Composition for changing plasticity of bone meal particles as well as preparation method and application of composition

Abstract

The invention provides a composition for changing plasticity of bone meal particles as well as a preparation method and application of the composition, and relates to the technical field of biomedical materials. The composition is prepared from the following raw material components: bioactive glass, dihydrazide modified sodium hyaluronate, sodium hyaluronate with the molecular weight of 50kDa to 500kDa, a polylactic acid-glycolic acid copolymer and carboxymethyl cellulose. The preparation method of the dihydrazide modified sodium hyaluronate comprises the following steps: carrying out carboxyl activation on sodium hyaluronate, carrying out a cross-linking reaction on the activated hyaluronic acid and adipic dihydrazide, and then terminating the reaction. The composition provided by the invention not only can provide support, but also can adjust the degradation rate to be matched with the bone regeneration period, and all the components synergistically improve the mechanical property and the viscoelastic property. The composition disclosed by the invention can be formed in situ at a bone defect part, provides a space for growth and proliferation of bone cells, improves the biological activity and osteoinductivity of an artificial bone, and promotes bone growth and repair.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomedical materials, and particularly relates to a composition for changing the plasticity of bone powder particles, a preparation method and application thereof. BACKGROUND

[0002] Granular artificial bone substitute material is a commonly used bone graft material for guided bone regeneration in the field of bone implantation. Current bone graft materials are generally based on animal-derived materials (bovine bone, porcine bone, etc.). After removing organic matter from animal-derived materials through different processes, the materials are crushed into particles within a certain particle size range. When used, the particles are rehydrated or re-blooded and filled into the defect site to form a stable three-dimensional scaffold structure, guide cell growth and vascular invasion, and ultimately achieve bone reconstruction. However, granular bone graft materials are easily washed away by physiological saline or blood in clinical use, lack cohesion, and are inconvenient to operate. By adding excipients to aggregate the bone graft materials into a mass, stable implantation can be achieved.

[0003] Therefore, the selection and modification of excipients are crucial. If the excipients are not used properly, it may lead to insufficient biological activity of artificial bone after implantation, difficulty in matching the degradation and tissue regeneration and repair speed, and difficulty in achieving true "biological fusion" with the host tissue. In addition, if some excipients have insufficient performance, they may limit the growth of artificial bone, have weak compressive strength, and be insufficient to support the effective growth of bone tissue.

[0004] Sodium hyaluronate (sodium hyaluronate, HA) is a naturally occurring polysaccharide substance in the human body, which has excellent biocompatibility and biological properties. It usually does not cause immune and rejection reactions. The main products of its degradation and metabolism in the human body are water and carbon dioxide, which are then excreted outside the body through the body's metabolic pathways. The molecular structure of sodium hyaluronate is more stable, and it has excellent viscoelastic properties. Mixing with inorganic bone powder particles will not cause chemical changes, and it gives artificial bone powder good physical plasticity. In the field of orthopedic and dental technology, sodium hyaluronate gel is often used as an excipient to take advantage of its high viscosity and physical properties to mix and stir with bone substitute materials to make them into a dough-like state.

[0005] Chinese invention patent CN102727424A discloses a sodium hyaluronate gel injection for use in the joint cavity and its preparation method. The method involves cross-linking sodium hyaluronate using a cross-linking agent, 1,4-butanediol diglycidyl ether (BDDE), in a sodium hydroxide and acetone solution. Deionized water is added for complete swelling, followed by purification with isotonic buffer for 6-10 hours to obtain cross-linked sodium hyaluronate. However, this cross-linking process inevitably leaves some residual organic solvents such as acetone, which may be harmful to the human body. Furthermore, using sodium hyaluronate alone can lead to bone loss due to the high viscoelasticity of hyaluronic acid. Excessive aqueous solution or significant blood leakage can cause bone particles encapsulated in the hyaluronic acid gel to slide. In some horizontal and vertical bone augmentation surgeries, the lack of sufficient bone wall support and the resulting bone particle movement can lead to instability in the bone graft space in the defect area, thus affecting new bone formation.

[0006] Chinese invention patent CN117752856A discloses a bone graft material, its preparation method, and its application. The bone graft material comprises bone particles and cholesterol-modified sodium hyaluronate, wherein the mass ratio of sodium hyaluronate to cholesterol in the cholesterol-modified sodium hyaluronate ranges from 100:1 to 20:1. The cholesterol-modified sodium hyaluronate is obtained by reacting a mixture of cholesterol and sodium hyaluronate with a catalyst selected from carbodiimide or p-toluenesulfonic acid. This invention uses cholesterol-modified sodium hyaluronate as a cross-linking raw material, which can significantly improve the mechanical properties of the product after cross-linking sodium hyaluronate and bone particles are combined, exhibiting strong plasticity and resistance to flow. However, the introduction of cholesterol may alter the degradation pathway and rate of sodium hyaluronate. If degradation is too slow, it may accumulate in the body for a long time, leading to foreign body reactions or hindering tissue repair. The cholesterol grafting rate needs to be controlled; excessively high rates may reduce the water solubility of the material, making it difficult to process and shape.

[0007] Furthermore, when repairing irregularly shaped bone defects, a highly malleable material is needed to avoid dead cavities caused by incompatibility between the bone powder particles and the bone defect, which could lead to nonunion or delayed healing. Currently, there is a lack of a malleable material with clearly defined raw materials, synergistic effects among its components, a simple preparation method, mechanical properties meeting osteogenic requirements, high compressive strength, promoting rapid new bone formation, and applicability to filling and repair various complex structures. Summary of the Invention

[0008] This invention addresses the problems existing in the prior art by providing a composition that alters the plasticity of bone powder particles, its preparation method, and its applications. Diazidamide-modified sodium hyaluronate, as an excipient in bone materials, provides support and regulates the degradation rate to match the bone regeneration cycle. Combined with sodium hyaluronate with a molecular weight of 50kDa-500kDa and carboxymethyl cellulose, it synergistically improves mechanical properties and exhibits excellent viscoelasticity, enabling in-situ molding at bone defect sites and enhancing the composition's plasticity. Furthermore, by incorporating polylactic acid-glycolic acid copolymer and bioactive glass in a specific ratio, it provides space for bone cell growth and proliferation, enhancing the bioactivity and osteoinductive properties of artificial bone, and promoting bone growth and repair. The composition of this invention can be widely applied in orthopedic materials, dental materials, and other technical fields.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows: First, the present invention provides a composition for modifying the plasticity of bone meal particles, wherein the raw material components, by weight, include: 40-70 parts of bioactive glass, 5-9 parts of dihydrazide-modified sodium hyaluronate, 5-10 parts of sodium hyaluronate with a molecular weight of 50kDa-500kDa, 8-20 parts of polylactic acid-glycolic acid copolymer and 4-8 parts of carboxymethyl cellulose.

[0010] Preferably, the composition comprises, by weight, 50-60 parts of bioactive glass, 6-8 parts of dihydrazide-modified sodium hyaluronate, 6-8 parts of sodium hyaluronate with a molecular weight of 50kDa-500kDa, 10-16 parts of polylactic acid-glycolic acid copolymer, and 5-7 parts of carboxymethyl cellulose.

[0011] More preferably, the composition, by weight, comprises: 55 parts bioactive glass, 7 parts dihydrazide-modified sodium hyaluronate, 7 parts sodium hyaluronate with a molecular weight of 50kDa-500kDa, 15 parts polylactic acid-glycolic acid copolymer, and 6 parts carboxymethyl cellulose.

[0012] Preferably, the sodium hyaluronate has a molecular weight of 50kDa-500kDa, and more preferably a molecular weight of 100kDa-300kDa.

[0013] Preferably, the preparation method of the dihydrazide-modified sodium hyaluronate includes the following steps: (1) Sodium hyaluronate was subjected to pH adjustment and carboxyl group activation to obtain activated sodium hyaluronate; (2) The activated hyaluronic acid and adipic acid dihydrazide (ADH) were cross-linked and then the reaction was terminated to obtain dihydrazide-modified sodium hyaluronate.

[0014] More preferably, in step (1), the sodium hyaluronate is sodium hyaluronate with a molecular weight of 50kDa-2000kDa; even more preferably, in step (1), the sodium hyaluronate is sodium hyaluronate with a molecular weight of 500kDa-1000kDa.

[0015] More preferably, in step (1), the pH is adjusted to 5-6; the reagent used to adjust the pH is a conventional acid-base reagent used in the art, and is not limited to hydrochloric acid, sulfuric acid, citric acid, or sodium hydroxide.

[0016] More preferably, in step (1), the pH is adjusted to 5.5.

[0017] More preferably, step (1) specifically involves: preparing a hyaluronic acid solution into a sodium hyaluronate solution with a mass concentration of 5%-10%, adjusting the pH, adding 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), stirring evenly, and obtaining activated sodium hyaluronate.

[0018] More preferably, step (1) specifically involves: preparing a sodium hyaluronate solution with a mass concentration of 8% by hyaluronic acid solution, adjusting the pH, adding 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), stirring evenly to obtain activated sodium hyaluronate.

[0019] More preferably, in step (1), the molar ratio of the carboxyl group in the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to that in sodium hyaluronate is 2-5:1; the molar ratio is preferably 4:1.

[0020] More preferably, in step (1), the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide is 1:1-2; the molar ratio is preferably 1:1.5.

[0021] More preferably, in step (1), the stirring is specifically: stirring at room temperature and in the dark for 15-30 minutes until the sodium hyaluronate is activated into an active ester intermediate, which is conducive to cross-linking reaction.

[0022] More preferably, in step (2), the molar ratio of the adipic acid dihydrazide to the carboxyl group in sodium hyaluronate is 1-3:1; the molar ratio is preferably 1.5-2.5:1; the most preferably molar ratio is 2:1.

[0023] More preferably, in step (2), the crosslinking reaction is specifically carried out at room temperature, away from light, and stirred for 8-16 hours; even more preferably, in step (2), the stirring time is preferably 12 hours.

[0024] More preferably, in step (2), the termination reaction is specifically performed by adding water and stirring for 20-40 minutes to terminate the reaction and obtain a reaction solution; then dialyzing the reaction solution for 3-7 days to remove unreacted substances or byproducts, centrifuging to obtain sodium hyaluronate modified with dihydrazide.

[0025] Preferably, in the polylactic acid-glycolic acid copolymer, the molar ratio of lactic acid to glycolic acid is 50-65:35-50.

[0026] More preferably, in the polylactic acid-glycolic acid copolymer, the molar ratio of lactic acid to glycolic acid is 60:40.

[0027] Then, the present invention provides a method for preparing the above composition, comprising the steps of: S1. Sodium hyaluronate with a molecular weight of 50kDa-500kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed to obtain mixture 1; S2, Mixture 1 is mixed with bioactive glass and dihydrazide-modified sodium hyaluronate to obtain a composition.

[0028] Preferably, in step S1, the mixing conditions are: room temperature, stirring for 30-40 minutes.

[0029] Preferably, in step S2, the mixing conditions are: room temperature, stirring for 30-40 minutes.

[0030] Furthermore, the present invention provides the application of the above composition in the preparation of bone graft materials.

[0031] Preferably, the bone graft material is not limited to bone graft materials used in orthopedics or dentistry.

[0032] Finally, the present invention provides a bone graft material comprising: artificial bone material and the above-described composition.

[0033] Preferably, the artificial bone material is a material commonly used in the art, and is not limited to animal-derived materials, bioceramic materials, metallic materials, or polyetheretherketone polymer materials.

[0034] Compared with the prior art, the present invention has the following beneficial effects: 1. In the composition of the present invention, dihydrazide-modified sodium hyaluronate serves as an excipient for bone materials, providing support and regulating the degradation rate to match the bone regeneration cycle. Combined with medium- to high molecular weight sodium hyaluronate and carboxymethyl cellulose, it enhances mechanical properties and plasticity. Combined with polylactic acid-glycolic acid copolymer and bioactive glass in a specific ratio, it provides space for the growth and proliferation of osteocytes, improves the bioactivity and osteoinductive properties of artificial bone, and promotes bone growth and repair.

[0035] 2. In the composition of the present invention, the sodium hyaluronate used is sodium hyaluronate modified with diazid and sodium hyaluronate with a molecular weight of 50kDa-500kDa. The above two different types of sodium hyaluronate and carboxymethyl cellulose interact with each other to regulate the strength and plasticity of the composition. Among them, the diacylhydrazine modified sodium hyaluronate is prepared under mild reaction conditions, without the use of organic solvents, and is safe to use. The cross-linked sodium hyaluronate (HA) gel has good plasticity and excellent viscoelasticity, and can be formed in situ at the bone defect site. Detailed Implementation

[0036] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0037] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0038] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels. Products of the same properties from different manufacturers have no significant impact on the technical effect. For example, in a specific embodiment of the present invention, the bioactive glass was purchased from Shenzhen Dazhou Medical Technology Co., Ltd., and is a bio-glass artificial bone, with national medical device registration number 20243172535; the sodium hyaluronate (molecular weight in the range of 100kDa-300kDa) was purchased from Hangzhou Xiehe Medical Supplies Co., Ltd.; the sodium hyaluronate (molecular weight in the range of 300kDa-500kDa) was purchased from Hangzhou Xiehe Medical Supplies Co., Ltd.; the sodium hyaluronate (molecular weight in the range of 50kDa-100kDa) was purchased from Hangzhou Xiehe Medical Supplies Co., Ltd.; the polylactic acid was purchased from Zhejiang Haizheng Biomaterials Co., Ltd.; the glycolic acid was purchased from Corbion Purac (Netherlands); the carboxymethyl cellulose was purchased from Corbion Purac (Netherlands); the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride was purchased from Adamas; the N-hydroxysuccinimide was purchased from Adamas; and the collagen was purchased from Nitta. Gelatin (Nitta Gelatin, Japan); the adipic acid dihydrazide was purchased from Adamas.

[0039] Example 1 A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 55 parts bioactive glass, 7 parts dihydrazide-modified sodium hyaluronate, 7 parts sodium hyaluronate with a molecular weight of 100kDa-300kDa, 15 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40), and 6 parts carboxymethyl cellulose.

[0040] The preparation method of the dihydrazide-modified sodium hyaluronate is as follows: (1) Sodium hyaluronate with a molecular weight of 500kDa-1000kDa was prepared into an 8% sodium hyaluronate aqueous solution. The pH was adjusted to 5.5 with dilute hydrochloric acid. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide were added in sequence and stirred evenly to obtain activated sodium hyaluronate. The molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to the carboxyl group in sodium hyaluronate was 4:1, and the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to N-hydroxysuccinimide was 1:1.5. (2) The activated hyaluronic acid and adipic dihydrazide were subjected to a cross-linking reaction. The molar ratio of the carboxyl groups in adipic dihydrazide to sodium hyaluronate was 2:1. The reaction was carried out at room temperature, in the dark, and stirred for 12 hours. Water was added and stirred for 30 minutes to terminate the reaction and obtain the reaction solution. The reaction solution was then dialyzed for 7 days to remove unreacted substances or byproducts. After centrifugation, sodium hyaluronate modified with dihydrazide was obtained.

[0041] The preparation method of the composition in this embodiment is as follows: S1. Sodium hyaluronate with a molecular weight of 100kDa-300kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 35 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0042] Example 2 A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 50 parts bioactive glass, 6 parts dihydrazide-modified sodium hyaluronate, 6 parts sodium hyaluronate with a molecular weight of 300kDa-500kDa, 10 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 65:35) and 5 parts carboxymethyl cellulose.

[0043] The preparation method of dihydrazide-modified sodium hyaluronate is the same as in Example 1.

[0044] The preparation method of the composition in this embodiment is as follows: S1. Sodium hyaluronate with a molecular weight of 300kDa-500kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 35 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0045] Example 3 A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 60 parts bioactive glass, 8 parts dihydrazide-modified sodium hyaluronate, 8 parts sodium hyaluronate with a molecular weight of 50kDa-100kDa, 16 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 50:50) and 7 parts carboxymethyl cellulose.

[0046] The preparation method of dihydrazide-modified sodium hyaluronate is the same as in Example 1.

[0047] The preparation method of the composition in this embodiment is as follows: S1. Sodium hyaluronate with a molecular weight of 50kDa-100kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 35 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0048] Example 4 A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 40 parts bioactive glass, 5 parts dihydrazide-modified sodium hyaluronate, 5 parts sodium hyaluronate with a molecular weight of 100kDa-300kDa, 8 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40) and 4 parts carboxymethyl cellulose.

[0049] The preparation method of the dihydrazide-modified sodium hyaluronate is as follows: (1) Sodium hyaluronate with a molecular weight of 500kDa-1000kDa was prepared into a 5% sodium hyaluronate aqueous solution. The pH was adjusted to 5 with dilute hydrochloric acid. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide were added in sequence and stirred evenly to obtain activated sodium hyaluronate. The molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to the carboxyl group in sodium hyaluronate was 5:1, and the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to N-hydroxysuccinimide was 1:2. (2) The activated hyaluronic acid and adipic dihydrazide were subjected to a cross-linking reaction. The molar ratio of the carboxyl groups in adipic dihydrazide to sodium hyaluronate was 1:1. The reaction was carried out at room temperature, in the dark, and stirred for 16 hours. Water was added and stirred for 20 minutes to terminate the reaction and obtain the reaction solution. The reaction solution was then dialyzed for 7 days to remove unreacted substances or byproducts. After centrifugation, sodium hyaluronate modified with dihydrazide was obtained.

[0050] The preparation method of the composition in this embodiment is as follows: S1. Sodium hyaluronate with a molecular weight of 100kDa-300kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 30 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 30 minutes to obtain the composition.

[0051] Example 5 A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 70 parts bioactive glass, 9 parts dihydrazide-modified sodium hyaluronate, 10 parts sodium hyaluronate with a molecular weight of 100kDa-300kDa, 20 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40) and 8 parts carboxymethyl cellulose.

[0052] The preparation method of the dihydrazide-modified sodium hyaluronate is as follows: (1) Sodium hyaluronate with a molecular weight of 500kDa-1000kDa was prepared into a 10% sodium hyaluronate aqueous solution. The pH was adjusted to 6 with dilute hydrochloric acid. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide were added in sequence and stirred evenly to obtain activated sodium hyaluronate. The molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to the carboxyl group in sodium hyaluronate was 2:1, and the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to N-hydroxysuccinimide was 1:1. (2) The activated hyaluronic acid and adipic dihydrazide were subjected to a cross-linking reaction. The molar ratio of adipic dihydrazide to the carboxyl group in sodium hyaluronate was 3:1. The reaction was carried out at room temperature, in the dark, and stirred for 8 hours. Water was added and stirred for 40 minutes to terminate the reaction and obtain the reaction solution. The reaction solution was then dialyzed for 7 days to remove unreacted substances or byproducts. After centrifugation, sodium hyaluronate modified with dihydrazide was obtained.

[0053] The preparation method of the composition in this embodiment is as follows: S1. Sodium hyaluronate with a molecular weight of 100kDa-300kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 40 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 40 minutes to obtain the composition.

[0054] Comparative Example 1 Unlike Example 1, sodium hyaluronate with a molecular weight of 100kDa-300kDa was replaced with collagen.

[0055] A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 55 parts bioactive glass, 7 parts dihydrazide-modified sodium hyaluronate, 7 parts collagen, 15 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40) and 6 parts carboxymethyl cellulose.

[0056] The preparation method of dihydrazide-modified sodium hyaluronate is the same as in Example 1; The method for preparing the composition is as follows: S1. Mix collagen, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer, and stir at room temperature for 35 min to obtain mixture 1; S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0057] Comparative Example 2 Unlike Example 1, the raw material component sodium hyaluronate with a molecular weight of 100kDa-300kDa was replaced with sodium hyaluronate with a low molecular weight of 5kDa-30kDa.

[0058] A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 55 parts bioactive glass, 7 parts dihydrazide-modified sodium hyaluronate, 7 parts sodium hyaluronate with a molecular weight of 5kDa-30kDa, 15 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40), and 6 parts carboxymethyl cellulose.

[0059] The preparation method of dihydrazide-modified sodium hyaluronate is the same as in Example 1.

[0060] The method for preparing the composition is as follows: S1. Sodium hyaluronate with a molecular weight of 5kDa-30kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 35 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0061] Comparative Example 3 Unlike Example 1, the polylactic acid-glycolic acid copolymer was replaced with a polylactic acid-polyvinylamine mixture in a molar ratio of 60:40.

[0062] A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 55 parts bioactive glass, 7 parts dihydrazide-modified sodium hyaluronate, 7 parts sodium hyaluronate with a molecular weight of 100kDa-300kDa, 15 parts polylactic acid-polyvinylamine mixture (molar ratio of polylactic acid to polyvinylamine is 60:40) and 6 parts carboxymethyl cellulose.

[0063] Everything else is the same as in Example 1.

[0064] Comparative Example 4 Unlike Example 1, carboxymethyl cellulose was replaced with chitosan.

[0065] The preparation method of dihydrazide-modified sodium hyaluronate is the same as in Example 1.

[0066] The method for preparing the composition is as follows: S1. Sodium hyaluronate with a molecular weight of 100kDa-300kDa, chitosan and polylactic acid-glycolic acid copolymer are mixed and stirred at room temperature for 35 minutes to obtain mixture 1. S2, Mixture 1, and dihydrazide-modified sodium hyaluronate and bioactive glass are mixed and stirred at room temperature for 35 minutes to obtain the composition.

[0067] Comparative Example 5 Unlike Example 1, the weight parts of each component in the composition are different.

[0068] A composition for modifying the plasticity of bone meal particles, comprising the following raw materials by weight: 35 parts bioactive glass, 2 parts dihydrazide-modified sodium hyaluronate, 12 parts sodium hyaluronate with a molecular weight of 100kDa-300kDa, 6 parts polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40) and 15 parts carboxymethyl cellulose.

[0069] Everything else is the same as in Example 1.

[0070] Comparative Example 6 Unlike Example 1, the preparation method of dihydrazide-modified sodium hyaluronate is different, specifically the preparation order is different, and the auxiliary activator N-hydroxysuccinimide is not used: (1) Prepare an 8% sodium hyaluronate aqueous solution with a molecular weight of 500kDa-1000kDa, adjust the pH to 5.5 with dilute hydrochloric acid, add adipic acid dihydrazide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride for cross-linking reaction, and stir at room temperature, away from light for 12 hours. The molar ratio of carboxyl groups in the adipate dihydrazide to sodium hyaluronate is 2:1; the molar ratio of carboxyl groups in the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to sodium hyaluronate is 4:1. (2) Add water and stir for 30 minutes to terminate the reaction and obtain the reaction solution; then dialyze the reaction solution for 7 days to remove unreacted substances or byproducts, centrifuge to obtain dihydrazide-modified sodium hyaluronate.

[0071] The composition was prepared using the dihydrazide-modified sodium hyaluronate prepared above, and the preparation method and formulation of the composition were the same as in Example 1.

[0072] Comparative Example 7 The repair material was prepared using the raw materials described in Example 1 of CN115337457A. (1) Add the raw materials calcium carbonate, sodium carbonate, phosphorus pentoxide and silicon dioxide in a mass percentage ratio of 25%:24%:6%:45%, mix them evenly and put them into a crucible, heat to 1200℃ to form a homogenized low-viscosity glass liquid, and then cool to 750℃ to obtain the glass liquid. (2) Remove the connective tissue from the surface of the bovine cancellous bone, cut it into strips, wash it thoroughly and dry it, load it into a stainless steel mesh basket with a handle, immerse it fully in the above glass liquid, vibrate it at a frequency of 5 times / second and a vibration amplitude of 5 cm for 30 min; then take it out, let it stand for 30 s, quench it in water to cool it, dry it and grind it into bone particles with a particle size of 0.1-20 μm; (3) After the 0.3 g / mL bovine Achilles tendon collagen gel aqueous solution and the 0.035 g / mL sodium hyaluronate (molecular weight 100kDa-300kDa) gel aqueous solution are fully dispersed, they are mixed evenly at a volume ratio of 1:1 to obtain a mixed solution; 100 mL of the mixed solution is taken, 50 g of bone particles are added and stirred evenly, and then freeze-dried to obtain a plastic composite active biological bone collagen repair material.

[0073] The comparative example composition was prepared as follows: 0.3 g / mL of bovine Achilles tendon collagen gel aqueous solution and 0.035 g / mL of sodium hyaluronate (molecular weight 100 kDa-300 kDa) gel aqueous solution were fully dispersed and then mixed evenly at a volume ratio of 1:1 to obtain a mixed solution; 100 mL of the mixed solution was taken, 10 g of glass melt was added and stirred evenly, and then freeze-dried in a mold to obtain the composition of comparative example 7.

[0074] Test 1 Compressive Strength Test The compositions of each embodiment and comparative example were filled into a cubic mold of the same size to ensure that the finished product dimensions were consistent (side length of 10mm) to obtain samples of each embodiment and comparative example; 10 samples from each group were taken for compressive strength testing, and the average value of the test results was taken.

[0075] The method for testing compressive strength is as follows: Place the sample on a mechanical strength tester. All samples are placed in the same direction and have the same contact area. Measure and record the loading force at the moment of fracture, and calculate the compressive strength of each sample. The results of the compressive strength are shown in Table 1.

[0076] Compressive strength σ = P / A; where σ is the compressive strength in MPa; P is the maximum failure load in N; and A is the cross-sectional area of ​​the specimen in mm. 2 .

[0077] Table 1

[0078] In Table 1, #This indicates that each comparative example group had a significant difference compared to the Example 1 group. # P < 0.05.

[0079] As can be seen from Table 1, the compositions of the embodiments of the present invention, under specific conditions of interaction between two specific molecular weights of sodium hyaluronate, composite polylactic acid-glycolic acid copolymer, and carboxymethyl cellulose, exhibit significantly improved mechanical strength. Furthermore, the dihydrazide-modified sodium hyaluronate in the compositions of the present invention, compared to the dihydrazide-modified sodium hyaluronate prepared by different modification methods described in Comparative Example 6, results in compositions with superior mechanical properties. This indicates that dihydrazide-modified sodium hyaluronate, as an excipient for bone materials, provides mechanical support, and, in combination with medium-to-high molecular weight sodium hyaluronate and carboxymethyl cellulose, enhances the mechanical properties of the composition.

[0080] Experiment 2 Stability The compositions of Examples 1-5 and Comparative Examples 1-6 were mixed with bone meal granules (bovine bone granules) at a mass ratio of 1:0.5, and an equal mass of water was added to mix and stir evenly. The mixtures were then placed in a mold and freeze-dried to obtain bone graft materials. The bone collagen repair material of Comparative Example 7 was prepared according to the method in Comparative Example 7.

[0081] Mix 2g of the above-mentioned bone graft material with 3mL of fresh rabbit blood and knead to form a ball (controlling the base area to 100mm²). 2 ) Observe the formability and viscoelasticity; then soak in rabbit blood for 24 hours, observe the shape, and evaluate whether the material easily disperses in blood by measuring the bottom area after 24 hours.

[0082] Mix 2g of collagen repair material from Comparative Example 7 with 3mL of fresh rabbit blood and knead to form a ball (controlling the base area to 100mm²). 2 ) Observe the formability and viscoelasticity; then soak in rabbit blood for 24 hours, observe the shape, and evaluate whether the material easily disperses in blood by measuring the bottom area after 24 hours.

[0083] The viscoelastic properties, shape, and bottom area after 24 hours of storage for each embodiment and comparative example are shown in Table 2.

[0084] Table 2

[0085] In Table 2, # and ## This indicates that each comparative example group had a significant difference compared to the Example 1 group. # P < 0.05 ## P < 0.01.

[0086] As shown in Table 2, the composition of the present invention, in which bioactive glass, dihydrazide-modified sodium hyaluronate, sodium hyaluronate with a molecular weight of 50kDa-500kDa, polylactic acid-glycolic acid copolymer (molar ratio of lactic acid to glycolic acid is 60:40), and carboxymethyl cellulose interact and are indispensable, not only has high mechanical properties under specific types and ratios, but also significantly improved plasticity. In contrast, the compositions of the comparative examples all showed varying degrees of scattering and collapse in rabbit blood environment over time, with a significant decrease in viscoelasticity and lower formability than the embodiments of the present invention.

[0087] Experiment 3: Detection of cartilage repair rate To verify the repair effect of the composition prepared in this invention on bone defects, the compositions prepared in the above examples and comparative examples were mixed with bone particles to obtain bone graft materials, and animal experiments were conducted, specifically as follows: Test samples: The compositions of Examples 1-5 and Comparative Examples 1-6 were mixed with bone meal particles (bovine bone particles) at a mass ratio of 1:0.5, and an equal mass of water was added and stirred evenly. The mixture was then placed in a mold and freeze-dried to obtain bone graft material. The repair material prepared in step (3) of Comparative Example 7 (bone graft material of Comparative Example 7).

[0088] New Zealand white rabbits were general anesthetized, and a cartilage defect (3.0 mm in diameter and 5.0 mm in depth) was created in the trochlear groove of the rabbit's right leg using a dental grinder. Bone graft material of the same size as the defect was then implanted into the defect. The control group did not receive bone graft material. Five rabbits were used in each experimental group. The rabbits were euthanized 12 weeks post-surgery, and tissue repair was observed to calculate the cartilage repair rate.

[0089] The results of animal bone cartilage repair rates are shown in Table 3.

[0090] Table 3

[0091] In Table 3, # and ## This indicates that each comparative example group had a significant difference compared to the Example 1 group. # P < 0.05 ## P < 0.01.

[0092] As shown in Table 3, the compositions and bone graft materials prepared in the embodiments of the present invention have a cartilage repair rate of 91%-96%, which is significantly better than the repair rate of the comparative examples (69%-81%). This indicates that using the compositions of the present invention, the defects in animal bone and cartilage are uniformly filled with cartilage tissue, and the newly formed tissue has good connectivity with the surrounding normal cartilage. The compositions and bone graft materials of the present invention have a good repair effect on bone defects.

[0093] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A composition for changing the plasticity of bone dust particles, characterized by, The raw material components include, in parts by weight: 40-70 parts of bioactive glass, 5-9 parts of sodium hyaluronate modified by dihydrazide, 5-10 parts of sodium hyaluronate with a molecular weight of 50 kDa-500 kDa, 8-20 parts of polylactic acid-glycolic acid copolymer, and 4-8 parts of carboxymethyl cellulose.

2. The composition of claim 1, wherein, The raw material components include, in parts by weight: 50-60 parts of bioactive glass, 6-8 parts of sodium hyaluronate modified by dihydrazide, 6-8 parts of sodium hyaluronate with a molecular weight of 50 kDa-500 kDa, 10-16 parts of polylactic acid-glycolic acid copolymer, and 5-7 parts of carboxymethyl cellulose.

3. The composition of claim 2, wherein, The raw material components include, in parts by weight: 55 parts of bioactive glass, 7 parts of sodium hyaluronate modified by dihydrazide, 7 parts of sodium hyaluronate with a molecular weight of 50 kDa-500 kDa, 15 parts of polylactic acid-glycolic acid copolymer, and 6 parts of carboxymethyl cellulose.

4. The composition according to any one of claims 1 to 3, characterized in that, The preparation method of the sodium hyaluronate modified by dihydrazide comprises the steps of: (1) adjusting the pH and activating the carboxyl group of sodium hyaluronate to obtain activated sodium hyaluronate; (2) crosslinking the activated sodium hyaluronate and adipic acid dihydrazide, and then performing termination reaction to obtain sodium hyaluronate modified by dihydrazide.

5. The composition of claim 4, wherein, In step (1) of the preparation method of the sodium hyaluronate modified by dihydrazide, the sodium hyaluronate has a molecular weight of 50 kDa-2000 kDa; preferably, in step (1), the sodium hyaluronate has a molecular weight of 500 kDa-1000 kDa. In step (1), the pH is adjusted to 5-6. In step (1), the sodium hyaluronate solution is prepared into a sodium hyaluronate solution with a mass concentration of 5%-10%, and after the pH is adjusted, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide are added, and stirring is performed until uniformity is achieved, to obtain activated sodium hyaluronate; the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to the carboxyl group in the sodium hyaluronate is 2-5:

1. In step (1), the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to N-hydroxysuccinimide is 1:1-2.

6. The composition of claim 4, wherein, In step (2) of the preparation method of the sodium hyaluronate modified by dihydrazide, the molar ratio of adipic acid dihydrazide to the carboxyl group in the sodium hyaluronate is 1-3:1; preferably, in step (2), the molar ratio of adipic acid dihydrazide to the carboxyl group in the sodium hyaluronate is 1.5-2.5:

1. In step (2), the crosslinking reaction is specifically performed at room temperature, in the dark, and with stirring for 8-16 h. In step (2), the termination reaction is specifically performed by adding water and stirring for 20-40 min for termination treatment to obtain a reaction solution; then the reaction solution is dialyzed for 3-7 days to remove unreacted substances or byproducts, centrifugation is performed, and sodium hyaluronate modified by dihydrazide is obtained.

7. The composition according to any one of claims 1 to 3, characterized in that, In the polylactic acid-glycolic acid copolymer, the molar ratio of lactic acid to glycolic acid is 50-65:35-50; preferably, the molar ratio of lactic acid to glycolic acid is 60:

40.

8. Process for the preparation of a composition according to any one of claims 1 to 7, characterized in that, The method comprises the steps of: S1, mixing sodium hyaluronate with molecular weight of 50kDa-500kDa, carboxymethyl cellulose and polylactic acid-glycolic acid copolymer to obtain mixture 1; S2, mixing mixture 1 with bioactive glass and dihydrazide modified sodium hyaluronate to obtain the composition.

9. Use of the composition of any one of claims 1-7 in the preparation of bone graft material.

10. A bone graft material, characterized by, The bone graft material comprises: artificial bone material and the composition of any one of claims 1-7.

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