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Alveolar bone repair material containing bmp-2 and its preparation method and application

A technology of BMP-2 and repair materials, applied in the field of regenerative medicine, can solve problems such as poor mechanical properties, fast degradation, and difficulty in achieving balance

Active Publication Date: 2021-02-26
ZHEJIANG RISING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Natural polymer chitosan material has become an important material for tissue engineering due to its good biocompatibility, degradation characteristics and benign interaction with surrounding tissues, and it is also a good carrier of BMP-2, but it is sometimes used as a bone repair material. The problem of poor mechanical properties and rapid degradation, even if it is combined with other materials, it is difficult to achieve a good balance. In addition, the allergy of chitosan is also a major obstacle in its clinical application

Method used

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  • Alveolar bone repair material containing bmp-2 and its preparation method and application
  • Alveolar bone repair material containing bmp-2 and its preparation method and application
  • Alveolar bone repair material containing bmp-2 and its preparation method and application

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Experimental program
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Effect test

Embodiment 1B

[0036] The preparation of embodiment 1BMP-2

[0037] In order to facilitate the experiment process and quality control, the BMP-2 used in the experiment was prepared by the applicant:

[0038] According to the BMP-2 amino acid and gene sequence in Genbank, the BMP-2 gene sequence suitable for expression in Escherichia coli is constructed, and its nucleotide sequence (containing partial restriction site) is SEQ ID NO.1 in the sequence table; Insert the gene fragment into the pET plasmid vector after digestion, and transform the E. coli host after ligation; screen the positive clones and induce the expression of BMP-2; ferment, disrupt the bacteria, collect the inclusion bodies, lyse the inclusion bodies and refold the protein; anion and cation exchange layer BMP-2 was obtained by analysis and purification, and electrophoresis showed a single band of about 12kDa (consistent with the reported BMP-2), and the amino acid sequence of the product was SEQ ID NO.2 in the sequence listi...

Embodiment 2

[0040] The preparation of embodiment 2 modified chitosan

[0041] Chitosan modified with allyltrimethylammonium chloride:

[0042] Dissolve 3g chitosan in 250ml 5% (volume) acetic acid solution; under argon protection, heat to 90°C, add 6ml 0.08mol / L cerium nitrate, react for 30min; add 9ml 50% (mass) allyltrimethyl Ammonium chloride aqueous solution, reacted for 2h; cooling, ethanol precipitation, washing, suction filtration, and vacuum drying to obtain allyltrimethylammonium chloride modified chitosan.

[0043] Product infrared spectrum detection shows 1415cm -1 (-NH), 1555cm -1 The characteristic absorption peak of (C=O) shows that allyltrimethylammonium chloride has been grafted on chitosan.

[0044]Chitosan modified with acrylamide:

[0045] Dissolve 3g chitosan solution in 250ml 5% (volume) acetic acid solution; under argon protection, heat to 50°C, add 5ml0.06mol / L cerium nitrate, react for 30min; add 8g acrylamide, react for 2h; cool, add Adjust the pH to 10 with ...

Embodiment 3

[0047] Preparation of Example 3 Alveolar Bone Restoration Material and Its Basic Properties

[0048] Alveolar bone repair materials of hydroxyapatite, allyltrimethylammonium chloride or acrylamide modified chitosan, hydroxypropyl methylcellulose, and BMP-2 polypeptide components (mass ratio 5000:2000 : 1500: 300) Store refrigerated at 4°C when not in use. Before use, mix the BMP-2 polypeptide with the calcium matrix evenly, dissolve the modified chitosan and hydroxypropyl methylcellulose in PBS buffer at pH 5 at a mass / volume concentration of 40%, and dissolve the BMP-2 polypeptide The mixture with the calcium base is added to the modified chitosan and hydroxypropyl methylcellulose solution, and mixed evenly to obtain the alveolar bone repair material. After the material is prepared, it needs to be molded and implanted within about 15 minutes.

[0049] The compressive strength of the newly prepared finished material is about 177MPa, the bending strength is about 250MPa, and ...

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Abstract

The application provides an alveolar bone repair material, comprising hydroxyapatite, allyltrimethylammonium chloride or acrylamide-modified chitosan, hydroxypropylmethylcellulose, and BMP-2 polypeptide . The material can be used for alveolar bone restoration and dental implant.

Description

technical field [0001] The invention belongs to the field of biomedical materials containing active protein and the field of regenerative medicine. Specifically, the application provides an alveolar bone repair material comprising hydroxyapatite, allyltrimethylammonium chloride or acrylamide modified Sexual chitosan, hydroxypropyl methylcellulose, and BMP-2 polypeptide. The material can be used for alveolar bone restoration and dental implant. Background technique [0002] Bone morphogenetic protein-2 (bone morphogenetic protein-2, BMP-2) is a key factor for promoting bone formation discovered by Marshall R. Urist in 1965 when conducting osteogenesis experiments. BMP-2 belongs to the TGF-α family, which is secreted by osteoblasts and acts on osteoblasts. BMP-2 is the main signaling molecule for cell differentiation into mineral deposition osteoblasts and plays a role in inducing osteoblast differentiation. . It is expressed during limb growth, endochondral ossification, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/22A61L27/20A61L27/12A61L27/54
CPCA61L27/12A61L27/20A61L27/227A61L27/54A61L2300/252A61L2300/412A61L2430/12C08L89/00C08L5/08C08L1/284
Inventor 曹建新
Owner ZHEJIANG RISING BIOTECH CO LTD
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