A through-hole bone repair material and a preparation method thereof

Abstract

The application belongs to the field of bone repair materials, and particularly relates to a bone repair material with a through-hole structure and a preparation method thereof. The application provides a preparation method of a through-hole bone repair material, and the preparation method comprises the following steps: 1) preparing a bone repair raw material slurry: uniformly mixing raw materials of the bone repair material with water to form a bone repair raw material slurry; 2) preparing a bone repair preform; and 3) sintering treatment. The through-hole composite bone repair / bone reconstruction material provided by the application has a loofah sponge reverse structure, contains magnesium hydrogen phosphate with good solubility and degradability, calcium hydrogen phosphate capable of slowly degrading, and calcium sulfate capable of rapidly degrading and absorbing, has osteogenic elements calcium and phosphorus and a magnesium element capable of promoting bone tissue formation, can gradually degrade from a side in contact with bone tissue, and can provide components in the whole process of repairing and reconstructing bone tissue from a wound surface, promote tissue repair and reconstruction, and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of bone repair materials, in particular to a bone repair material with through-hole structure and a preparation method thereof. BACKGROUND

[0002] Bone tissue is the support system of the human body, bearing the core system of human weight, movement and activity. Bone is divided into compact bone and cancellous bone. Compact bone is formed by closely arranged bone plates, which are formed by parallel arrangement of bone collagen fibers embedded in calcified matrix, with uniform thickness. Between the two bone plates, there is a series of regularly arranged lacunae, which have bone cells with multiple protrusions connected to each other by capillaries. The bone plates arranged on the bone surface are the outer ring bone plates, and the bone plates arranged around the bone marrow cavity are the inner ring bone plates. Between the inner and outer ring bone plates, there are many concentrically arranged Harris bone plates, with the central tube being the Haversian canal, which is parallel to the long axis of the bone and has branches connected into a network, with blood vessels and nerves passing through the tube. Cancellous bone is a reticular structure formed by bone plates with many large voids, with bone marrow in the reticular pores. Cancellous bone exists in the femoral end of long bone, short bone and the interior of irregular bone. Bone tissue is the main component of various bones that make up the skeletal system. The skeleton is the support of the body, protecting soft organs, with muscles attached to it, acting as a lever for the movement organs. The functions of bone tissue in the human body are (1) support and protect the body; (2) concentrate 99% of the body's calcium, which is an organ for maintaining blood calcium balance, and appropriate blood calcium concentration is necessary for the normal functioning of the heart; (3) hematopoietic function, with a large number of bone marrow stem cells in the bone marrow that can be induced to differentiate into various blood cells into the blood.

[0003] Therefore, protecting the stability of bone tissue structure is one of the most important factors to ensure human health. However, its structure is caused by diseases, developmental failure, accidents and aging, etc. resulting in necrosis, damage and loss of function of bone tissue, which requires a large amount of biological materials as repair and replacement of bone tissue. Bone tissue is the basic system that supports and maintains the normal physiological functions of the human body.

[0004] According to the structure and function requirements of bone tissue, the repair material matching the bone tissue should have the same or similar structure as the original bone tissue, so that blood vessels can quickly grow into it, tissue fluid can diffuse into it, and bone cells can grow inside the material, gradually completing the repair and reconstruction of bone tissue. Therefore, the bone repair material should have through pores, matching porosity and relatively matching pore size, so as to meet the growth of blood vessels, the diffusion of tissue and the growth and proliferation of cells inside the material, forming bone tissue. According to this requirement, the pore through rate of porous sponge, physically foamed and chemically foamed materials is relatively low, which is difficult to match the structure of bone tissue, and the repair effect is limited. SUMMARY

[0005] Based on the aforementioned technical deficiencies, this invention uses natural loofah sponge as a template. By selecting multi-component composite materials and forming a composite material with a loofah sponge structure, and then removing the loofah sponge from the composite material through ablation thermal decomposition, a bone tissue repair material with fully interconnected pores is finally formed. This invention can also control the pore size by processing and controlling the diameter of the loofah sponge fibers, and control the porosity and density by using the amount of loofah sponge in the calcium-phosphorus composite material and the molding strength. This prepares a bone repair material with fully interconnected pores and adjustable pore size and porosity, providing sufficient channels for blood, blood vessels and tissue fluid, and promoting the repair and reconstruction of bone tissue.

[0006] The technical solution of this invention:

[0007] The first technical problem to be solved by the present invention is to provide a method for preparing a porous bone repair material, the method comprising the following steps:

[0008] 1) Preparation of bone repair material slurry: Mix the raw materials of bone repair material with water to form bone repair material slurry;

[0009] 2) Preparation of bone repair preform: The bone repair raw material slurry is placed in a loofah sponge and fully immersed to fill the loofah sponge with the bone repair slurry, and then allowed to solidify to obtain the bone repair preform;

[0010] 3) Sintering treatment: The bone repair preform is sintered to remove the loofah sponge, thus obtaining the bone repair material with a through-pore structure.

[0011] Further, in step 1), the raw materials of the bone repair material include the following components in the following proportions: 35-55 parts by weight of calcium-phosphorus-magnesium complex salt, 25-55 parts by weight of calcium sulfate hemihydrate, and 0-5 parts by weight of magnesium pyrophosphate (preferably 1-5 parts by weight); wherein, the molar ratio of magnesium to calcium in the calcium-phosphorus-magnesium complex salt is 0.1-0.5:1.

[0012] Furthermore, in step 1), the ratio of the raw material of the bone repair material to water is 1:0.5-1.5 (g / mL).

[0013] Further, in step 1), the calcium-phosphorus-magnesium complex salt is prepared by the following method: magnesium salt, calcium salt and phosphorus-containing substance are mixed and ground finely, then water is added to make a paste, and then vacuum dried to obtain the calcium-phosphorus-magnesium complex salt; wherein, the phosphorus-containing substance is at least one of calcium dihydrogen phosphate (Ca(H2PO4)2), adenosine triphosphate (ATP) or cyclic adenosine monophosphate (cAMP).

[0014] Furthermore, the magnesium salt is at least one of MgO or MgCO3, and the calcium salt is at least one of CaO or CaCO3.

[0015] Furthermore, in step 1), the bone repair material slurry is prepared using the following method:

[0016] (1) Take 40-50 g of magnesium oxide, 50-60 g of calcium oxide, 450-470 g of calcium phosphate, mix and grind to 200 mesh, add 100-150 ml of deionized water, stir for 25-35 minutes, mix evenly, dry at 70-80°C for 5-10 hours, form calcium magnesium phosphorus compound 1;

[0017] Take 20-40 g of magnesium oxide, 14-28 g of calcium oxide, 493-553 g of phosphorus-containing material, mix and grind to 200 mesh, add 140-160 ml of deionized water, stir for 25-35 minutes, mix evenly, dry at 70-80°C for 5-10 hours, form calcium magnesium phosphorus compound 2;

[0018] Take 20-40 g of magnesium oxide, 14-28 g of calcium oxide, 493-553 g of phosphorus-containing material, mix and grind to 200 mesh, add 140-160 ml of deionized water, stir for 25-35 minutes, mix evenly, dry at 70-80°C for 5-10 hours, form calcium magnesium phosphorus compound 2;

[0019] 2) Take 450-550 g of calcium sulfate dihydrate and hydrothermally treat it at 110-130°C and 0.1-0.5 MPa for 5-10 hours to form calcium sulfate hemihydrate, grind to 200 mesh or more as component B;

[0020] 3) Take magnesium pyrophosphate and grind to 200 mesh or more to form component C;

[0021] 4) Mix 200-210 g of component A, 100-120 g of component B, and 5-10 g of component C, add 150-250 ml of deionized water to make a slurry, and form a bone repair raw material slurry.

[0022] Further, in step 2), the loofah is pretreated before use, so that the fiber diameter of the loofah is 200-1000 μm.

[0023] Further, in step 2), the pretreatment method is: first, according to the selected loofah, the loofah is cut into a regular cube with a length of 10-200 mm, a width of 10-50 mm, and a height of 0.5-20 mm; then soaked in a treatment solvent, and then dried for standby; the treatment solvent is selected from: sodium hydroxide-hydrogen peroxide solution, lithium chloride-dimethylacetamide solution, lithium bromide-dimethylacetamide solution, lithium chloride-dimethylformamide solution, lithium bromide-dimethylformamide solution or copper ammonia solution; the soaking treatment makes the fiber diameter of the loofah decrease, and the loofah becomes finer.

[0024] Further, the method for preparing the bone repair preform in step 2) is as follows: placing the pretreated loofah sponge in a metal mold, pouring the bone repair raw material slurry obtained in step 1) into the loofah sponge in the mold, controlling the density and strength of the loofah sponge cuboid after pouring the slurry according to the viscosity of the slurry by using different pressures, and adjusting the solidification time in the mold according to the change of the composition of the slurry to form a loofah sponge cuboid filled with the slurry; after complete solidification, the bone repair preform is prepared, and the mold is opened to take out the bone repair preform for standby.

[0025] Further, in step 3), the process of the sintering treatment is as follows: placing the bone repair preform into a muffle furnace, gradually increasing the temperature to 100-120℃ for 30-45 minutes, then increasing the temperature to 200-250℃ for 30-60 minutes, and then increasing the temperature to 500-550℃ for 5-12 hours. In this process, the loofah sponge has been decomposed at high temperature and removed from the preform in the form of carbon dioxide and water. The space left after the decomposition of the loofah sponge fibers forms fiber channels similar to the original loofah sponge, and the final product is a through composite bone repair and reconstruction material with a reversed loofah sponge structure.

[0026] Further, the method for preparing the through-hole bone repair material further comprises step 4): post-treatment of the through-hole bone repair material according to clinical requirements, such as cutting and polishing the shape and size, washing with water for injection, air-drying, using radiation sterilization or epoxy sterilization, and finally packaging for standby.

[0027] The second technical problem to be solved by the present application is to provide a through-hole bone repair material prepared by the above preparation method.

[0028] The present application has the following advantages:

[0029] The through composite bone repair / bone reconstruction material with a reversed loofah sponge structure prepared by the present application contains magnesium hydrogen phosphate with good solubility and degradability, calcium hydrogen phosphate capable of slow degradation, and calcium sulfate capable of rapid degradation and absorption after pyrolysis at 550℃, has osteogenic elements calcium, phosphorus and magnesium elements that promote bone tissue formation, can gradually degrade from the side in contact with the bone tissue, provide components in the whole process of repair and reconstruction of the bone tissue from the wound surface, and promote tissue repair and reconstruction. In addition, the obtained material has full-through pores after the decomposition of the loofah sponge fibers, can provide sufficient space for tissue fluid and blood vessels to grow, can make the formed new bone through, can quickly form continuous through bone tissue, and can accelerate the degradation and absorption of the material. Moreover, the degradation rate can be controlled according to the composition and matched with bone tissue regeneration and reconstruction. The material can be widely used in the repair and reconstruction of bone trauma and defects. DETAILED DESCRIPTION

[0030] The first step of the application selects luffa sponge as a template. Luffa sponge is a filamentous vascular bundle interwoven into a long prism or a long cylindrical shape, slightly curved, 30-70 cm long, 7-10 cm in diameter. The surface is light yellow white. Light in weight, tough in quality, elastic, cannot be broken. The cross section can see 3 chambers of the ovary, hollow, showing a relatively regular three-dimensional structure. The diameter of the luffa sponge fiber bundle is generally 2000-5000 microns, and the diameter can be from the thicker bundle fiber (2000 microns), generally tubular, not consistent with the pore size required by capillary blood vessels and bone cells. The luffa sponge obtained from the luffa is cleaned and naturally dried. Then according to the luffa sponge network, it is cut into a regular cube with a length of 10-200 mm, a width of 10-50 mm, and a height of 0.5-20 mm; then the fiber diameter of the luffa sponge is reduced by using a pretreatment solvent to obtain fine and finer luffa sponge. Then, dry for standby. The application changes the luffa sponge fiber bundle into a diameter of 200-1000 microns, which can adopt three schemes: (1) using alkali, hydrogen peroxide combined treatment method to pretreat the natural luffa sponge fiber, using 5-15% strong sodium oxide solution, 10-15% hydrogen peroxide solution mixture. Soak the luffa sponge in the mixed solution, shake at 37℃ for 15-90 minutes, and use deionized water to ultrasonic clean three times for standby; (2) using lithium chloride dimethyl acetamide solution, lithium chloride dimethyl formamide solution, lithium bromide dimethyl acetamide solution, lithium bromide dimethyl formamide solution. 50-100g of anhydrous lithium chloride (60-120g of bromide) dried at 150℃ for 6 hours is added to 1000ml of dimethyl acetamide (dimethyl formamide), ultrasonic dispersion is used for 15-30 minutes, a transparent solution is formed, the luffa sponge is soaked in the mixed solution, and is shaken at 37℃ for 5-30 minutes. Use deionized water to ultrasonic clean three times for standby; (3) use copper ammonia solution to treat luffa sponge fiber bundle, add 1L, 0.2mol·L of copper sulfate solution in the container, then add 8mol / L of ammonia water, then add ammonia water until the precipitate is completely dissolved, at this time, a deep blue solution containing copper ammonia complex ions is obtained, the end point of the copper ammonia solution preparation is marked by the complete disappearance of the precipitate and the transparent blue solution. Soak the luffa sponge in the copper ammonia solution, shake at 37℃ for 5-30 minutes, and use deionized water to ultrasonic clean three times for standby.

[0031] The second step of the present application selects calcium sulfate hemihydrate, magnesium oxide, magnesium carbonate, calcium oxide, calcium carbonate, adenosine triphosphate, cyclic adenosine monophosphate, calcium dihydrogen phosphate or magnesium pyrophosphate as raw materials, as the main component of the preparation of porous bone repair material. Calcium sulfate is a necessary component, magnesium oxide and magnesium carbonate are selected or used together, calcium oxide and calcium carbonate are selected or used together, calcium dihydrogen phosphate, adenosine triphosphate and cyclic adenosine monophosphate are selected or used together, and finely ground (200 mesh and above) magnesium pyrophosphate is used as an optional item. The present application mixes magnesium oxide / magnesium carbonate / calcium oxide / calcium carbonate with calcium dihydrogen phosphate or adenosine triphosphate and cyclic adenosine monophosphate in a certain proportion, grinds them (200 mesh and above), then adds deionized water to make a paste, dries it at 0-20℃ under vacuum for 8-15 hours, grinds it (200 mesh and above) to form component (A); calcium sulfate dihydrate is hydrothermally treated at 120℃, 0.4MPa steam, and then grinded (200 mesh and above) to form calcium sulfate hemihydrate (B), then A and B are mixed in a certain proportion, deionized water is added, magnesium pyrophosphate (C) is added, and the slurry (D) is prepared for use. The composition of the composite D is adjusted according to the proportion of A, B and C, and the D is grouted into the loofah sponge to form the required shape and size, and then it is solidified (the slurry D is solidified within 20-45 minutes according to the proportion of A, B and C and the amount used) to prepare a preform.

[0032] Magnesium oxide / magnesium carbonate / calcium oxide / calcium carbonate and calcium dihydrogen phosphate or adenosine triphosphate and cyclic adenosine monophosphate undergo the following component changes in the presence of water, and the following is a schematic diagram of the changes (not a complete chemical reaction equation):

[0033]

[0034] Among them, MgO can be replaced by CaO, CaCO3, MgCO3. Calcium dihydrogen phosphate (Ca(H2PO4)2 can be replaced by adenosine triphosphate (ATP) or cyclic adenosine monophosphate (cAMP), and the products formed in the presence of water are different (but they will tend to be the same after high-temperature calcination at 550℃):

[0035]

[0036] Adenosine triphosphate can be replaced by cyclic adenosine monophosphate (cAMP):

[0037]

[0038] Among them, is an intermediate product, which can enhance the elasticity and forming performance of the composite during the solidification process.

[0039] The ratio of magnesium oxide or magnesium carbonate to calcium dihydrogen phosphate is (mol): 0.5-1.0:1.0; the ratio of calcium oxide or calcium carbonate to calcium dihydrogen phosphate is (mol): 0.5-1.0:1.0; the ratio of magnesium oxide or magnesium carbonate to adenosine triphosphate (ATP) is (mol) 1.0-2.0:1.0; the ratio of calcium oxide or calcium carbonate to adenosine triphosphate (ATP) is (mol) 1.0-2.0:1.0; the ratio of magnesium oxide or magnesium carbonate to cyclic adenosine monophosphate (cAMP) is (mol) 0.5-1.5:1.0; the ratio of calcium oxide or calcium carbonate to cyclic adenosine monophosphate (cAMP) is (mol) 0.5-1.5:1.0; magnesium oxide, magnesium carbonate, calcium oxide, and calcium carbonate can be used alone or in combination, but the total ratio of magnesium and calcium (mol) is (Mg:Ca) 0.1-0.5:1.0. Calcium dihydrogen phosphate, adenosine triphosphate, and cyclic adenosine monophosphate are selected from one or three or used in combination, and the ratio follows the magnesium-calcium ratio (Mg:Ca) 0.1-0.5:1.0.

[0040] The third step of the present application is to place the pretreated luffa sponge in a metal mold, pour the prepared slurry D into the luffa sponge in the mold, and immerse the luffa sponge in the slurry D, with the height of the slurry D being 1-5 mm higher than the luffa sponge. A mold cover is used to press and compact the slurry D, with a pressure of 2-10 MPa, and the mold is opened after 20-45 minutes to take out the luffa sponge filled with slurry D, forming a solidified solid cuboid. According to the thickness of the dense layer between the upper layer of the cuboid and the luffa sponge, the surface is polished to keep the pure composite D thickness layer at 0.0-1.0 mm, and 0.0 mm is exactly the luffa sponge exposed. After grouting, molding, and shaping, the ABC composite filled dense cuboid luffa sponge preformed product E is formed.

[0041] The fourth step of the present application is to sinter the ABC compound filled dense cuboid loofah preform product E. The different cuboid loofah preform products E are placed in a muffle furnace, the oxygen flow of the oxygen inlet of the muffle furnace is 1.0-5.0 ml / min, gradually heated to 100-120℃ for 30-45 minutes, then heated to 200℃-250℃, kept at this temperature for 30-60 minutes, then heated to 500-550℃, kept at this temperature for 5-10 hours. After that, the oxygen valve is closed, the heating is turned off, gradually cooled to room temperature, the muffle furnace is opened, and the preform product E after complete burning is taken out. A white porous cuboid is obtained, in which the loofah has been decomposed at high temperature, in the form of carbon dioxide and water, and ATP-Mg, ATP-Ca, cAMP-Mg and cAMP-Ca have been decomposed, carbon and nitrogen elements have been decomposed in the form of gas, and the loofah has also been decomposed into carbon dioxide and water at this temperature. The space left after the decomposition of the loofah fibers forms fiber channels as the original loofah, and the final product is a through composite bone repair and reconstruction material F with a reversed loofah structure.

[0042] The highest heating temperature selected in the present application is 550℃, because MgHPO4, CaHPO4 and CaSO4 can remain stable in performance and composition after being kept at 550℃ for 5-12 hours, and will not be converted into calcium phosphate, magnesium phosphate, etc. or decomposed. ATP-Mg, ATP-Ca, cAMP-Mg and cAMP-Ca will be decomposed, and carbon and nitrogen elements will be decomposed in the form of gas, forming inorganic substances calcium hydrogen phosphate or magnesium hydrogen phosphate. The present application strictly limits the ratio of calcium oxide / calcium carbonate, magnesium oxide / calcium carbonate to ATP, cAMP, ensuring that the organic matter decomposed at 550℃ is MgHPO4, CaHPO4, not other components such as Mg3(PO4)2, Ca3(PO4)2, etc.

[0043]

[0044] After pyrolysis at a high temperature of 500-550℃ for a long time, the organic components in the product are decomposed into ammonia, carbon dioxide and water. At no higher than 550℃, the product is mainly calcium hydrogen phosphate (CaHPO4), magnesium hydrogen phosphate (MgHPO4) and calcium sulfate (CaSO4). The original in-situ channels left after the decomposition of the loofah fibers form through pores, the ammonia, carbon dioxide and water produced by decomposition move out of the channels to form escape pores, and the final product is a degradable multi-porous bone repair and reconstruction material with through multi-pores, mainly composed of

[0045] CaSO4-CaHPO4-MgHPO4, with a reversed loofah structure.

[0046] Further, the present application is to obtain a through-porous composite bone repair and reconstruction material F with a reticulated sponge reverse structure according to clinical requirements, and to perform type modification on the four sides of the rectangular parallelepiped to form the following shapes: (1) all four sides are through-porous structures, (2) three sides are through-porous structures, (3) two sides are through-porous structures, and (4) cut into a cylindrical shape. After completing the type modification, the product is washed with water for injection, dried, sterilized by irradiation or epoxy, and packaged for use.

[0047] The through-porous composite bone repair and reconstruction material F with a reticulated sponge reverse structure after sterilization is tested for porosity and pore connectivity, and evaluated for degradation, mechanical properties, and biological properties to obtain properties such as safety, non-toxicity, high activity, easy blood vessel growth through pores to promote bone tissue growth, and accelerated material degradation. According to the T16886 standard requirements, the through-porous product F after sterilization is implanted into the back of New Zealand white rabbits to observe its in vivo degradation, inflammation, and other biological safety, and implanted into the femur of a rabbit to observe the bone repair function and efficacy. The product with a size of 10mm*5mm*5mm and a morphology of all four sides being through-porous structures and two sides being through-porous structures is selected for implantation and observation. The animal experiment is carried out in accordance with relevant ethical requirements, and a professional doctor performs the clinical surgical procedure.

[0048] Example 1

[0049] (1) Treatment of reticulated sponge: Mix 500ml of 10% sodium hydroxide solution and 500ml of 10% hydrogen peroxide solution. Cut the dried reticulated sponge into a rectangular parallelepiped with a length of 50mm, a width of 25mm, and a thickness of 10mm, immerse it in the mixed solution, shake at 37°C for 30 minutes, and then ultrasonically clean it with deionized water three times. Dry and reserve for use.

[0050] (2) Preparation of calcium-phosphorus-magnesium composite salt component A:

[0051] Weigh 40g of magnesium oxide, 56g of calcium oxide, and 468g of calcium phosphate, mix and grind to 200 mesh, add 100ml of deionized water, stir for 30 minutes, and then mix uniformly. Dry at 80°C for 8 hours to form a calcium-magnesium-phosphorus composite (i);

[0052] Weigh 40g of magnesium oxide, 28g of calcium oxide, and 507g of ATP, mix and grind to 200 mesh, add 150ml of deionized water, stir for 30 minutes, and then mix uniformly. Dry at 80°C for 8 hours to form an ATP-calcium-magnesium-phosphorus composite (ii);

[0053] Mix 150g of (i) and 50g of (ii) to form component A.

[0054] Take 500g of calcium sulfate dehydrate and hydrothermal treatment at 120℃, 0.4MPa for 9 hours to form calcium sulfate hemihydrate, which is ground (200 mesh and above). Take 100g of calcium sulfate hemihydrate to form component B.

[0055] Take 10g of magnesium pyrophosphate and grind (200 mesh and above) to form component C.

[0056] Mix component A, component B and component C, add 200ml of deionized water to form slurry D in the form of slurry. To prevent slurry D from solidifying, pour it into the loofah as soon as possible.

[0057] (3) Put the loofah treated in process (1) into a metal mold, pour the prepared slurry D into the loofah in the mold, and make sure that the slurry D completely covers the loofah, with the height of the slurry D being 2mm higher than the loofah. Use the mold cover to press and compact, with the pressure being 5MPa, and keep for 15 minutes. Then open the mold and take out the loofah filled with slurry D to form a solidified solid cuboid. Grind the surface to expose the loofah exactly, and form a dense cuboid loofah preform product E filled with ABC composite.

[0058] (4) Prepare a bone repair product with through multi-hole and reverse structure of loofah: put the cuboid loofah preform product E obtained in process (3) into a muffle furnace, and gradually heat to 120℃ for 35 minutes, then to 200℃, and keep at this temperature for 45 minutes. Then heat to 550℃ and keep at this temperature for 8 hours. Then close the oxygen valve and turn off the heating, and gradually cool to room temperature. Open the muffle furnace and take out the completed burning product. A white porous cuboid is obtained, which forms the final product, a through composite bone repair and reconstruction material F with reverse structure of loofah.

[0059] (5) Shape the through composite bone repair and reconstruction material F with reverse structure of loofah obtained in process (4), and grind the four sides of the cuboid to form a through porous structure on all four sides. Cut part of it into 10mm*5mm*5mm as rabbit femur implant material, and cut part of it into 10mm*10mm*8mm as mechanical sample test, and use the particles or small pieces left after cutting as samples for porosity test, cytotoxicity test, degradation test and muscle implant test. After shaping, wash with water for injection, dry and sterilize the samples for cytotoxicity test and implantation.

[0060] Performance test of the through composite bone repair and reconstruction material with reverse structure of loofah obtained by the present application:

[0061] 1. Porosity and through-hole chlorine test: tested by professional testing agency by density determination drainage method;

[0062] 2, Pore size was tested by scanning electron microscope;

[0063] 3, Compression test method: according to national standard test, loading speed 1 mm / min;

[0064] 4, Degradation test method: take injection molding material in PBS solution for degradation test: the preparation method of PBS is as follows: accurately take KH2PO4(0.544 g), Na2HPO4·12H2O(7.16 g), NaCl(16 g) and KCl(0.402 g) and dissolve in 2 L deionized water, and then use a volumetric flask to make up the volume, and the pH is determined to be between 7.2 and 7.4; put the sample into a centrifuge tube and add a certain amount of PBS solution, the volume of PBS solution to sample mass ratio is 1 g / 30 mL; then put the centrifuge tube containing the sample in a constant temperature shaking incubator with a temperature of 37°C and a rotation speed of 80 rpm / min; take out the sample and calculate the weight loss rate at 1(1D), 1w, 4w, 12w and 24w days. Three control groups are set up for each sample.

[0065] 5, pH determination: use a pH meter to determine the supernatant of the sample, and replace PBS every 7 days.

[0066] 6, Cytotoxicity and cell proliferation rate test: prepare the leaching solution according to the relevant provisions of T16886 on biological materials, and calculate the proliferation rate compared with the blank.

[0067] 7: Functional test of through composite bone repair and reconstruction material with reticular counter structure. According to the requirements of T16886, implant into the back of New Zealand white rabbits to observe the in vivo degradation, inflammation and other biological safety, the specific operation is as follows: implant two 2.5 g products into the muscle on both sides of the spine of rabbits, three rabbits in each group, observe the effect, and the operation is in accordance with the requirements of T16886.

[0068] 8, implant 10mm*5mm*5mm specifications into rabbit femur, and take samples for embedding, sectioning and observation at 4 weeks and 12 weeks.

[0069] Test results:

[0070] Porosity (%): 63; Pore through rate (%): 81.

[0071] Pore size distribution (μm): 350-750.

[0072] Compression strength (MPa): 7.5.

[0073] Degradation results: 1w: 20.5%; 4w: 39.4%; 12w: 65.3%; 24w: 92.1%.

[0074] pH value change: 1w: 7.19: 4w: 7.31; 12w: 7.29; 24w: 7.33.

[0075] Cytotoxicity and cell proliferation rate: 0 level; the standard extract compared with the blank, the proliferation rate was 126%, and after dilution, the proliferation rate was 139%; relative to the blank, i.e. the culture solution, the extract of the through composite bone repair and reconstruction material with the reverse structure of loofah could provide more nutrients, cell differentiation and proliferation.

[0076] Muscle wound after surgery: 1 day, 1 week, 4 weeks, no redness, exudation, effusion phenomenon, 1 week wound healing, 4 weeks completely healed.

[0077] Femur repair effect: 4 weeks, a large number of blood vessels and blood vessels pass through the through hole, 12 weeks, the through hole is covered with new bone, most of the material is degraded, 24 weeks, the new bone is basically connected into a whole and there is no interface with the original tissue bone.

[0078] Example 2 (same as Example 1, the treatment scheme of loofah uses lithium chloride-dimethylacetamide solution)

[0079] (1) Treatment of loofah: weigh 60g of lithium chloride, dissolve in 1000ml of dimethylacetamide to form a transparent solution. Cut the dried loofah into a cuboid with a length of 50mm, a width of 25mm and a thickness of 10mm, soak in the mixed solution, shake at 37℃ for 10 minutes, ultrasonic cleaning with deionized water for three times, and dry for standby.

[0080] (2)-(5) same as Example 1.

[0081] Test results:

[0082] Porosity (%): 66. Through hole rate (%): 85.

[0083] Pore size distribution (μm): 300-800.

[0084] Compressive strength (MPa): 7.2.

[0085] Degradation results: 1w: 18.6%; 4w: 35.1%; 12w: 61.8%; 24w: 90.9%.

[0086] pH value change: 1w: 7.21: 4w: 7.32; 12w: 7.31; 24w: 7.34.

[0087] Cytotoxicity and cell proliferation rate: 0 level; the standard extract compared with the blank, the proliferation rate was 126%, and after dilution, the proliferation rate was 139%; relative to the blank, i.e. the culture solution, the extract of the through composite bone repair and reconstruction material with the reverse structure of loofah could provide more nutrients, cell differentiation and proliferation.

[0088] Muscle wound surgery: 1 day, 1 week, 4 weeks without redness, exudation, effusion phenomenon, 1 week wound healing, 4 weeks completely healed.

[0089] Femur repair effect: 4 weeks of a large number of blood vessels and blood vessels through the through hole, 12 weeks of new bone covering the through hole, most of the material degrading, 24 weeks of new bone basically connected into a whole without interface with the original bone tissue.

[0090] Example 3 (same as Example 1, treatment scheme of loofah using copper ammonia solution)

[0091] (1) Treatment of loofah: add 1L, 0.2mol / L copper sulfate solution to a beaker, then add 8mol / L ammonia water, and a light blue basic copper sulfate precipitate is formed. Continue to add ammonia water until the precipitate is completely dissolved, at which time a deep blue solution containing copper ammonia complex ions is obtained. The end point of the preparation of the copper ammonia solution is marked by the complete disappearance of the precipitate and the formation of a transparent blue solution. Dry loofah is cut into a cuboid with a length of 50mm, a width of 25mm, and a thickness of 10mm, soaked in the mixed solution, shaken for 5 minutes at 37℃, washed with deionized water three times, and dried for standby.

[0092] (2)-(5) Same as Example 1.

[0093] Test results:

[0094] Porosity (%): 70. Pore connectivity rate (%): 88.

[0095] Pore size distribution (μm): 280-790.

[0096] Compressive strength (MPa): 6.8.

[0097] Degradation results: 1w: 18.2%; 4w: 36.3%; 12w: 60.4%; 24w: 90.2%.

[0098] pH value change: 1w: 7.21; 4w: 7.28; 12w: 7.33; 24w: 7.33.

[0099] Cytotoxicity and cell proliferation rate: grade 0; the standard extract has a proliferation rate of 126% compared to the blank, and the proliferation rate is 139% after dilution; compared to the blank, the extract of the through-porous composite bone repair and reconstruction material with reversed structure of loofah can provide more nutrients, cell differentiation and proliferation.

[0100] Muscle wound surgery: 1 day, 1 week, 4 weeks without redness, exudation, effusion phenomenon, 1 week wound healing, 4 weeks completely healed.

[0101] The femur repair effect: 4 weeks of a large number of blood vessels and blood vessels through the through hole, 12 weeks of new bone covering in the through hole, most of the material degrades, and 24 weeks of new bone basically connects into a whole and has no interface with the original bone.

[0102] Example 4 (same as Example 1, different calcium-phosphorus complex composition, cAMP instead of ATP)

[0103] (1) Same as Example 1

[0104] (2) Preparation of calcium-phosphorus-magnesium complex salt component A: 40 g of magnesium oxide, 56 g of calcium oxide, and 468 g of calcium phosphate are weighed, mixed and ground to 200 mesh, 100 ml of deionized water is added, stirred for 30 minutes, and after uniform mixing, dried at 80°C for 8 hours to form a calcium-magnesium-phosphorus complex (i). 20 g of magnesium oxide, 14 g of calcium oxide, and cAMP 493 g are weighed, mixed and ground to 200 mesh, 150 ml of deionized water is added, stirred for 30 minutes, and after uniform mixing, dried at 80°C for 8 hours to form a cAMP-calcium-magnesium-phosphorus complex (ii); 150 g of (i) and 50 g of (ii) are mixed to form component A.

[0105] Take 500 g of calcium sulfate dihydrate and hydrothermally treat it at 120°C and 0.4 MPa of water vapor for 9 hours to form calcium sulfate hemihydrate, which is ground (200 mesh and above). Take 100 g of calcium sulfate hemihydrate to form component B.

[0106] Take 10 g of magnesium pyrophosphate and grind it (200 mesh and above) to form component C.

[0107] Mix components A, B and C, add 200 ml of deionized water, and mix into a slurry to form slurry D. To prevent slurry D from solidifying, pour it into a loofah as soon as possible.

[0108] (3)-(5) Same as Example 1.

[0109] Test results:

[0110] Porosity (%): 60, pore connectivity rate (%): 85.

[0111] Pore size distribution (μm): 310-770.

[0112] Compressive strength (MPa): 7.2.

[0113] Degradation results: 1w: 15.8%; 4w: 32.%; 12w: 60.1%; 24w: 85.9%.

[0114] pH value change: 1w: 7.32; 4w: 7.31; 12w: 7.33; 24w: 7.34.

[0115] Cytotoxicity and cell proliferation rate: 0 level; the standard extract has a proliferation rate of 132% compared with the blank, and the proliferation rate is 145% after dilution by one time; the extract of the through-penetrating composite bone repair and reconstruction material with the reticulated structure of loofah sponge can provide more nutrients, cell differentiation and proliferation compared with the blank, i.e. the culture solution.

[0116] Muscle wound after surgery: no redness, exudation and effusion occurred at 1 day, 1 week and 4 weeks; the wound was basically healed at 1 week, and completely healed at 4 weeks.

[0117] Femur repair effect: a large number of blood vessels and blood vessel penetration through the through hole at 4 weeks, new bone covered in the through hole at 12 weeks, most of the material degraded, and the new bone was basically connected into a whole and had no interface with the original bone at 24 weeks.

[0118] Example 5 (same as Example 1, different calcium-phosphorus composite composition, cAMP and ATP are used together)

[0119] (1) Same as Example 1

[0120] (2) Preparation of calcium-phosphorus-magnesium composite salt component A: 40 g of magnesium oxide, 56 g of calcium oxide and 468 g of calcium phosphate are weighed, mixed and finely ground to 200 mesh, 100 ml of deionized water is added, stirred for 30 minutes, uniformly mixed, dried at 80°C for 8 hours to form a calcium-magnesium-phosphorus composite (i). 40 g of magnesium oxide, 28 g of calcium oxide, 335 g of ATP and 218 g of cAMP are weighed, mixed and finely ground to 200 mesh, 150 ml of deionized water is added, stirred for 30 minutes, uniformly mixed, dried at 80°C for 8 hours to form an ATP-calcium-magnesium-phosphorus composite (ii); 150 g of (i) and 50 g of (ii) are mixed to form component A.

[0121] 500 g of calcium sulfate dihydrate is hydrothermally treated at 120°C and 0.4 MPa water vapor for 9 hours to form calcium sulfate hemihydrate, which is finely ground (200 mesh and above). 100 g of calcium sulfate hemihydrate is weighed to form component B.

[0122] 10 g of magnesium pyrophosphate is finely ground (200 mesh and above) to form component C.

[0123] Components A, B and C are mixed, 200 ml of deionized water is added, and a slurry D is formed. In order to prevent the slurry D from solidifying, it is quickly poured into the loofah sponge.

[0124] (3)-(5) Same as Example 1.

[0125] Test results:

[0126] Porosity (%): 68, pore penetration rate (%): 89.

[0127] Pore size distribution (μm): 300-790.

[0128] Compressive strength (MPa): 6.8.

[0129] Degradation results: 1w: 21.3%; 4w: 38.7%; 12w: 66.4%; 24w: 92.5%.

[0130] pH value changes: 1w: 7.31; 4w: 7.32; 12w: 7.33; 24w: 7.33.

[0131] Cytotoxicity and cell proliferation rate: 0 level; the standard extract has a proliferation rate of 131% compared with the blank, and the proliferation rate is 143% after dilution; compared with the blank, i.e. the culture solution, the extract of the through-porous composite bone repair and reconstruction material with the reticulated structure of loofah sponge can provide more nutrients, cell differentiation and proliferation.

[0132] Muscle wound after surgery: no redness, exudation and effusion occurred at 1 day, 1 week and 4 weeks, the wound was basically healed at 1 week, and completely healed at 4 weeks.

[0133] Femur repair effect: a large number of blood vessels and blood vessels pass through the through holes at 4 weeks, new bone covers the through holes at 12 weeks, most of the material degrades, and new bone is basically connected into a whole at 24 weeks, and there is no interface with the original bone tissue.

[0134] Example 6 (same as Example 1, the ratio of calcium phosphate salt is different from Example 1)

[0135] (1) Same as Example 1

[0136] (2) Preparation of calcium, magnesium and phosphorus composite salt component A: weigh 40g of magnesium oxide, 56g of calcium oxide and 468g of calcium phosphate, mix and grind to 200 mesh, add 100ml of deionized water, stir for 30 minutes, mix uniformly, dry at 80°C for 8 hours to form a calcium magnesium phosphorus composite (i). Weigh 40g of magnesium oxide, 28g of calcium oxide and 507g of ATP, mix and grind to 200 mesh, add 150ml of deionized water, stir for 30 minutes, mix uniformly, dry at 80°C for 8 hours to form an ATP-calcium magnesium phosphorus composite (ii); mix 180g of (i) and 30g of (ii) to form component A.

[0137] Weigh 500g of calcium sulfate dihydrate, hydrothermally treat at 120°C and 0.4MPa water vapor for 9 hours to form calcium sulfate hemihydrate, and grind (200 mesh and above). Weigh 120g of calcium sulfate hemihydrate to form component B.

[0138] Weigh 10g of magnesium pyrophosphate, grind (200 mesh and above) to form component C.

[0139] Mixing component A, component B and component C, add 200ml deionized water, mix into slurry, form slurry D, ready for use. To prevent slurry D from solidifying, pour into loofah as soon as possible.

[0140] (3)-(5) Same as example 1.

[0141] Test results:

[0142] Porosity (%): 71, pore penetration rate (%): 89.

[0143] Pore size distribution (μm): 300-800.

[0144] Compressive strength (MPa): 5.9.

[0145] Degradation results: 1w: 22.7%; 4w: 41.3%; 12w: 67.4%; 24w: 95.5%.

[0146] pH value change: 1w: 7.05: 4w: 7.211; 12w: 7.30; 24w: 7.32.

[0147] Cytotoxicity and cell proliferation rate: 0 level; standard extract compared with blank, proliferation rate 132%, after dilution, proliferation rate 146%; relative to the blank, i.e. culture solution, the extract of the through-penetrating composite bone repair and reconstruction material with loofah reverse structure can provide more nutrients, cell differentiation and proliferation.

[0148] Muscle wound surgery: 1 day, 1 week, 4 weeks, no redness, exudation, effusion phenomenon, 1 week wound healing, 4 weeks completely healed.

[0149] Femur repair effect: 4 weeks, a large number of blood vessels and blood vessels pass through the through-hole, 12 weeks, new bone covers the through-hole, most of the material degrades, 24 weeks, new bone is basically connected into a whole and has no interface with the original bone tissue.

[0150] Example 7 (same as example 1, different amount of magnesium pyrophosphate component)

[0151] (1) Same as example 1.

[0152] (2) Preparation of calcium-magnesium-phosphorus composite salt component A: weigh 40g magnesium oxide, 56g calcium oxide, 468g calcium phosphate, mix and grind to 200 mesh, add 100ml deionized water, stir for 30 minutes, mix uniformly, dry at 80°C for 8 hours to form calcium-magnesium-phosphorus composite (i). Weigh 40g magnesium oxide, 28g calcium oxide, ATP 507g, mix and grind to 200 mesh, add 150ml deionized water, stir for 30 minutes, mix uniformly, dry at 80°C for 8 hours to form ATP-calcium-magnesium-phosphorus composite (ii); mix 150g of (i) and 50g of (ii) to form component A.

[0153] Take 500 g of calcium sulfate dihydrate and hydrothermally treat it at 120°C and 0.4 MPa for 9 hours to form calcium sulfate hemihydrate, which is finely ground (200 mesh or more). Take 100 g of the calcium sulfate hemihydrate to form component B.

[0154] Take 5 g of magnesium pyrophosphate and finely grind it (200 mesh or more) to form component C.

[0155] Mix components A, B and C, add 200 ml of deionized water, and mix to form slurry D. Pour the slurry D into a loofah as soon as possible to prevent it from solidifying.

[0156] (3)-(5) Same as in Example 4.

[0157] Test results:

[0158] Porosity (%): 60, pore connectivity rate (%): 80.

[0159] Pore size distribution (μm): 300-710.

[0160] Compressive strength (MPa): 7.5.

[0161] Degradation results: 1w: 21.1%; 4w: 39.7%; 12w: 66.5%; 24w: 93.4%.

[0162] pH value changes: 1w: 7.22; 4w: 7.31; 12w: 7.33; 24w: 7.33.

[0163] Cytotoxicity and cell proliferation rate: Grade 0; the standard extract has a proliferation rate of 131% compared to the blank, and after being diluted by one time, the proliferation rate is 145%; compared to the blank, the extract of the through-porous composite bone repair and reconstruction material with the reverse structure of loofah has more nutrients, cell differentiation and proliferation.

[0164] Muscle wound surgery: no redness, exudation or effusion occurred at 1 day, 1 week and 4 weeks after surgery; the wound was basically healed at 1 week, and completely healed at 4 weeks.

[0165] Femur repair effect: a large number of blood vessels and blood vessels pass through the through-pores at 4 weeks, new bone covers the through-pores at 12 weeks, most of the material degrades, and the new bone is basically integrated into a whole at 24 weeks, with no interface with the original bone tissue.

[0166] Example 8 (same as Example 5, different calcium-phosphorus salt ratio)

[0167] (1) Same as Example 1.

[0168] (2) Preparation of calcium-phosphorus-magnesium composite salt component A: 40 g of magnesium oxide, 56 g of calcium oxide, and 468 g of calcium dihydrogen phosphate were weighed, mixed, and ground to 200 mesh, 100 ml of deionized water was added, stirred for 30 minutes, and after uniform mixing, dried at 80°C for 8 hours to form a calcium-magnesium-phosphorus composite (i). 40 g of magnesium oxide, 28 g of calcium oxide, 335 g of ATP, and 218 g of cAMP were weighed, mixed, and ground to 200 mesh, 150 ml of deionized water was added, stirred for 30 minutes, and after uniform mixing, dried at 80°C for 8 hours to form an ATP-calcium-magnesium-phosphorus composite (ii); 150 g of (i) and 50 g of (ii) were mixed to form component A.

[0169] Calcium sulfate dihydrate 500 g was hydrothermally treated at 120°C and 0.4 MPa for 9 hours to form calcium sulfate hemihydrate, which was ground (200 mesh and above). 120 g of calcium sulfate hemihydrate was weighed to form component B.

[0170] Magnesium pyrophosphate 10 g was ground (200 mesh and above) to form component C.

[0171] Components A, B, and C were mixed with 200 ml of deionized water to form slurry D in the form of a slurry, which was ready for use. To prevent the solidification of slurry D, it was quickly poured into the loofah sponge.

[0172] (3)-(5) The same as in Example 4.

[0173] Test results:

[0174] Porosity (%): 72; pore connectivity rate (%): 90.

[0175] Pore size distribution (μm): 300-800.

[0176] Compressive strength (MPa): 6.5.

[0177] Degradation results: 1w: 21.7%; 4w: 39.9%; 12w: 66.8%; 24w: 93.3%.

[0178] pH value change: 1w: 7.15; 4w: 7.26; 12w: 7.30; 24w: 7.33.

[0179] Cytotoxicity and cell proliferation rate: grade 0; the standard extract had a proliferation rate of 136% compared to the blank, and after dilution, the proliferation rate was 150%; compared to the blank, the extract of the through-composite bone repair and reconstruction material with a reversed structure of loofah sponge could provide more nutrients, cell differentiation, and proliferation.

[0180] Muscle wound surgery: no redness, exudation, and effusion occurred at 1 day, 1 week, and 4 weeks after surgery, the wound was basically healed at 1 week, and completely healed at 4 weeks.

[0181] The femur repair effect: 4 weeks of a large number of blood vessels and blood vessels through the through hole, 12 weeks of new bone covering in the through hole, most of the material degrades, 24 weeks of new bone basically connected into a whole and no interface with the original bone tissue.

[0182] Example 9 (same as Example 1, grouting mold preparation loofah rectangle pressure difference)

[0183] (1)-(2) Same as Example 1.

[0184] (3) Place the loofah treated in process (1) in a metal mold, pour the prepared slurry D into the loofah in the mold, the slurry D completely submerges the loofah, and the height is 4 mm higher than the loofah, use the mold cover plate to press and compact, the pressure is 10 MPa, keep for 15 minutes, then open the mold, take out the loofah filled with slurry D, form a solidified solid cuboid, polish the surface to expose the loofah, form a dense loofah rectangle preformed product E filled with ABC composite.

[0185] (4)-(5) Same as Example 1.

[0186] Test results:

[0187] Porosity (%): 60. Pore connectivity (%): 80.

[0188] Pore size distribution (μm): 300-700.

[0189] Compressive strength (MPa): 9.1.

[0190] Degradation results: 1w: 18.3%; 4w: 36.1%; 12w: 62.2%; 24w: 89.4%.

[0191] pH value change: 1w: 7.20: 4w: 7.29; 12w: 7.33; 24w: 7.34.

[0192] Cytotoxicity and cell proliferation rate: 0 level; the standard extract compared with the blank has a proliferation rate of 131%, and after dilution, the proliferation rate is 144%; relative to the blank, the culture solution with the through composite bone repair and reconstruction material with reversed structure of loofah can provide more nutrients, cell differentiation and proliferation.

[0193] Muscle wound surgery: 1 day, 1 week, 4 weeks, no redness, exudation, effusion phenomenon, 1 week wound healing, 4 weeks completely healed.

[0194] The femur repair effect: 4 weeks of a large number of blood vessels and blood vessels through the through hole, 12 weeks of new bone covering in the through hole, most of the material degrades, 24 weeks of new bone basically connected into a whole and no interface with the original bone tissue.

[0195] Example 10 (same as Example 1, different pyrolysis temperature in muffle furnace)

[0196] (1) - (3) same as Example 1

[0197] (4) Preparation of a bone repair product with through multi-porous, with reticulated sponge reverse structure. The cuboid reticulated sponge preform product E obtained from process (3) was placed in a muffle furnace, the oxygen flow rate at the oxygen inlet of the muffle furnace was 2.5 ml / min, gradually heated to 120°C for 35 minutes, then heated to 200°C, and kept at this temperature for 45 minutes, then heated to 525°C, and kept at this temperature for 10 hours. After that, the oxygen valve was closed, the heating was turned off, and the temperature was gradually lowered to room temperature. The completed burning product was taken out of the muffle furnace. A white multi-porous cuboid was obtained, forming the final product of the through composite bone repair and reconstruction material F with reticulated sponge reverse structure.

[0198] (5) same as Example 1.

[0199] Test results:

[0200] Porosity (%): 60. Hole through rate (%): 85.

[0201] Pore size distribution (μm): 310-900

[0202] Compressive strength (MPa): 6.5.

[0203] Degradation results: 1w: 19.4%; 4w: 38.8%; 12w: 64.9%; 24w: 91.8%.

[0204] pH value change: 1w: 7.20; 4w: 7.30; 12w: 7.32; 24w: 7.33.

[0205] Cytotoxicity and cell proliferation rate: Grade 0; the standard extract has a proliferation rate of 137% compared to the blank, and the proliferation rate is 149% after dilution; compared to the blank, the extract of the through composite bone repair and reconstruction material with reticulated sponge reverse structure can provide more nutrients, cell differentiation and proliferation.

[0206] Muscle wound surgery: no redness, exudation, and effusion occurred at 1 day, 1 week, and 4 weeks after surgery; the wound was basically healed at 1 week, and completely healed at 4 weeks.

[0207] Femur repair effect: at 4 weeks, a large number of blood vessels and blood vessels passed through the through holes, at 12 weeks, new bone covered the through holes, the material was mostly degraded, and at 24 weeks, the new bone was basically integrated into a whole and had no interface with the original bone tissue.

[0208] Comparative Example 1, calcium hydrogen phosphate / calcium sulfate composite

[0209] Take 60 g of calcium sulfate hemihydrate, 40 g of calcium hydrogen phosphate, mix them evenly, then add 50 ml of water for injection to stir into a paste, mix evenly, put the mixed product into a mold to make a test sample, take it out after solidification and put it into a muffle furnace. The oxygen flow rate at the oxygen inlet of the muffle furnace is 2.5 ml / min, gradually heated to 120°C and kept for 35 minutes, then heated to 200°C and kept for 45 minutes, then heated to 550°C and kept for 10 hours. Then, close the oxygen valve and turn off the heating, gradually cool to room temperature, open the muffle furnace and take out the completed burning product.

[0210] Test results:

[0211] Porosity (%): 20. Pore connectivity (%): 15.

[0212] Pore size distribution (μm): 100-300.

[0213] Compressive strength (MPa): 15.

[0214] Degradation results: 1w: 15.4%; 4w: 28.8%; 12w: 45.3%; 24w: 59.8%.

[0215] pH value change: 1w: 7.13; 4w: 7.21; 12w: 7.29; 24w: 7.30.

[0216] Cytotoxicity and cell proliferation rate: Grade 0; the standard extract has a proliferation rate of 115% compared with the blank, and after dilution, the proliferation rate is 126%, and the cell proliferation is slower than the relative example.

[0217] Muscle wound surgery: no redness, exudation, and effusion occurred at 1 day, 1 week, and 4 weeks, most of the wound healed at 1 week, and basically healed at 4 weeks.

[0218] Femur repair effect: blood vessels are difficult to pass through at 4 weeks, new bone surrounds the material at 12 weeks, the material partially degrades, and one-third of the material is wrapped in new bone at 24 weeks, and the effect is relatively poor compared with the relative example.

[0219] Comparative Example 2: Calcium sulfate calcium hydrogen phosphate hydrogen peroxide foaming compound

[0220] Take 60 g of calcium sulfate hemihydrate, 40 g of calcium hydrogen phosphate, mix them well, add 45 ml of water for injection, and 5 ml of 10% hydrogen peroxide, mix well, put the mixed product into a mold to make a test sample, after curing, take it out and put it into a muffle furnace. The oxygen flow rate at the oxygen inlet of the muffle furnace is 2.5 ml / min, gradually increase the temperature to 120°C and keep it for 35 minutes, then increase the temperature to 200°C and keep it for 45 minutes, then increase the temperature to 550°C and keep it for 10 hours. Then, close the oxygen valve and turn off the heating, gradually cool to room temperature, open the muffle furnace and take out the completed burning product.

[0221] Test results:

[0222] Porosity (%): 75. Hole penetration rate (%): 50; the through-hole rate is lower than that of the comparative example.

[0223] Pore size distribution (μm): 100-1500, the pore size distribution range is wider than that of the comparative example.

[0224] Compressive strength (MPa): 3.5. The strength is lower than that of the comparative example.

[0225] Degradation results: 1w: 19.4%; 4w: 32.7%; 12w: 65.3%; 24w: 85.8%.

[0226] pH value change: 1w: 7.16; 4w: 7.20; 12w: 7.25; 24w: 7.30.

[0227] Cytotoxicity and cell proliferation rate: 0 level; the standard extract has a proliferation rate of 118% compared with the blank, and the proliferation rate is 130% after dilution, the cell proliferation is slower than that of the comparative example.

[0228] Muscle wound after surgery: no redness, exudation, effusion occurred at 1 day, 1 week and 4 weeks, most of the wound healed at 1 week, and basically healed at 4 weeks.

[0229] Femur repair effect: part of the blood vessels pass through the pores to form new bone at four weeks, more new bone formation passes through the pores at 12 weeks, the material partially degrades, part of the new bone links through at 24 weeks, the effect is relatively poor compared with the comparative example.

[0230] Comparative example 3: the same as example 1, but without ATP calcium phosphate salt.

[0231] Test results:

[0232] Porosity (%): 55. Hole penetration rate (%): 70; the through-hole rate is lower than that of the comparative example.

[0233] Pore size distribution (μm): 300-1000, the pore size distribution range is wider than that of the comparative example.

[0234] Compressive strength (MPa): 4.1. Lower strength compared to the previous embodiment.

[0235] Degradation results: 1w: 20.1%; 4w: 33.6%; 12w: 66.1%; 24w: 89.4%.

[0236] pH changes: 1w: 7.21; 4w: 7.30; 12w: 7.33; 24w: 7.33.

[0237] Cytotoxicity and cell proliferation rate: Grade 0; the standard extract showed a proliferation rate of 119% compared to the blank, and 134% after dilution by one time, indicating slightly slower cell proliferation compared to the example.

[0238] After muscle wound surgery: no redness, swelling, oozing, or fluid accumulation occurred at 1 day, 1 week, and 4 weeks. The wound was mostly healed at 1 week and basically healed at 4 weeks.

[0239] Femoral bone repair effect: Around the periphery, some blood vessels pass through the pores to form new bone. At 12 weeks, more new bone forms and passes through the pores. The material is partially degraded. At 24 weeks, most of the new bone is connected and connected. The effect is relatively poor compared to the previous example.

[0240] Comparative Example 4 is the same as Example 1, but the sintering temperature is 650°C, which is 100°C higher than that of Example 1.

[0241] Test results:

[0242] Porosity (%): 66. Pore penetration rate (%): 80.

[0243] Pore ​​size distribution (μm): 300-1500, which is a wider range than that of the previous embodiment.

[0244] Compressive strength (MPa): 2.7. Lower strength compared to the previous embodiment.

[0245] Degradation results: 1w: 12.9%; 4w: 25.8%; 12w: 57.7%; 24w: 65.3%. At excessively high pyrolysis temperatures, dicalcium phosphate is converted to calcium phosphate, which is more difficult to degrade.

[0246] pH value changes: 1w: 7.23; 4w: 7.31; 12w: 7.33; 24w: 7.34.

[0247] Cytotoxicity and cell proliferation rate: Grade 0; the standard extract showed a proliferation rate of 120% compared to the blank, and 135% after being diluted by one time.

[0248] After muscle wound surgery: no redness, swelling, oozing, or fluid accumulation occurred at 1 day, 1 week, and 4 weeks. The wound was mostly healed at 1 week and basically healed at 4 weeks.

[0249] The femur repair effect: after four weeks, some blood vessels pass through the pores and form new bone, after 12 weeks, more new bone forms through the pores, the material partially degrades, and after 24 weeks, part of the new bone links through, and the effect is relatively poor compared to the example.

[0250] Comparative Example 5: The same as Example 1, all components are the same as Example 1. No loofah sponge is used as a template, and no high-temperature pyrolysis is performed.

[0251] Product D formed by curing the ABC of Example 1 was tested.

[0252] Test results:

[0253] Porosity (%): 28. Hole penetration rate (%): 11; both the porosity and the hole penetration rate are lower than those of the example.

[0254] Pore size distribution (μm): 100-300, the pore size is very small, which is not conducive to cell growth.

[0255] Compressive strength (MPa): 15.6.

[0256] Degradation results: 1w: 20.1%; 4w: 31.4%; 12w: 51.6%; 24w: 64.6%. The surface is wrapped with deposited calcium phosphate salt, and the degradation is slow in the later stage

[0257] pH value change: 1w: 7.19; 4w: 7.27; 12w: 7.31; 24w: 7.33.

[0258] Cytotoxicity and cell proliferation rate: 0 level; the standard extract has a proliferation rate of 125% compared with the blank, and after being diluted by one time, the proliferation rate is 140%.

[0259] Muscle wound surgery: no redness, exudation, and effusion occurred after 1 day, 1 week, and 4 weeks, most of the wound healed after 1 week, and basically healed after 4 weeks.

[0260] Femur repair effect: after four weeks, blood vessels cannot pass through the material, new bone is formed around the material, after 12 weeks, more new bone is formed around the material, the material partially degrades but cannot pass through the interior of the material, after 24 weeks, part of the new bone links through around the material, and part of the material is wrapped in the new bone. The repair effect is relatively poor compared to the example.

[0261] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing a porous bone repair material, characterized in that, The preparation method includes the following steps: 1) Preparation of bone repair material slurry: The raw materials of the bone repair material are mixed with water to form a bone repair material slurry; the raw materials of the bone repair material include the following components in the following proportions: 35-55 parts by weight of calcium-phosphorus-magnesium complex salt, 25-55 parts by weight of calcium sulfate hemihydrate, and 0-5 parts by weight of magnesium pyrophosphate; wherein, the molar ratio of magnesium to calcium in the calcium-phosphorus-magnesium complex salt is 0.1-0.5:1; and the calcium-phosphorus-magnesium complex salt is prepared by the following method: weighing a portion of magnesium salt, calcium salt, and phosphorus-containing substance, mixing and grinding them finely, adding deionized water and stirring to mix evenly to form calcium-magnesium-phosphorus complex 1; weighing the remaining magnesium salt, calcium salt, and phosphorus-containing substance, mixing and grinding them finely, adding deionized water and stirring to mix evenly to form calcium-magnesium-phosphorus complex 2; weighing complex 1 and complex 2 and mixing them to form calcium-phosphorus-magnesium complex salt; the phosphorus-containing substance is at least one of calcium dihydrogen phosphate, adenosine triphosphate, or cyclic adenosine monophosphate; the magnesium salt is at least one of MgO or MgCO3; and the calcium salt is at least one of CaO or CaCO3. 2) Preparation of bone repair preform: The bone repair raw material slurry is placed in a loofah sponge and fully immersed to fill the loofah sponge with the bone repair slurry, and then allowed to solidify to obtain the bone repair preform; 3) Sintering treatment: The bone repair preform is sintered to remove the loofah sponge, thus obtaining a bone repair material with a through-pore structure; wherein, the sintering treatment process is as follows: the bone repair preform is placed in a muffle furnace, the temperature is gradually raised to 100-120℃ and held for 30-45 minutes, then the temperature is raised to 200-250℃ and held for 30-60 minutes, and then the temperature is raised to 500-550℃ and held for 5-12 hours.

2. The method for preparing a porous bone repair material according to claim 1, characterized in that, In step 1), the ratio of raw materials to water in the bone repair material is 1:0.5 to 1.5 (g / mL).

3. The method for preparing a porous bone repair material according to claim 1, characterized in that, In step 1), the bone repair material slurry is prepared using the following method: (1) Weigh 40-50 g magnesium oxide, 50-60 g calcium oxide and 450-470 g calcium dihydrogen phosphate, mix and grind to 200 mesh, add 100-150 ml deionized water, stir for 25-35 minutes, mix evenly, and dry at 70-80℃ for 5-10 hours to form calcium magnesium phosphate complex 1. Weigh 20-40 g of magnesium oxide, 14-28 g of calcium oxide, and 493-553 g of phosphorus-containing substances, mix and grind to 200 mesh, add 140-160 ml of deionized water, stir for 25-35 minutes, mix evenly, and dry at 70-80℃ for 5-10 hours to form calcium magnesium phosphorus complex 2. Weigh out 1 / 150-180g of complex 1 and 2 / 30-50g of complex 2 and mix them to form component A; 2) Weigh 450-550g of calcium sulfate dihydrate and hydrothermally treat it in steam at 110-130℃ and 0.1-0.5 MPa for 5-10 hours to form calcium sulfate hemihydrate. Grind it to 200 mesh or finer as component B. 3) Weigh magnesium pyrophosphate and grind it to 200 mesh or finer to form component C; 4) Mix 200-210 g of component A, 100-120 g of component B and 5-10 g of component C, add 150-250 ml of deionized water to form a paste, thus forming a bone repair raw material paste.

4. The method for preparing a porous bone repair material according to claim 1, characterized in that, In step 2), the loofah sponge is pretreated before use so that the fiber diameter of the loofah sponge is 200-1000 μm.

5. The method for preparing a porous bone repair material according to claim 4, characterized in that, The pretreatment method is as follows: First, the selected loofah sponge is divided according to its veins and cut into regular cubes with a length of 10mm to 200mm, a width of 10mm to 50mm, and a height of 0.5mm to 20mm; then, it is soaked in a treatment solvent and then dried for later use; the treatment solvent is selected from: sodium hydroxide-hydrogen peroxide solution, lithium chloride-dimethylacetamide solution, lithium bromide-dimethylacetamide solution, lithium chloride-dimethylformamide solution, lithium bromide-dimethylformamide solution, or copper ammonia solution.

6. The method for preparing a porous bone repair material according to claim 1, characterized in that, The preparation method of the perforated bone repair material also includes step 4): post-processing of the perforated bone repair material according to clinical requirements: cutting and polishing the shape and size, washing with water for injection, air drying, sterilizing with irradiation or epoxy, and finally packaging for later use.

7. A porous bone repair material, characterized in that, The bone repair material is prepared using the preparation method described in any one of claims 1 to 6.

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