A bone cement composition and its use

Abstract

The application discloses a bone cement composition and application thereof, and relates to the technical field of bone cement. The bone cement composition comprises a solid phase component and a liquid phase component, wherein the solid phase component comprises: 60-75 parts of a composite calcium phosphate salt, 10-20 parts of strontium glass powder, 8-12 parts of a reinforcing agent, and 3-7 parts of a coagulation reaction regulator; and the liquid phase component comprises: 20-30 parts of a phosphate buffer, 3-8 parts of a sodium citrate solution, 5-15 parts of an anti-inflammatory agent, and 1-5 parts of a hyaluronic acid solution. The reinforcing agent is prepared by reacting 1,10-phenanthroline-2,9-dicarboxylic acid with 2-aminomethylfuran to obtain an intermediate 1, and then reacting the intermediate 1 with poly(ethylene glycol) maleimide. The bone cement composition prepared by the application has good mechanical properties, self-repairing properties and anti-inflammatory properties, and has a good application prospect in the field of bone repair.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of bone cement, in particular to a bone cement composition and application thereof. BACKGROUND

[0002] Bone cement is a key material for filling bone defects, fixing implants or strengthening vertebral bodies in orthopedic surgery. At present, the most widely used in clinical application is polymethyl methacrylate (PMMA) bone cement and calcium phosphate cement. PMMA bone cement has the advantages of high mechanical strength and fast curing, but has the problems of biological inertness, high exothermic temperature and mismatched elastic modulus. Calcium phosphate cement gradually replaces PMMA due to its biodegradability and bone conduction, but its inherent brittleness and insufficient mechanical strength limit its application in load-bearing bone repair; at the same time, traditional calcium phosphate cement lacks active bone induction and anti-inflammatory function, and the bone integration rate is slow.

[0003] The Chinese invention patent with publication number CN112533651A discloses a bone cement composition, which comprises: a powder component comprising at least one acrylic acid polymer, a liquid component comprising a monomer, an antibiotic and an acid-functionalized polymer, wherein the powder component and the liquid component react to form bone cement. The bone cement formed by the bone cement composition has good antibacterial activity and compressive modulus, but its anti-inflammatory performance is still insufficient. SUMMARY

[0004] In view of the deficiencies of the prior art, the purpose of the present application is to provide a bone cement composition and application thereof.

[0005] To achieve the above-mentioned purpose, the present application realizes the following technical solutions:

[0006] A bone cement composition comprises a solid phase component and a liquid phase component;

[0007] The solid phase component comprises the following components in parts by weight: composite calcium phosphate salt 60-75 parts, strontium glass powder 10-20 parts, reinforcing agent 8-12 parts, and coagulation reaction modifier 3-7 parts;

[0008] The liquid phase component comprises the following components in parts by weight: phosphate buffer 20-30 parts, sodium citrate solution 3-8 parts, anti-inflammatory agent 5-15 parts, and hyaluronic acid solution 1-5 parts.

[0009] The reinforcing agent is prepared by the following method:

[0010] S1: under nitrogen protection, 1,10-phenanthroline-2,9-dicarboxylic acid was mixed with DMF, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, N-hydroxysuccinimide and triethylamine were added and activated, then 2-aminomethylfuran was added in batches, and the mixture was reacted at 25-30℃ for 8-9h to obtain intermediate 1;

[0011] S2: under nitrogen protection, intermediate 1 and poly(ethylene glycol) maleimide were mixed with anhydrous toluene, and the mixture was reacted at 100-110℃ for 13-14h to obtain the reinforcing agent. In this step, the carbon-carbon double bond of the furan ring of intermediate 1 and the carbon-carbon double bond of poly(ethylene glycol) maleimide undergo Diels-Alder reaction.

[0012] In step S1, the molar ratio of 1,10-phenanthroline-2,9-dicarboxylic acid to 2-aminomethylfuran is 1:(2.1-2.3).

[0013] In step S2, the molar ratio of intermediate 1 to poly(ethylene glycol) maleimide is 1:(2.05-2.2).

[0014] The anti-inflammatory agent is prepared by the following method:

[0015] N1: under nitrogen protection, 5-oxovaleric acid, dicyclohexyl carbodiimide, 4-dimethylaminopyridine were mixed with anhydrous DMF and activated, then curcumin was added in batches, and the mixture was reacted at 80-95℃ for 6-8h to obtain intermediate A;

[0016] N2: under nitrogen protection, intermediate A and 4-aminophenylphosphorylcholine were mixed with anhydrous ethanol and PBS buffer, and the mixture was reacted at 35-40℃ for 10-12h to obtain the anti-inflammatory agent.

[0017] In step N1, the molar ratio of curcumin to 5-oxovaleric acid is 1:(2.1-2.3).

[0018] In step N2, the molar ratio of intermediate A to 4-aminophenylphosphorylcholine is 1:(2.05-2.2).

[0019] The composite calcium phosphate salt comprises α-tricalcium phosphate and tetracalcium phosphate, wherein the weight fraction of α-tricalcium phosphate is 70%, and the weight fraction of tetracalcium phosphate is 30%; the coagulation reaction regulator is one of calcium hydrogen phosphate and calcium carbonate; the concentration of the sodium citrate solution is 0.1M; and the concentration of the hyaluronic acid solution is 0.2w / v%.

[0020] A bone cement composition is prepared by the following steps:

[0021] (1) Weigh by weight parts: composite calcium phosphate salt 60-75 parts, strontium glass powder 10-20 parts, reinforcing agent 8-12 parts, coagulation reaction regulator 3-7 parts, phosphate buffer 20-30 parts, sodium citrate solution 3-8 parts, anti-inflammatory agent 5-15 parts, hyaluronic acid solution 1-5 parts;

[0022] (2) Mix the composite calcium phosphate salt, strontium glass powder, reinforcing agent and coagulation reaction regulator in a three-dimensional mixer to obtain a solid phase component;

[0023] (3) Mix the phosphate buffer, sodium citrate solution, anti-inflammatory agent and hyaluronic acid solution uniformly, degas by ultrasonic, to obtain a liquid phase component;

[0024] (4) Mix the solid phase component and the liquid phase component uniformly according to a mass ratio of 2:1, stir, and the bone cement composition is obtained.

[0025] Due to the adoption of the above technical scheme, the beneficial effects of the present application include:

[0026] (1) The reinforcing agent prepared in the present application combines the aromatic ring rigid core, Diels-Alder dynamic bond and oligoether flexible chain to form a rigid-dynamic-flexible synergistic structure, which can simultaneously improve the rigidity and toughness of the bone cement composition, thereby improving the mechanical properties of the bone cement composition, and can also endow the bone cement composition with certain self-repairing ability and enhance the fatigue resistance.

[0027] (2) The anti-inflammatory agent prepared in the present application comprises a curcumin anti-inflammatory core-pH sensitive Schiff base bond-phosphorylcholine structure, which can improve the anti-inflammatory performance and interfacial bonding capacity of the bone cement composition, and achieve long-acting and sustained-release anti-inflammatory effect. DETAILED DESCRIPTION

[0028] The present application will be further described below in conjunction with examples, but the present application is not limited to these examples.

[0029] Example 1: Preparation of reinforcing agent

[0030] S1: Under nitrogen protection, 0.1 mol 1,10-phenanthroline-2,9-dicarboxylic acid was added to 400 ml DMF at room temperature, stirred and mixed, 0.21 mol 1-ethyl-(3-dimethylaminopropyl) carbonyldiimidazole hydrochloride, 0.2 mol N-hydroxysuccinimide and 0.2 mol triethylamine were added, activated for 45 min, then 0.21 mol 2-aminomethyl furan (added in 3 batches with an interval of 10 min) was added, reacted at 25℃ for 9 h, the mixture was poured into 800 ml ice water, filtered, the filter cake was washed with deionized water (3×200 ml), purified by silica gel column chromatography (eluent: dichloromethane / methanol V / V=10:1), rotary evaporation at 60℃ for 5 h, and the intermediate 1 was obtained; the reaction equation is as follows:

[0031]

[0032] The nuclear magnetic hydrogen spectrum data thereof are as follows:

[0033] 1 H NMR (500 MHz, DMSO- d 6) δ 9.22 (s, 2H), 8.35 (d, J = 0.5 Hz, 2H),8.10 (s, 2H), 8.05 (d, J = 0.6 Hz, 2H), 7.46 (s, 2H), 6.32 (s, 2H), 6.24 (s,2H), 4.49 (d, J = 0.5 Hz, 4H)。

[0034] S2: 0.1 mol of intermediate 1 and 0.205 mol of poly(ethylene glycol) maleimide are added to 500 ml of anhydrous toluene under nitrogen protection, stirred and uniformly mixed, and reacted at 100°C for 14 h; after the reaction is completed, rotary evaporation is performed at 60°C for 4 h, the residue is dissolved in 1000 ml of anhydrous ethanol / water (the volume ratio of anhydrous ethanol to water is 3:1), dialyzed with deionized water (the molecular weight cut-off is 1000 Da) for 48 h (water is replaced every 6 hours, and 2000 ml of water is replaced each time), and freeze-dried at -50°C for 24 h to obtain the enhancer; the reaction equation is as follows:

[0035]

[0036] Example 2 Preparation of the enhancer:

[0037] S1: 0.1 mol of 1,10-phenanthroline-2,9-dicarboxylic acid is added to 400 ml of DMF under nitrogen protection at room temperature, stirred and uniformly mixed, 0.21 mol of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 0.2 mol of N-hydroxysuccinimide and 0.2 mol of triethylamine are added, activated for 45 min, then 0.22 mol of 2-aminomethylfuran (added in 3 batches with an interval of 10 min) is added, and reacted at 30°C for 8 h; the mixture is poured into 800 ml of ice water, filtered, the filter cake is washed with deionized water (3×200 ml), purified by silica gel column chromatography (eluent: dichloromethane / methanol V / V=10:1), and rotary evaporation is performed at 60°C for 5 h to obtain intermediate 1;

[0038] S2: Under nitrogen protection, 0.1 mol of intermediate 1 and 0.21 mol of poly(ethylene glycol) maleimide were added into 500 ml of anhydrous toluene, stirred and mixed, reacted at 110°C for 13 h, rotary evaporated at 60°C for 4 h, the residue was dissolved in 1000 ml of anhydrous ethanol / water (the volume ratio of anhydrous ethanol to water was 3:1), dialyzed with deionized water (the molecular weight cut-off was 1000 Da) for 48 h (the water was changed every 6 h, and 2000 ml of water was used each time), and freeze-dried at -50°C for 24 h to obtain the enhancer.

[0039] Example 3: Preparation of an enhancer

[0040] S1: Under nitrogen protection, 0.1 mol of 1,10-phenanthroline-2,9-dicarboxylic acid was added into 400 ml of DMF at room temperature, stirred and mixed, 0.21 mol of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 0.2 mol of N-hydroxysuccinimide and 0.2 mol of triethylamine were added, activated for 45 min, then 0.23 mol of 2-aminomethylfuran (added in three batches with an interval of 10 min) was added, reacted at 30°C for 8 h, the mixture was poured into 800 ml of ice water, filtered, the filter cake was washed with deionized water (3×200 ml), purified by silica gel column chromatography (eluent: dichloromethane / methanol V / V=10:1), and rotary evaporated at 60°C for 5 h to obtain intermediate 1.

[0041] S2: Under nitrogen protection, 0.1 mol of intermediate 1 and 0.22 mol of poly(ethylene glycol) maleimide were added into 500 ml of anhydrous toluene, stirred and mixed, reacted at 110°C for 13 h; after the reaction was completed, rotary evaporated at 60°C for 4 h, the residue was dissolved in 1000 ml of anhydrous ethanol / water (the volume ratio of anhydrous ethanol to water was 3:1), dialyzed with deionized water (the molecular weight cut-off was 1000 Da) for 48 h (the water was changed every 6 h, and 2000 ml of water was used each time), and freeze-dried at -50°C for 24 h to obtain the enhancer.

[0042] Example 4: Preparation of an anti-inflammatory agent

[0043] N1: Under nitrogen protection, 0.21 mol of 5-oxovaleric acid, 0.21 mol of dicyclohexyl carbodiimide and 0.04 mol of 4-dimethylaminopyridine were added into 300 ml of anhydrous DMF, stirred and mixed, activated at room temperature for 45 min, 0.1 mol of curcumin was added in batches (divided into three batches with an interval of 10 min), reacted at 80°C for 8 h, filtered, the filtrate was rotary evaporated at 65°C for 3 h, purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate V / V=1:2), and rotary evaporated at 50°C for 2 h to obtain intermediate A; the reaction equation is shown as follows:

[0044]

[0045] The nuclear magnetic hydrogen spectrum data thereof are as follows:

[0046] 1 H NMR (500 MHz, DMSO- d 6) δ 9.59 (s, 2H), 7.52 (t, J = 0.9 Hz, 2H),7.37 (dd, J = 2.0, 1.0 Hz, 2H), 7.27 (ddd, J = 7.5, 1.9, 0.9 Hz, 2H), 7.23(d, J = 7.5 Hz, 2H), 7.00 (s, 2H), 3.96 (d, J = 12.4 Hz, 2H), 3.87 (s, 6H),2.58 (s, 4H), 2.54 (s, 4H), 1.79 (s, 4H).

[0047] N2: 0.1 mol of intermediate A, 0.205 mol of 4-aminophenyl phosphorylcholine and 450 ml of anhydrous ethanol and 150 ml of PBS buffer (the concentration of the PBS buffer is 0.1 M, and the pH is 7.2) are uniformly mixed, and the mixture is reacted at 35°C for 12 h. After being cooled to room temperature, the reaction solution is transferred to a dialysis bag (the molecular weight cut-off is 1000 Da), and dialysis is performed for 24 h (the solution is replaced every 6 h, and each time 2000 ml of solution is used; the first two times of dialysis are performed using PBS buffer with a concentration of 0.1 M and a pH of 7.2, and the last two times of dialysis are performed using deionized water), and the solution is freeze-dried at -50°C for 24 h to obtain the anti-inflammatory agent. The reaction equation is as follows:

[0048]

[0049] The nuclear magnetic hydrogen spectrum data thereof are as follows:

[0050] 1 H NMR (500 MHz, DMSO- d6) δ 7.52 (t, J = 0.9 Hz, 2H), 7.37 (dd, J = 2.0, 1.0 Hz, 2H), 7.36 (s, 2H), 7.35-7.32 (m, 4H), 7.30-7.21 (m, 4H), 7.17-7.11 (m, 4H), 7.00 (s, 2H), 6.30 (s, 2H), 4.26 (s, 4H), 3.96 (d, J = 12.4 Hz, 2H), 3.87 (s, 6H), 3.60 (s, 4H), 3.21 (s, 18H), 2.58 (s, 4H), 2.21 (s, 4H), 1.74 (s, 4H).

[0051] Example 5 Preparation of Anti-inflammatory agent:

[0052] N1: Under nitrogen protection, 0.22 mol of 5-oxopentanoic acid, 0.22 mol of dicyclohexyl carbodiimide and 0.04 mol of 4-dimethylaminopyridine were added into 300 ml of anhydrous DMF, stirred and mixed, activated at room temperature for 45 min, 0.1 mol of curcumin was added in batches (divided into three batches, each batch interval of 10 min), reacted at 90°C for 7 h, filtered, the filtrate was rotary evaporated at 65°C for 3 h, purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate V / V = 1:2), rotary evaporated at 50°C for 2 h to obtain intermediate A;

[0053] N2: Under nitrogen protection, 0.1 mol of intermediate A, 0.21 mol of 4- aminophenyl phosphorylcholine and 450 ml of anhydrous ethanol and 150 ml of PBS buffer (PBS buffer concentration is 0.1 M, pH is 7.2) were mixed, reacted at 35°C for 12 h, cooled to room temperature, the reaction solution was transferred to a dialysis bag (molecular weight cut-off 1000 Da), dialyzed for 24 h (every 6 hours, 2000 ml was replaced, the first two times were dialyzed with 0.1 M, pH 7.2 PBS buffer, and the last two times were dialyzed with deionized water), freeze-dried at -50°C for 24 h to obtain the anti-inflammatory agent.

[0054] Example 6 Preparation of Anti-inflammatory agent:

[0055] N1: Under nitrogen protection, 0.23 mol of 5-oxopentanoic acid, 0.22 mol of dicyclohexyl carbodiimide and 0.04 mol of 4-dimethylamino pyridine were added into 300 ml of anhydrous DMF, stirred and mixed, activated at room temperature for 40 min, 0.1 mol of curcumin was added in batches (divided into three batches, each batch was separated by 10 min), reacted at 95°C for 6 h, filtered, the filtrate was rotary evaporated at 65°C for 3 h, purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate V / V = 1:2), rotary evaporated at 50°C for 2 h, to obtain intermediate A;

[0056] N2: Under nitrogen protection, 0.1 mol of intermediate A, 0.22 mol of 4-aminophenyl phosphorylcholine and 450 ml of anhydrous ethanol and 150 ml of PBS buffer solution (the concentration of PBS buffer solution was 0.1 M, and the pH was 7.2) were mixed, reacted at 40°C for 10 h; after the reaction was completed, it was cooled to room temperature, the reaction solution was transferred to a dialysis bag (the molecular weight cut-off was 1000 Da), dialyzed for 24 h (every 6 hours, 2000 ml was replaced, the first two times were dialyzed by 0.1 M PBS buffer solution with a pH of 7.2, and the last two times were dialyzed by deionized water), and freeze-dried at -50°C for 24 h, to obtain the anti-inflammatory agent.

[0057] Example 7 Preparation of bone cement composition:

[0058] (1) The following were weighed by weight: composite calcium phosphate salt 60 g (α-tricalcium phosphate 42 g, tetracalcium phosphate 18 g), strontium glass powder 10 g, reinforcing agent (prepared in Example 1) 8 g, coagulation reaction modifier (calcium hydrogen phosphate) 3 g, phosphate buffer solution 20 g, sodium citrate solution 3 g, anti-inflammatory agent (prepared in Example 4) 5 g, hyaluronic acid solution 1 g;

[0059] (2) The composite calcium phosphate salt, strontium glass powder, reinforcing agent and coagulation reaction modifier were mixed in a three-dimensional mixer at 60 rpm for 2 h to obtain a solid phase component;

[0060] (3) The phosphate buffer solution, sodium citrate solution, anti-inflammatory agent and hyaluronic acid solution were mixed, and degassed by ultrasonic at 20 kHz for 10 min to obtain a liquid phase component;

[0061] (4) The solid phase component and the liquid phase component were mixed in a mass ratio of 2:1, and stirred for 5 min to obtain the bone cement composition.

[0062] Example 8 Preparation of bone cement composition:

[0063] (1) Weigh by weight: composite calcium phosphate salt 70 g (α-tricalcium phosphate 49 g, tetracalcium phosphate 21 g), strontium glass powder 15 g, reinforcing agent (prepared in Example 2) 10 g, coagulation reaction modifier (calcium carbonate) 5 g, phosphate buffer 25 g, sodium citrate solution 5 g, anti-inflammatory agent (prepared in Example 5) 10 g, hyaluronic acid solution 3 g;

[0064] (2) The composite calcium phosphate salt, strontium glass powder, reinforcing agent and coagulation reaction modifier are mixed in a three-dimensional mixer at 60 rpm for 2 h to obtain a solid phase component;

[0065] (3) The phosphate buffer, sodium citrate solution, anti-inflammatory agent and hyaluronic acid solution are mixed uniformly, degassed by ultrasonic at 20 kHz for 10 min to obtain a liquid phase component;

[0066] (4) The solid phase component and the liquid phase component are mixed uniformly at a mass ratio of 2:1, and stirred for 5 min to obtain a bone cement composition.

[0067] Example 9 Preparation of bone cement composition:

[0068] (1) Weigh by weight: composite calcium phosphate salt 70 g (α-tricalcium phosphate 49 g, tetracalcium phosphate 21 g), strontium glass powder 15 g, reinforcing agent (prepared in Example 2) 10 g, coagulation reaction modifier (calcium carbonate) 5 g, phosphate buffer 25 g, sodium citrate solution 5 g, anti-inflammatory agent (prepared in Example 5) 10 g, hyaluronic acid solution 3 g;

[0069] (2) The composite calcium phosphate salt, strontium glass powder, reinforcing agent and coagulation reaction modifier are mixed in a three-dimensional mixer at 60 rpm for 2 h to obtain a solid phase component;

[0070] (3) The phosphate buffer, sodium citrate solution, anti-inflammatory agent and hyaluronic acid solution are mixed uniformly, degassed by ultrasonic at 20 kHz for 10 min to obtain a liquid phase component;

[0071] (4) The solid phase component and the liquid phase component are mixed uniformly at a mass ratio of 2:1, and stirred for 5 min to obtain a bone cement composition.

[0072] Comparative Example 1

[0073] The raw material composition and preparation method of the bone cement composition are basically the same as those of Example 8, except that no reinforcing agent is added in the composition.

[0074] Comparative Example 2

[0075] The raw material composition and preparation method of the bone cement composition are basically the same as those of Example 8, except that no anti-inflammatory agent is added in the composition.

[0076] Comparative Example 3

[0077] The raw material composition and the preparation method of the bone cement composition of Example 8 are basically the same, except that the reinforcing agent is replaced by an equal weight of a reinforcing agent prepared by the following method:

[0078] The preparation method of the reinforcing agent is basically the same as that of Example 2, except that 1,10-phenanthroline-2,9-dicarboxylic acid in step S1 is replaced by an equal molar amount of phthalic acid.

[0079] Comparative Example 4

[0080] The raw material composition and the preparation method of the bone cement composition are basically the same as those of Example 8, except that the reinforcing agent is replaced by an equal weight of a reinforcing agent prepared by the following method:

[0081] The preparation method of the reinforcing agent is basically the same as that of Example 2, except that poly(ethylene glycol) maleimide in step S2 is replaced by an equal molar amount of N-(2-hydroxyethyl) maleimide.

[0082] Comparative Example 5

[0083] The raw material composition and the preparation method of the bone cement composition are basically the same as those of Example 8, except that the anti-inflammatory agent is replaced by an equal weight of intermediate A prepared in Example 5.

[0084] Comparative Example 6

[0085] The raw material composition and the preparation method of the bone cement composition are basically the same as those of Example 8, except that the anti-inflammatory agent is replaced by an equal weight of an anti-inflammatory agent prepared by the following method:

[0086] The preparation method of the anti-inflammatory agent is basically the same as that of Example 5, except that 5-oxovaleric acid in step N1 is replaced by 0.105 mol of 5-oxovaleric acid.

[0087] The strontium glass powder used in the examples and comparative examples of the present application is QZ-0020 produced by Jiangsu Qizheng New Material Technology Co., Ltd.; the number average molecular weight of hyaluronic acid is 10 kDa; and the number average molecular weight of poly(ethylene glycol) maleimide is 625 Da. The concentration of the phosphate buffer used in Examples 7-9 and Comparative Examples of the present application is 0.2 M, and the pH is 7.2; the concentration of the sodium citrate solution is 0.1 M, and the pH is 6.5; and the concentration of the hyaluronic acid solution is 0.2 w / v%.

[0088] Preparation of phosphate buffer solution (0.2 M, pH 7.2): weigh 35.814 g of sodium hydrogen phosphate dihydrate (Na2HPO4·H2O) and 13.609 g of potassium dihydrogen phosphate (KH2PO4), respectively, dissolve in 400 ml of deionized water, combine and transfer into a 1000 ml volumetric flask after complete dissolution, adjust the pH to 7.20 ± 0.02 with 1 M NaOH or HCl, and finally dilute to 1000 ml. Thus, a 0.2 M, pH = 7.2 phosphate buffer solution is obtained.

[0089] The bone cement compositions prepared in Examples 7-9 and Comparative Examples 1-6 were subjected to compressive strength, self-repairing performance and anti-inflammatory performance tests. The test results are shown in Table 1.

[0090] The compressive strength of the bone cement composition was tested according to the YY 0459-2003 standard, and a universal testing machine was used to test the compressive strength at a loading speed of 20 mm / min. The sample was kept at 23°C for 24 h before testing.

[0091] Preparation of samples for compressive strength test: 100 g of bone cement prepared in the comparative examples and examples was poured into a polytetrafluoroethylene cylindrical mold, and cured at 37°C for 1 h to obtain the sample (sample size: diameter 6 mm, length 12 mm).

[0092] Sample damage test: a knife was used to make a damage cut (0.5 mm deep and 10 mm long) at the center of the gauge section of the sample, and then the sample was immersed in 50 ml of PBS buffer solution at 37°C, 0.1 M and pH = 7.4. After 24 h of immersion, the repaired bone cement was obtained.

[0093] The anti-inflammatory performance of the bone cement composition was determined by macrophage inflammatory factor inhibition experiment, and the inflammatory factor inhibition rate was used as the characterization.

[0094] Sample preparation: 100 g of bone cement composition prepared in Comparative Examples 7-9 and Examples 1-6 was poured into a polytetrafluoroethylene cylindrical mold, and cured at 37°C for 1 h to obtain the sample (sample size: diameter 6 mm, length 12 mm). At the same time, the bone cement composition of Comparative Example 2 was used as a control.

[0095] The samples were sterilized by irradiation with a dose of 15 kGy of γ-rays, and the sterile samples were placed in a 96-well plate, and RAW264.7 cells were inoculated at a cell concentration of 5 × 10 3 2 ​The RAW264.7 cells were inoculated in a 6-well culture plate at a concentration of 1x106 cells / mL, and the culture plate was placed in a 95% humidity, 37°C and 5% CO2 environment for expansion culture, and the fresh complete culture medium was replaced every other day. The culture solution used in the culture process was composed of 90vol.% H-DMEM and 10vol.% FBS. The RAW264.7 cells used in the experiment were cells in the 3rd to 6th generation. After 3 days of culture, the cells growing on the sample were digested using 0.25wt% trypsin / EDTA, and then centrifuged at 1000r / min for 5min. The cells were blocked using a 1wt% BSA PBS solution, washed with PBS for 3 times after 30min of blocking, and then diluted CD11c (eBioscience) and CD206 (eBioscience) straight antibodies were added and incubated at room temperature for 10min. The cells were then washed with PBS for 3 times, placed in a serum-free DMEM culture medium (containing 1wt% penicillin-streptomycin), and then 1μg / ml lipopolysaccharide (E. coli O111:B4) was added to induce inflammation for 2h. A blank control group (only culture medium) and a lipopolysaccharide control group (lipopolysaccharide+ culture medium) were set. After 24h of culture, the supernatant was taken, and the concentrations of TNF-α and IL-6 were detected using an ELISA kit. The inflammation factor inhibition rate = (lipopolysaccharide control group concentration-experimental group concentration) / lipopolysaccharide control group concentration x 100%.

[0096] Table 1

[0097]

[0098] As can be seen from Table 1, the bone cement composition prepared in Examples 7-9 has excellent compressive strength, self-repairing performance and anti-inflammatory performance.

[0099] As can be seen from Table 1, when no reinforcing agent is added in Comparative Example 1, the tensile and compressive strength and self-repairing performance of the bone cement composition prepared are poor. When no anti-inflammatory agent is added in Comparative Example 2, the inflammation factor inhibition rate is significantly lower than that in the examples.

[0100] The compressive strength of the bone cement composition prepared in Comparative Example 3 is lower than that in the examples, mainly because the rigid structure of phthalic acid used is poorer than that of 1,10-phenanthroline-2,9-dicarboxylic acid, resulting in a decrease in the rigidity of the reinforcing agent, thereby reducing the compressive strength of the bone cement composition.

[0101] The compressive strength of the bone cement composition prepared in Comparative Example 4 is lower than that in the examples, mainly because the N-(2-hydroxyethyl) maleimide used is a short-chain structure, lacking a polyethylene glycol segment structure, and the prepared reinforcing agent has increased brittleness, resulting in a decrease in the compressive strength of the bone cement composition.

[0102] The anti-inflammatory performance of the bone cement composition prepared in Example 5 is reduced compared to the examples, mainly because the anti-inflammatory agent is replaced by an equal weight of intermediate A, the pH-sensitive Schiff base bond and phosphorylcholine structure are missing in the anti-inflammatory agent molecule, resulting in a decrease in long-acting sustained-release anti-inflammatory effect, thereby reducing the anti-inflammatory performance.

[0103] The anti-inflammatory performance of the bone cement composition prepared in Comparative Example 6 is reduced compared to the examples, because the prepared anti-inflammatory agent contains only one aldehyde group, the Schiff base site is reduced, and the anti-inflammatory agent drug loading is reduced, resulting in a decrease in anti-inflammatory performance.

[0104] The above is only a preferred embodiment of the present application and is not intended to limit the present application; however, for those of ordinary skill in the art, some minor changes, modifications, and equivalent variations of the above disclosed technical content can be made without departing from the scope of the technical solutions of the present application, and all such equivalent embodiments are within the scope of the present application; at the same time, any equivalent changes, modifications, and variations of the above embodiments according to the essential technology of the present application are still within the protection scope of the technical solutions of the present application.

Claims

1. A bone cement composition, characterized in that, It comprises a solid phase component and a liquid phase component; The solid phase component comprises the following components in parts by weight: composite calcium phosphate salt 60-75 parts, strontium glass powder 10-20 parts, reinforcing agent 8-12 parts, coagulation reaction modifier 3-7 parts; The liquid phase component comprises the following components in parts by weight: phosphate buffer 20-30 parts, sodium citrate solution 3-8 parts, anti-inflammatory agent 5-15 parts, hyaluronic acid solution 1-5 parts; The reinforcing agent is prepared by the following method: S1: Under nitrogen protection, 1,10-phenanthroline-2,9-dicarboxylic acid is mixed with DMF, 1-ethyl-(3-dimethylaminopropyl) carbonyl diimide hydrochloride, N-hydroxysuccinimide and triethylamine are activated, and then 2-aminomethyl furan is added in batches, and the reaction is carried out at 25-30℃ for 8-9h to obtain intermediate 1; S2: Under nitrogen protection, intermediate 1 and poly(ethylene glycol) maleimide are mixed with anhydrous toluene, and the reaction is carried out at 100-110℃ for 13-14h to obtain the reinforcing agent; The anti-inflammatory agent is prepared by the following method: N1: Under nitrogen protection, 5-oxovaleric acid, dicyclohexyl carbodiimide, 4-dimethylamino pyridine and anhydrous DMF are mixed and activated, and then curcumin is added in batches, and the reaction is carried out at 80-95℃ for 6-8h to obtain intermediate A; N2: Under nitrogen protection, intermediate A and 4-aminophenyl phosphocholine are mixed with anhydrous ethanol and PBS buffer, and the reaction is carried out at 35-40℃ for 10-12h to obtain the anti-inflammatory agent.

2. A bone cement composition according to claim 1, wherein, In step S1, the molar ratio of 1,10-phenanthroline-2,9-dicarboxylic acid to 2-aminomethyl furan is 1:(2.1-2.3).

3. A bone cement composition according to claim 1, wherein, In step S2, the molar ratio of intermediate 1 to poly(ethylene glycol) maleimide is 1:(2.05-2.2).

4. A bone cement composition according to claim 1, wherein In step N1, the molar ratio of curcumin to 5-oxovaleric acid is 1:(2.1-2.3).

5. A bone cement composition according to claim 1, wherein, In step N2, the molar ratio of intermediate A to 4-aminophenyl phosphocholine is 1:(2.05-2.2).

6. A bone cement composition according to claim 1, wherein, The composite calcium phosphate salt comprises α-tricalcium phosphate and tetracalcium phosphate, wherein the weight fraction of α-tricalcium phosphate is 70%, and the weight fraction of tetracalcium phosphate is 30%; the coagulation reaction modifier is one of calcium hydrogen phosphate and calcium carbonate; the concentration of the sodium citrate solution is 0.1M; and the concentration of the hyaluronic acid solution is 0.2w / v%.

7. The bone cement composition according to any one of claims 1 to 6, characterized in that, It is prepared by the following steps: (1) The following components are weighed in parts by weight: composite calcium phosphate salt 60-75 parts, strontium glass powder 10-20 parts, reinforcing agent 8-12 parts, coagulation reaction modifier 3-7 parts, phosphate buffer 20-30 parts, sodium citrate solution 3-8 parts, anti-inflammatory agent 5-15 parts, and hyaluronic acid solution 1-5 parts; (2) The composite calcium phosphate salt, strontium glass powder, reinforcing agent and coagulation reaction modifier are mixed in a three-dimensional mixer to obtain a solid phase component; (3) The phosphate buffer, sodium citrate solution, anti-inflammatory agent and hyaluronic acid solution are mixed and degassed by ultrasonic to obtain a liquid phase component; (4) The solid phase component and the liquid phase component are mixed in a mass ratio of 2:1, and stirred to obtain a bone cement composition.

8. Use of the bone cement composition according to any one of claims 1 to 6 for the preparation of a bone repair material.

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