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A kind of degradable tissue repair material and preparation method

A tissue repair and raw material technology, applied in medical science, prosthesis, etc., can solve the problems of unsuitable spider silk protein, long degradation time, slow degradation of spider silk protein, etc.

Active Publication Date: 2021-07-06
SUZHOU BAIYUAN GENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the natural spider silk protein degrades very slowly, and usually requires the assistance of microorganisms. For example, making it into a degradable plastic bag and burying it in the soil requires microorganisms to ferment and degrade it for several years. The degradation time is too long, so it is not suitable It is used in materials that require a short degradation time, such as tissue repair materials implanted in the human body. Therefore, for tissue repair materials, natural spider silk proteins are not suitable as raw materials for degradable tissue repair materials

Method used

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  • A kind of degradable tissue repair material and preparation method
  • A kind of degradable tissue repair material and preparation method
  • A kind of degradable tissue repair material and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Artificially synthesized recombinant spidroin gene MaSp1

[0031] The recombinant spidroin protein sequence described in this example is shown in SEQ ID NO.1. According to the recombinant spidroin protein sequence selected above and the principle of using preferred codons in Escherichia coli, a coding sequence consistent with the amino acid sequence of the recombinant spidroin protein was designed. For the nucleotide sequence of the recombinant spidroin protein, see the nucleotide sequence shown in SEQ ID NO.2 in the sequence listing.

Embodiment 2

[0032] Example 2. High expression of the gene MaSp1 of artificially synthesized recombinant spidroin in Escherichia coli

[0033] (1) The MaSp1 gene was synthesized by a gene synthesis company, cloned into the prokaryotic expression vector pET-28a+ containing a strong T7 promoter, and constructed into a recombinant plasmid pET-28a+-MaSp1 containing the artificially synthesized MaSp1 gene;

[0034] (2) Prepare Escherichia coli BL21(DE3) competent cells, and transform the recombinant plasmid pET-28a+-MaSp1 into the host cell BL21(DE3) by heat shock method (heat shock at 42°C for 45 seconds) to obtain the engineered plasmid containing the recombinant plasmid strains;

[0035] (3) Inoculate the engineered strain into the LB culture medium solution, culture it on a shaker at 37°C at 220rpm, when the concentration OD600 of the engineered strain reaches 0.6-0.8, add 0.5mmol / L IPTG, induce expression at 37°C for 6 hours, and then Lysis was carried out, and the resulting cell lysate w...

Embodiment 3

[0036] Embodiment 3 degradable tissue repair material

[0037] This embodiment provides a degradable tissue repair material, including the following raw materials in parts by weight:

[0038]

[0039] This embodiment also provides a method for preparing the above-mentioned degradable tissue repair material, comprising the following steps:

[0040] S1. Weigh 5 g of recombinant spidroin protein and add it to 80 ml of HFIP solvent to dissolve, then add 20 g of chitosan and 40 ml of acetic acid solution with a volume concentration of 70%, heat and stir at a heating temperature of 60° C. to obtain a mixture;

[0041] S2. Adjust the pH value of the mixture A to 6.8, then add 10 g of phytic acid weighed therein, heat and stir, first control the heating temperature to 80°C, keep it for 1 hour, then lower the temperature to 45°C, and keep the constant temperature to obtain B mixture, spare;

[0042] S3. Add 5 g of carbon fibers to the mixture of B, stir, cool down to room temperat...

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Abstract

The degradable tissue repair material provided by the present invention, the raw material of the degradable tissue repair material includes recombinant spidroin protein; the amino acid sequence of the recombinant spidroin protein is shown in SEQ ID NO.1; the recombinant spidroin protein, By inserting a protease cleavage site between two adjacent spidroin protein domain monomers, when the recombinant spidroin protein is used in the human body, the protease cleavage site on the recombinant spidroin protein can be eliminated by the human body. The enzymatic hydrolysis of the recombinant spidroin protein can make the biomaterials made of the recombinant spidroin protein gradually degrade in the human body, so using the above-mentioned recombinant spidroin protein as raw material to prepare tissue repair materials can ensure the mechanical stability of the original spidroin protein. While ensuring performance or biological quality, it can also guarantee its degradability and meet clinical requirements.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to a degradable tissue repair material and a preparation method. Background technique [0002] Tissue engineering technology is a new method for treating tissue and organ defects, which consists of biological scaffolds, cells, growth factors and other elements. Among them, biological scaffolds play an important role in tissue engineering as the physical support for cell growth, proliferation, and new tissue formation. In the medical field, a large number of synthetic materials are needed to repair and treat tissue damage. Specifically, in the direction of bone defect materials, current tissue replacement materials are mainly composed of calcium phosphate materials, hydroxyapatite, etc. However, the inherent shortcomings of these materials limit their applications. Calcium phosphate materials have good bioactivity and osteoconductivity, but their poor mechanical prop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/22A61L27/20A61L27/08A61L27/58
CPCA61L27/08A61L27/20A61L27/227A61L27/58
Inventor 李琳李春车锐
Owner SUZHOU BAIYUAN GENT CO LTD
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