Bone cement solid-phase composition, bone cement and use method of bone cement

Abstract

The invention provides a bone cement solid-phase composition, bone cement and a use method of the bone cement, and belongs to the technical field of biological materials. The bone cement solid-phase composition provided by the invention contains a certain amount of polyglutamic acid, and when the bone cement solid-phase composition is used for bone cement, abundant carboxyl groups in a side chain of the polyglutamic acid and free calcium ions in calcium hydroxide generate chelation to form a stable PGA-Ca < 2 + > compound; the speed of generating hydroxyapatite precipitate by reaction of calcium ions in calcium hydroxide and sodium polyphosphate is obviously reduced, so that the time for injection is prolonged. The result of the embodiment shows that the initial setting time of the bone cement prepared from the bone cement solid-phase composition provided by the invention can reach 17.8 + / -1.51 min.

Description

Technical Field

[0001] This invention relates to the field of biomaterials technology, and in particular to a bone cement solid composition, bone cement, and a method of using the same. Background Technology

[0002] Osteomyelitis is a common orthopedic disease caused by pathogenic microorganisms infecting bone tissue. It is considered one of the most challenging orthopedic diseases due to its difficult treatment, long course, and high recurrence rate. The key to treating osteomyelitis lies in effective anti-infection measures and bone loss repair.

[0003] Current technology uses calcium hydroxide phosphate bone cement, composed of polyphosphates and calcium hydroxide, to treat osteomyelitis. This cement is injected into the bone loss site caused by osteomyelitis, inhibiting bacterial growth and promoting osteoblast growth. However, the initial setting time of calcium hydroxide phosphate bone cement is 4.5–9.3 minutes, resulting in a short injection time, making it unsuitable for more complex myelitis treatments requiring longer injection times. Summary of the Invention

[0004] The purpose of this invention is to provide a bone cement solid composition, bone cement, and a method of using the same. The bone cement prepared from the bone cement solid composition provided by this invention has a long injection time.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: A bone cement solid composition comprising sodium polyphosphate (NaPO3). n The sodium polyphosphate contains calcium hydroxide and polyglutamic acid; the molar ratio of phosphate ions in the sodium polyphosphate to calcium ions in the calcium hydroxide is 1:(1~10); the sodium polyphosphate (NaPO3) n The value of n is 3-40; the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). n It is 5-20% of the total mass of calcium hydroxide.

[0006] Preferably, the polyglutamic acid is sodium polyphosphate (NaPO3) by mass. n And 10-20% of the total mass of calcium hydroxide.

[0007] Preferably, the molar ratio of phosphate ions in the sodium polyphosphate to calcium ions in the calcium hydroxide is 1:(2~5).

[0008] Preferably, the sodium polyphosphate (NaPO3) n n in the range is 15 to 30.

[0009] The present invention also provides a bone cement comprising a solid phase composition and a liquid phase component, wherein the solid phase composition adopts the bone cement solid phase composition described in the above technical solution.

[0010] Preferably, the volume ratio of the liquid phase component to the mass ratio of the solid phase composition is 0.25~0.7 mL / g.

[0011] Preferably, the liquid phase component includes one or more of pure water, phosphate buffer, disodium hydrogen phosphate aqueous solution, sodium dihydrogen phosphate aqueous solution, SBF simulated body fluid, and physiological saline.

[0012] Preferably, the particle size of the solid component is 20-400 mesh.

[0013] The present invention also provides a method for using the bone cement described in the above technical solution, comprising: The solid-phase composition and liquid-phase components are mixed and then self-cured.

[0014] Preferably, the self-curing temperature is 20~40℃ and the self-curing humidity is 40~100%.

[0015] This invention provides a bone cement solid composition comprising sodium polyphosphate (NaPO3). n The sodium polyphosphate contains calcium hydroxide and polyglutamic acid; the mass ratio of phosphate ions in the sodium polyphosphate to calcium ions in the calcium hydroxide is 1:(1~10); the sodium polyphosphate (NaPO3) n The value of n is 3-40; the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). n The amount of polyglutamic acid in the bone cement solid composition provided by this invention is 5-20% of the total mass of calcium hydroxide. When used in bone cement, the abundant carboxyl groups in the side chains of polyglutamic acid chelate with free calcium ions in calcium hydroxide to form stable PGA-Ca. 2+ The complex significantly reduces the rate at which calcium ions in calcium hydroxide react with sodium polyphosphate to form hydroxyapatite precipitate, thereby prolonging the time available for injection. Examples show that the initial setting time of bone cement prepared from the bone cement solid composition provided by this invention can reach 17.8 ± 1.51 min. Attached Figure Description

[0016] Figure 1 The bar chart shows the initial setting time and final setting time of the bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention. Figure 2 The pH changes of bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention in SBF simulated body fluid after 2, 4, 6 and 8 days are shown. Figure 3The bar chart shows the weight loss ratio of bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 after 8 days in SBF simulated body fluid. Figure 4 The images show the XRD patterns of bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 after curing for 3 hours. Figure 5 The images show the XRD patterns of bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention after solidification in SBF simulated body fluid for 8 days. Figure 6 The diagram shows the anti-collapse properties of the bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention. Figure 7 The erosion resistance diagrams are for the bone cements prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention. Figure 8 The graphs show the antibacterial properties of bone cement and blank control prepared in Examples 2, 4, 6 and Comparative Example 2 of this invention against Staphylococcus aureus. Figure 9 The graphs show the antibacterial properties of bone cement prepared in Examples 1, 2, and 3 of this invention and Comparative Example 1 against Escherichia coli and the blank control. Figure 10 A bar chart showing the biocompatibility of bone cement prepared in different amounts in Examples 2, 4, 6 and Comparative Example 2 with rat bone marrow mesenchymal stem cells. Figure 11 Add a bar chart to the present invention showing the biocompatibility of bone cement prepared in Example 2 and Comparative Example 2 with rat bone marrow mesenchymal stem cells and a blank control at 1, 3, and 5 days. Figure 12 Add cell staining images of rat bone marrow mesenchymal stem cells incubated with bone cement prepared in Example 2 and Comparative Example 2, and a blank control, to the present invention; Figure 13 Alkaline phosphatase staining images of rat bone marrow mesenchymal stem cells incubated with bone cement prepared in Example 2 and Comparative Example 2, and a blank control, are added to this invention. Detailed Implementation

[0017] This invention provides a bone cement solid composition comprising sodium polyphosphate (NaPO3). n Calcium hydroxide and polyglutamic acid.

[0018] In this invention, the bone cement solid composition comprises sodium polyphosphate (NaPO3). n In this invention, the sodium polyphosphate (NaPO3) n It reacts with calcium hydroxide to form hydroxyapatite, which stimulates osteoblast differentiation and growth. In this invention, the sodium polyphosphate (NaPO3) is used. nIn this context, n is 3 to 40. As one embodiment of the present invention, the sodium polyphosphate (NaPO3) n In this context, n can be 15 to 30. In an embodiment of the invention, the sodium polyphosphate (NaPO3) n The 'n' in the equation can specifically be 3, 5, 8, 10, 12, 15, 18, 20, 25, 28, 30, 25, or 40. This invention defines the sodium polyphosphate (NaPO3) as such. n The n in the formula ensures that the reaction with calcium hydroxide forms hydroxyapatite, which stimulates osteoblast differentiation and growth, while also ensuring that the resulting bone cement has high compressive strength.

[0019] In this invention, the bone cement solid composition includes calcium hydroxide. In this invention, the calcium hydroxide makes the bone cement highly alkaline in the early stages, inhibiting bacterial growth; subsequently, it reacts with sodium polyphosphate (NaPO3). n The reaction forms hydroxyapatite, which stimulates osteoblast differentiation and growth.

[0020] In this invention, the sodium polyphosphate (NaPO3) n The molar ratio of phosphate ions to calcium ions in calcium hydroxide is 1:(1~10). As one embodiment of the present invention, the sodium polyphosphate (NaPO3) is... n The molar ratio of phosphate ions to calcium ions in calcium hydroxide is 1:(2~5). This invention specifies sodium polyphosphate (NaPO3). n The molar ratio of phosphate ions to calcium ions in calcium hydroxide further ensures the quality of sodium polyphosphate (NaPO3). n It reacts with calcium hydroxide to form hydroxyapatite, which stimulates osteoblast differentiation and growth.

[0021] In this invention, the bone cement solid composition comprises polyglutamic acid. In this invention, the abundant carboxyl groups in the side chains of the polyglutamic acid chelate with free calcium ions in calcium hydroxide to form a stable PGA-Ca... 2+ The complex significantly reduces the rate at which calcium ions in calcium hydroxide react with sodium polyphosphate to form hydroxyapatite precipitate, thereby prolonging the injection time. In this invention, the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). n The polyglutamic acid comprises 5-20% of the total mass of calcium hydroxide. In one embodiment of the invention, the molecular weight of the polyglutamic acid can be 300,000-1,000,000. In specific embodiments of the invention, the molecular weight of the polyglutamic acid can be 300,000, 500,000, 800,000, or 1,000,000. In one embodiment of the invention, the mass of the polyglutamic acid can be sodium polyphosphate (NaPO3). nThe amount is 10-20% of the total mass of calcium hydroxide. In embodiments of the present invention, the mass of the polyglutamic acid may specifically be sodium polyphosphate (NaPO3). n The content of polyglutamic acid is 5%, 10%, 15%, or 20% of the total mass of calcium hydroxide. This invention further ensures that the polyglutamic acid chelates with free calcium ions in calcium hydroxide to form a stable PGA-Ca complex by limiting the mass of polyglutamic acid. 2+ The complex reduces the rate at which calcium ions in calcium hydroxide react with sodium polyphosphate to form hydroxyapatite precipitate, thereby prolonging the time available for injection.

[0022] The bone cement solid composition provided by this invention contains a certain amount of polyglutamic acid. When used in bone cement, the abundant carboxyl groups in the side chains of polyglutamic acid chelate with free calcium ions in calcium hydroxide to form stable PGA-Ca. 2+ The complex significantly reduces the rate at which calcium ions in calcium hydroxide react with sodium polyphosphate to form hydroxyapatite precipitate, thereby prolonging the time available for injection.

[0023] The present invention also provides a bone cement comprising separately packaged solid and liquid components, wherein the solid component is the bone cement solid composition described in the above technical solution.

[0024] In one embodiment of the present invention, the particle size of the solid phase component can be 20-400 mesh. In specific embodiments of the present invention, the particle size of the solid phase component can be 20 mesh, 40 mesh, 80 mesh, 100 mesh, 200 mesh, 300 mesh, or 400 mesh. In one embodiment of the present invention, the liquid phase component can be one or more of pure water, phosphate buffer, disodium hydrogen phosphate aqueous solution, sodium dihydrogen phosphate aqueous solution, SBF simulated body fluid, and physiological saline. In one embodiment of the present invention, the concentrations of the phosphate buffer, disodium hydrogen phosphate aqueous solution, sodium dihydrogen phosphate aqueous solution, SBF simulated body fluid, and physiological saline can be independently 0.9-5%. In specific embodiments of the present invention, the concentrations of the phosphate buffer, disodium hydrogen phosphate aqueous solution, sodium dihydrogen phosphate aqueous solution, SBF simulated body fluid, and physiological saline can be specifically independently 0.9%, 1%, 2%, 3%, 4%, or 5%. The present invention limits the composition of the liquid phase component to ensure that the solid phase component can be formed into a paste, filling three-dimensional space and achieving in-situ solidification. In one embodiment of the present invention, the volume ratio of the liquid phase component to the mass ratio of the solid phase component is 0.25~0.7 mL / g. In specific embodiments of the present invention, the volume ratio of the liquid phase component to the mass ratio of the solid phase component can be 0.25 mL / g, 0.3 mL / g, 0.5 mL / g, 0.6 mL / g, or 0.7 mL / g. The present invention, by limiting the volume ratio of the liquid phase component to the mass ratio of the solid phase component, further ensures that the solid phase component is in a paste-like state and is not too thin, thus affecting its curing effect and mechanical properties.

[0025] The bone cement provided by this invention has a long injection time.

[0026] The present invention also provides a method for using the bone cement described in the above technical solution, comprising: The solid and liquid components, which are packaged separately, are mixed and then self-cured.

[0027] In one embodiment of the present invention, the solid components may be ground before mixing. In another embodiment, the grinding time may be 1 to 5 minutes. Specifically, in embodiments of the present invention, the grinding time may be 1 minute, 2 minutes, 3 minutes, 4 minutes, or 5 minutes.

[0028] In one embodiment of the present invention, the mixing of the solid-phase composition and the liquid-phase component can be carried out under stirring. In one embodiment of the present invention, the stirring rate can be 80-180 rpm. In one embodiment of the present invention, the stirring time can be stopped once the solid-phase composition and the liquid-phase component have been stirred into a paste.

[0029] In one embodiment of the present invention, the self-curing temperature can be 20~40℃, and the self-curing humidity can be 40~100%. In embodiments of the present invention, the self-curing humidity can specifically be 40%, 60%, 80%, or 100%. The present invention ensures that bone cement fully cures in a near-physiological environment by limiting the self-curing temperature and humidity.

[0030] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0031] Example 1 A bone cement solid composition comprising sodium polyphosphate (NaPO3). 20 It is composed of calcium hydroxide and polyglutamic acid (γ-PGA); the sodium polyphosphate (NaPO3) is also present. 20 The molar ratio of phosphate ions to calcium ions in calcium hydroxide is 1:2; the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). 20 The total mass of the polyglutamic acid is 10% of that of calcium hydroxide; the polyglutamic acid has a molecular weight of 980,000 and a particle size of 100 mesh.

[0032] Example 2 A bone cement, designated PCDC20-2+10%PGA, is composed of separately packaged solid and liquid phase components; the solid phase component is the bone cement solid phase composition of Example 1; the liquid phase component is pure water; the volume ratio of the pure water to the mass of the solid phase component is 0.4 mL / g.

[0033] The above-mentioned bone cement is used as follows: after grinding the solid component for 3 minutes, it is sieved to obtain a solid component with a particle size of ~. Then, it is stirred with pure water at 120 rpm to form a paste, which is then poured into a stainless steel mold (6 mm in diameter and 12 mm in height) and flattened with a trowel. Then, it is allowed to self-cur in an environment of 37°C and 100% relative humidity.

[0034] Example 3 The difference between this embodiment and Embodiment 1 is that the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). 20 The total mass of calcium hydroxide is 20%; the rest is the same as in Example 1.

[0035] Example 4 The difference between this embodiment and embodiment 2 is that the bone cement solid composition of embodiment 3 is used; the resulting bone cement is denoted as PCDC20-2+20%PGA.

[0036] Example 5 The difference between this embodiment and Embodiment 1 is that the mass of the polyglutamic acid is sodium polyphosphate (NaPO3). 20 The total mass of calcium hydroxide is 5%; the rest is the same as in Example 1.

[0037] Example 6 The difference between this embodiment and embodiment 2 is that the bone cement solid composition of embodiment 5 is used; the resulting bone cement is denoted as PCDC20-2+5%PGA.

[0038] Example 7 The difference between this embodiment and Embodiment 2 is that the liquid phase component is a 4% sodium dihydrogen phosphate aqueous solution, while the rest is the same as in Embodiment 2.

[0039] Comparative Example 1 The difference between this comparative example and Example 1 is that polyglutamic acid is omitted; otherwise, they are the same as in Example 1.

[0040] Comparative Example 2 The difference between this comparative example and Example 2 is that the bone cement solid composition described in Comparative Example 1 was used to obtain bone cement, denoted as PCDC20-2.

[0041] The initial and final setting times of the bone cement in Examples 2, 4, 6 and Comparative Example 2 were determined using the initial and final setting needles of a Vicat apparatus. The test results are as follows:Figure 1 As shown in the figure, the initial setting times of the bone cement prepared in Examples 2, 4, and 6 of this invention are 14.2±0.8, 17.8±1.51, and 10.1±0.52, respectively, while the initial setting time of the bone cement prepared in Comparative Example 2 is 5±0.9. Therefore, the bone cement prepared by this invention has a longer injection time.

[0042] The cured bone cements from Examples 2, 4, 6, and Comparative Example 2 were impregnated in SBF simulated body fluid, with a mass ratio of bone cement to SBF simulated body fluid of 0.05 g∙mL. -1 The SBF simulated body fluid was updated every 48 hours. Then, at 2, 4, 6, and 8 days later, the pH of the SBF simulated body fluid was measured using a pH meter. The results are as follows: Figure 2 As shown in the figure, the bone cement of Examples 2, 4, 6 and Comparative Example 2 had a pH of 13 after 2 days, which decreased to 7.5 after 4 days and remained stable. This is due to the calcium hydroxide and sodium polyphosphate (NaPO3) in the bone cement. n As the reaction progresses, the alkaline components decrease. On the other hand, the ionic composition of SBF-mimicked body fluid is very similar to that of human blood plasma, and it also has a certain pH buffering effect. This weakly alkaline environment can significantly inhibit the growth of osteoclasts, promote osteogenic differentiation of stem cells, and mineralization of osteoblasts.

[0043] The bone cement from Examples 2, 4, 6, and Comparative Example 2, after being impregnated for 8 days, was removed, rinsed with deionized water, and dried at 60°C for 12 hours. The mass was recorded as M. t According to the formula, the weight loss ratio = (M0 - M...) t ) / M t The experiment was repeated three times for each sample, and the average value was calculated. The results are as follows: Figure 3 As shown in the figure, the weight loss ratios of bone cement in Examples 2, 4, 6, and Comparative Example 2 were 20.5±7.6%, 17.4±4.7%, 30.8±1.28%, and 31.8±2.3%, respectively. This indicates that the weight loss ratio decreased with increasing polyglutamic acid content, suggesting that the introduction of polyglutamic acid slows down the degradation rate of bone cement. The main mechanism lies in the formation of an interpenetrating network structure with a higher ion cross-linking density. As a polyanionic electrolyte, polyglutamic acid can compete with sodium polyphosphate for calcium ion binding, achieving not only its own cross-linking but, more importantly, serving as an additional cross-linking site, interacting with sodium polyphosphate segments via Ca2+. 2+ The formation of heterogeneous crosslinks significantly increases the crosslinking density of the three-dimensional network, reduces the swelling rate of the matrix, and effectively hinders the diffusion of water molecules and the process of ion exchange, thus macroscopically exhibiting a slowdown in degradation kinetics.

[0044] The phase composition of bone cements prepared in Examples 2, 4, 6, and Comparative Example 2 after curing for 3 hours and 8 days in SBF simulated body fluid was characterized using X-ray diffraction. The results are shown below. Figures 4-5 As shown in the figure. From the figure, we can see that: Figure 4 In the XRD patterns of bone cement in Examples 2, 4, 6 and Comparative Example 2 after curing for 3 days, there were characteristic peaks of calcium hydroxide, indicating that the bone cement after curing for 3 hours did not have characteristic peaks of hydroxyapatite. It mainly generated amorphous calcium phosphate precursors, and the nucleation and growth process of hydroxyapatite had not yet reached the detection limit of XRD. Figure 5 In the XRD patterns of bone cement from Examples 2, 4, 6 and Comparative Example 2 after 8 days of curing, obvious hydroxyapatite characteristic peaks were observed.

[0045] The bone cements from Examples 2, 4, 6, and Comparative Example 2 were placed in petri dishes, submerged in water, sealed with sealing film, and shaken on a shaker at 120 rpm for 1 hour. The anti-collapse properties of the bone cements in the aqueous environment were then observed. Figure 6 As shown in the figure, the bone cements of Examples 2, 4, 6 and Comparative Example 2 exhibit good anti-collapse properties.

[0046] The bone cements from Examples 2, 4, 6, and Comparative Example 2 were filled into syringes and then directly injected into culture dishes containing deionized water. The mixtures were left to stand for one day, and the erosion resistance of the bone cements was observed. The results are as follows: Figure 7 As shown in the figure, the bone cements of Examples 2, 4, 6 and Comparative Example 2 exhibit good erosion resistance.

[0047] Bone cements from Examples 2, 4, 6, and Comparative Example 2, respectively, were mixed with a concentration of 5 × 10⁻⁶ after curing for 3 hours and after curing in SBF simulated body fluid for 8 days. 6 Staphylococcus aureus and Escherichia coli were co-incubated for 12 hours, then 100 μL was plated, photographed after 12 hours, and colony counts were performed. The results are as follows: Figures 8-9 As shown in the figure. From the figure, we can see that: Figure 8 In comparison with the blank control, the bone cement of Examples 2, 4, 6 and Comparative Example 2, after curing for 3 hours, all showed better bactericidal effects against Staphylococcus aureus. This is because the OH- in the alkaline microenvironment... - The generated free radicals can react with bacterial DNA, inhibiting DNA replication and reducing bacterial activity, thereby causing mutations. In addition, the alkaline microenvironment can also disrupt the components of the bacterial cell membrane, inhibiting bacterial growth by inhibiting ATP synthesis and inducing oxidative stress. However, the bone cements of Examples 2, 4, 6 and Comparative Example 2, after solidification in SBF-simulated body fluid for 8 days, did not exhibit antibacterial effects. These results indicate that during the degradation process, calcium hydroxide is gradually degraded to form hydroxyapatite, reducing alkalinity and thus gradually eliminating the antibacterial effect. Figure 9In the study, the bone cements of Examples 2, 4, 6 and Comparative Example 2, after curing for 3 hours, all showed good bactericidal effects against Escherichia coli, while the bone cements of Examples 2, 4, 6 and Comparative Example 2, after curing for 8 days in SBF simulated body fluid, did not have antibacterial effects.

[0048] Bone cement extracts prepared in Examples 2, 4, and 6 and Comparative Example 2 at concentrations of 25 mg / mL, 50 mg / mL, and 100 mg / mL, respectively, were co-incubated with rat bone marrow mesenchymal stem cells (rBMSCs) at 37°C and 5% CO2. After co-incubation for 1 day and 7 days, the rat bone marrow mesenchymal stem cells were treated with CCK-8 reagent, and the results were analyzed as follows: Figure 10 As shown in the figure, the cell proliferation effect was best when the extract of bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 was 50 mg / mL. Furthermore, the bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 all showed good cell viability after co-incubation for 1 day and 7 days, indicating that the bone cement prepared in Examples 2, 4, 6 and Comparative Example 2 has good cell compatibility and can be used for a long time.

[0049] Bone cement extracts prepared in Example 2 and Comparative Example 2 at a concentration of 50 mg / mL were co-incubated with rat bone marrow mesenchymal stem cells (rBMSCs) at 37°C and 5% CO2. After co-incubation for 1, 3, and 5 days, the rat bone marrow mesenchymal stem cells were treated with CCK-8 reagent. Cells were seeded in well plates, with the group without material serving as a blank control. The results were analyzed as follows: Figure 11 As shown in the figure, there was no difference in the three groups of rat bone marrow mesenchymal stem cells when the co-incubation time was 1 day. However, at 3 and 5 days, the CCK8 results of the bone cement extract prepared in Example 2 for incubating rat bone marrow mesenchymal stem cells were significantly higher than those in Comparative Example 1 and the blank control group. This may be because: the abundant hydrophilic carboxyl groups on the polyglutamic acid molecular chain first improve the surface properties of the material, making it hydrophilic instead of hydrophobic, which is conducive to the adsorption of cell adhesion proteins and the initial attachment of cells; secondly, polyglutamic acid can be enzymatically hydrolyzed in vivo, and its degradation can form a porous structure inside the material, promoting cell migration, nutrient exchange and three-dimensional tissue ingrowth.

[0050] Bone cement extracts prepared in Example 2 and Comparative Example 2 at a concentration of 50 mg / mL were co-incubated with rat bone marrow mesenchymal stem cells (rBMSCs) in α-MEM medium for 3 days. The rat bone marrow mesenchymal stem cells were then extracted and treated with the FITC phalloidin kit. Cells were seeded in well plates, with the group without material serving as a blank control. The results are as follows: Figure 12As shown in the figure: the first row of red represents the skeleton of rat bone marrow mesenchymal stem cells, the second row of blue represents the nuclei of rat bone marrow mesenchymal stem cells, and the third row represents rat bone marrow mesenchymal stem cells. The results show that the rat bone marrow mesenchymal stem cells (rBMSCs) after co-incubation with the bone cement extract prepared in Example 2 and Comparative Example 2 have normal cell morphology, without any signs of curling, shrinkage, or breakage, and can adhere well to the well plate, indicating that the bone cement cells prepared in Example 2 and Comparative Example 2 have good cell compatibility.

[0051] Two groups of bone cement extracts (50 mg / mL each) prepared in Example 2 and Comparative Example 2 were incubated with rat bone marrow mesenchymal stem cells (rBMSCs) in α-MEM medium for 12 h. The α-MEM medium was then replaced with osteogenic induction medium (prepared by adding 0.5 mL of 50 nM ascorbic acid, 62.5 μL of 0.1 μM dexamethasone, and 1 μL of 10 mM β-glycerophosphate per 50 mL of complete medium). After culturing for 5 and 10 days respectively, the rat bone marrow mesenchymal stem cells were harvested. The 24-well plates containing the rat bone marrow mesenchymal stem cells (rBMSCs) were washed with phosphate buffer, fixed with 4% paraformaldehyde (FDA) at room temperature for 30 min, stained with a BCIP / NBT alkaline phosphatase staining kit, and then completely washed with phosphate buffer to terminate staining. A blank control was used without the added extract. The results are as follows: Figure 13 As shown in the figure: The first row of the figure shows the staining pattern after 5 days of incubation. It can be seen that there is no significant difference between Example 2, Comparative Example 2, and the blank control. The second row of the figure shows the staining pattern after 10 days of incubation. It can be seen that after 10 days of incubation, the ALP staining intensity and the proportion of positive staining area in Example 2 are significantly higher than those in Comparative Example 2. This indicates that the bone cement prepared in Example 2 of this invention significantly enhances ALP activity when incubated with rat bone marrow mesenchymal stem cells.

[0052] In summary, the bone cement prepared from the solid phase composition of the bone cement provided by the present invention has a long injection time.

[0053] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A bone cement solid phase composition comprising sodium polyphosphate (NaPO3)n), calcium hydroxide and polyglutamic acid; wherein the molar ratio of phosphate ions in the sodium polyphosphate (NaPO3)n) to calcium ions in the calcium hydroxide is 1 : (1-10); wherein n in the sodium polyphosphate (NaPO3)n) is 3-40; and wherein the polyglutamic acid is present in an amount of 5-20% by mass of the total mass of the sodium polyphosphate (NaPO3)n) and the calcium hydroxide. n n n n ​​​​ 2. The bone cement solid phase composition according to claim 1, characterized in that, The mass of the polyglutamic acid is 10-20% of the total mass of the sodium polyphosphate (NaPO3) n and calcium hydroxide.

3. The bone cement solid phase composition according to claim 1 or 2, characterized in that, The ratio of the amount of substance of phosphate ions in the sodium polyphosphate to the amount of substance of calcium ions in the calcium hydroxide is 1: (2-5).

4. The bone cement solid phase composition according to claim 1, characterized in that, Said sodium polyphosphate (NaP03) n n is 15-30.

5. A bone cement comprising a solid phase component and a liquid phase component separately packaged, characterized in that, The solid phase component is the bone cement solid phase composition according to any one of claims 1-4.

6. The bone cement of claim 5, wherein, The ratio of the volume of the liquid phase component to the mass of the solid phase component is 0.25-0.7 mL / g.

7. Bone cement as claimed in claim 5 or 6, characterized in that The liquid phase component comprises one or more of pure water, a phosphate buffer, a sodium phosphate dibasic aqueous solution, a sodium phosphate monobasic aqueous solution, a simulated body fluid (SBF) and physiological saline.

8. The bone cement of claim 5 wherein, The particle size of the solid phase component is 20-400 mesh.

9. A method for using the bone cement according to any one of claims 5-8, comprising: mixing the separately packaged solid phase component and liquid phase component and then self-curing.

10. The method of use of claim 9, wherein, The self-curing temperature is 20-40 DEG C and the self-curing humidity is 40-100%.

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