Preparation method of functionalized polymer composite bone cement microspheres
By modifying amino acid polymers on polymer composite bone cement microspheres, the problem of slow degradation speed and lack of biological activity of traditional bone cement microspheres is solved, and cell adhesion and improvement of degradation performance are achieved, and new bone formation during bone repair is promoted.
Patent Information
- Application Number
- CN202510233705.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-28
- Publication Date
- 2025-05-23
AI Technical Summary
When traditional bone cement microspheres repair bone tissue in the body, they degrade slowly, easily bleed, lack biological activity and cell adhesion ability, and cannot effectively promote the formation and integration of new bones.
By directly modifying the amino acid polymer to the surface and interior of the polymer composite bone cement microspheres, its biological activity and cell adhesion promotion function are enhanced, and degradation speed and anti-collapse performance are improved.
The cell adhesion-promoting function of polymer composite bone cement microspheres is realized, its degradation and anti-collapse properties are improved, and new bone formation and integration during bone repair is promoted.
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Abstract
Description
Technical Field
[0001] The invention relates to a method for preparing functionalized polymer composite bone cement microspheres. Background Art
[0002] Bone tissue defects are common diseases, which may be caused by trauma, infection and tumors. The traditional clinical treatment method is to use autologous bone and allogeneic bone for repair, but this also brings many problems, such as limited sources and immune rejection, which restrict their application. Therefore, it is particularly important to develop biomaterials with good biocompatibility and promote the formation of new bone tissue.
[0003] Bone cement microspheres are an important research direction in bone cement composites. Bone cement microspheres have good fluidity and can easily fill bone defects of various shapes to ensure that the defect area is fully repaired. They have the characteristics of not having a rejection reaction with surrounding tissues and can be safely used for in vivo repair. The structure of bone cement microspheres can also promote the growth and vascularization of new bone tissue and accelerate the repair process of bone defects. However, their degradation rate is slow, they are easy to disintegrate, lack biological activity and cell adhesion ability, and cannot effectively promote the formation and integration of new bones. To solve these problems, the present invention functionalizes polymer composite bone cement microspheres. Amino acid polymers are directly modified to the surface and interior of polymer composite bone cement microspheres, so that the polymer composite bone cement microspheres have good biological activity and have a cell adhesion promoting function, while improving the degradation rate of bone cement microspheres, anti-disintegration performance, promoting in situ osteogenesis, and providing space for bone tissue to grow inward.
[0004] In view of the above background, the present invention provides a functionalized polymer composite bone cement microsphere, which realizes the functions of the polymer composite bone cement microsphere in bone repair in vivo and cell expansion in vitro. Summary of the invention
[0005] The purpose of the present invention is to provide a method for preparing functionalized polymer composite bone cement microspheres.
[0006] The object of the present invention is achieved through the following scheme: a method for preparing functionalized polymer composite bone cement microspheres, comprising the following steps.
[0007] (1) preparing a polymer solution with a concentration of 0-50% (w / v), adding bone cement powder at a concentration of 0.01%-200% of the mass of the solution to obtain a mixed solution; (2) preparing the mixed solution into polymer composite bone cement microspheres; (3) Modifying the microspheres with cell adhesion promoting amino acid polymers to obtain cell adhesion promoting function;
[0008] In step (1), the polymer is one or more of gelatin, chitosan, polyacrylamide, polyvinyl alcohol, polyethylene glycol, polyacrylic acid, polyhydroxyethyl methacrylate and its derivatives, sodium alginate, hyaluronic acid, collagen, silk fibroin, and agarose.
[0009] In step (1), the bone cement is one or more of polymethyl methacrylate (PMMA) bone cement, calcium phosphate bone cement, calcium sulfate bone cement, hydroxyapatite bone cement, carbonated apatite bone cement, and strontium hydroxyapatite.
[0010] In step (2), the preparation method is one or more of ion gel method, thermal gel method, chemical gel method, electrochemical gel method, droplet generation method, spray drying method, batch emulsification method, microfluidic emulsion method, photolithography method, electrospray method, mechanical crushing method, oil-water mixing method, electrospray method, and centrifugation method.
[0011] In step (3), the amino acid polymer is one or more of α-amino acid, β-amino acid, and γ-amino acid.
[0012] The present invention provides a functionalized polymer composite bone cement microsphere, which is prepared according to any of the above methods.
[0013] The present invention provides an application of functionalized polymer composite bone cement microspheres in bone repair in vivo and cell expansion in vitro.
[0014] The advantages of the present invention are: 1. The polymer composite bone cement microspheres are functionalized with cell adhesion promoting amino acid polymers to make the polymer composite bone cement microspheres have the function of promoting cell adhesion.
[0015] 2. While maintaining the bone repair-promoting ability of bone cement microspheres, improve the degradation and anti-collapse properties of bone cement microspheres.
[0016] 3. After degradation, the microspheres form bones in situ in the bone cement, providing space for the ingrowth of bone tissue and having the potential to promote bone repair. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the morphology of the microspheres. Figure 2 shows the cell adhesion morphology on the microspheres after Live-Dead staining under an inverted microscope at 10x magnification. DETAILED DESCRIPTION
[0017] The following examples are implemented based on the technical solutions of the invention, and provide detailed implementation methods and specific operation processes, but the protection scope of the present invention is not limited to the following examples. Example 1
[0018] A functionalized polymer composite bone cement microsphere is prepared according to the following steps: (1) Dissolve 2 mg of sodium alginate in 100 ul of water to prepare a 2% sodium alginate aqueous solution, and add 50 mg of α-TCP to make a mixed solution; (2) Add 100ul of 15mg / ml calcium chloride to the centrifuge tube, add the mixed solution to the syringe, and then place the syringe in the centrifuge tube. Place the centrifuge tube in a centrifuge and centrifuge at 1200rpm for 10 minutes to produce polymer composite bone cement microspheres; (3) Wash the polymer composite bone cement microspheres three times with normal saline.
[0019] (4) Dissolve 4 mg of DbaYKY-DMCO polymer in 100 ul of Tris-HCl to prepare an amino acid polymer solution. Mix the polymer composite bone cement microspheres with the amino acid polymer solution for 12 hours to prepare functionalized polymer composite bone cement microspheres.
[0020] Spherical polymer composite bone cement microspheres with a diameter of 200-400 microns are prepared, such as Figure 1 The morphology of microspheres under white field is shown. Example 2
[0021] Cell adhesion test with polymer composite bone cement microspheres:
[0022] The cells used in this experiment were preosteoblast MC3T3 cells. The cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 100 µg / mL streptomycin, 100 U / mL penicillin, and 2 mmol L-glutamine in a 37 °C incubator (5% CO). 2 MC3T3 osteoblast precursor cells were cultured in a 95% humidity (10% CO, 20% CO). When the cell coverage in the culture dish reached about 90%, the cells were diluted to 4 × 10 6 cell / mL for later use. The functionalized polymer composite bone cement microspheres in Example 1 were mixed with 1 ml of the diluted cell solution mentioned above. After that, the mixed solution was placed on a shaker and shaken rapidly for 30 minutes, and then placed in a 37°C constant temperature incubator for 30 minutes. The above steps were repeated 3 times. The mixed solution was transferred to a 24-well plate for culture. After 24 hours, the well plate was taken out, the α-MEM culture medium in the well plate was aspirated with a pipette, and then rinsed twice with physiological saline. The adhesion and growth of cells on the surface were observed using a Live-Dead Cell Staining Kit. 30 µL of the prepared live-dead cell staining reagent was added dropwise to the surface of the microspheres and incubated for 10 min. After the incubation, the surface was rinsed twice with physiological saline, and the cell staining was observed directly under an inverted fluorescence microscope.
[0023] After staining and observation, the pre-osteoblast MC3T3 cells evenly covered the microspheres, as Figure 2 shown by the cell adhesion on the microspheres after Live-Dead staining under 10X magnification.
Claims
1. A method for preparing functionalized polymer composite bone cement microspheres, characterized in that: The following steps are involved: (1) preparing a polymer solution with a concentration of 0-50% (w / v), adding bone cement powder at a concentration of 0.01%-200% of the mass of the solution to obtain a mixed solution; (2) preparing the mixed solution into polymer composite bone cement microspheres; (3) The microspheres are modified with cell adhesion promoting amino acid polymers to obtain cell adhesion promoting function.
2. The method for preparing functionalized polymer composite bone cement microspheres according to claim 1, characterized in that: In step (1), the hydrogel is one or more of gelatin, chitosan, polyacrylamide, polyvinyl alcohol, polyethylene glycol, polyacrylic acid, polyhydroxyethyl methacrylate and its derivatives, sodium alginate, hyaluronic acid, collagen, silk fibroin, and agarose.
3. The method for preparing functionalized polymer composite bone cement microspheres according to claim 1, characterized in that: In step (1), the bone cement is one or more of polymethyl methacrylate (PMMA) bone cement, calcium phosphate bone cement, calcium sulfate bone cement, hydroxyapatite bone cement, carbonated apatite bone cement, and strontium hydroxyapatite.
4. The method for preparing functionalized polymer composite bone cement microspheres according to claim 1, characterized in that: In step (2), the preparation method is one or more of ion gel method, thermal gel method, chemical gel method, electrochemical gel method, droplet generation method, spray drying method, batch emulsification method, microfluidic emulsion method, photolithography method, electrospray method, mechanical crushing method, oil-water mixing method, electrospray method, and centrifugation method.
5. The method for preparing functionalized polymer composite bone cement microspheres according to claim 1, characterized in that: In step (3), the amino acid polymer is one or more of α-amino acid, β-amino acid, and γ-amino acid.
6. A functionalized polymer composite bone cement microsphere, characterized in that: Prepared according to any one of claims 1 to 5.
7. An application of the method for preparing functionalized polymer composite bone cement microspheres according to claim 6 in bone repair in vivo and cell expansion in vitro.