Zinc-doped high-toughness injectable organic-inorganic composite bioactive material and preparation method thereof

Abstract

The invention discloses a zinc-doped high-toughness injectable organic-inorganic composite bioactive material and a preparation method thereof.The composite bioactive material comprises a water aqua and powder, the water aqua comprises water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid and deionized water, and the powder comprises water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid and deionized water. The powder comprises calcium hydrophosphate, tetracalcium phosphate and a zinc source. The zinc element is added, so that proliferation, differentiation and the like of osteoblasts are remarkably promoted, unexpected osteogenic activity is shown, all excellent mechanical properties in an existing bioactive material are completely reserved, and the properties include but not limited to coagulation time, compressive strength, toughness, injectability and the like; furthermore, the addition of zinc has no negative effects.

Description

A zinc-doped, highly tough, injectable organic-inorganic composite bioactive material and its preparation method Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a zinc-doped, highly tough, injectable organic-inorganic composite bioactive material and its preparation method. Background Technology

[0002] Bone cement is a type of bone filling material that can be injected into the human body and solidify and shape in situ. Clinically, it is mainly used in vertebral body repair surgery and to fill large bone defects. It has a wide range of clinical needs and huge market value. During its development, bone cement has formed a variety of systems, among which the two most important systems are polymethyl methacrylate (PMMA) bone cement and calcium phosphate cement (CPC).

[0003] The existing patent, "Organic-Inorganic Composite Bioactive Material Based on Calcium Phosphate and its Preparation Method" (ZL202111220731.3, hereinafter referred to as the calcium phosphate biomaterial patent), provides a high-performance bioactive material formulation. This formulation combines calcium phosphate nanoclusters, an organic polymer calcium stabilizer, and an inorganic small molecule calcium crosslinking agent to form an organic-inorganic crosslinked bioactive material. It effectively solves the problem of insufficient mechanical strength and stability of CPC in a liquid environment, allowing it to retain not only the good biocompatibility of CPC, but also self-curing properties with minimal heat release during curing, and malleability before complete curing, as well as good toughness and long-term mechanical stability, making it an excellent bone repair material. However, this patent does not modify the osteogenic activity of CPC.

[0004] Therefore, existing technologies regarding the osteogenic activity of CPC still need improvement and development. Summary of the Invention

[0005] The purpose of this invention is to provide a zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material, which solves the problem of poor osteogenic activity of existing organic-inorganic composite bioactive materials in CPC.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: a zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material, comprising an aqueous solution and a powder, wherein the aqueous solution comprises water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water, and the powder comprises dicalcium phosphate, tetracalcium phosphate, and a zinc source.

[0007] Preferably, the amount of zinc source added is 0.1% to 10% of the total mass of the powder.

[0008] Preferably, the amount of zinc source added is 0.1% to 4.0% of the total mass of the powder.

[0009] Preferably, the mass ratio of dicalcium phosphate to tetracalcium phosphate in the powder is 1:(2~5).

[0010] Preferably, the zinc source is zinc sulfate.

[0011] Preferably, the volume ratio of the aqueous solution to the mass ratio of the powder is 0.65~35mg of powder per 1μl of aqueous solution; the mass percentage of polyvinylpyrrolidone in the aqueous solution is 1.2%~20%; the mass percentage of citric acid in the aqueous solution is 0.5%~20%; the water-soluble calcium salt is calcium chloride; the concentration of water-soluble calcium chloride in the aqueous solution is not greater than 0.1mol / L; and the molar ratio of Ca in the water-soluble calcium salt to P in the water-soluble phosphate is 0.5~1.5:1.

[0012] Preferably, the water-soluble phosphate is one or more of orthophosphate, monohydrogen phosphate, and dihydrogen phosphate.

[0013] Preferably, the mass ratio of the water-soluble calcium salt to the polyaspartic acid is 1:1 to 10.

[0014] Preferably, the mass ratio of the water-soluble phosphate to the polyacrylic acid is 1:1 to 10.

[0015] Another technical solution of the present invention is implemented as follows: a method for preparing the above-mentioned zinc-doped high-toughness injectable organic-inorganic composite bioactive material, the preparation method comprising: mixing dicalcium phosphate, tetracalcium phosphate and a zinc source to obtain a powder; mixing water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid and deionized water to obtain an aqueous solution; mixing the powder and the aqueous solution, stirring, and allowing to stand for curing to obtain the zinc-doped high-toughness injectable organic-inorganic composite bioactive material.

[0016] Compared with existing technologies, this invention, by adding zinc, not only significantly promotes osteoblast proliferation and differentiation, but also exhibits unexpected osteogenic activity. Simultaneously, it fully retains all the excellent mechanical properties of existing bioactive materials, including but not limited to solidification time, compressive strength, toughness, and injectability, thus confirming that the addition of zinc has no negative effects. Furthermore, from a safety and economic perspective, this invention avoids the use of expensive growth factors, achieving activation using safe and reliable zinc, resulting in low cost, ease of industrialization, and broad market prospects. Attached Figure Description

[0017] Figure 1 is a schematic diagram of the cured state of the injectable organic-inorganic composite bioactive materials obtained by adding zinc sulfate of different concentrations in an embodiment of the present invention; Figure 2 is a schematic diagram of the alkaline phosphatase activity test results of the different injectable organic-inorganic composite bioactive materials obtained in this embodiment; Figure 3 is a schematic diagram of the water absorption performance of the injectable organic-inorganic composite bioactive material obtained in Example 3; Figure 4 is a schematic diagram of the stress-strain curve of the injectable organic-inorganic composite bioactive material obtained in Example 3. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0019] This invention provides a zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material, comprising an aqueous solution and a powder. The aqueous solution includes water-soluble calcium salts, water-soluble phosphates, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water. The powder comprises dicalcium phosphate, tetracalcium phosphate, and a zinc source.

[0020] In this embodiment, the amount of zinc source added is 0.1% to 10% of the total mass of the solid powder; preferably 0.1% to 4.0%; the mass ratio of dicalcium phosphate to tetracalcium phosphate in the powder is 1:(2~5); the zinc source is zinc sulfate; the ratio between the volume of the aqueous solution and the mass of the powder is 0.1% to 4.0% per 1 μl of aqueous solution. The aqueous solution contains 0.65~35mg powder; the mass percentage of polyvinylpyrrolidone in the aqueous solution is 1.2%~20%; the mass percentage of citric acid in the aqueous solution is 0.5%~20%; the water-soluble calcium salt is calcium chloride; the concentration of water-soluble calcium chloride in the aqueous solution is not greater than 0.1mol / L; the water-soluble phosphate is one or more of orthophosphate, monohydrogen phosphate, and dihydrogen phosphate; the molar ratio of Ca in the water-soluble calcium salt to P in the water-soluble phosphate is 0.5~1.5:1; the mass ratio of the water-soluble calcium salt to the polyaspartic acid is 1:1~10; the mass ratio of the water-soluble phosphate to the polyacrylic acid is 1:1~10.

[0021] This invention also provides a method for preparing the above-mentioned zinc-doped high-toughness injectable organic-inorganic composite bioactive material. The preparation method includes: mixing dicalcium phosphate, tetracalcium phosphate, and a zinc source to obtain a powder; mixing water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water to obtain an aqueous solution; mixing the powder and the aqueous solution, stirring, and allowing to stand for curing to obtain the zinc-doped high-toughness injectable organic-inorganic composite bioactive material.

[0022] This invention, by incorporating zinc, not only significantly promotes osteoblast proliferation and differentiation but also exhibits unexpected osteogenic activity. Simultaneously, it fully retains all the excellent mechanical properties of existing bioactive materials, including but not limited to solidification time, compressive strength, toughness, and injectability, thus confirming that the addition of zinc has no negative effects. Furthermore, from a safety and economic perspective, this invention avoids the use of expensive growth factors, achieving activation solely through safe and reliable zinc, resulting in low cost, ease of industrialization, and broad market prospects.

[0023] The present invention will be further explained and illustrated below through specific embodiments. Embodiment 1

[0024] The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material provided in Example 1 is obtained through the following steps: (1) 100 mg of dicalcium phosphate, 300 mg of tetracalcium phosphate, and 9 mg of zinc sulfate are mixed to obtain 409 mg of powder; (2) water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water are mixed to obtain 400 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2%, and the mass percentage of citric acid is 2%; (3) the powder and the aqueous solution are mixed, stirred, and allowed to stand for curing to obtain the organic-inorganic composite bioactive material (see the morphology corresponding to concentration 1 in Figure 1 for details). Example 2

[0025] The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material provided in Example 2 is obtained through the following steps: (1) 100 mg of dicalcium phosphate, 300 mg of tetracalcium phosphate, and 12.5 mg of zinc sulfate are mixed to obtain 412.5 mg of powder; (2) water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water are mixed to obtain 400 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2%, and the mass percentage of citric acid is 2%; (3) the powder and the aqueous solution are mixed, stirred, and allowed to stand to solidify to obtain the organic-inorganic composite bioactive material (see the morphology corresponding to concentration 2 in Figure 1 for details). Example 3

[0026] The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material provided in Example 3 is obtained through the following steps: (1) 100 mg of dicalcium phosphate, 300 mg of tetracalcium phosphate, and 4 mg of zinc sulfate are mixed to obtain 404 mg of powder; (2) water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water are mixed to obtain 400 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2%, and the mass percentage of citric acid is 2%; (3) the powder and the aqueous solution are mixed, stirred, and allowed to stand to solidify to obtain the organic-inorganic composite bioactive material (see the morphology corresponding to concentration 3 in Figure 1 for details). Example 4

[0027] The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material provided in Example 4 is obtained through the following steps: (1) 100 mg of dicalcium phosphate, 300 mg of tetracalcium phosphate, and 15 mg of zinc sulfate are mixed to obtain 415 mg of powder; (2) water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water are mixed to obtain 400 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2%, and the mass percentage of citric acid is 2%; (3) the powder and the aqueous solution are mixed, stirred, and allowed to stand for curing to obtain the organic-inorganic composite bioactive material (see the morphology corresponding to concentration 5 in Figure 1 for details). Example 5

[0028] The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material provided in Example 5 is obtained through the following steps: (1) 100 mg of dicalcium phosphate, 300 mg of tetracalcium phosphate, and 9 mg of zinc sulfate are mixed to obtain 409 mg of powder; (2) water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water are mixed to obtain 380 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2.2%, and the mass percentage of citric acid is 2.2%; (3) the powder and the aqueous solution are mixed, stirred, and allowed to stand for curing to obtain the organic-inorganic composite bioactive material. Comparative Example

[0029] (1) Mix 100 mg of dicalcium phosphate and 300 mg of tetracalcium phosphate to obtain 400 mg of powder; (2) Mix water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid and deionized water to obtain 400 μl of aqueous solution; wherein, the mass percentage of polyvinylpyrrolidone is 2% and the mass percentage of citric acid is 2%; (3) Mix the powder and the aqueous solution, stir, and let stand to solidify to obtain organic-inorganic composite bioactive material.

[0030] Analysis of Figure 1 shows that bioactive materials were produced by adding different concentrations of zinc sulfate to the powder and then mixing it with an aqueous solution.

[0031] To verify the performance of the zinc-doped high-toughness injectable organic-inorganic composite bioactive material obtained in the embodiments of the present invention, the zinc-doped high-toughness injectable organic-inorganic composite bioactive materials obtained in Examples 1 and 2, as well as the organic-inorganic composite bioactive material obtained in the comparative example, were tested. The specific testing method is as follows: Referring to Figure 2, the control group, the comparative example, and the zinc-doped high-toughness injectable organic-inorganic composite bioactive materials prepared in Examples 1 and 2 of the present invention were respectively immersed in the extraction solution. After immersion for 24 hours, human bone marrow mesenchymal stem cells (hMSCs) were cultured using the extraction solution. On the 3rd and 7th days of culture, the cells were lysed and the cell lysate was extracted as the alkaline phosphatase detection sample. According to the requirements of the alkaline phosphatase detection kit, the sample to be tested, buffer solution, and matrix solution were added to the 96-well microplate in sequence, incubated at 37°C for 15 minutes, and then the colorimetric solution was added for color development. Alkaline phosphatase breaks down disodium phenyl phosphate, producing free phenol and phosphoric acid. The phenol reacts with 4-aminoantipyrrolidone in an alkaline solution and is oxidized by potassium ferricyanide to form a red quinone derivative. The intensity of the red color indicates the enzyme activity. The values ​​were measured using a microplate reader and calculated; the specific results are shown in Figure 2.

[0032] As shown in Figure 2, in in vitro cell experiments, the composite bioactive material obtained by this invention can significantly enhance the alkaline phosphatase activity of human bone marrow mesenchymal stem cells on day 7, thereby promoting the proliferation and differentiation of osteoblasts and exhibiting unexpected osteogenic activity.

[0033] As shown in Figure 3, the composite bioactive material obtained by the present invention has excellent water absorption capacity.

[0034] As shown in Figure 4, the composite bioactive material obtained by the present invention has excellent mechanical properties. It did not break when compressed to 50% strain, and its compressive strength increased with the increase of the compression distance, showing good fracture resistance and good toughness.

[0035] In summary, this invention, by adding zinc, not only enhances bioactivity but also fully retains all the excellent mechanical properties of existing bioactive materials. These properties include, but are not limited to, solidification time, compressive strength, toughness, and injectability, thus proving that the addition of zinc has no negative effects. Furthermore, from a safety and economic perspective, this invention avoids the use of expensive growth factors, achieving activation using safe and reliable zinc, resulting in low cost, ease of industrialization, and broad market prospects.

[0036] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material, characterized in that, The product includes aqueous solutions and powders. The aqueous solution includes water-soluble calcium salts, water-soluble phosphates, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid, and deionized water. The powder includes dicalcium phosphate, tetracalcium phosphate, and a zinc source.

2. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 1, characterized in that, The amount of zinc source added is 0.1% to 10% of the total mass of the powder.

3. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 2, characterized in that, The amount of zinc source added is 0.1% to 4.0% of the total mass of the powder.

4. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 3, characterized in that, The mass ratio of dicalcium phosphate to tetracalcium phosphate in the powder is 1:(2~5).

5. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to any one of claims 1-4, characterized in that, The zinc source is zinc sulfate.

6. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 5, characterized in that, The volume ratio of the aqueous solution to the mass of the powder is 0.65~35mg of powder per 1μl of aqueous solution; the mass percentage of polyvinylpyrrolidone in the aqueous solution is 1.2%~20%; the mass percentage of citric acid in the aqueous solution is 0.5%~20%; the water-soluble calcium salt is calcium chloride; the concentration of water-soluble calcium chloride in the aqueous solution is not greater than 0.1mol / L; the molar ratio of Ca in the water-soluble calcium salt to P in the water-soluble phosphate is 0.5~1.5:

1.

7. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 6, characterized in that, The water-soluble phosphate is one or more of orthophosphate, monohydrogen phosphate, and dihydrogen phosphate.

8. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 6, characterized in that, The mass ratio of the water-soluble calcium salt to the polyaspartic acid is 1:1 to 10.

9. The zinc-doped, high-toughness, injectable organic-inorganic composite bioactive material according to claim 6, characterized in that, The mass ratio of the water-soluble phosphate to the polyacrylic acid is 1:1 to 10.

10. A method for preparing a zinc-doped, highly tough, injectable organic-inorganic composite bioactive material as described in any one of claims 1 to 9, characterized in that, The preparation method includes: mixing dicalcium phosphate, tetracalcium phosphate and a zinc source to obtain a powder; mixing water-soluble calcium salt, water-soluble phosphate, polyacrylic acid, polyaspartic acid, polyvinylpyrrolidone, citric acid and deionized water to obtain an aqueous solution; mixing the powder and the aqueous solution, stirring, and allowing to stand and solidify to obtain a zinc-doped high-toughness injectable organic-inorganic composite bioactive material.

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