide mineral recombination components for the redeposition, transformation and metabolism of bone minerals, which may stimulate the formation of new bones and have some degree of potential osteoinductivity; artificial bone Or the degradation rate of bone tissue engineering material is too fast to provide sufficient space-time support and guidance for the bone repair process, and too slow degradation will hinder the formation, replacement and shaping of new bone. The degradation of inorganic bone graft materials in vivo mainly through two pathways: humoral-mediated dissolution and cell-mediated degradation. The process of dissolution is the process of hydrolysis of materials and binders under the action of body fluids, and the material gradually dissociates into physical dissolution processes of particles, molecules and ions. The cell-mediated degradation process is mainly the biodegradation process of phagocytosis of macrophages and osteoclasts. The degradation process of inorganic bone grafting materials in vitro is related to its composition. The degradation rate is closely related to the particle size, porosity, specific surface area, crystallinity and solubility of the material. Solubility is the most important factor. Among the most common bone graft replacement materials in clinical practice, calcium sulfate has the fastest degradation rate [calcium sulfate completely degraded in the body for 45-72d, more than twice as fast as autologous bone], hydroxyapatite in calcium phosphate material Has the slowest degradation rate [non-porous massive hydroxyapatite can not be completely degraded in the body for 10 years], far greater than the rate of new bone formation, other calcium and phosphorus components such as tricalcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate The degradation rate of polyhydrogen phosphate, calcium pyrophosphate, etc. is in between, and has a relatively moderate degradation rate. In order to overcome the obvious shortcomings of calcined bovine cancellous bone porous hydroxyapatite, in the past 20 years, scientists have attempted to convert calcined bovine cancellous porous hydroxyapatite into tricalcium phosphate or complex phase phosphorus containing tricalcium phosphate. Gray stone ceramics. Commonly known as Paris Cement Calcium sulphate mud paste and cement granules are the oldest materials applied to bone defect filling, because of its good biotolerance; 2 good space filling characteristics; 3 faster absorption speed and complete bioabsorption; 4. Potential Osteogenic activity; 5 good bone conduction and extended to date because faster absorption provides space for bone repair. Calcium sulphate dissolves about 0.2 grams at room temperature in 100 ml of water. Calcium sulphate forms a high calcium environment when it degrades in vivo, providing a calcium source for bone formation of new bone tissue, and binding to phosphate in body fluids to promote new bone minerals. The potential osteoinductive activity is related to the local high calcium and acidity microenvironment during the dissolution of calcium sulfate; the degradation of calcium sulfate in the body forms a calcium source for the formation of new bone during localized high calcium environment, and promotes it to varying degrees. Osteoblast formation and differentiation; the degradation of calcium sulfate in the body to form a local acid micro-environment may promote the micro-dissolution of human bone minerals, resulting in osteogenic active protein exposure, which is conducive to the formation of new bone; but the clinical application of calcium sulfate mud paste Or the common disadvantage of the same synthetic material is that it is difficult to have the ideal three-dimensional interconnected mesh structure, and the complete degradation time of calcium sulfate in the body is 45-72d, which is more than twice as fast as the autogenous bone. It is difficult to make the new bone formation continuous and stable. Bone conduction support, can not provide three-dimensional porous micro-structure for bone tissue regeneration, that is, lack of good osteoconductive structural basis repair cells, blood vessels In the graft, it is not conducive to the formation of new bones; even if the calcium sulfate elemental scaffold with high porosity and high specific surface area of three-dimensional interconnected mesh structure is successfully prepared, the degradation rate will be faster and the strength will be worse; the calcium sulfate material dissolves. After the drop, the local microenvironment may be acidified and may cause an inflammatory reaction. The advantage of synthetic composite materials is that the composition of different degradation characteristics and the composition ratio of the components can be selected to achieve a certain balance between the degradation rate and pH of the composite material, and it is possible to adsorb more active proteins (signal proteins) in the human body. Improve the biological activity of materials and more meet the ideal requirements for bone graft replacement materials.

In addition, the biological activity of calcium phosphate and other bio-based materials may be enhanced by the incorporation of biologically active ions. Studies have shown that these biologically active ions can effectively stimulate protein activity and promote fine Cell growth and bone growth. The human body contains about 25 g of magnesium, which plays an important role in human bone formation and all growth processes, maintenance of bone cell structure and function, bone metabolism and remodeling. Calcium magnesium phosphate-based bone cement with low magnesium content can significantly improve cell adhesion. Calcium-doped calcium phosphate cement has become an increasingly important new bone repair biomaterial because it can promote the interface between implant materials and bone tissue: magnesium-doped bone cement is easier to prepare, and 73% b-tricalcium phosphate in western countries such as New Zealand. The bone cement of /21% calcium dihydrogen phosphate / 5% magnesium hydrogen phosphate formula is clinically applied. The composite formula contains magnesium bone cement with degradability, which can release beneficial elements such as calcium, phosphorus and magnesium. It can be degraded and ion exchanged in the body after transplantation, and does not have three-dimensional interpenetrating mesh structure to hinder repair of cells and blood vessels. Early in-depth grafts, lacking good three-dimensional interconnected mesh structure of bone conduction.

We successfully converted calcined bovine porous hydroxyapatite into apatite-calcium sulfate composite scaffold, and converted calcined bovine porous hydroxyapatite into degradable magnesium-containing multiphase apatite porous ceramic, while maintaining calcined cattle The ideal three-dimensional interconnected mesh structure of bone porous hydroxyapatite and its good mechanical strength have successfully improved its degradation characteristics. The latter also successfully incorporated the crystal lattice of magnesium ions into soluble calcium phosphate, forming good degradation characteristics. Calcium-doped calcium phosphate such as tri-magnesium phosphate, etc., the material releases successful active ionized magnesium, beneficial ionized calcium, etc. when it is dissolved; we now try to further integrate their advantages to incorporate sulfur, phosphorus, zinc and magnesium into calcined bovine bone. The porous scaffold transforms the elemental hydroxyapatite porous scaffold into a composite porous bioscaffold containing calcium and zinc containing calcium and zinc with good degradation characteristics.

Disclosure of invention

The object of the present invention is to solve the problems that the existing bone graft substitute materials have good three-dimensional interpenetrating mesh structure, mechanical strength, degradability and biological activity, and the sulfur, phosphorus, magnesium and zinc are simultaneously incorporated into the natural bone mineral complex. In the exquisite three-dimensional interconnected mesh structure of the bull calcined cancellous bone and bone porous support, the calcined bovine cancellous elemental porous hydroxyapatite can be transformed into the doped bone active ion magnesium and zinc. Calcium phosphate-calcium sulfate composite scaffold material, the magnesium-calcium-containing calcium phosphate-calcium sulfate porous composite biological scaffold of the invention has good three-dimensional interpenetrating mesh structure and osteoconductivity, degradability, good mechanical strength and biological Compatibility; at the same time, the growth of calcium sulfate whiskers with larger aspect ratio in the mesh can increase the specific surface area of the material and may improve cell adhesion. The composite bioscaffold may have potential osteoinductivity due to the effective incorporation of the osteogenic active ion magnesium, zinc and calcium sulfate which can produce a local high calcium environment upon degradation. The composite bioscaffold may more satisfy the ideal conditions for bone graft replacement materials or bone tissue engineering scaffold materials.

The technical solution adopted by the present invention to solve the technical problem thereof is:

A calcium phosphate-calcium sulfate porous composite biological scaffold capable of degrading magnesium and zinc, and removing the porous scaffold of bovine calcined cancellous bone and bone through a quaternary system containing magnesium source, zinc source, sulfur source and phosphorus source After drying, it is obtained by calcination at a high temperature.

X-ray powder diffraction analysis of the composite bioscaffold material is a calcium phosphate-calcium sulfate porous composite bioscaffold material containing active ions of magnesium and zinc, such as calcium sulfate / zinc pyrophosphate / magnesium zinc phosphate, calcium sulfate / zinc pyrophosphate / magnesium magnesium phosphate / Hydroxyapatite, calcium sulfate / zinc phosphate / magnesium magnesium phosphate, calcium sulfate / magnesium zinc phosphate / magnesium pyrophosphate. The composite stent contains magnesium and zinc apatite components with good degradation rate, such as tricalcium phosphate, magnesium pyrophosphate, magnesium zinc phosphate, zinc pyrophosphate, tri-zinc phosphate, etc., which have a fast absorption rate and complete bioabsorption. The potential osteogenic calcium sulphate and some materials also contain the slowest rate of dissolution of hydroxyapatite in the phosphorus compound; due to the obvious difference in the rate of dissolution between the various components of the composite bioscaffold material, the composite bioscaffold material can be realized Gradient degradation; because we change the composition ratio and mass ratio of the composite bioscaffold material by changing the mass ratio of the reactants in the compound formula, concentration, impregnation and hydrothermal reaction time, calcination temperature and time, the composite bioscaffold material The degradation rate can thus be effectively controlled, such as in the case of other conditions within the experimental conditions, with the solution The increase of sulfur content can gradually increase the content of calcium sulfate; with the increase of phosphorus in the unit solution, the hydroxyapatite with a ratio of calcium to phosphorus of 1.67 gradually becomes zinc phosphorus, (calcium + magnesium) phosphorus, (zinc). + Magnesium phosphate with a phosphorus ratio of 1.5, such as tri-zinc phosphate, tri-magnesium phosphate, magnesium zinc phosphate, zinc-phosphorus, magnesium-phosphorus, and zinc pyrophosphate, we can effectively control the composition and mass ratio of the scaffold components. Therefore, the dissolution rate of the composite biological scaffold material can be effectively regulated. The composite bioscaffold material can form a high calcium environment favorable for new bone formation in the early stage of simulated body fluid environment, and has sustained release of calcium, magnesium and zinc ions, which may support the potential osteogenic activity of the composite bioscaffold material. The composite biological scaffold material retains the exquisite three-dimensional interconnected mesh microstructure of the bovine natural bone mineral and its good mechanical strength, and at the same time, whisker growth with a large aspect ratio in the growth of the mesh can increase the material ratio. Surface area improves cell and protein adhesion. In the animal bone cancellous bone defect area, bone repair cells were observed to adhere, proliferate, differentiate, and secrete bone matrix in the scaffold. Very early blood vessel formation was observed in the scaffold, and the osteogenesis process was similar to the physiological state of intramembranous osteogenesis; No obvious immunological rejection and inflammatory reaction were observed during the observation period, suggesting that the composite bioscaffold material has good biocompatibility. The rapid and good bone repair in the animal bone defect area also suggests the potential osteogenic activity of the composite bioscaffold material. .

Preferably, the quaternary system for treating the bovine calcined cancellous bone ore porous scaffold containing magnesium source, zinc source, sulfur source and phosphorus source is selected as one of the following schemes:

Scheme 1: The porous scaffold of the calcined cancellous bone ore is first immersed in the magnesium source-zinc source composite solution and evaporated to dryness, and then enters the hydrothermal reaction in the sulfur source-phosphorus source composite solution;

Scheme 2: The hydrothermal reaction of the porous scaffold of the calcined cancellous bone and bone mineral in the magnesium source-zinc source-sulfur source-phosphorus source composite solution.

Preferably, the hydrothermal reaction is carried out by a constant temperature hydrothermal method, and the temperature is controlled at 60-70 ° C for 36-72 hours.

Preferably, in the first scheme, the ratio of the liquid of the bovine calcined cancellous bone ore porous stent to the magnesium source-zinc source composite solution is 10g: 40-60mL, and the bovine calcined cancellous bone and bone porous support and the sulfur source-phosphorus source The solid solution ratio of the composite solution was 10 g: 50-100 mL.

Preferably, in the second embodiment, the ratio of the liquid of the bovine calcined cancellous bone mineral porous scaffold to the magnesium source-zinc source-sulfur source-phosphorus source complex solution is 10 g: 50-100 mL.

Preferably, the high temperature calcination parameter is calcined at 750-900 ° C for 2-9 hours.

Preferably, the magnesium source is magnesium sulfate; the zinc source is zinc nitrate; the sulfur source is a combination of sulfuric acid and soluble sulfate, the soluble sulfate is magnesium sulfate or a combination of magnesium sulfate and sodium sulfate; The phosphorus source is a combination of phosphoric acid or phosphoric acid and diammonium hydrogen phosphate.

Magnesium sulphate in the magnesium source is both a source of magnesium ions and a source of sulfur.

Preferably, the final concentration of magnesium ions in the quaternary system containing the magnesium source, the zinc source, the sulfur source and the phosphorus source is 0.05-0.2 mol/L, and the final concentration of zinc ions is 0.2-0.8 mol/L.

Preferably, the final concentration of the sulfuric acid in the quaternary system containing the magnesium source, the zinc source, the sulfur source and the phosphorus source is 0.1-0.2 mol/L, and the final concentration of the sulfate provided by the sulfate is 0.05-0.2 mol/L. The final concentration of phosphoric acid is 1.7-3.4 wt%, and the final phosphorus concentration provided by diammonium hydrogen phosphate is 0.1-0.8 mol/L. Sulfate contains all of the sulfates in the magnesium source and sulfur source.

Preferably, the calcium phosphate-zinc calcium phosphate-calcium porous composite bioscaffold has a material composition of one of the following composite components: calcium sulfate/zinc pyrophosphate/magnesium magnesium phosphate, calcium sulfate/zinc pyrophosphate / Magnesium calcium phosphate / hydroxyapatite, calcium sulfate / zinc phosphate / magnesium magnesium phosphate, calcium sulfate / magnesium zinc phosphate / magnesium pyrophosphate.

Preferably, the magnesium phosphate-calcium phosphate porous composite bioscaffold demagnetizes the molar percentage of magnesium ions to total cations in the range of 1-15%, and the molar percentage of zinc ions to the total cations is 10-85%.

Preferably, the bovine calcined cancellous bone mineral porous scaffold has a porosity of 70-85% and a pore diameter of 400-1400 μm.

Preferably, the calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite bioscaffold can be degraded by a gradient; the degradable calcium phosphate-calcium phosphate porous composite bioscaffold retains the calcined pine The three-dimensional interconnected mesh structure and mechanical strength of the porous bone ore bone support, and the whisker growth with large aspect ratio in the mesh, the whisker aspect ratio is 8-25:1, which can effectively increase the specific surface area of the material. .

Preferably, the preparation method of the bovine calcined cancellous bone ore porous scaffold is:

(1) cutting the bovine cancellous bone into a bone strip or a bone piece having a thickness of 0.5-1 cm to obtain a raw material bone;

(2) The raw material bone is placed in distilled water for cooking for 40-60 minutes in an autoclave, and then washed with drinking water at 40-60 ° C, and this step is repeated 4-6 times;

(3) The raw material bone treated in the step (2) is dried in a constant temperature oven at 80-120 ° C for 12-24 hours, then placed in a calcining furnace, calcined at 900-1200 ° C for 8-12 hours, and cooled to obtain a calcined pine. Porous bone mineral bone support.

The beneficial effects of the invention are:

The invention can stably and effectively convert the calcined cancellous porous hydroxyapatite elemental scaffold (bovine calcined cancellous bone and bone mineral porous scaffold) into a degradable magnesium-calcium-calcium phosphate-calcium sulfate composite scaffold material which is rich in conversion components. Such as calcium sulfate / zinc pyrophosphate / magnesium zinc phosphate, calcium sulfate / zinc pyrophosphate / calcium magnesium phosphate / hydroxyapatite, calcium sulfate / zinc phosphate / magnesium magnesium phosphate, calcium sulfate / magnesium zinc phosphate / magnesium pyrophosphate. The invention effectively incorporates osteogenic active ion magnesium ions, zinc ions, sulfate ions, phosphorus ions, etc.; magnesium-containing components are tricalcium phosphate, magnesium magnesium phosphate, magnesium pyrophosphate, and magnesium-containing components having good degradation characteristics. Material The total mass of the material is 9.5-80%, the molar ratio of magnesium ion content to total cation is 1.0-15%; the zinc-containing component is magnesium zinc phosphate, zinc pyrophosphate, tri-zinc phosphate, etc. with good degradation characteristics, and zinc-containing component 10-88% of the total mass of the material, the molar ratio of zinc ion content to total cation is 10-85%; the composite scaffold contains calcium sulfate, sulfuric acid which has a faster degradation rate, can be completely degraded and has potential osteoinductive active ions. Calcium accounts for 12-60% of the total mass of the composite; some composite scaffolds also contain hydroxyapatite.

The composite bio-scaffold material maintains the three-dimensional interpenetrating micro-structure of natural bone and bone and its good mechanical strength. At the same time, there is a bundle of larger aspect ratio whiskers in the mesh, and the whisker aspect ratio is 8-25. : 1, can effectively increase the specific surface area of the material to improve cell adhesion. Due to the large difference in the dissolution rate of the components, the composite bioscaffold material can achieve stepwise dissolution; and the composition and mass ratio of the composite bioscaffold material can be effectively regulated according to the formulation and the production process, as in the experiment. Under the condition that other conditions in the condition range are unchanged, the content of calcium sulfate can be gradually increased with the increase of sulfur in the solution; with the increase of phosphorus in the unit solution, the hydroxyapatite with a ratio of calcium to phosphorus of 1.67 Zinc pyrophosphate, magnesium pyrophosphate, which has a ratio of zinc, (calcium + magnesium), (zinc + magnesium) to phosphorus of 1.5, magnesium triphosphate, zinc phosphate, zinc triphosphate, zinc phosphate, magnesium phosphate Transformation, we can effectively regulate the composition and mass ratio of the scaffold components, so the dissolution rate of the composite bioscaffold material can be effectively regulated. Calcium sulfate degradation can form an early high calcium environment, containing magnesium, zinc degradable calcium phosphate such as magnesium tricalcium phosphate, magnesium zinc phosphate, zinc triphosphate and zinc pyrophosphate, magnesium pyrophosphate degradation sustainable release of osteogenic active ion magnesium , zinc and calcium ions, etc., are conducive to the formation of new bone and provide further space for bone repair; the material can maintain good mechanical strength and mesh structure after a large proportion of dissolution, these can be simulated in the body fluid dissolution test. It was confirmed that the dissolution and redeposition of the magnesium-containing, zinc-calcium phosphate-calcium sulfate composite scaffold can be seen under electron microscope. The calcined bovine cancellous bone mineral of the invention transforms the magnesium-containing, zinc-calcium phosphate-calcium sulfate composite scaffold material into the cancellous bone defect region of the animal, and the repaired cells can be well recruited, adhered, proliferated and differentiated, secreted matrix and rapidly The formation of vascular network, the material can achieve intra-membranous osteogenesis similar to physiological state, suggesting good bone conduction and potential osteoinductive activity of calcined bovine cancellous bone minerals transformed with magnesium-doped, calcium-calcium phosphate-calcium sulfate composite scaffold; No immunological rejection and obvious inflammation were found during the process, suggesting that the calcined bovine cancellous bone mineral was transformed into a magnesium-calcium-calcium-calcium phosphate-calcium sulfate composite scaffold with good biocompatibility.

In summary, the calcined bovine cancellous bone mineral is transformed into a calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite scaffold material which has good three-dimensional interconnected mesh structure and osteoconductivity, degradability, good mechanical strength and biocompatibility. At the same time, there is a large aspect ratio calcium sulfate whisker growth in the mesh, which can increase the specific surface area of the material to improve cell adhesion. The composite bioscaffold material may have potential osteoinductivity due to the effective incorporation of the osteogenic active ion magnesium, zinc and calcium sulfate which can produce a local high calcium environment upon degradation. The composite bioscaffold may more satisfy the ideal conditions for bone graft replacement materials or bone tissue engineering scaffold materials.

DRAWINGS

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagram showing the XRD analysis of a component of the product of the present invention.

Figure 2 is a scanning electron micrograph of one of the products of the present invention.

Figure 3 is a graph showing the calcium value of the product of the present invention in the early stage of simulated body fluid dissolution experiments (n = 3, human serum calcium ion reference value 2-2.67 mmol / l).

Figure 4 is a scanning electron micrograph of a material after simulating a body fluid dissolution test of the product of the present invention.

Figure 5 is an early histological illustration of a transplantation experiment of the product of the present invention.

Figure 6 is an early histological illustration of another transplantation experiment of the product of the present invention.

Figure 7 is a histological illustration of a later stage of a transplant experiment of the product of the present invention.

Best way to implement the invention

The technical solution of the present invention will be further specifically described below through specific embodiments.

In the present invention, the materials and equipment used, etc., are commercially available or commonly used in the art unless otherwise specified. The methods in the following examples, unless otherwise stated, are all conventional methods in the art.

Preparation Example 1 of Pork Calcined Cancellous Bone Mineral Porous Scaffold

(1) cutting the bovine cancellous bone (the bovine femur cancellous bone) into a 0.5 cm thick bone strip to obtain the raw bone;

(2) The raw material bone is placed in distilled water and cooked in a pressure cooker for 40 minutes, then washed with 40 ° C water, and this step is repeated 6 times;

(3) The raw material bone treated in the step (2) is dried in a constant temperature oven at 80 ° C for 24 hours, then placed in a calciner, calcined at 900 ° C (heating rate 10 ° C / min) for 12 hours, and then calcined with the furnace. Calcined cancellous bone ore porous stent.

Preparation Example 2 of Pork Calcined Cancellous Bone Mineral Porous Scaffold

(1) cutting the bovine cancellous bone (bovine femoral condyle cancellous bone) into a bone piece having a thickness of 1 cm to obtain a raw material bone;

(2) The raw material bone is placed in distilled water and cooked in a pressure cooker for 60 minutes, then washed with 60 ° C water, and this step is repeated 4 times;

(3) The raw material bone treated in the step (2) is dried in a constant temperature oven at 120 ° C for 12 hours, then placed in a calcining furnace, calcined at 1200 ° C (heating rate 10 ° C / min) for 8 hours, and then calcined with the furnace. Calcined cancellous bone ore porous stent.

Preparation Example 3 of Pork Calcined Cancellous Bone Mineral Porous Scaffold

(1) cutting the bovine cancellous bone (the bovine femur cancellous bone) into a 0.8 cm thick bone strip to obtain the raw bone;

(2) The raw bone is placed in distilled water and cooked in a pressure cooker for 50 minutes, then washed with 50 ° C water, and this step is repeated 5 times;

(3) The raw material bone treated in the step (2) is dried in a constant temperature oven at 100 ° C for 18 hours, then placed in a calcining furnace, calcined at 1000 ° C (heating rate 10 ° C / min) for 10 hours, and then calcined with the furnace. Calcined cancellous bone ore porous stent.

General Embodiment 1:

A: The final concentration of magnesium ions in the quaternary system containing the magnesium source, the zinc source, the sulfur source and the phosphorus source is 0.05-0.2 mol/L, and the final concentration of zinc ions is 0.2-0.8 mol/L; The concentration is 0.1-0.15mol/L, the final concentration of sulfate provided by sulfate is 0.05-0.3mol/L, the final concentration of phosphoric acid is 1.7-3.4wt%, and the final concentration of phosphorus provided by diammonium hydrogen phosphate is 0.1-0.8. Mol/L for ingredients.

B: Weigh zinc nitrate, magnesium sulfate first to prepare zinc nitrate, magnesium sulfate solution, bovine calcined cancellous bone and bone porous support and magnesium source-zinc source composite solution solid-liquid ratio of 10g: 40-60 ml; calcined cattle The porous bone ore porous stent is immersed in a magnesium sulfate solution, immersed for 15-30 minutes and then microwave dried: microwave output power 300-500w, time 15-24 minutes.

C: Prepare the sulfur source and the phosphorus source composite solution according to the solid-liquid ratio of the bovine calcined cancellous bone and bone porous support with the sulfur source and the phosphorus source composite solution of 10g: 50-100 ml, and the calcined cancellous bovine after the step B treatment The bone and bone mineral porous support is immersed and immersed and hydrothermally reacted. The hydrothermal reaction adopts a constant temperature hydrothermal reaction, and the reaction temperature is controlled at 60-70 ° C, and the reaction time is 36-72 hours.

D: Take out the porous support in a constant temperature oven and dry at 70-90 ° C for 24-48 hours.

E: The bovine calcined cancellous bone ore porous support after the treatment in step D is placed in a calciner, calcined at 750-900 ° C (heating rate 2.5 ° C / min) for 2-9 hours, and calcined pine after cooling with the furnace. Porous bone mineral porous scaffold conversion material.

General Embodiment 2:

A: The final concentration of magnesium ions in the quaternary system containing the magnesium source, the zinc source, the sulfur source and the phosphorus source is 0.05-0.2 mol/L, and the final concentration of zinc ions is 0.2-0.8 mol/L; The concentration is 0.1-0.15 mol/L, the final concentration of sulfate provided by sulfate is 0.05-0.3 mol/L, and the final concentration of phosphoric acid is 1.7-3.4 wt%. The final concentration of phosphorus provided by diammonium hydrogen phosphate is 0.1-0.8 mol/L.

B: According to the ratio of material to liquid of magnesium source-zinc source-sulfur source-phosphorus source composite solution of 10g:50-100mL, a magnesium source-zinc source-sulfur source-phosphorus source composite solution is prepared, and the bovine calcined cancellous bone ore is porous. The immersion impregnation and hydrothermal reaction of the support, the hydrothermal reaction takes a constant temperature hydrothermal reaction, the reaction temperature is controlled at 60-70 ° C, and the reaction time is 36-72 hours.

C: Take out the porous support in a constant temperature oven and dry at 70-90 ° C for 24-48 hours.

D: The bovine calcined cancellous bone ore porous support after the step C treatment is placed in a calciner, calcined at 750-900 ° C (heating rate 2.5 ° C / min) for 2-9 hours, and the calcined pine is obtained after cooling with the furnace. Porous bone mineral porous scaffold conversion material.

Each sample was subjected to general observation for X-ray diffraction (XRD) analysis; part of the sample was selected for observation by microscopy with scanning electron microscopy; animal experiments for simulating body fluid dissolution and bone defect repair were performed. The general shape and strength of the material were observed generally, and some samples were tested for compressive strength using INSTRON-5566. The simulated body fluid elution experiment uses medical sodium chloride injection as a simulated body fluid, and the solid-liquid mass ratio of the test material to the simulated body fluid is 1 gram: 100 ml, placed in a covered beaker, and simulated body fluid is carried out under the constant temperature of 37 ° C. In the dissolution test, 30 days after the dissolution test, the AU5800 automatic biochemical analyzer was used to measure the calcium, phosphorus and magnesium ions in the simulated body fluid every 3 days. The simulated body fluid was replaced 40% every 3 days before the dissolution. The simulated body fluid was not replaced in the later stage; the mass of the sample after 30 days of the dissolution test was measured by a domestic electronic balance and the degradation rate was calculated; XRD analysis and scanning electron microscope observation of the material before and after the start of the dissolution test were performed. Animal bone defect repair test selected 48 healthy New Zealand white rabbits, which resulted in bone defects of 8 mm in diameter in rabbit femoral condyles. They were randomly divided into experimental group (porous composite biomaterial) and control group (imported synthetic calcium phosphate material). The same artificial bone defects were made to the experimental group and the control group, respectively, and then the rabbits in the experimental group were treated with porous composite organisms. The bone defect was repaired by the material. The rabbits in the control group were treated with imported synthetic calcium phosphate bone substitute material for bone defect repair. The experimental animals were sacrificed at 1, 2, 4 and 8 weeks after operation to perform histological examination of bone defect repair.

Example 1

604153

According to the amount of 0.2 mol/L magnesium sulfate hexahydrate, 0.4 mol/L zinc nitrate hexahydrate, 3.4 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.4 mol/L diammonium hydrogen phosphate, 2.4 g of magnesium sulfate hexahydrate, Liushui 5.9 g of zinc nitrate, 2 ml of phosphoric acid (85 wt%), 5 ml of sulfuric acid, and 2.64 g of diammonium hydrogen phosphate. Prepare 50 ml of magnesium and zinc complex solution with 2.4 g of magnesium sulfate and 5.9 g of zinc nitrate, and immerse 10 g of bovine calcined cancellous bone and bone porous support, and dry it with microwave; take 5 ml of sulfuric acid, 2 ml of phosphoric acid, and phosphoric acid. Hydrogen diammonium 2.64 g was prepared into 50 ml of phosphorus and sulfur composite solution, and the bovine calcined cancellous bone and bone porous support was put into the 70 ° C reaction for 60 hours, and taken out and dried at 70 ° C for 48 hours; the temperature was raised from 2.5 ° C to 750 ° C per minute to maintain After 2 hours, the furnace was cooled to room temperature to obtain 604153A;

604153A

CaSO ₄ 43.3%

(Zn _0.73 Mg _0.27 ) ₃ (PO ₄ ) ₂ 46.1%

Mg ₂ (P ₂ O ₇ ) 10.6%.

Example 2

602154

According to 0.1 mol/L magnesium sulfate hexahydrate, 0.6 mol/L zinc nitrate hexahydrate, 1.7 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.4 mol/L diammonium hydrogen phosphate, 1.2 g of magnesium sulfate hexahydrate, Liushui 8.93 g of zinc nitrate, 1 ml of phosphoric acid, 5 ml of sulfuric acid, and 2.64 g of diammonium hydrogen phosphate. First use 1.2 g of magnesium sulfate, 5.9 g of zinc nitrate Prepare 50 ml of magnesium and zinc composite solution, immerse 10 g of bovine calcined cancellous bone and bone porous support, and dry it with microwave; 5 ml of sulfuric acid, 1 ml of phosphoric acid and 2.64 g of diammonium phosphate to prepare phosphorus and sulfur composite solution 50 ML, constant temperature 70 ° C reaction for 60 hours, after removal, 70 ° C drying for 48 hours; temperature rise 2.5 ° C to 750 ° C per minute, after 2 hours, with the furnace down to room temperature to get 602154A;

602154A

CaSO ₄ 18.1%

Zn ₂ (P ₂ O ₇ ) 21.7%

(Zn _0.73 Mg _0.27 ) ₃ (PO ₄ ) ₂ 60.2%.

Example 3

602155

1.2 g of magnesium sulfate hexahydrate, Liushui, in an amount of 0.1 mol/L magnesium sulfate hexahydrate, 0.5 mol/L zinc nitrate hexahydrate, 1.7 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.4 mol/L diammonium hydrogen phosphate. 6.94 g of zinc nitrate, 1 ml of phosphoric acid, 5 ml of sulfuric acid, and 2.64 g of diammonium hydrogen phosphate. Prepare 50 ml of magnesium and zinc complex solution with 1.2 g of magnesium sulfate and 8.93 g of zinc nitrate, and immerse 5 g of bovine calcined cancellous bone and bone porous support, and dry it with microwave; 5 ml of sulfuric acid, 1 ml of phosphoric acid, hydrogen phosphate Diammonium 2.64 g preparation of 50 ml of phosphorus and sulfur composite solution, reacted at 70 ° C for 60 hours, and dried at 70 ° C for 48 hours; heated at 2.5 ° C to 750 ° C per minute for 2 hours and then cooled to room temperature to obtain 602155 A;

602155A

CaSO ₄ 37.8%

(Zn _0.73 Mg _0.27 ) ₃ (PO ₄ ) ₂ 46.1%

Zn ₂ (P ₂ O ₇ ) 14.6%.

Example 4

604156

According to the amount of 0.05 mol/L magnesium sulfate hexahydrate, 0.6 mol/L zinc nitrate hexahydrate, 1.7 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.6 mol/L diammonium hydrogen phosphate, 0.6 g of magnesium sulfate hexahydrate, Liushui 8.93 g of zinc nitrate, 1 ml of phosphoric acid, 5 ml of sulfuric acid, and 3.96 g of diammonium hydrogen phosphate. Prepare 50 ml of magnesium and zinc complex solution with 0.6 g of magnesium sulfate and 8.93 g of zinc nitrate, and immerse 5 g of bovine calcined cancellous bone and bone porous support, and dry it with microwave; use 5 ml of sulfuric acid, 1 ml of phosphoric acid, phosphoric acid Hydrogen diammonium (3.96 g) prepared sulfur and phosphorus complex solution 50 ml, constant temperature 70 ° C reaction 60 hours, 70 ° C drying 48 hours after removal; temperature 2.5 ° C to 750 ° C per minute, maintained 2 hours after the furnace down to room temperature to obtain 604156A;

604156A

CaSO ₄ 12.4%

Zn ₂ (P ₂ O ₇ ) 22.5%

(Zn _0.73 Mg _0.27 ) ₃ (PO ₄ ) ₂ 55.1%.

Example 5

604157

According to the amount of 0.05 mol/L magnesium sulfate hexahydrate, 0.8 mol/L zinc nitrate hexahydrate, 1.7 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.8 mol/L diammonium hydrogen phosphate, 0.6 g of magnesium sulfate hexahydrate, Liushui 8.93 g of zinc nitrate, 1 ml of phosphoric acid, 5 ml of sulfuric acid, and 5.28 g of diammonium hydrogen phosphate. Prepare 50 ml of magnesium and zinc complex solution with 0.6 g of magnesium sulfate and 8.93 g of zinc nitrate, and immerse 10 g of bovine calcined cancellous bone and bone porous support, and dry it with microwave; use 5 ml of sulfuric acid, 1 ml of phosphoric acid, phosphoric acid 5.28 g of diammonium hydroxide was prepared into 50 ml of sulfur and phosphorus complex solution, reacted at 70 ° C for 60 hours, and dried at 70 ° C for 48 hours after taking out; the temperature was raised from 2.5 ° C to 750 ° C per minute, and after maintaining for 2 hours, the furnace was cooled to room temperature to obtain 604157 A;

604157A

CaSO ₄ 11.8%

Zn ₂ (P ₂ O ₇ ) 46.3%

(Zn _0.73 Mg _0.27 ) ₃ (PO ₄ ) ₂ 41.9%.

Example 6

601154

According to 0.05 mol/L magnesium sulfate hexahydrate, 0.2 mol/L zinc nitrate, 1.7 w% phosphoric acid, 0.15 mol/L sulfuric acid, 0.6 g of magnesium sulfate hexahydrate, 2.97 g of zinc nitrate hexahydrate, 1 ml of phosphoric acid, 5 ml of sulfuric acid . First, 0.6 g of magnesium sulfate hexahydrate and 2.97 g of zinc nitrate were prepared, and 50 ml of a magnesium and zinc composite solution was prepared, and 5 g of the porous scaffold of the calcined cancellous bone and bone was impregnated and microwave dried; 5 ml of 1 mol/L sulfuric acid was used. Prepare 50 ml of sulfur and phosphorus complex solution in 1 ml of phosphoric acid, put the bovine calcined cancellous bone and bone porous support into it, and heat the reaction at a constant temperature of 60 ° C for 72 hours; increase the temperature by 2.5 ° C to 750 ° C per minute, and keep it for 2 hours. 601154A to room temperature;

601154A

Example 7

605093

According to 0.05 mol/L magnesium sulfate hexahydrate, 0.3 mol/L zinc nitrate, 1.7 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.6 g of magnesium sulfate hexahydrate, 4.46 g of zinc nitrate, 1 ml of phosphoric acid, and 5 ml of sulfuric acid were taken. Take 0.6 g of magnesium sulfate and 4.46 of zinc nitrate to prepare 50 ml of magnesium and zinc, and immerse 5 g of bovine calcined cancellous bone and bone porous support with a porosity of about 85%, and dry it with microwave; use 5 ml of sulfuric acid and 1.5 ml of phosphoric acid. 2.64 g of diammonium hydrogen phosphate was prepared into 50 ml of sulfur and phosphorus complex solution. The bovine calcined cancellous bone and bone porous support was put into the 70 ° C reaction for 60 hours, and after removal, it was dried at 90 ° C for 20 hours and weighed 6.21 g; the temperature was raised by 2.5 ° C per minute. 750 ° C, after 2 hours, the furnace was cooled to room temperature to obtain 605093;

605093

Example 8

605095

According to 0.05 mol/L magnesium sulfate hexahydrate, 0.2 mol/L zinc nitrate, 2.55 wt% phosphoric acid, 0.1 mol/L sulfuric acid, 0.4 diammonium phosphate, 0.6 g of magnesium sulfate hexahydrate, 2.97 g of zinc nitrate, 1.5 ml of phosphoric acid. 5 ml of sulfuric acid and 2.64 g of diammonium hydrogen phosphate. Prepare 50 ml of magnesium and zinc complex solution by taking 0.6 g of magnesium sulfate and 2.97 g of zinc nitrate, and immerse 5 g of bovine calcined cancellous bone and bone porous support with a porosity of about 85%, microwave drying; 5 ml of sulfuric acid, 1.5 with phosphoric acid Prepare 50 ml of sulfur and phosphorus complex solution in milliliters and diammonium hydrogen phosphate 2.64 g. Put the bovine calcined cancellous bone and bone porous support into the 70 ° C reaction for 60 hours, remove and then dry at 90 ° C for 20 hours and weigh 6.14 g; °C to 750 °C, maintain 2 After the hour, the furnace is cooled to room temperature to obtain 605095;

605095

Test results:

Material observation, strength measurement, XRD component analysis and scanning electron microscopy observation of various products intact to maintain the prefabricated morphology of bovine cancellous bone, without fragmentation, collapse or powdering, etc., with good mechanical strength; 10 × 10 × 10mm The compressive strength of the No. 1-5 specimens tested with INSTRON-5566 cancellous bone is shown in Table 1. X-ray diffraction (XRD) confirmed that the calcined bovine pine ore suspension impregnated with magnesium and zinc was hydrothermally reacted with the sulfur source-phosphorus source composite solution. After drying, it was converted into magnesium-doped and zinc-degradable apatite-sulfuric acid by calcination. Calcium porous composite bioscaffold materials such as calcium sulfate / zinc pyrophosphate / magnesium zinc phosphate, calcium sulfate / zinc pyrophosphate / magnesium magnesium phosphate / hydroxyapatite, calcium sulfate / zinc phosphate / magnesium magnesium phosphate, calcium sulfate / magnesium zinc phosphate A composite such as /magnesium pyrophosphate is shown in Fig. 1. The magnesium-containing component is tricalcium phosphate, magnesium magnesium phosphate, magnesium pyrophosphate, etc., and the magnesium component accounts for 9.5-80% of the total mass of the material, and the molar ratio of magnesium ion content to total cation is 1.0-15. : 100; the zinc-containing component is magnesium zinc phosphate, zinc pyrophosphate, tri-zinc phosphate, etc. having good degradation characteristics, the zinc-containing component accounts for 10-88% of the total mass of the material, and the molar ratio of zinc ion content to total cation is 10 -85:100; composite scaffold contains calcium sulfate with faster degradation rate, complete degradation and potential osteoinductive active ions, calcium sulfate accounts for 12-60% of the total mass of the composite; some composite scaffolds also contain hydroxyapatite . Scanning by electron microscopy (see Figure 2), the product maintains the cow The main structure of the three-dimensional interconnected mesh microstructure of the cancellous bone natural bone and bone, while the calcium sulfate whisker with larger aspect ratio in the mesh grows, which can increase the specific surface area of the material and possibly improve cell adhesion.

Table 1 compressive strength

Numbering Compressive strength (MPa) 1 12.41 2 9.38 3 5.96 4 8.71 5 10.5

2. In vitro dissolution experiments of materials

The simulated body fluid elution experiment was carried out by using medical sodium chloride injection as a simulated body fluid. The solid-liquid mass ratio of the test material to the simulated body fluid was 1-2 g: 100 ml, placed in a covered beaker, and simulated at 37 ° C under constant temperature. The body fluid dissolution test was carried out for 4 weeks. The AU5800 automatic biochemical analyzer was used to measure the calcium, phosphorus and magnesium ions in the simulated body fluid. The mass of the material was measured by the domestic electronic balance for 4 weeks and the degradation rate was calculated. XRD analysis and scanning electron microscopy of the material before and at the end of the experiment. The experiment found that the material has a good degradation rate (Table 2). In the early (half-month) simulation of the body fluid experiment, most samples have a higher calcium ion concentration in the simulated body fluid, which is maintained at the normal reference value of human serum (2-2.67mmol/l). Between 1-4 times the median value (Fig. 3); there are also active ion magnesium and zinc release. At the end of the dissolution test, the mesh of the stent became large and the mechanical strength was good (Table 3). XRD analysis of the simulated body fluid test materials revealed that the composition and mass ratio of the material changed with time, and the calcium sulfate and magnesium-containing, zinc-phosphorus apatite components gradually decreased or disappeared, and the scaffold material gradually changed to hydroxyapatite. Scanning electron microscopy revealed material dissolution and redeposition of mineral components (Figure 4).

Table 2: Degradation rate of materials immersed in simulated body fluid for 4 weeks (n=3)

Grouping Degradation rate Hydroxyapatite 1.56% Product A of the invention 35.67% Product B of the invention 29.63% Product of the invention C 25.32% Product of the invention D 42.38%

Table 3 Compressive strength after dissolution test

3. Animal bone defect repair test

In the early stage of transplantation of the porous composite biomaterial group (1 week), the cells and blood vessels can be seen in the entire space of the scaffold, and the repaired cells can proliferate, differentiate, and secrete the bone matrix (Fig. 5, Fig. 6); trabecular bone formation occurs in two weeks. New bone tissue is perfectly integrated with the scaffold (Figure 7). No immunological rejection and obvious inflammation were observed during the observation, and the material had good biocompatibility.

The above-mentioned embodiments are only a preferred embodiment of the present invention, and are not intended to limit the present invention in any way, and other variations and modifications are possible without departing from the technical solutions described in the claims.

Claims (14)

A calcium phosphate-calcium sulfate porous composite biological scaffold capable of degrading magnesium and zinc, characterized in that: a porous scaffold containing a magnesium source, a zinc source, a sulfur source and a phosphorus source is passed through a porous scaffold of a calcined cancellous bone ore. The system is treated, taken out and dried, and calcined at a high temperature. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1, wherein the bovine calcined cancellous bone ore porous scaffold comprises a magnesium source, a zinc source and sulfur. The choice of source and phosphorus source for the quaternary system treatment is one of the following options: Scheme 1: The porous scaffold of the calcined cancellous bone ore is first immersed in the magnesium source-zinc source composite solution and evaporated to dryness, and then enters the hydrothermal reaction in the sulfur source-phosphorus source composite solution; Scheme 2: The hydrothermal reaction of the porous scaffold of the calcined cancellous bone and bone mineral in the magnesium source-zinc source-sulfur source-phosphorus source composite solution. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 2, wherein the hydrothermal reaction adopts a constant temperature hydrothermal method to control the temperature of 60-70 ° C, time 36 -72 hours. The calcium phosphate-calcium phosphate porous composite biological scaffold capable of degrading magnesium and zinc according to claim 2, wherein in the first scheme, the bovine calcined cancellous bone and bone porous support is combined with the magnesium source-zinc source. The ratio of the solution to the solution is 10g: 40-60mL, and the ratio of the liquid mixture of the bovine calcined cancellous bone mineral porous support and the sulfur source-phosphorus source composite solution is 10g: 50-100mL. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 2, wherein in the second embodiment, the bovine calcined cancellous bone and bone porous support and the magnesium source-zinc source- The solid-liquid ratio of the sulfur source-phosphorus source composite solution is 10 g: 50-100 mL. A porous composite of calcium phosphate-calcium phosphate degrading magnesium and zinc according to claim 1 or 2 The bioscaffold is characterized in that the high temperature calcination parameter is calcined at 750-900 ° C for 2-9 hours. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the magnesium source is magnesium sulfate; the zinc source is zinc nitrate; The sulfur source is a combination of sulfuric acid and soluble sulfate, which is magnesium sulfate or a combination of magnesium sulfate and sodium sulfate; the phosphorus source is a combination of phosphoric acid or phosphoric acid and diammonium hydrogen phosphate. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 7, wherein the magnesium in the quaternary system containing a magnesium source, a zinc source, a sulfur source and a phosphorus source The final concentration of ions is 0.05-0.2 mol/L, and the final concentration of zinc ions is 0.2-0.8 mol/L. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 7, characterized in that the sulfuric acid in the quaternary system containing a magnesium source, a zinc source, a sulfur source and a phosphorus source The final concentration is 0.1-0.15mol/L, the final concentration of sulfate provided by sulfate is 0.05-0.3mol/L, the final concentration of phosphoric acid is 1.7-3.4wt%, and the final concentration of phosphorus provided by diammonium hydrogen phosphate is 0.1- 0.8 mol/L. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold is degradable The material composition is one of the following composite components: calcium sulfate/magnesium magnesium phosphate/magnesium pyrophosphate, calcium sulfate/zinc pyrophosphate/magnesium magnesium phosphate, calcium sulfate/zinc phosphate/magnesium phosphate/hydroxyapatite, sulfuric acid Calcium / zinc phosphate / magnesium magnesium phosphate, calcium sulfate / zinc pyrophosphate / calcium magnesium phosphate / hydroxyapatite, calcium sulfate / zinc pyrophosphate / magnesium magnesium phosphate. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold is degradable The molar percentage of medium magnesium ions to total cations is 1-15%, and the molar percentage of zinc ions to total cations is 10-85%. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the porous calcined cancellous bone ore porous scaffold has a porosity of 70-85%. , aperture 400-1400μm. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold is degradable The material can be degraded by gradient; the calcium phosphate-zinc calcium phosphate-calcium porous composite bioscaffold can maintain the three-dimensional interpenetrating mesh structure and mechanical strength of the bovine calcined cancellous bone ore porous scaffold, and the mesh has Larger aspect ratio whiskers grow with a whisker aspect ratio of 8-25:1, which effectively increases the specific surface area of the material. The calcium phosphate-zinc calcium phosphate-calcium sulfate porous composite biological scaffold according to claim 1 or 2, wherein the preparation method of the bovine calcined cancellous bone ore porous scaffold is: (1) cutting the bovine cancellous bone into a bone strip or a bone piece having a thickness of 0.5-1 cm to obtain a raw material bone; (2) The raw material bone is placed in distilled water for cooking for 40-60 minutes in an autoclave, and then washed with drinking water at 40-60 ° C, and this step is repeated 4-6 times; (3) The raw material bone treated in the step (2) is dried in a constant temperature oven at 80-120 ° C for 12-24 hours, then placed in a calcining furnace, calcined at 900-1200 ° C for 8-12 hours, and cooled to obtain a calcined pine. Porous bone mineral bone support.

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