Modified bio-mineralized scaffold incorporated with lithium, magnesium, and phosphorus as base

Abstract

Provided is a modified bio-mineralized scaffold incorporated with lithium, magnesium, and phosphorus as a base. The scaffold is prepared by immersing a calcined cancellous bone mineral porous scaffold from cattle or swine into a composite solution containing active metal ions (magnesium, lithium, and one or more of strontium, zinc, iron, and calcium) combined with a phosphorus source, followed by hydrothermal reaction treatment, baking, drying, and high-temperature calcination. The modified bio-mineralized scaffold can effectively stabilize the incorporated magnesium, lithium, phosphorus, and other osteogenic-active ions (one or more of strontium, zinc, iron, and calcium), and retains the three-dimensional interconnected porous network structure and natural crystal structure of the calcined cancellous bone mineral porous scaffold from cattle or swine, while enabling the growth of calcium phosphate crystals containing active ions. The scaffold exhibits excellent hydrophilicity, is capable of partial dissolution, and can improve cell adhesion and osteogenic activity.

Description

A modified biological bone mineral scaffold based on lithium magnesium phosphorus Technical Field

[0001] The invention relates to the technical field of bone transplant materials, in particular to a modified biological bone mineral scaffold doped with lithium magnesium phosphorus as the basis. Background Art

[0002] Bone transplantation is a common tissue transplantation, second only to blood transfusion. Autologous bone transplantation is the gold standard for bone defect repair because the three essential elements of autologous bone repair include cells, active proteins, and a mineralized extracellular matrix scaffold (primarily composed of hydroxyapatite, but also including other ions such as Mg, Fe, Si, Zn, Mn, Cu, Na, K, F, S, C, and Cl). The research and development of bone graft replacement materials is currently a key focus of medical research. Human bone mineralization undergoes extensive homogeneous replacement, and human bone possesses a complex composition and structure. The design of bone graft replacement materials (artificial bone) or bone tissue engineering scaffolds critically considers the complex composition and structure (unique three-dimensional interconnected pore structure and natural nanocrystalline structure) as well as the physicochemical properties (such as good hydrophilicity, degradation properties, and mechanical strength) of this heavily mineralized tissue.

[0003] Human bone cannot be completely replaced by the limited properties provided by a single material. More importantly, the ideal artificial bone or bone tissue engineering scaffold must provide a three-dimensional porous microstructure for bone tissue regeneration and provide space for the adhesion and physiological activities of osteoblast-related cells. At the same time, the scaffold must have a composition similar to that of human bone mineral. The dissolution of the scaffold [including physical, chemical and cellular aspects] can provide calcium, phosphorus ions and other osteogenic active ions such as magnesium ions, just like the dissolution of autologous bone mineralized extracellular matrix scaffold, forming a high-calcium and osteogenic active ion-rich microenvironment.

[0004] Calcium and phosphate are the main components of human bone mineral, and calcium-phosphate materials are currently the most common bone graft substitute materials used clinically. Currently, there is a lack of ideal bone graft substitute materials in clinical practice, primarily due to the inability to combine ideal three-dimensional interconnected mesh structures, high porosity and specific surface area, appropriate degradability, and good hydrophilicity, osteoconductivity, osteoinductivity, and mechanical strength.

[0005] An ideal three-dimensional interconnected pore structure is a fundamental requirement for bone tissue engineering scaffolds or bone graft substitutes. Clinically used artificial bones with relatively ideal three-dimensional interconnected pore structures are all derived from animal materials. Among these, porous hydroxyapatite ceramic bone (including the best-selling Swiss Geistlich artificial bone material in my country) derived from bovine cancellous bone via a high-temperature sintering process is characterized by its composition, structure, and pore size highly similar to the mineralized extracellular matrix of human cancellous bone, resulting in excellent biocompatibility, osteoconductivity, and relatively high mechanical strength. Implantation in the body facilitates the recruitment of bone repair cells, the adhesion of active factors, the exchange of oxygen and tissue fluid, and the formation and entry of blood vessels, providing excellent adhesion support and physiological activity for bone repair cells. However, a significant drawback is that the bone mineral derived from high-temperature sintering of bovine cancellous bone—"elemental" hydroxyapatite—is extremely stable, undergoes slow degradation in vivo, and has the lowest solubility among calcium-phosphate bone graft materials. Consequently, it lacks sufficient osteogenic activity and is not conducive to bone repair and remodeling.

[0006] Another key requirement for artificial bone or bone tissue engineering scaffolds is an ideal degradation rate. Ideally, artificial bone degradation should match the rate of new bone formation, gradually degrading to provide space for new bone replacement while guiding new bone formation. The calcium ions and other osteogenic ions continuously released during degradation provide mineral reorganization components for bone mineral redeposition, remodeling, and metabolism. This process may stimulate new bone formation, thus possessing a degree of potential osteoinductivity and forming the biochemical basis for the osteogenic activity of mineral materials. Excessively rapid degradation of artificial bone or bone tissue engineering scaffolds hinders the provision of sufficient temporal and spatial support and guidance for bone repair, while excessively slow degradation can hinder new bone formation, replacement, and remodeling. The in vitro degradation of mineral bone graft materials is related to their composition, as well as to particle size, porosity, specific surface area, crystallinity, and solubility, with solubility being a key influencing factor. In the past 20 years, scientists have attempted to convert calcined bovine cancellous bone porous hydroxyapatite (calcium-phosphorus molar ratio of 5:3) doped with phosphorus into tricalcium phosphate (calcium-phosphorus molar ratio of 3:2), calcium pyrophosphate (calcium-phosphorus molar ratio of 1:1), or a composite calcium-phosphorus porous scaffold containing tricalcium phosphate, calcium pyrophosphate, and hydroxyapatite, with the aim of improving the material's degradation properties and osteogenic activity.

[0007] Another hot topic in artificial bone research over the past decade has been the study of the effects of metal ion doping on osteoblastic activity. A review summarizes how metal ions regulate bone formation by promoting or inhibiting various key processes involved in bone formation and repair, including stem cell proliferation, osteogenic differentiation, osteoblast physiological activity and differentiation, bone formation and mineralization, and inhibition of osteoclast differentiation, as well as vascular endothelial cells. Among them, lithium silver aluminum ions have a promoting effect on stem cell proliferation; copper ions have a suppressing effect on stem cell proliferation; silver calcium lithium magnesium vanadium zinc ions have a promoting effect on osteoblast differentiation; aluminum cobalt copper iron manganese ions have a suppressing effect on osteoblast differentiation; silver calcium gallium lithium magnesium vanadium ions have a promoting effect on the physiological activity and differentiation of osteoblasts; aluminum cobalt copper iron manganese ions have a suppressing effect on the physiological activity and differentiation of osteoblasts; silver calcium cobalt copper lithium magnesium strontium zinc has a promoting effect on formation; aluminum iron ions have a suppressing effect on bone formation; silver calcium cobalt copper lithium magnesium strontium zinc has a promoting effect on mineralization; aluminum copper cobalt iron ions have a suppressing effect on mineralization; lithium gallium zinc strontium ions have a suppressing effect on osteoclast differentiation; iron ions have a promoting effect on osteoclast differentiation; cobalt copper manganese gallium ions promote endothelial cell proliferation and revascularization; silver copper gallium zinc ions have a suppressing effect on biofilm.

[0008] Calcium is the most common mineral in the human body and is primarily stored in bone. Calcium homeostasis is tightly regulated by parathyroid hormone (PTH) and calcitonin, which regulate serum calcium levels by stimulating (PTH) or inhibiting (calcitonin) bone resorption by osteoclasts. During bone remodeling, data indicate that bone-resorbing osteoclasts generate local concentrations of extracellular calcium ions up to 40 mol / L. This increased calcium in the microenvironment inhibits osteoclast resorption activity and promotes the proliferation and differentiation of mesenchymal stromal cells and osteoblasts. In the 1980s, extracellular calcium was shown to activate an extracellular calcium protein-coupled receptor, known as the calcium sensor receptor (CaSR). Concerning the hyperresponsiveness of osteocytes to extracellular calcium, elevated calcium levels promote osteocyte proliferation, chemotaxis, and osteogenic differentiation, activating bone marrow-derived mesenchymal stromal cells in a dose-dependent manner via CaSR activation. Tricalcium phosphate (calcium-phosphorus molar ratio 3:2) and calcium pyrophosphate (calcium-phosphorus molar ratio 1:1) have better degradation properties than hydroxyapatite (calcium-phosphorus molar ratio 5:3). Artificial bone materials containing tricalcium phosphate and calcium pyrophosphate are more conducive to the formation of a microenvironment with a higher concentration of calcium ions in the transplantation area than simple hydroxyapatite.

[0009] The most actively researched divalent cation currently being used clinically is magnesium. Magnesium is widely present in nature, with an adult human containing approximately 25g of magnesium. It plays a crucial role in bone formation and all growth processes, maintaining bone cell structure and function, bone metabolism, and remodeling. Low-magnesium magnesium phosphate-based bone cement can significantly enhance cell adhesion. Magnesium-doped calcium phosphate bone cement is gaining increasing attention as a novel bone repair biomaterial because it promotes the formation of a bony interface between implant materials and bone tissue. Magnesium-doped bone cement is relatively easy to formulate, and a formula of 73% β-tricalcium phosphate / 21% monocalcium phosphate / 5% magnesium hydrogen phosphate is already in clinical use in Western countries such as New Zealand. Magnesium-containing composite bone cements are biodegradable, releasing calcium, phosphorus, and magnesium, which are beneficial elements for bone formation. After implantation, they undergo degradation and ion exchange within the body. However, they lack a three-dimensional interconnected mesh structure, hindering the early penetration of repair cells and blood vessels into the graft. Magnesium ions promote osteoblast differentiation, osteoblast physiological activity and differentiation, bone formation, and mineralization. Our previous studies have suggested that magnesium ion-doped modified bio-bone mineral scaffolds can improve osteogenesis and vascularization.

[0010] Lithium is a non-essential trace element and, therefore, has no known biological function in humans. However, due to its beneficial effects on psychotherapy, lithium has been widely introduced for medical use. Among various proposed mechanisms of action for lithium, stimulation of neural progenitor cell proliferation through the Wnt / β-catenin pathway, leading to increased gray matter in the brain, is widely accepted. Interestingly, the proliferation of other cell types, such as mesenchymal stem cells, is also regulated by the Wnt / β-catenin pathway, suggesting that lithium may also regulate cell proliferation. Indeed, a recent study reported that lithium-mediated Wnt / β-catenin signaling increased the proliferation of hMSCs in vitro. Furthermore, previous studies have shown that lithium is a major regulator of osteoblastogenesis, making its application in tissue engineering even more attractive. Rats receiving a lithium solution by oral gavage showed increased bone density, the amount of newly formed mature bone tissue, and bone regeneration compared to those receiving a lithium saline solution, suggesting that lithium accelerates callus ossification and bone healing. Preliminary studies investigating lithium release, toxicity, and osteoblast activity in lithium-doped bone cement suggest that lithium is a promising metal ion. In vivo application significantly improves bone formation and defect repair rates, and exhibits superior osteoconductivity compared to pure calcium phosphate cement. Lithium appears to directly regulate and promote all key aspects of bone formation and repair, including stem cell proliferation, osteogenic differentiation, osteoblast physiological activity and differentiation, bone formation, mineralization, and inhibition of osteoclast differentiation, thereby promoting bone formation.

[0011] Silver can benefit bone formation by promoting stem cell proliferation, osteoblast differentiation, osteoblast physiological activity and differentiation, bone formation, mineralization, and biofilm inhibition. However, due to the difficulty in assessing the harmful effects of silver on the human body, silver ions are not used as a doping element in our products. For the same reason, copper, manganese, cobalt, and aluminum are not selected as doping elements.

[0012] Our previous efforts have been to effectively incorporate two or more of the osteogenic active ions magnesium, magnesium zinc, magnesium strontium and phosphate, sulfate into the calcined bovine bone porous hydroxyapatite bone mineral scaffold; while maintaining the ideal three-dimensional interconnected mesh structure and good mechanical strength of the calcined bovine bone porous hydroxyapatite, we have attempted to improve its degradation characteristics and osteogenic activity. Summary of the Invention

[0013] The purpose of the present invention is to solve the above problems and provide a modified biological bone mineral scaffold based on lithium magnesium phosphorus to obtain a bone tissue engineering scaffold and bone transplant substitute material with good three-dimensional interconnected mesh structure, mechanical strength, degradability and osteogenic activity.

[0014] To achieve the above objectives, the present invention provides a modified biological bone mineral scaffold based on lithium, magnesium and phosphorus, which is obtained by immersing a porous scaffold of calcined bovine or porcine cancellous bone mineral in a composite solution containing one or more active metal ions of magnesium, lithium, strontium, zinc, iron, and calcium and a phosphorus source, undergoing a hydrothermal reaction treatment, baking and drying, and then calcining at a high temperature. The metal ion calcium mentioned herein refers to supplementary calcium and does not include the calcium inherent in the biological bone mineral scaffold.

[0015] The method comprises immersing a porous scaffold of calcined bovine or porcine cancellous bone mineral into a composite solution containing active metal ions of magnesium, lithium, strontium, zinc, iron, and calcium and a phosphorus source for hydrothermal reaction treatment. The method comprises: first immersing the porous scaffold of calcined bovine or porcine cancellous bone mineral into a metal ion source solution containing active ions of magnesium and lithium and one or more of strontium, zinc, iron, and calcium and a white sugar source solution, drying the liquid by microwave or constant temperature oven, baking and drying at 96° C.-198° C., and then immersing the scaffold into the phosphorus source composite solution for hydrothermal reaction; the phosphorus source composite solution is a binary system of phosphorus source, thereby forming a binary system containing osteogenic active metal ions and phosphorus source.

[0016] The hydrothermal reaction adopts a constant temperature hydrothermal method, controlling the temperature at 60-100° C. and the time for 24-48 hours.

[0017] The present invention provides a modified biological bone mineral scaffold based on lithium magnesium phosphorus. The material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the metal ion source solution can be 15-50 g:100 mL, and the material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the phosphorus source composite solution can be 15-50 g:100 mL.

[0018] In the metal ion source solution, the magnesium source is one of magnesium acetate, magnesium sulfate, magnesium hydrogen phosphate, etc.; the lithium source is lithium chloride; the supplementary calcium source is one of calcium chloride and calcium hydroxide; the zinc source is a soluble zinc salt such as zinc nitrate and zinc acetate; the strontium source is a soluble strontium salt such as strontium nitrate, strontium acetate, and strontium sulfate; the iron source is a soluble iron salt such as ferrous sulfate, ferrous chloride, ferric chloride, and ferric acetate; the phosphorus source binary system is a composite solution of phosphoric acid and a soluble phosphate; the soluble phosphate is selected from one or a combination of diammonium hydrogen phosphate, ammonium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, and magnesium hydrogen phosphate.

[0019] The final concentration of magnesium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.05-0.20 mol / L; the final concentration of lithium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.06-0.6 mol / L; the final concentration of zinc ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.1-0.6 mol / L; the final concentration of strontium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-0.9 mol / L; the final concentration of ferrous ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.1-0.6 mol / L; the final concentration of calcium ions supplemented by the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-1.5 mol / L; preferably, the final concentration of phosphate provided by phosphoric acid in the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-0.9 mol / L. ; The final concentration of phosphate provided by soluble phosphate in the binary system containing osteogenic active metal ions and phosphorus sources is 0.06-0.6 mol / L; preferably, the total molar concentration of osteogenic active cations including calcium (the molar number of lithium ions of monovalent ions is halved, and the molar concentration of trivalent iron ions is multiplied by 1.5) in the reaction system to the molar concentration ratio of phosphorus ions is 1.1-1.6:1.

[0020] After the hydrothermal reaction, the liquid is dried at a constant temperature and baked to dryness, and the baking and drying temperature is preferably 75°C-198°C.

[0021] During high-temperature calcination, the parameters of the high-temperature calcination are preferably 750° C.-1200° C., and the calcination time is 6 hours-24 hours.

[0022] The present invention provides a modified biological bone mineral scaffold doped with lithium and magnesium as the basis. The preparation method of the porous scaffold of calcined cancellous bone mineral of cattle or pigs is as follows:

[0023] (1): Porous scaffolds made of calcined cancellous bone or porcine bone mineral in the form of bone strips or blocks

[0024] (1-1): Cut the cancellous bone of cattle or pigs into cancellous bone strips or bone blocks with a thickness of 0.5-5 cm to obtain the raw bone; cylindrical cancellous bone strips can also be obtained using a ring saw;

[0025] (1-2): Place the raw bones in distilled water in a pressure cooker and cook for 36-60 minutes, then rinse with 50-75℃ drinking water, and repeat this step 5-6 times;

[0026] (1-3): The raw bone treated in step (1-2) 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 6-12 hours, and then slowly cooled to room temperature to obtain a calcined cancellous bone mineral porous scaffold. The powder diffraction composition of the bovine or porcine cancellous bone mineral material is hydroxyapatite;

[0027] (2): Granular bovine or porcine cancellous bone mineral porous scaffold

[0028] (2-1): Cut the cancellous bones of cattle or pigs into bone strips or blocks with a thickness of 0.5-6 cm to obtain raw bones;

[0029] (2-2): Place the raw bone in distilled water in a pressure cooker and cook for 36-60 minutes, then rinse with 50-75℃ drinking water, and repeat this step 5-6 times;

[0030] (2-3): The raw bone treated in step (2-2) 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 6-12 hours, and then slowly cooled to room temperature to obtain a bovine or porcine calcined cancellous bone mineral porous scaffold;

[0031] (2-4): The bovine cancellous bone mineral porous scaffold obtained in step (2-3) is crushed with a food crusher, and various specifications of particles such as 0.2-1mm, 1-3mm, 3-5mm, and 5-7mm of granular bovine cancellous bone mineral porous cell scaffold are screened with a stainless steel sieve for use; the porcine cancellous bone mineral porous scaffold obtained in step (2-3) is crushed with a food crusher, and 0.2-0.8mm and 0.33-1mm of granular porcine cancellous bone mineral porous cell scaffold are screened with a stainless steel sieve for use.

[0032] The present invention continues to address the problems that existing bone graft substitute materials have difficulty in achieving a good three-dimensional interconnected mesh structure, mechanical strength, degradability and osteogenic activity. It uses bovine (porcine) cancellous bone mineral, which is highly similar to the structure and composition of the mineralized cell matrix of human cancellous bone, as a precursor and the main source of calcium and phosphorus, and incorporates active ions magnesium, lithium and phosphorus as a basis. At the same time, one or more other osteogenic active ions such as strontium, zinc, iron, and calcium are also incorporated. White sugar is used as an adhesive and pore-forming agent to form a modified biological bone mineral scaffold incorporated with lithium, magnesium and phosphorus as a basis, in order to select more ideal bone tissue engineering scaffolds and bone graft substitute materials.

[0033] The modified biological bone mineral scaffold, obtained by the present invention and based on lithium and magnesium, can effectively stabilize the incorporated magnesium, lithium, and other osteogenic active ions such as strontium, zinc, iron, and calcium. It also maintains the three-dimensional interconnected mesh structure and natural crystal structure and relatively good mechanical strength of the porous scaffold of bovine calcined cancellous bone mineral. At the same time, the scaffold wall has honeycomb-like and Ganoderma lucidum-like calcium phosphate crystals containing active ions, which can effectively increase the material's specific surface area and improve cell adhesion and osteogenic activity. The modified biological bone mineral scaffold, provided by the present invention, based on lithium and magnesium, has a good microporous structure in its formed particles, such as particles of 0.25-1 mm, 1-2 mm, 2-4 mm, and 4-6 mm, with good porosity between the particles. The small particles and microparticles have good hydrophilicity, can be partially dissolved, and can serve as a good carrier for drugs such as antibiotics. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG1 is a general photograph of two types of modified stents of the present invention;

[0035] FIG2 is an XRD analysis diagram of the material of the present invention;

[0036] FIG3 is a scanning electron microscope image of a product according to an embodiment of the present invention;

[0037] FIG4 is a scanning electron micrograph of calcium phosphate crystals doped with bone-forming active ions according to an embodiment of the present invention;

[0038] FIG5 is an energy spectrum diagram of a modified material of the present invention;

[0039] FIG6 is a diagram illustrating bacterial culture of the product of the present invention as a carrier of antimicrobial agents;

[0040] FIG7 is a diagram of a combined cell culture product of the present invention;

[0041] FIG8 is a diagram illustrating the product of the present invention in an animal skull defect model experiment;

[0042] FIG9 is a diagram showing the mechanical strength of the product of the present invention.

[0043] Type your technical solution description paragraph here. DETAILED DESCRIPTION

[0044] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention and are not used to limit the present invention. Example 1:

[0045] The present embodiment provides a modified biological bone mineral scaffold based on lithium and magnesium, which is obtained by immersing a porous scaffold of calcined cancellous bone mineral from cattle or pigs in a composite solution containing active metal ions of magnesium, lithium, and one or more of strontium, zinc, iron, and calcium and a phosphorus source, and performing a hydrothermal reaction treatment, baking and drying, and then calcining at a high temperature; the porous scaffold of calcined cancellous bone mineral from cattle or pigs is subjected to a hydrothermal reaction treatment with a composite solution of a metal ion source containing active ions of magnesium, lithium, and one or more of calcium, strontium, zinc, and iron and a phosphorus source, and the following scheme is selected: Or the porcine calcined cancellous bone mineral porous scaffold is first immersed in a metal ion source solution containing active ions of magnesium and lithium and one or more of strontium, zinc, iron, and calcium, and a white sugar source solution, the liquid is dried by microwave or constant temperature oven, baked and dried at 96°C-198°C, and then placed in a phosphorus source composite solution for hydrothermal reaction; the phosphorus source composite solution is a phosphorus source binary system, thereby forming a binary system containing osteogenic active metal ions and phosphorus source; the hydrothermal reaction adopts a constant temperature hydrothermal method, with the temperature controlled at 60-100°C and the reaction time being 24-48 hours.

[0046] This embodiment provides a modified biological bone mineral scaffold based on lithium and magnesium. The material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the metal ion source solution can be 15-50 g:100 mL, and the material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the phosphorus source composite solution can be 15-50 g:100 mL.

[0047] This embodiment provides a modified bio-bone mineral scaffold based on lithium magnesium phosphate. In the metal ion source solution, the magnesium source is one of magnesium acetate, magnesium sulfate, magnesium hydrogen phosphate, etc.; the lithium source is lithium chloride; the supplementary calcium source is one of calcium chloride and calcium hydroxide; the zinc source is a soluble zinc salt such as zinc nitrate or zinc acetate; the strontium source is a soluble strontium salt such as strontium nitrate, strontium acetate, or strontium sulfate; the iron source is a soluble iron salt such as ferrous sulfate, ferrous chloride, ferric chloride, or ferric acetate; the phosphorus source binary system is a composite solution of phosphoric acid and a soluble phosphate; the soluble phosphate is selected from one or a combination of diammonium hydrogen phosphate, ammonium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, and magnesium hydrogen phosphate.

[0048] The final concentration of magnesium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.05-0.20 mol / L; the final concentration of lithium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.06-0.6 mol / L; the final concentration of zinc ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.1-0.6 mol / L; the final concentration of strontium ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-0.9 mol / L; the final concentration of ferrous ions in the binary system containing osteogenic active metal ions and phosphorus sources is 0.1-0.6 mol / L; the final concentration of calcium ions supplemented by the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-1.5 mol / L; preferably, the final concentration of phosphate provided by phosphoric acid in the binary system containing osteogenic active metal ions and phosphorus sources is 0.15-0.9 mol / L. ; The final concentration of phosphate provided by soluble phosphate in the binary system containing osteogenic active metal ions and phosphorus sources is 0.06-0.6 mol / L; preferably, the total molar concentration of osteogenic active cations including calcium (the molar number of lithium ions of monovalent ions is halved, and the molar concentration of trivalent iron ions is multiplied by 1.5) in the reaction system to the molar concentration ratio of phosphorus ions is 1.1-1.6:1.

[0049] After the hydrothermal reaction, the liquid is dried at a constant temperature and baked to dryness, and the baking and drying temperature is preferably 75°C-198°C.

[0050] During high-temperature calcination, the parameters of the high-temperature calcination are preferably 750° C.-1200° C., and the calcination time is 6 hours-24 hours.

[0051] This embodiment provides a modified biological bone mineral scaffold based on lithium and magnesium. The preparation method of the porous scaffold of calcined cancellous bone mineral of bovine or porcine is as follows:

[0052] (1): Porous scaffolds made of calcined cancellous bone or porcine bone mineral in the form of bone strips or blocks

[0053] (1-1): Cut the cancellous bone of cattle or pigs into cancellous bone strips or bone blocks with a thickness of 0.5-5 cm to obtain the raw bone; cylindrical cancellous bone strips can also be obtained using a ring saw;

[0054] (1-2): Place the raw bones in distilled water in a pressure cooker and cook for 36-60 minutes, then rinse with 50-75℃ drinking water, and repeat this step 5-6 times;

[0055] (1-3): The raw bone treated in step (1-2) 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 6-12 hours, and then slowly cooled to room temperature to obtain a calcined cancellous bone mineral porous scaffold. The powder diffraction composition of the bovine or porcine cancellous bone mineral material is hydroxyapatite; the compressive strength is 0.82-2 MPA / cm2 (Figure 1);

[0056] (2): Granular bovine or porcine cancellous bone mineral porous scaffold

[0057] (2-1): Cut the cancellous bones of cattle or pigs into bone strips or blocks with a thickness of 0.5-6 cm to obtain raw bones;

[0058] (2-2): Place the raw bone in distilled water in a pressure cooker and cook for 36-60 minutes, then rinse with 50-75℃ drinking water, and repeat this step 5-6 times;

[0059] (2-3): The raw bone treated in step (2-2) 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 6-12 hours, and then slowly cooled to room temperature to obtain a bovine or porcine calcined cancellous bone mineral porous scaffold;

[0060] (2-4): The bovine cancellous bone mineral porous scaffold obtained in step (2-3) is crushed with a food crusher, and various specifications of particles such as 0.2-1mm, 1-3mm, 3-5mm, and 5-7mm of granular bovine cancellous bone mineral porous cell scaffold are screened with a stainless steel sieve for use; the porcine cancellous bone mineral porous scaffold obtained in step (2-3) is crushed with a food crusher, and 0.2-0.8mm and 0.33-1mm of granular porcine cancellous bone mineral porous cell scaffold are screened with a stainless steel sieve for use.

[0061] The powder diffraction composition analysis of bovine and porcine cancellous bone minerals and the elemental analysis by inductively coupled plasma atomic emission spectrometry were performed.

[0062] Powder diffraction results showed that the mineral compositions of cattle and pig bones were all “elemental” hydroxyapatite (Ca5 ( P O4 )3 OH);

[0063] Inductively coupled plasma atomic emission spectrometry element detection results:

[0064] Ca39.2; P18.1; Li0.016, Mg0.72; Sr0.058; Zn0.021; Fe0.03;

[0065] In the specific implementation process of this embodiment, a specific bovine or porcine calcined cancellous bone porous bone mineral is immersed in a composite solution containing one or more active metal ions of magnesium, lithium, strontium, zinc, iron, and calcium and a phosphorus source for hydrothermal reaction treatment, and then baked, dried, and calcined at high temperature to obtain a sample. The sample will be further described as follows: Example 2:

[0066] [Sample of lithium-magnesium-phosphorus: No. 2104211] Prepare 0.15 mol / L magnesium sulfate heptahydrate, 0.2 mol / L lithium chloride, 0.30 mol / L 300 ml of a complex solution of diammonium hydrogen phosphate and 0.30 mol / L orthophosphoric acid was completely dissolved in a microwave at medium heat for 3 minutes, and 50 g of a porous scaffold of calcined bovine cancellous bone mineral with a porosity of approximately 65-80% was immersed in the solution. The total cation / phosphorus molar ratio in the reaction system was 1.3582. A hydrothermal reaction was carried out at 100°C in a 1000 ml beaker for 24 hours. The beaker was removed and 10.76 g of 0.105 mol / L white sugar was added to the solution. The reaction system was transferred to a 1000 ml beaker and dried in a constant temperature oven. After drying at 100°C for 14 hours (with repeated stirring at the beginning), the temperature was increased by 2.5°C per minute to 900°C, maintained for 360 minutes, and then cooled. The temperature was raised to 400°C after 180 minutes and then cooled to room temperature after 90 minutes. Sample No. 2104211 weighed 61.52 g and had good texture and strength. A small amount of the material contained unvolatile carbon black.

[0067] Analysis and testing of No. 2104211:

[0068] Ca9 Mg Li ( P O4 )7 46.7%

[0069] Ca2 ( P2 O7 52.0%

[0070] Li Mg P O4 1.3%

[0071] Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0072] Li 0.53, Mg 1.64, Ca 33.6, P 22.4;

[0073] The molar ratio of lithium to total cations is 7.77%;

[0074] The molar ratio of magnesium to total cations is 6.87%.

[0075] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium phosphorus was effectively incorporated. Example 3:

[0076] [Sample with Lithium, Magnesium, Calcium, and Phosphorus: No. 2105262] Prepare a 200ml solution of 0.1mol / L anhydrous calcium chloride, 0.05mol / L magnesium sulfate heptahydrate, 0.1388mol / L lithium chloride, and 0.04mol / L white sugar. Add 50g of a bovine cancellous bone mineral scaffold with a porosity of approximately 60-80%. Microwave on medium-low heat [stirring repeatedly] until the solution is reduced. Transfer the scaffold to a 1000ml beaker and bake in an incubator at 198°C for 300 minutes [stirring repeatedly during the initial period]. Once the scaffold turns dark brown and is completely dry, set aside. A 0.225 mol / L orthophosphoric acid (3 ml of orthophosphoric acid) and 200 ml of a 0.0909267 mol / L diammonium hydrogen phosphate complex solution were prepared, and a porous scaffold of calcined bovine cancellous bone mineral treated with magnesium, lithium, calcium, and white sugar was immersed therein [the molar ratio of total bone-forming active metal ions to phosphorus in the reaction system was 1.4975]. The reaction was hydrothermally reacted at 85°C [protected in a 1000 ml beaker] for 24 hours, and the solution was dried at 198°C [stirred repeatedly], and then baked at 198°C for 5 hours [stirred repeatedly in the early stages]. After the surface of the material was carbonized [weighing 58.37 g], ​​the temperature was raised by 2.5°C per minute to 900°C, maintained for 360 minutes, and then the furnace temperature was raised to 400°C for 180 minutes. The temperature was then lowered to room temperature at 70°C for 180 minutes to obtain a lithium-doped magnesium-calcium sample: No. 2105262 [53.76 g, with excellent appearance, texture, strength, and air permeability].

[0077] Lithium Calcium Magnesium Phosphate Ca9 Mg Li ( P O4 )7

[0078] Hydroxylapatite, syn Ca5 ( P O4 )3 OH

[0079] Calcium diphosphate - β| Calcium Phosphate Ca2 ( P2 O7 )

[0080] Sample No. 2105262:

[0081] Ca9 Mg Li ( P O4 )7 59.7%

[0082] Ca5 ( P O4 )3 OH 28.9%

[0083] Ca2 ( P2 O7 ) 11.4%

[0084] Powder diffraction composition analysis suggests that lithium magnesium calcium phosphate is effectively incorporated. Example 4:

[0085] [Sample with lithium, magnesium, calcium, and phosphorus: No. 2105202] Prepare a 200ml solution of 0.1mol / L anhydrous calcium chloride, 0.05mol / L magnesium sulfate heptahydrate, 0.06mol / L lithium chloride, and 0.04mol / L white sugar in distilled water. Immerse 50g of bovine cancellous bone mineral granules (2-4mm in diameter) in this solution. Microwave on medium heat for 2 minutes at a time, stirring repeatedly, until the liquid is dry. Transfer the scaffold to a 1000ml beaker and bake in a 198°C incubator for 300 minutes, stirring repeatedly. Once the scaffold turns dark brown and is completely dry, set aside. Prepare 200 ml of a composite solution of 0.225 mol / L orthophosphoric acid [add 3 ml of orthophosphoric acid] and 0.06466 mol / L diammonium hydrogen phosphate, immerse the porous scaffold of bovine calcined cancellous bone mineral treated with magnesium, lithium, calcium, etc. [the total divalent ion / phosphorus ion molar ratio of the reaction system is 1.49749], conduct a hydrothermal reaction at 85°C [protected in a 1000 ml beaker] for 24 hours, dry the solution at 98°C without protection [stirred repeatedly], and then bake at 198°C for 2 hours; carbonize the surface of the material, increase the temperature by 2.5°C per minute to 1075°C, maintain for 360 minutes, then wait for 400 minutes until the furnace temperature reaches 400°C, and then heat to room temperature to obtain a lithium-doped magnesium-calcium sample: No. 2105202 [weighing 52.79 grams, with good appearance, texture, and strength].

[0086] Powder diffraction results of sample 2105202:

[0087] Ca9 Mg Li ( P O4 )7 46.9%

[0088] Ca5 ( P O4 )3 OH 53.1%

[0089] Elemental detection results of sample 2105202 by inductively coupled plasma atomic emission spectrometry (unit: wt%):

[0090] Li 0.19, Mg1.42, Ca 35.8, P 20.2.

[0091] Magnesium / total cation molar ratio ≈5.96%

[0092] Lithium / total cation molar ratio ≈2.84%

[0093] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium phosphorus was effectively incorporated. Example 5:

[0094] [Sample of lithium magnesium strontium phosphate: No. 2104191] was prepared with a concentration of 0.05 mol / L magnesium sulfate heptahydrate, 0.10 mol / L strontium nitrate [6.3489 g of strontium nitrate], 0.25 mol / L lithium chloride [3.17925 g], 0.15 mol / L Prepare 300 ml of a composite solution of diammonium hydrogen phosphate [add 5.9427 g of diammonium hydrogen phosphate] and 0.3 mol / L orthophosphoric acid [add 6 ml of orthophosphoric acid] [the total metal ion / phosphorus ion molar ratio in the reaction system is 1.3391]. Immerse 50 g of a porous scaffold of bovine calcined cancellous bone mineral with a porosity of about 60-80% in it, and the solution scale is 325 ml; hydrothermally react at 70°C for 12 hours, then remove the protection, add 0.165 mol / L white sugar [10.76 g], and intermittently absorb the turbid solution at the bottom and pour it on the scaffold] for 12 hours; dry at 100°C for 12 hours to carbonize the surface of the material [stir several times at the beginning]. The temperature was raised by 2.5°C per minute to 900°C, maintained for 360 minutes, then raised to 400°C over 180 minutes and returned to room temperature over 90 minutes to obtain lithium-doped magnesium strontium sample No. 2104191 [58.97 g, excellent appearance, texture, and strength, with well-preserved microporous structure as shown in Figure 1].

[0095] 2104191 powder diffraction test results:

[0096] Ca9 Mg Li ( P O4 )7 88.4%

[0097] Ca2 (P2O7) 11.6%

[0098] 2104191 Inductively coupled plasma atomic emission spectrometry elemental analysis results (unit: wt%):

[0099] Li 0.77, Mg 1.57, Ca 32.6, P / Sr 4.08.

[0100] Lithium / total cation molar ratio ≈10.74%

[0101] Magnesium / total cation molar ratio ≈5.98%

[0102] Strontium / total cation molar ratio ≈ 4.51

[0103] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium strontium phosphide was effectively incorporated. Example 6:

[0104] [Sample with lithium magnesium strontium calcium: No. 2105261] Prepare a complex solution of 0.1 mol / L anhydrous calcium chloride, 0.05 mol / L magnesium sulfate heptahydrate, 0.1388 mol / L lithium chloride, 0.12 mol / L strontium nitrate, and 0.04 mol / L white sugar with distilled water, and immerse 50 g of a bone mineral scaffold containing approximately 75-90% bovine cancellous bone in the solution; microwave on medium-low heat to dry the liquid, and bake at a constant temperature of 198°C in the original 500 ml beaker for 300 minutes until the scaffold turns dark brown and is ready for use.

[0105] Prepare a 200 ml composite solution of 0.225 mol / L orthophosphoric acid and 0.1709267 g diammonium hydrogen phosphate. Immerse the previously treated bovine calcined cancellous bone mineral porous scaffold in this solution (the molar ratio of diosteogenic active ions to phosphorus in the reaction system is 1.4961). Hydrothermally react at 85°C (protected in a 1000 ml beaker for 24 hours, then unprotected at 198°C to dry the solution). Then bake at 198°C for 5 hours. Carbonize the material surface (weighing 60.40 g). Raise the temperature by 2.5°C / min to 900°C, maintain for 360 minutes, then raise the furnace temperature to 400°C for 180 minutes, and then return to room temperature at 70°C for 180 minutes. Sample number 2105261: [56.31 g, excellent appearance, texture, and strength; a small amount of material contains unvolatile carbon black].

[0106] 2105261 powder diffraction test results:

[0107] Ca9 Mg Li ( P O4 )7 61.7%

[0108] Ca5 ( P O4 )3 OH 30.1%

[0109] Ca2 ( P2 O7 ) 8.2% Example 7:

[0110] [Sample No. 2106151: Lithium-magnesium-strontium-calcium solution added] Prepare 200 ml of a complex solution containing 0.10 mol / L anhydrous calcium chloride, 0.055 mol / L magnesium sulfate heptahydrate, 0.576 mol lithium chloride trihydrate, 0.12 mol / L strontium nitrate, and 0.06 mol white sugar. Immerse 50 g of a bovine cancellous bone mineral scaffold (calcined at 988°C) in this solution. Microwave on medium heat and defrost to dry the solution. Dry the scaffold in the original 500 ml beaker in a 198°C incubator for 600 minutes (stirring repeatedly). The scaffold (weighing 62.14 g) will turn black and be set aside.

[0111] Prepare 0.45 mol / L orthophosphoric acid [add 6 ml of orthophosphoric acid] and 200 ml of 0.17733 mol / L diammonium hydrogen phosphate composite solution, immerse the bovine calcined cancellous bone mineral porous scaffold previously treated with lithium magnesium strontium calcium [the reaction system has a molar ratio of calcium and other osteogenic active ions [on a divalent basis] to phosphorus of 1.335418], hydrothermally react at 70°C [protected in a 1000 ml beaker] for 24 hours, microwave on medium heat, thaw to dry [stir repeatedly], and then bake at 198°C [stir repeatedly] for 6 hours; carbonize the surface of the material [weighing 63.05 grams], raise the temperature of the scaffold at 2.5°C per minute to 900°C, maintain for 360 minutes, then raise the furnace temperature to 400°C for 180 minutes, and cool to room temperature of 70°C for 180 minutes to obtain lithium magnesium strontium calcium sample: No. 2106151 [57.26 grams, excellent appearance, texture, and strength].

[0112] Sample 2106151 was sent for powder diffraction and elemental analysis, and the results were:

[0113] Ca9 Mg Li ( P O4 )7 78.4%

[0114] Ca5 ( P O4 )3 ( OH ) 11.2%

[0115] Li Ca ( P O4 ) 7.4%

[0116] Li3 P O4 3.1%

[0117] Inductively coupled plasma atomic emission spectrometry elemental analysis results for sample 2106151 (unit: wt%):

[0118] Li 1.18; Mg 1.24; Ca 32.8; P 20.9; Sr 2.74.

[0119] Magnesium / total cation molar ratio ≈5.17%

[0120] Lithium / total cation molar ratio ≈17.2%

[0121] Strontium / total cation molar ratio ≈ 3.17%

[0122] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium phosphorus was effectively incorporated. Example 8:

[0123] [Sample number 2106152: Lithium-magnesium-strontium-calcium solution added] Prepare 200 ml of a solution containing 0.10 mol / L anhydrous calcium chloride, 0.055 mol / L magnesium sulfate heptahydrate, 0.5769 mol / L lithium chloride trihydrate, 0.12 mol / L strontium nitrate, and 0.06 mol / L white sugar. Immerse 50 g of bovine cancellous bone mineral scaffold in the solution. Microwave on medium heat and defrost on medium heat (stirring repeatedly) until the liquid is reduced. Bake in the original 500 ml beaker in a 198°C incubator for 300 minutes until the scaffold turns brownish-black (weighing 67.18 g). Dry thoroughly and set aside.

[0124] Prepare 200 ml of a 0.45 mol / L orthophosphoric acid (add 6 ml of orthophosphoric acid) and 0.17733 mol / L diammonium hydrogen phosphate (DAHPO) complex solution; immerse the porous bovine cancellous bone mineral scaffold, previously added with calcium, magnesium, lithium, strontium, and sugar, in this solution (set the ratio of the metal ion molar concentration (divalent) to the phosphorus ion molar concentration in the reaction system to 1.35). Hydrothermal reaction was carried out at 70°C [protected in a 1000 ml beaker] for 24 hours, microwaved on medium heat, thawed to dryness, and then dried at 198°C [stirred repeatedly] for 6 hours; the carbonized scaffold [weighing 66.63 grams] on the surface of the material was heated at 2.5°C per minute to 900°C, maintained for 360 minutes, then the furnace temperature was raised to 400°C for 360 minutes, and then cooled to room temperature at 70°C for 360 minutes to obtain lithium magnesium strontium calcium sample: No. 2106152 [59.35 grams, with good appearance and pore structure, and excellent strength (significantly higher than its precursor)].

[0125] 2106152 sent powder diffraction and inductively coupled plasma atomic emission spectrometry element detection results:

[0126] Ca9 Mg Li ( P O4 )7 79.5%

[0127] Li3 P O4 8.5%

[0128] Ca2 (P2O7 12.0%)

[0129] 2106152 Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0130] Li 1.15, Mg 1.21, Ca 32.1, P 21.1, Sr 2.66.

[0131] Magnesium / total cations molar ratio 5.21%

[0132] Lithium / total cations molar ratio 16.57%

[0133] Molar ratio of strontium to total cations: 3.71%

[0134] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, strontium, calcium and phosphorus were effectively incorporated. Example 9:

[0135] [Sample number 2106281: Lithium-magnesium-strontium-calcium solution added] Prepare 200 ml of a complex solution of 0.10 mol / L anhydrous calcium chloride, 0.055 mol / L magnesium sulfate heptahydrate, 0.36 mol lithium chloride trihydrate, 0.3 mol / L strontium nitrate, and 0.06 mol white sugar. Immerse 50 g of a bovine cancellous bone mineral scaffold (calcined at 988°C) in this solution. Microwave on medium-low heat until the solution is dried. Bake in a 198°C incubator in the original 500 ml beaker (stirring repeatedly) for 600 minutes until the scaffold turns completely black (weight 70.27 = 71.68 - 1.41 g powder). Set aside.

[0136] Prepare a concentration of 0.45 mol / L orthophosphoric acid [add 6 ml of orthophosphoric acid), 200 ml of 0.18836 mol / L diammonium hydrogen phosphate complex solution, immerse the bovine calcined cancellous bone mineral previously added with calcium, magnesium, lithium, strontium and white sugar [the molar ratio of total metal ions to phosphorus in the reaction system is 1.389] in it, hydrothermally react at 60°C [protected in a 1000 ml beaker] for 24 hours, microwave on medium heat, thaw to dry, and then bake at 198°C [stirred repeatedly] for 6 hours; carbonize the surface of the material [weight 69.35 g = 73.02-3.67 powder] and then raise the temperature by 2.5°C per minute to 900°C, maintain for 360 minutes, then raise the furnace temperature to 400°C for 180 minutes, and then cool to room temperature of 70°C for 180 minutes to obtain lithium magnesium strontium phosphate calcium doped sample: No. 2106281: the bracket weighs 62.8 grams.

[0137] The product of this embodiment is combined with cell culture, as shown in Figure 8. Through staining of cells and scaffolds and analysis under a confocal microscope, it was found that the bioactivity and surface nanolattice structure of the artificial bone scaffold can promote cell adhesion and spreading on the surface of the artificial bone, thereby promoting cell bioactivity.

[0138] The powder diffraction and elemental analysis of sample No. 2106281 yielded the following results:

[0139] Ca9 Mg Li ( P O4 )7 69.7%

[0140] Ca10.132 (PO4)5.958 (OH)3.258 30.3%

[0141] Inductively coupled plasma atomic emission spectrometry elemental analysis results for sample 2106281 (unit: wt%):

[0142] Li0.64, Mg 1.04, Ca 29.1, P 18.6, Sr6.9.

[0143] Magnesium / cation molar ratio 4.7829%

[0144] Lithium / cation molar ratio 10.3%

[0145] Strontium / cation molar ratio 8.767%

[0146] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium phosphorus strontium was effectively incorporated. Example 10:

[0147] [Lithium-magnesium-strontium-calcium added sample: No. 2106081] Prepare 200 ml of a complex solution containing 0.1 mol / L anhydrous calcium chloride, 0.05 mol / L magnesium sulfate heptahydrate, 0.1388 mol / L lithium chloride, 0.12 mol / L strontium nitrate, and 0.05 mol / L white sugar. Immerse 50 g of the bovine cancellous bone mineral scaffold in this solution. Microwave on medium heat (stirring repeatedly) and incubate at 136°C for 120 minutes to dry the liquid. Bake in a 198°C oven for 300 minutes to completely dry. The scaffold will turn into a dark brown powder, ready for use [57.31 g].

[0148] Prepare a 0.225 mol / L orthophosphoric acid solution (add 3 ml of orthophosphoric acid) and 200 ml of a 0.098 mol / L diammonium phosphate composite solution, pour it into a beaker containing a porous scaffold of calcined bovine cancellous bone mineral treated with calcium, magnesium, lithium, and white sugar (the molar ratio of total cations / phosphorus in the reaction system is 1.556). Hydrothermal reaction was performed at 85°C (protected in a 1000 ml beaker) for 27 hours, followed by drying at 108°C for 5 hours without protection (intermittently absorbing the turbid solution at the bottom and pouring it on the scaffold). Bake at 198°C for 5 hours until the surface of the material is completely carbonized (weight 58.10 g), with a temperature increase of 2.5°C per minute. 385 minutes to 800℃, maintained for 360 minutes, then the furnace temperature was raised to 400℃ for 360 minutes, and then it was lowered to room temperature of 35℃ for 360 minutes to obtain lithium-magnesium strontium calcium phosphate doped sample: No. 2106081: 54.22 grams, with excellent appearance, texture and strength, and a small part of the material contains unvolatile carbon black.

[0149] Hydroxylapatite, syn Ca5 ( P O4 )3 ( OH )

[0150] Whitlockite magnesian, syn Ca18 Mg2 H2 ( P O4 )1

[0151] Sample 2106081 was sent for material powder diffraction and elemental analysis, and the results were:

[0152] Ca5 ( P O4 )3 ( OH ) 75.3%

[0153] Ca18 Mg2 H2 ( P O4 )14 24.7% Example 11:

[0154] [Sample with lithium magnesium strontium calcium phosphorus: No. 2111231] Prepare 100 ml of a composite solution of 1.8 mol / L anhydrous calcium chloride (19.98 g), 0.2 mol / L magnesium acetate tetrahydrate (24.29 g), 0.36 mol lithium chloride trihydrate (1.526 g), 0.45 mol / L strontium nitrate (9.52 g), and 0.06 mol / L white sugar (4.1 g), and immerse 50 g of bovine cancellous bone mineral scaffold in it; microwave on low heat [stir repeatedly] to dry the liquid, and bake at a constant temperature of 176°C in the original 1000 ml beaker for 300 minutes. The surface of the scaffold will turn into a dark brown sugar mud-like state. Bake at 196°C for 360 minutes [powder and small particles will fall off] for later use.

[0155] Assuming the reaction solution is 100 ml, the molar ratio of calcium phosphate and active ions to phosphorus in the reaction system is 1.54676, then the phosphate concentration of the reaction system except the scaffold is 2.086666 mol / L, 6 ml of orthophosphoric acid and 15.66 of diammonium hydrogen phosphate are needed to prepare 0.9 mol / orthophosphoric acid, 1.18667 mol / diammonium hydrogen phosphate composite solution, the bovine calcined cancellous bone mineral porous scaffold previously treated with lithium magnesium calcium strontium, etc. is immersed in it; 75℃ hydrothermal reaction [2000 ml beaker protection, half an hour later, evenly distributed white oval neoplasia can be seen around the scaffold, 24 hours [23 hours later, the solution is seen to be dry, the last hour is not protected], a small part is taken out [divide into two parts 2111231, 21112312] and the rest is not protected and continues to be dried at 126℃ for two hours under unprotected conditions, after taking out a small part [2111232] the rest is dried at 198℃ for another two hours [2111233, 2111234]. Take 2111231 and 2111232 and heat them at 5°C per minute until the temperature reaches 1075°C [set the room temperature to 20°C], maintain it for 6 hours and then cool it down [set it to drop to 150°C in 360 minutes] [excellent strength, the elastic modulus may be quite good, the brittleness is small, the appearance and ventilation are good, 2111231 weighs 9.19 grams, 2111232 weighs 13.64 grams; take 2111231 and 2111233 and heat them at 5°C per minute for 132 minutes to 750°C, maintain it for 720 minutes, then drop the furnace temperature to 100°C in 360 minutes, and cool it down to room temperature one minute later to obtain lithium-doped magnesium strontium calcium phosphate samples: No. 2111231: (7.63 grams); No. 2111233 (19.02 grams); [excellent appearance texture and strength; a small part of the material contains unvolatile carbon black, but the strength is higher than the sample calcined at 1075°C].

[0156] Take 2111234 and raise the temperature by 5°C per minute for 172 minutes to 900°C. After maintaining for 480 minutes, the furnace temperature is increased to 100°C for 360 minutes and then to room temperature of 10°C for 180 minutes to obtain 2111234; [22.05 grams, excellent appearance, texture and strength; a small part of the material contains unvolatile carbon black] [total weight of the scaffold is 71.53 grams, precursor is 50 grams, 1.4306 times]; sent for powder diffraction and elemental analysis; 211123 was subjected to scanning electron microscopy and energy spectrum detection, and there was good formation of calcium phosphate crystals containing active ions in the shape of ant nests, which may further increase the necessary surface area of ​​the material and facilitate the adhesion of bone repair cells. 2111234 Powder diffraction: Magnesium Phosphate (Ca2.589 Mg0.411) (P O4)2; Calcium diphosphate - β | Calcium Phosphate Ca2 (P2 O7) Dicalcium diphosphate(V) - α | Calcium Phosphate Ca2 P2 O7

[0157] 2111231【1075℃】

[0158] (Ca2.589 Mg0.411) (PO4)2 73.7%

[0159] Ca2 ( P2 O7 ) 26.3%

[0160] 2111232【1075℃】

[0161] (Ca2.589 Mg0.411) (PO4)2 90.8%

[0162] Ca2 ( P2 O7 ) 9.2%

[0163] 2111233【750℃】

[0164] Ca9 Mg Li ( P O4 )7 23.1%

[0165] Ca2 ( P2 O7 ) 27.5%

[0166] Ca5 ( P O4 )3 ( OH ) 49.5%

[0167] 2111234【900℃】

[0168] (Ca2.589 Mg0.411) (PO4) 34.7%

[0169] Ca2 ( P2 O7 ) 21.9%

[0170] Ca2 P2 O7 43.4%

[0171] Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0172] 2111234 Lithium 0.51; Magnesium 0.99; Strontium 4.41; Calcium 33.0; Phosphorus 21.7.

[0173] Magnesium / total cation molar concentration ≈ 4.166%

[0174] Lithium / total cation molar concentration ≈7.42%

[0175] Strontium / total cation molar concentration ≈ 5.0532%

[0176] Powder diffraction composition and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, strontium, calcium and phosphorus were effectively incorporated. Example 12:

[0177] [Sample with lithium magnesium, strontium, calcium and phosphorus: No. 2111291] Prepare 100 ml of a complex solution of 0.9 mol / L anhydrous calcium chloride, 0.2 mol / L magnesium acetate tetrahydrate, 0.36 mol lithium chloride trihydrate, 0.45 mol / L strontium nitrate and 0.06 mol / L white sugar, and immerse 50 g of bovine cancellous bone mineral scaffold in it; microwave on low heat [stir repeatedly] to dry the liquid, dry at a constant temperature of 75°C in the original 500 ml beaker for 180 minutes, and bake at 196°C in a crucible for 300 minutes [the scaffold turns dark brown with sand-like particles falling off] for later use.

[0178] Prepare 100 ml of reaction solution in a new 500 ml beaker (set the molar ratio of calcium and active ions to phosphorus in the reaction system to 1.55). The phosphate concentration in the reaction system, excluding the scaffold, should be 1.342 mol / L. Prepare a composite solution of 0.6 mol / orthophosphoric acid and 0.7342 mol / diammonium hydrogen phosphate. Microwave on medium heat for 4 minutes to completely dissolve it. Immerse the porous scaffold of bovine calcined cancellous bone mineral treated with lithium, magnesium, calcium, strontium, etc. previously. Hydrothermally react at 75°C [protected by a 2000 ml beaker, evenly distributed oval white growths can be seen around the scaffold in half an hour] for 24 hours. After the liquid is dried at 75°C, bake it in a crucible at 196°C [the scaffold is uniformly dark brown, weighing 84.42 g, and is evenly divided into two parts 2111291 and 2111292].

[0179] 2111291 was heated at 5°C / min for 172 minutes to 900°C, held for 720 minutes, then heated to 100°C over 360 minutes. After one minute, the material returned to room temperature. [32.53 / 42.21 / 25g, excellent appearance, texture, and strength; new particles were observed on the support wall under a magnifying glass]. 2111292 was heated at 5°C / min for 148 minutes (at 72°C) to 750°C, held for 720 minutes, then heated to 100°C over 360 minutes. After cooling to room temperature, the material returned to room temperature. [Excellent appearance, texture, and strength; a small amount of unvolatile carbon black was observed].

[0180] The powder elements of materials 2111291 and 2111292 indicate the effective incorporation of elements such as lithium, magnesium, strontium, and phosphorus, as shown in Figure 2.

[0181] Scanning electron microscopy and energy spectrum analysis were performed on sample No. 2111291, which showed the formation of uniform calcium phosphate crystals containing active ions. The calcium phosphate crystals were in the shape of Ganoderma lucidum clusters, which may further increase the specific surface area of ​​the material and facilitate the adhesion and spreading of bone repair cells. Scanning electron microscopy was performed, as shown in Figures 3, 4, and 5.

[0182] 2111291【900℃】

[0183] (Ca2.589 Mg0.411) (PO4)2 55.7%

[0184] Ca5 ( P O4 )3 OH 38.0%

[0185] Sr ( P O3 )2 6.3%

[0186] 2111292【750℃】

[0187] Ca9 Mg Li ( P O4 )7 27.1%

[0188] Ca5 ( P O4 )3 ( OH ) 53.2%

[0189] Li Mg P O47.1%

[0190] Sr ( P O3 )2 12.6%

[0191] Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0192] 2111291 Lithium 0.55; Magnesium 1.02; Strontium 4.64; Calcium 34.5; Phosphorus 20.4.

[0193] The powder diffraction composition and inductively coupled plasma atomic emission spectrometry elemental analysis of the material indicated that lithium magnesium strontium calcium and phosphorus were effectively incorporated. Example 13:

[0194] [Sample with lithium magnesium strontium: No. 2112121] Prepare 100 ml of a composite solution of 0.225 mol / L, 0.1 mol / L magnesium acetate tetrahydrate, 0.18 mol lithium chloride trihydrate, 0.225 mol / L strontium nitrate (4.762), and 0.06 mol / L white sugar with distilled water, and immerse 25 g of a bracket of porcine cancellous bone mineral particles [φ0.2-0.8 mm] in it; microwave on low heat to dry the liquid [stir repeatedly], dry it at a constant temperature of 75°C in the original 500 ml beaker for 120 minutes, and bake it in a crucible for 196 minutes [the bracket turns dark brown] for later use.

[0195] The molar ratio of calcium and active ions to phosphorus ions in the reaction system is set to 1.55, so the phosphate concentration in the reaction system excluding the scaffold is 0.5258. A 100 ml composite solution of 0.3 mol / orthophosphoric acid and 0.2258 mol / diammonium hydrogen phosphate is prepared in a new 500 ml beaker. The pig calcined cancellous bone mineral particles previously treated with lithium magnesium calcium strontium and white sugar are poured into the new beaker, and the scaffold is immersed in the composite liquid (the molar ratio of calcium phosphorus and active ions to phosphorus in the reaction system is about 1.35); after hydrothermal reaction at 75°C [protected in a 1000 ml beaker] for 24 hours [maintained at 37°C for 24 hours], after the liquid is dried at 75°C, it is baked in a crucible at 196°C [with intermittent stirring. The scaffolds are finally uniformly dark brown and weigh 38.46 g]. They are divided into two parts of 19.23 g each, namely 2112121 and 2112122.

[0196] Take 2112121, raise the temperature by 3°C / min to 750°C, hold for 180 minutes, then reduce the oven temperature to 10°C over 480 minutes, then return to room temperature after 1 minute. [Weight: 16.56 g, excellent appearance, texture, and strength; surface appears to have a coating]. Take 2112122, raise the temperature by 5°C / min to 1200°C [126 minutes], hold for 3 hours, then reduce the temperature [set to 15°C over 636 minutes] to obtain 2112122 [Weight: 15.89 g, excellent strength, possibly good elastic modulus, low brittleness, good appearance, and good ventilation].

[0197] Submitted for inspection:

[0198] 2112121【750℃】

[0199] Ca9 Mg Li ( P O4 )7 45.7%

[0200] Ca5 ( P O4 )3 ( OH ) 54.3%

[0201] 2112122【1200℃】

[0202] Ca2.86 Mg0.14 ( P O4 )2 100% Example 14:

[0203] [Sample with lithium magnesium strontium calcium phosphate: No. 2112151] Prepare 100 ml of a complex solution of 0.35 mol / L anhydrous calcium chloride, 0.1 mol / L magnesium acetate tetrahydrate, 0.18 mol lithium chloride trihydrate, 0.225 mol / L strontium sulfate, and 0.06 mol / L white sugar, and immerse 25 g of a scaffold of porcine cancellous bone mineral particles [φ0.3-0.8 mm, composed of hydroxyapatite] in it; microwave on low heat to dry the liquid [stir repeatedly], dry it at a constant temperature of 75°C in the original 500 ml beaker for 120 minutes, and bake it in a crucible for 196 minutes [the scaffold turns dark brown with sand-like falling particles] for later use.

[0204] The molar ratio of calcium and active ions to phosphorus ions in the reaction system was set at 1.5. The phosphate concentration in the reaction system, excluding the scaffold, should be 0.6766666. A 100ml solution of 0.3mol / orthophosphoric acid and 0.37666mol / diammonium hydrogen phosphate was prepared in a new 500ml beaker. The porcine calcined cancellous bone mineral microparticle scaffold, previously added with lithium magnesium calcium strontium and white sugar, was immersed in the solution. The reaction was hydrothermally reacted at 75°C for 42 hours. After drying the liquid at 75°C, the scaffold was baked in a crucible at 196°C with intermittent stirring. The scaffolds were uniformly black and weighed a total of 37.05g. The scaffolds were divided into two equal parts: 2112151 [17.52] and 2112152 [17.53].

[0205] Take 2112151, raise the temperature by 3 degrees Celsius per minute to 900°C, maintain for 180 minutes, then reduce the furnace temperature to 10°C after 480 minutes, and return to room temperature after 1 minute. Obtain 2112151: [15.44 g, excellent appearance, texture, and strength; the surface appears to have a coating].

[0206] Take 2112152 and heat it up by 5°C per minute to 1200°C [heating takes 396 minutes], maintain the temperature for 3 hours, and then cool it down [set to 15°C for 536 minutes] to obtain 2112152 [excellent strength, possibly quite good elastic modulus, low brittleness, good appearance and ventilation, weighing 17.43 grams].

[0207] Both powder element testing and inductively coupled plasma atomic emission spectrometry element detection indicate that the material is doped with effective elements.

[0208] 2112151【900℃】

[0209] Ca9 Mg Li ( P O4 )7 52.4%

[0210] Ca5 ( P O4 )3 OH 47.6%

[0211] 2112152【1200℃】

[0212] Ca5 ( P O4 )3 OH 17.4%

[0213] Ca2.86 Mg0.14 (P O4 )2 82.6%2112152Elemental detection results by inductively coupled plasma atomic emission spectrometry (unit: wt%):

[0214] Lithium 0.73, magnesium 1.24, strontium 5.58, calcium 33.2, phosphorus 20.7.

[0215] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, strontium, calcium and phosphorus were effectively incorporated. Example 15:

[0216] [Sample with lithium, magnesium, zinc, calcium, and phosphorus: No. 2109061] Prepare 200 ml of a complex solution of 0.10 mol / L anhydrous calcium chloride, 0.08 mol / L magnesium acetate tetrahydrate, 0.18 mol lithium chloride, and 0.18 mol zinc acetate dihydrate. Immerse 50 g of bovine cancellous bone mineral particles [0.4-1 mm, composed of hydroxyapatite, calcined at 988°C] in the solution. Microwave on medium heat or defrost until the liquid is dried (add 4.02 g of granulated sugar at appropriate times). Dry in the original 500 ml beaker in a 176°C incubator for 180 minutes. Stir repeatedly until the powder changes from caramel color to a brown, sandy consistency. Set aside.

[0217] The total cation / phosphorus ion molar ratio of the reaction system was set to 1.48; 200 ml of a complex solution of 0.3 mol / L orthophosphoric acid and 0.21836 mol / L 12-water disodium hydrogen phosphate was prepared, and black sand-like powdered bone mineral particles with calcium, magnesium, strontium and white sugar were immersed in it. After hydrothermal reaction at 65°C for 24 hours, the liquid was dried at low temperature in a microwave oven. Bake at 176°C for 6 hours, stirring every half hour. The particles gradually turn from caramel color to brown and are set aside [weight 65.64 g]. Then, heat at 5.8°C per minute to 900°C [set room temperature to 30°C, heat for 150 minutes], maintain for 180 minutes, and then cool to 188°C for three hours [weight 30.77 g]. The volume is measured with a measuring cup to be 30 ml. The relative density of tricalcium phosphate is about 3.2 g / cm³, and its porosity is calculated to be about 67%. The particles are magnified and photographed and then observed on a computer. It is found that the modified bioscaffold particles with a diameter of 0.4-1 mm have micropores as shown in Figure 1.

[0218] The powder diffraction and inductively coupled plasma atomic emission spectrometry elemental detection of the material indicated that lithium magnesium phosphorus and other elements were effectively incorporated into the material.

[0219] 2109061

[0220] Ca9 Mg Li ( P O4 )7 72.7%

[0221] Na Ca P O4 12.1%

[0222] Ca8.8 (PO4)6 (OH)1.92 15.2%

[0223] Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0224] Li 0.47, Mg 1.14, Ca 28.29, P 20.5, Zn2.39.

[0225] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium magnesium zinc calcium phosphorus were effectively incorporated into the microparticle scaffold. Example 16:

[0226] [Sample with lithium-magnesium-zinc addition: No. 2108121] Prepare 200 ml of a composite solution of 0.6 mol / L zinc acetate dihydrate, 0.12 mol lithium chloride trihydrate, 0.1 mol magnesium acetate tetrahydrate, and 0.06 mol / L white sugar; add 60 g of the scaffold prepared at 1075°C, microwave the solution on medium heat to completely dry it, and dry it in the original 500 ml beaker in a constant temperature box at 196°C for 300 minutes [stir more in the early stage] to evenly carbonize the surface of the scaffold for use.

[0227] Prepare 200 ml of a 0.45 mol / L phosphoric acid / 0.6 mol / L diammonium phosphate solution using 6 ml of phosphoric acid and 15.8472 g of diammonium hydrogen phosphate. After complete dissolution, add the solution to a beaker containing a bone mineral scaffold containing zinc, lithium, magnesium, and sugar (solid-to-liquid ratio of 30:100 ml, total cation / phosphorus ion molar ratio of 1.298). Hydrothermal reaction was carried out at 65°C for 24 hours, with the 1000 ml beaker covered. After stopping the hydrothermal reaction, microwave drying was performed, followed by baking at 196°C for 5 hours (stirring repeatedly). The temperature was then increased by 5°C per minute to 1075°C (setting the room temperature at 25°C for 210 minutes). Maintain the temperature for 6 hours, then cool down to 300°C over 400 minutes. [The scaffold exhibited excellent strength, good elasticity, and appearance, but poor ventilation. The scaffold weighed 77.40 g, and surface nodules were visible under a magnifying glass.]

[0228] The powder diffraction and inductively coupled plasma atomic emission spectrometry elemental detection of the material indicated that lithium magnesium phosphorus and other elements were effectively incorporated into the material.

[0229] 2108121

[0230] Ca9 Mg Li ( P O4 )7 40.2%

[0231] Ca2.86 Mg0.14 ( P O4 )2 41.8%

[0232] Zn2 P2 O7 17.1%

[0233] The elemental analysis results of sample 2108121 by inductively coupled plasma atomic emission spectrometry (unit: wt%) are as follows:

[0234] Mg0.88; Ca26.7; P 19.6; Zn9.6.

[0235] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, zinc and phosphorus were effectively incorporated. Example 17:

[0236] [Lithium Magnesium Zinc Calcium 2107191] Prepare a 200ml solution containing 0.10mol / L anhydrous calcium chloride, 0.06mol / L magnesium acetate tetrahydrate, 0.24mol lithium chloride trihydrate, 0.1mol / L zinc acetate, and 0.06mol / L white sugar. Add 50g of bovine cancellous bone mineral particles [0.4-1mm, calcined at 988°C]. Microwave on medium heat or thaw to dry the liquid. Dry in a 198°C incubator in the original 500ml beaker for 300 minutes. Stir repeatedly to disperse the particles until the powder surface gradually and evenly turns black. Set aside.

[0237] Prepare 200 ml of a composite solution of 0.3 mol / L orthophosphoric acid and 0.165 mol / L diammonium hydrogen phosphate, immerse the black sand-like bone mineral particles previously added with calcium magnesium lithium zinc and white sugar in it [the total cation / phosphorus ion molar ratio of the reaction system is 1.38], conduct a hydrothermal reaction at 52°C for 25 hours and 28 minutes, dry at 72°C and stir repeatedly to dry the liquid into a sand-like state, bake at 196°C for 6 hours, the scaffold weighs 64.45 grams, and the temperature is raised by 2.5°C per minute to 1075°C [the room temperature is set at 25°C and the temperature is raised for 420 minutes], maintained for 6 hours, and then cooled [the weight is 57.28 grams, the particles are white and uniform, and many of the particles have micropores under a magnifying glass].

[0238] After testing 2107191, it was found that:

[0239] Ca9 Mg Li ( P O4 )7 78.3%

[0240] Ca5 ( P O4 )3 OH 21.7% Example 18:

[0241] [Add lithium magnesium zinc calcium phosphorus 2109071] Prepare 200 ml of a complex solution of 0.1 mol anhydrous calcium chloride, 0.08 mol magnesium acetate tetrahydrate, 0.09 mol lithium chloride, and 0.18 mol zinc acetate dihydrate, immerse 50 g of porcine cancellous bone mineral particles [0.33-1 mm, hydroxyapatite, calcined at 988°C] in it, microwave on medium heat and thaw on heat to dry the liquid (add 4.1 g of white sugar at appropriate times); dry in the original 500 ml beaker in a constant temperature box at 176°C for 180 minutes, stir the powder repeatedly, and the powder will gradually change from caramel color to brown sand-like powder and be completely dry for use.

[0242] Prepare a 200 ml solution of 0.3 mol / L orthophosphoric acid and 0.21836 mol / L disodium hydrogen phosphate. Add it to the beaker containing the brown, sand-like bone mineral particles treated with calcium, magnesium, sodium, strontium, and sugar (the total cation / phosphorus molar ratio in the reaction system is 1.4642). Hydrothermally react at 65°C for 24 hours. Microwave the liquid to dryness, then bake at 176°C for 3 hours, stirring every half hour. Gradually transform the powder from caramel color to brown, sand-like powder, and set aside.

[0243] The material was divided into two equal portions, 36.92 grams each, designated 2109071 and 21090711. 2109071 was heated at 5.8°C per minute to 900°C (150 minutes), maintained for 180 minutes, and then cooled to 188°C over a three-hour period. [Weight: 33.99 grams] The material was sintered into a hard-to-disperse mass similar to the beaker.

[0244] 21090711 Raise the temperature by 5°C per minute to 1075°C [set the room temperature to 25°C and heat it up in 210 minutes], maintain it for 180 minutes, set it at 220 and drop it to 188°C [weight 32.42 grams, good color], [the material is sintered into a block with the same shape as the beaker that is difficult to disperse, which may be well used in the block support].

[0245] 2109071【900℃】

[0246] Ca9 Mg Na ( P O4 )7 97.6%

[0247] Ca H2 P2 O7 2.40%

[0248] 21090711【1075℃】

[0249] Ca9 Mg Li ( P O4 )7 97.4%

[0250] Na Ca PO4 2.60%

[0251] 2109071 Inductively coupled plasma atomic emission spectrometry elemental analysis results Unit: wt%:

[0252] Li 0.26; Mg 1.78; Ca 29..6; P 19.45; Zn4.71.

[0253] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, zinc, calcium and phosphorus were effectively incorporated. Example 19:

[0254] [Sample with Lithium, Magnesium, Iron, and Phosphorus: No. 2108291] Prepare 200 ml of a complex solution containing 0.0675 mol / L magnesium acetate tetrahydrate, 0.12 mol / L lithium chloride trihydrate, 0.30 mol / L anhydrous ferric chloride, and 0.06 mol / L white sugar. Add 50 g of bovine cancellous bone mineral scaffold. Microwave on medium heat or defrost until the liquid is reduced to a dry consistency. Bake in a 198°C incubator in the original 500 ml beaker for 300 minutes. Stir repeatedly until the powder changes from caramel color to a dark, sandy consistency. Set aside.

[0255] The ratio of total metal ions to phosphorus in the characteristics was set to 1.1:1; 6 ml of 0.45 mol / L orthophosphoric acid and 200 ml of a composite solution of 0.71136 mol ammonium hydrogen phosphate were prepared, and the black-brown bone mineral scaffold previously treated with calcium magnesium lithium iron and white sugar was added [protected in a 1000 ml beaker] and hydrothermally reacted at 70°C for 24 hours; microwave thawed and dried, and then baked at 196°C for 3 hours [stirred repeatedly]; the temperature was increased by 5°C per minute for 175 minutes to 900°C, maintained for 360 minutes, and then the furnace temperature was increased to 188°C for 420 minutes to obtain 2108291 [73.44 g, the color became more uniform light green, the strength was stronger, and the ventilation was poor] and the sample was sent for powder diffraction. 2108291 was calcined at 1175°C for 8 hours to obtain 21082911, which had a very beautiful color. Scanning electron microscopy and energy spectrum of 21082911 revealed that calcium phosphate crystals containing active ions were formed on the wall of the stent, and the new material was honeycomb-shaped.

[0256] Li Fe ( P2 O7 ) Lithium iron(III) diphosphate | Lithium Iron Phosphate

[0257] 2108291

[0258] Ca9 Mg Li ( P O4 )7 15.7%

[0259] Li Fe ( P2 O7 ) 14.3%

[0260] Ca2 ( P2 O7 ) 56.8%

[0261] Na2 Mg ( S O4 )2 ( H2 O )4 1.5%

[0262] Ca10.042 (PO4)5.952 (OH)2.292 11.7%

[0263] 21082911

[0264] Ca9 Mg Li ( P O4 )7 65.2%

[0265] Ca2 ( P2 O7 ) 44.61% Example 20:

[0266] [Sample number 2111161: Lithium-magnesium-strontium-iron-phosphorus Added] Prepare a 200ml solution of 0.08mol magnesium acetate tetrahydrate, 0.18mol lithium chloride trihydrate, 0.15mol / L anhydrous ferric chloride, 0.15mol / L strontium nitrate, and 0.06mol / L white sugar. Immerse 50g of bovine cancellous bone mineral scaffold in the solution. Microwave on a freezer setting to dry the solution (the scaffold turns orange-red). Bake at 176°C for 5 hours to carbonize the resulting scaffold (63.2g).

[0267] Set the molar ratio of divalent metal ions to phosphorus in the reaction system to 1.5 [molar concentration of lithium chloride ÷ 2, molar concentration of ferric chloride × 1.5]; if 4 ml of phosphoric acid and 5.5 g of 0.20833 mol of ammonium hydrogen phosphate are used to prepare 200 ml of a composite solution, immerse the brown bone mineral scaffold previously added with lithium magnesium strontium iron and white sugar [protected by a 1000 ml beaker] in it, hydrothermally react at 75°C for 24 hours, thaw in a microwave oven until dry, and then bake at 176°C for 5 hours [stirring repeatedly];

[0268] Heat up 5℃ to 1075℃ per minute, maintain for 6 hours and then cool down [set to 75℃ in 5 hours] [excellent strength, elastic modulus may be quite good, less brittle, good appearance and ventilation.

[0269] Sample No. 2111161 received:

[0270] (Ca2.589 Mg0.411) (PO4)2 97.42%

[0271] Ca9.868 (PO4)5.586 (OH)4.006 3.69%

[0272] Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0273] Lithium 0.37, magnesium 1.12, calcium 31.5, phosphorus 19.5, strontium 3.88, iron 1.53.

[0274] Lithium / total cation molar concentration ≈ 5.72%

[0275] Magnesium / total cation molar concentration ≈ 4.943%

[0276] Strontium / total cation molar concentration ≈ 4.784%

[0277] Iron / total cation molar concentration ≈ 2.94%

[0278] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated the effective incorporation of lithium magnesium strontium iron phosphorus. Example 21:

[0279] [Sample No. 2107192 with Lithium, Magnesium, Zinc, Iron, Calcium, and Phosphorus:] Prepare 200 ml of a complex solution of 0.10 mol / L anhydrous calcium chloride, 0.06 mol / L magnesium acetate tetrahydrate, 0.18 mol lithium chloride tetrahydrate, 0.09 mol / L zinc acetate, 0.09 mol ferric chloride, and 0.06 mol / L white sugar. Add 50 g of porcine cancellous bone mineral microparticles [0.33-1 mm, hydroxyapatite, calcined at 988°C]. Microwave on medium heat and thaw until the liquid is dry. Dry in a 198°C incubator in the original 500 ml beaker for 300 minutes. Stir repeatedly until the powder gradually turns into a black sand-like consistency and set aside.

[0280] A 200ml composite solution containing 0.35mol / L orthophosphoric acid and 0.21836mol / L diammonium hydrogen phosphate was prepared. Black sand-like bone mineral powder, previously treated with calcium, magnesium, lithium, strontium, and sugar, was immersed in this solution (total cation concentration in the reaction system: (monovalent lithium ion concentration divided by 2, trivalent iron ion concentration × 1.5) / phosphorus molar ratio of 1.4219). The reaction was hydrothermally reacted at 52°C for 25 hours and 28 minutes, uncovered in a larger beaker. The reaction ended just as the solution approached the plane of the support. After baking at 196°C (with repeated stirring during the initial stage), the particles turned dark brown after 6 hours (weighing 65.36g). The temperature was then raised by 2.5°C per minute to 1075°C (at room temperature of 25°C, with a 420-minute heating period). After 6 hours of this, the temperature was lowered. The particles were white, weighing 58.15g, and exhibited numerous micropores under a magnifying glass.

[0281] Sample No. 2107192 was tested and found to be:

[0282] Ca9 Mg Li ( P O4 )7 100%

[0283] 2107192 Inductively coupled plasma atomic emission spectrometry element detection results (unit: wt%):

[0284] Magnesium 1.11; Lithium 0.35; Calcium 33.1; Phosphorus 21.8; Zinc 1.71; Iron 1.46.

[0285] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental analysis indicated that lithium, magnesium, strontium, iron, calcium, and phosphorus were effectively incorporated. Example 22:

[0286] [Sample with lithium, magnesium, zinc, strontium, iron, calcium and phosphorus: No. 2107011] Prepare a complex solution of 0.10 mol / L anhydrous calcium chloride, 0.055 mol / L magnesium sulfate heptahydrate, 0.18 mol lithium chloride trihydrate, 0.2 mol / L zinc acetate, 0.09 mol / L anhydrous ferric chloride, 0.18 mol / L strontium nitrate, and 0.10 mol / L white sugar, and add 50 g of bovine cancellous bone mineral particles [0.4-1 mm, calcined at 988°C]; bake in a 500 ml beaker at 198°C in a constant temperature oven for 300 minutes, repeatedly stir the powder until it gradually becomes uniform and black sand-like and ready for use.

[0287] The total cation / phosphorus molar ratio in the reaction system was set at 1.3538. A 200 ml solution of 0.45 mol / L orthophosphoric acid and 0.18836 mol / L diammonium hydrogen phosphate was prepared and poured into a 1000 ml beaker containing the bone mineral particles (presumably a 1000 ml beaker) containing calcium, magnesium, lithium, zinc, strontium, iron, and sugar. The reaction was hydrothermally reacted at 65°C for 24 hours. The solution was then microwaved on medium heat, thawed, and dried at 198°C for 9 hours (with repeated stirring). After carbonization of the material surface, the temperature was increased by 2.5°C / min over 350 minutes to 900°C. This temperature was maintained for 360 minutes, then increased to 400°C in the oven for 180 minutes, and then returned to room temperature at 70°C for 180 minutes to yield 2107011 (62.45 g, brown). A sample of 210711 was submitted for powder diffraction and elemental analysis. The volume of the powder scaffold was measured with a measuring cup. Based on the measurement, the porosity of the powder scaffold was calculated to be approximately 65%.

[0288] Sample No. 2107011 test:

[0289] Ca9 Mg Li ( P O4 )7 82.0%

[0290] Ca5 ( P O4 )3 ( OH ) 10.4%

[0291] Ca3 ( P O4 )2 7.6%

[0292] Elemental detection results of sample 2107011 by inductively coupled plasma atomic emission spectrometry (unit: wt%):

[0293] Magnesium 1.04, lithium 0.29, calcium 27.7, phosphorus 17.6, zinc 2.24, strontium 1.8, iron 0.73.

[0294] Powder diffraction composition analysis and inductively coupled plasma atomic emission spectrometry elemental methods indicate that lithium, magnesium, strontium, iron, calcium, and phosphorus are effectively incorporated. Example 23:

[0295] Stent dissolution test in simulated body fluids:

[0296] 2000 ml of sodium lactate-Ringer's solution was prepared with sodium lactate, sodium chloride, potassium chloride, and calcium chloride, with the pH controlled between 6.0 and 7.0. Three portions of each of the modified scaffolds (5 g each of lithium magnesium phosphate, lithium magnesium phosphate strontium, lithium magnesium phosphate zinc, and lithium magnesium phosphate iron) were added. The modified biomineralized scaffolds were placed in a 200 ml medical plastic bottle at a solid-to-liquid ratio of 5 g:100-200 ml. The sodium lactate-Ringer's solution was replaced four times weekly. The replaced solution was analyzed using a biochemical analyzer for elements such as calcium, phosphorus, lithium, magnesium, potassium, sodium, strontium, zinc, and iron. The modified scaffolds were removed and dried at 80°C for 24 hours, then weighed to calculate the dissolution rate. The material was then submitted for powder diffraction analysis. The four-week dissolution rate of the modified scaffolds ranged from 3 to 15%, indicating that the scaffold material exhibits dissolution properties in simulated body fluids. Example 24:

[0297] Tests as drug carriers:

[0298] 5 grams each of the 2104041 [Lithium Magnesium Strontium], 210423 [Lithium Magnesium], and [Lithium Magnesium Iron 2108291] block scaffolds were weighed and placed in the center of a glass dish. 5 grams each of the [Lithium Magnesium Zinc Iron Calcium 2107192] and [Lithium Magnesium Zinc Strontium Iron Calcium 2107011] microparticle scaffolds were weighed and stacked in a circular pattern in the center of a glass dish. A 3-ml pipette was used to drip-feed a physiological saline injection onto the scaffolds to assess their hydrophilicity and ability to retain simulated body fluids. Results showed that all scaffolds and microparticle scaffolds exhibited good hydrophilicity. The mass-to-volume ratios of physiological saline content for the block scaffolds were 5 grams:10 milliliters, 5 grams:6 milliliters, and 5 grams:10 milliliters, respectively. The mass-to-volume ratios of physiological saline content for the microparticle scaffolds were 5 grams:5.5 milliliters and 5 grams:5 milliliters, respectively. 5 grams each of 2104041 [lithium magnesium strontium], 210423 [lithium magnesium], and [lithium magnesium iron 2108291] block scaffolds were weighed and placed in the center of a glass dish; 5 grams each of [lithium magnesium zinc iron calcium 2107192] and [lithium magnesium zinc strontium iron calcium 2107011] microparticle scaffolds were weighed and stacked in a circular shape in the center of a glass dish; human plasma was aspirated with a 3 ml pipette and repeatedly drip-irrigated into the scaffolds. The results showed that various scaffold materials and microparticle scaffold materials had good hydrophilicity, and the scaffolds and microparticles were evenly stained light yellow, the same color as plasma. The mass-to-volume ratios of human plasma contained in each block scaffold were 5 grams:10 milliliters, 5 grams:6 milliliters, and 5 grams:10 milliliters, respectively. The mass-to-volume ratios of human plasma contained in various microparticle scaffolds were 5 grams:6-10 milliliters and 5 grams:6-10 milliliters, respectively. Cut the block stents 2104041 [lithium magnesium strontium], 210423 [lithium magnesium] and [lithium magnesium iron 2108291] into discs that meet the requirements; weigh 0.33 grams each of the [lithium magnesium zinc iron calcium 2107192] and [lithium magnesium zinc strontium iron calcium 2107011] microparticle stents; drip-irrigate the stent material with levofloxacin injection. The material has good hydrophilicity and solution retention capacity, and is transferred to a culture dish for antibacterial [Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli] tests, which show good antibacterial effects, as shown in Figure 6. Example 25:

[0299] Study on cell adhesion on scaffold surface:

[0300] MC3T3-E1 cells were co-cultured with 2104041 (lithium magnesium strontium), 210423 (lithium magnesium), and 2108291 (lithium magnesium iron) block scaffolds. Adhesion focal formation was observed using confocal laser scanning microscopy (CLSM). Immunofluorescence staining of the adhesion-related protein (vinculin) on the surface cells was performed. Adherent surface cells were digested, and the expression of intracellular adhesion-related proteins (integrin, vinculin) was quantitatively analyzed by Western blot. In vitro co-culture of cells with the modified scaffolds confirmed that the nanowhisker structure promoted cell adhesion, spreading, and focal formation on the material surface, promoting osteogenesis, as shown in Figure 7. Example 26:

[0301] In vivo studies as a filler material for repairing bone defects:

[0302] A skull defect model (7.5 and 10 mm in diameter) was created in rabbits. The defect area was filled with sheet-shaped modified scaffold material or bone powder of the same diameter and thickness. The mice were observed for two and four months, and imaging, histological, and histochemical analyses were performed. The modified scaffolds were able to effectively promote the repair of bone defects and had significant osteogenesis and angiogenesis activities, as shown in Figure 8.

[0303] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein with equivalents. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention. Example 27:

[0304] Mechanical strength testing:

[0305] The modified cylindrical scaffolds No. 2105202, 210419, 2106281 with a diameter of nearly 1 cm and a height of 1 cm and 2-3 of each of the precursor bovine cancellous scaffolds under the same conditions were selected for mechanical strength testing. The mechanical test results of the modified bio-bone mineral scaffold No. 210520 were: 1.25 MPa / cm2, 1.86 MPa / cm2, 2.47 MPa / cm2, average value = 1.86 MPa / cm2; the mechanical test results of the modified bio-bone mineral scaffold No. 210419 were: 3.20 MPa / cm2, 1.38 / cm2, average value = 2.29 MPa / cm2; the mechanical test results of the modified bio-bone mineral scaffold No. 2106281 were: 1.97 MPa / cm2, 4.57 MPa / cm2, The average mechanical strength of the modified scaffolds was 1.45 MPa, with an average value of 2.66 MPa / cm2. The strengths of the precursor bovine cancellous scaffolds were 0.82 MPa / cm2, 0.89 MPa / cm2, and 2.19 MPa / cm2, respectively, with an average value of 1.30 MPa / cm2. The average mechanical strength of the modified scaffolds was better than that of the latter, as shown in the corresponding chart data in Figure 9.

[0306] Elastic modulus (automatic Young's) (MPa) Maximum compressive stress (MPa) Compressive strain at maximum compressive stress (%) Diameter (mm) Height of compression plate (mm) 1149.60 2793.20 2994.69317 12.00000 14.00000 21198.93 2931.3750 10.24767 13.00000 13.50000

Claims

1. A modified bioactive bone mineral scaffold doped with lithium, magnesium and phosphorus, characterized in that: It is obtained by subjecting a bovine or porcine calcined cancellous bone mineral porous scaffold to hydrothermal reaction treatment by immersing it in a composite solution containing one or more of the active metal ions magnesium, lithium, strontium, zinc, iron, and calcium and a phosphorus source, followed by baking and drying and then high-temperature calcination.

2. The modified bioactive bone mineral scaffold doped with lithium, magnesium and phosphorus according to claim 1, wherein: Regarding the method of subjecting a bovine or porcine calcined cancellous bone mineral porous scaffold to hydrothermal reaction treatment by immersing it in a composite solution containing one or more of the active metal ions magnesium, lithium, strontium, zinc, iron, and calcium and a phosphorus source, the selected scheme is as follows: First, immerse the bovine or porcine calcined cancellous bone mineral porous scaffold in a metal ion source solution containing one or more of the active metal ions magnesium, lithium, strontium, zinc, iron, and calcium and a white sugar source solution, dry the liquid by microwave or in an incubator, bake and dry at 96 °C - 198 °C, and then carry out hydrothermal reaction in the phosphorus source composite solution; the phosphorus source composite solution is a phosphorus source binary system.

3. A modified bioactive bone mineral scaffold doped with lithium, magnesium and phosphorus according to claim 2, characterized in that: The hydrothermal reaction is carried out in a constant-temperature hydrothermal manner, controlling the temperature at 60 - 100 °C and the time at 24 - 48 hours.

4. A modified biological bone mineral scaffold doped with lithium, magnesium, and phosphorus according to claim 3, characterized in that: The material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the metal ion source solution is 15 - 50 g : 100 mL, and the material-liquid ratio of the bovine or porcine calcined cancellous bone mineral porous scaffold to the phosphorus source composite solution is 15 - 50 g : 100 mL.

5. A modified bio-bone mineral scaffold doped with lithium, magnesium, and phosphorus according to claim 4, characterized in that In the metal ion source solution, the magnesium source is one of magnesium acetate, magnesium sulfate, magnesium hydrogen phosphate, etc.; the lithium source is lithium chloride; the calcium source is one of calcium chloride and calcium hydroxide; the zinc source is one of soluble zinc salts such as zinc nitrate and zinc acetate; the strontium source is one of soluble strontium salts such as strontium nitrate, strontium acetate, and strontium sulfate; the iron source is one of soluble iron salts such as ferrous sulfate, ferrous chloride, ferric chloride, and iron acetate; the phosphorus source binary system is a composite solution of phosphoric acid and soluble phosphate; the soluble phosphate is selected from one or a combination of ammonium dihydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, and magnesium hydrogen phosphate.

6. The modified biological bone mineral scaffold doped based on lithium, magnesium and phosphorus according to claim 5, wherein: The final concentration of magnesium ions in the binary system containing osteogenic active metal ions and phosphorus source is 0.05 - 0.20 mol / L; the final concentration of lithium ions in the binary system containing osteogenic active metal ions and phosphorus source is 0.06 - 0.6 mol / L; the final concentration of zinc ions in the binary system containing osteogenic active metal ions and phosphorus source is 0.1 - 0.6 mol / L; the final concentration of strontium ions in the binary system containing osteogenic active metal ions and phosphorus source is 0.15 - 0.9 mol / L; the final concentration of ferrous ions in the binary system containing osteogenic active metal ions and phosphorus source is 0.1 - 0.6 mol / L; the final concentration of the binary system containing osteogenic active metal ions and phosphorus source as a supplement to calcium ions is 0.15 - 1.5 mol / ; preferably, the final concentration of phosphate groups provided by phosphoric acid in the binary system containing osteogenic active metal ions and phosphorus source is 0.15 - 0.9 mol / L; the final concentration of phosphate groups provided by soluble phosphate in the binary system containing osteogenic active metal ions and phosphorus source is 0.06 - 0.6 mol / L; preferably, the molar ratio of the total molar concentration of osteogenic active cations including calcium to the molar concentration of phosphate ions in the reaction system is 1.1 - 1.6:1; the concentration of the white sugar source solution is 0.03 - 0.2 mol / L.

7. A modified biological bone mineral scaffold doped with lithium, magnesium, and phosphorus according to claim 1 or 2, characterized in that After the hydrothermal reaction, select to keep the temperature constant to dry the liquid and then bake and dry it. The baking and drying temperature is 75°C - 198°C.

8. A modified biological bone mineral scaffold doped with lithium, magnesium, and phosphorus according to claim 1 or 2, characterized in that: The parameters of the high-temperature calcination are 750°C - 1200°C, and the calcination time is 6 hours - 24 hours.

9. A modified bio-bone mineral scaffold doped with lithium and magnesium according to claim 1 or 2, characterized in that, The preparation method of the bovine or porcine calcined cancellous bone mineral porous scaffold is as follows: (1): Bone strip, bone block bovine or porcine calcined cancellous bone mineral porous scaffold (1-1): Cut bovine or porcine cancellous bone into cancellous bone strips or blocks with a thickness of 0.5 - 5 cm to obtain raw bones; cylindrical cancellous bone strips can also be obtained by using a hole saw. (1-2): Place the raw bones in distilled water and cook them in a pressure cooker for 36 - 60 min, then wash them clean with drinking water at 50 - 75°C, and repeat this step 5 - 6 times. (1-3): Dry the raw bones processed in step (1-2) in a constant-temperature oven at 80 - 120°C for 12 - 24 hours, then place them in a calcination furnace, calcine them at 900 - 1200°C for 6 - 12 hours, and then slowly cool them to room temperature to obtain the calcined cancellous bone mineral porous scaffold. The powder diffraction components of the bovine or porcine cancellous bone mineral materials are all hydroxyapatite. (2): Granular bovine or porcine cancellous bone mineral porous scaffold (2-1): Cut bovine or porcine cancellous bone into bone strips or blocks with a thickness of 0.5 - 6 cm to obtain raw bones. (2-2): Place the raw bones in distilled water and cook them in a pressure cooker for 36 - 60 min, then wash them clean with drinking water at 50 - 75°C, and repeat this step 5 - 6 times. (2-3): Dry the raw bones processed in step (2-2) in a constant-temperature oven at 80 - 120°C for 12 - 24 hours, then place them in a calcination furnace, calcine them at 900 - 1200°C for 6 - 12 hours, and then slowly cool them to room temperature to obtain the bovine or porcine calcined cancellous bone mineral porous scaffold. (2-4): Crush the bovine cancellous bone mineral porous scaffold processed in step (2-3) with a food crushing machine, and use a stainless steel sieve to screen various specifications of particles such as 0.2-1 mm, 1-3 mm, 3-5 mm, and 5-7 mm of granular bovine cancellous bone mineral porous cell scaffolds for standby; crush the porcine cancellous bone mineral porous scaffold processed in step (2-3) with a food crushing machine, and use a stainless steel sieve to screen 0.2-0.8 mm and 0.33-1 mm of granular porcine cancellous bone mineral porous cell scaffolds for standby.

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