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A kind of mbmp2 protein composite hydrogel and preparation method and application thereof

A composite hydrogel, T4L-BMP2 technology, applied in the field of biomedical materials, can solve the problems such as the inability to achieve sustained release of rH-BMP2, the inability to fill the defect site well, and the short half-life of BMP-2 protein. Effects of biological safety, enhanced solubility and stability, strong mechanical strength

Active Publication Date: 2022-07-29
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the most used bioactive factor is recombinant bone morphogenetic protein (BMP-2). However, BMP-2 has the following problems in clinical application: 1) The half-life of BMP-2 protein is short, and it is easily hydrolyzed by enzymes or rapidly diluted by body fluids; 2) The operation of carrier biomaterials is difficult, surgery is required for implantation, and the defect cannot be well filled, which affects bone healing and regeneration; 3) The carrier is expensive, has limited biocompatibility, and degrades too quickly, making BMP -2 fast drain
However, it cannot achieve the sustained release of rH-BMP2, and the therapeutic effect quickly fades away
[0005] Since the repair of bone defects and bone regeneration are still important problems in clinical medicine, the current clinical application of BMP-2 also has the problems of high price and low stability.

Method used

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  • A kind of mbmp2 protein composite hydrogel and preparation method and application thereof
  • A kind of mbmp2 protein composite hydrogel and preparation method and application thereof
  • A kind of mbmp2 protein composite hydrogel and preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example

[0061] In the following preparation examples, examples, comparative examples and effect tests, the DNA fragment of T4L-BMP2 was synthesized by GenScript;

[0062] BMP-2, T4 lysozyme protein, agarose microspheres, type A pigskin gelatin, and ELISA kit were purchased from sigma.

[0063] The pET28a expression vector was purchased from GE Healthcare;

[0064] BL21(DE3) was purchased from GE Healthcare.

preparation example 1

[0066] This preparation example provides an agarose-MBMP2 composite microsphere, and the preparation method is as follows:

[0067] (1) Plasmid construction: The DNA fragment of T4L-BMP2 was subcloned into pET28a expression vector through NdeI and BamHI sites to obtain MBMP2 recombinant plasmid;

[0068] (2) Transformation: Competent cells of BL21 (DE3) were used, and the MBMP2 recombinant plasmid was introduced into them, and then the bacterial liquid was cultured on LB solid plate (containing 100 μg / mL ampicillin) at 37°C overnight, and the transformed monoclonal plate was successful. spare;

[0069] (3) Expanded culture: Pick out small bulge-like bacteria with neat edges, translucent, and smooth surface on the LB plate and transfer them to 5 mL of LB liquid medium, shake them at 37°C for 4 hours, and then change to 800 mL Continue to cultivate in LB liquid medium for about 5 hours. When the OD600 of the bacterial liquid reaches between 0.4 and 0.6, reduce the culture tempe...

preparation example 2

[0079] This preparation example provides an agarose-MBMP2 composite microsphere, and the preparation method is as follows:

[0080] Steps (1)-(10) are consistent with Preparation Example 1;

[0081] (11) Mix the agarose microspheres with MBMP2, the concentration of the agarose microspheres is 50 mg / mL, and the concentration of MBMP2 is 2 mg / mL, to obtain the agarose-MBMP2 composite microspheres, the mixing temperature is 4°C, and the mixing time is 4h.

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Abstract

The present invention provides an MBMP2 protein composite hydrogel and a preparation method and application thereof. The preparation materials of the MBMP2 protein composite hydrogel include gelatin, agarose-MBMP2 composite microspheres and TG enzyme, and the preparation method includes the following Steps: (1) dissolving gelatin in water and adjusting pH to obtain a gelatin solution; (2) mixing the gelatin solution obtained in step (1) with agarose-MBMP2 composite microspheres, and then mixing with TG enzyme solution, and incubating to obtain The MBMP2 protein complex hydrogel. The MBMP2 protein composite hydrogel provided by the invention has good stability, simple preparation process, high biological safety, low cost, easy clinical operation, and can realize the effect of long-term sustained release of MBMP2.

Description

technical field [0001] The invention belongs to the field of biomedical materials, in particular to an MBMP2 protein composite hydrogel and a preparation method and application thereof, in particular to a high stability MBMP2 protein composite hydrogel and its preparation method and application. Background technique [0002] The repair and regeneration of bone defects is still an important problem in clinical medicine. Currently, patients with large bone defects are mainly repaired and healed by means of autologous bone transplantation, allogeneic bone transplantation and tissue engineering material transplantation. In recent years, tissue engineering materials have attracted much attention, which can avoid the problems of immune rejection and insufficient donor sources, but also have the disadvantages of low biological activity and poor osteogenic induction ability. Therefore, effective bone tissue engineering requires the effective combination of scaffold materials and bio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/22A61L27/20A61L27/50A61L27/52A61L27/54C07K14/51C12N15/12C12N15/70
CPCA61L27/222A61L27/20A61L27/227A61L27/50A61L27/52A61L27/54C07K14/51C12N15/70A61L2300/252A61L2300/602A61L2300/412A61L2300/258A61L2300/414A61L2430/02C08L5/12C08L89/00
Inventor 彭松林陈欣谭宝玉唐榕泽杨大志
Owner SHENZHEN PEOPLES HOSPITAL
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