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Self-assembled polypeptide, polypeptide hydrogel and preparation method and application thereof

A hydrogel and self-assembly technology, applied in the field of biomedicine, can solve the problems of delayed wound healing and difficulty in endothelial cell adhesion, and achieve the effects of promoting proliferation and adhesion extension, large clinical transformation prospects, and promoting migration

Active Publication Date: 2021-08-10
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, excessive inflammation in chronic wounds causes damage to the extracellular matrix (ECM), making it difficult for endothelial cells to adhere, migrate, and proliferate, which also delays wound healing.

Method used

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  • Self-assembled polypeptide, polypeptide hydrogel and preparation method and application thereof
  • Self-assembled polypeptide, polypeptide hydrogel and preparation method and application thereof
  • Self-assembled polypeptide, polypeptide hydrogel and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Preparation and Morphological Characterization of Polypeptide Hydrogel

[0055] Preparation of polypeptide hydrogel: weigh 1 mg of polypeptide freeze-dried powder (Fmoc-LFKFFK-NH 2 ) into the container, add 3 μL of dimethyl sulfoxide (DMSO) to completely dissolve the peptide, then add 1 mL of PBS buffer solution with pH=6, and then use a pipette to fully mix the peptide solution to make the peptide in the The solution is uniformly distributed, and finally the mixed polypeptide solution is left to stand for more than 10 minutes to obtain a stable polypeptide hydrogel with a mass fraction of 0.1 wt.%. Such as figure 1 , the inverted vial experiment proved the formation of polypeptide hydrogel. Such as figure 2 , the polypeptide hydrogel prepared by this method is transparent and relatively stable.

[0056] Morphology (TEM) characterization of polypeptide hydrogel: prepare 0.1wt.% polypeptide hydrogel according to the above method, after thixotropy, take out 10 μ L an...

Embodiment 2

[0059] Effects of Hydrogels on Cell Growth Behavior

[0060] Prepare polypeptide hydrogels with mass fractions of 0.15wt.%, 0.2wt.%, and 0.3wt.% according to the hydrogel preparation method in Example 1, that is, weigh 1.5mg, 2mg, and 3mg of polypeptide freeze-dried powders respectively (Fmoc-LFKFFK-NH2) into the EP tube, add 3 μL of dimethyl sulfoxide (DMSO) to completely dissolve the peptide, then add 1 mL of PBS buffer with pH=6, and then use a pipette to fully pipette the peptide solution Mix evenly to make the polypeptide evenly distributed in the solution, and sterilized and stable polypeptide hydrogels with mass fractions of 0.15wt.%, 0.2wt.%, and 0.3wt.% can be obtained respectively. Finally, the amount of 10 μL per well was added to a 96-well plate, and finally the 96-well plate was sterilized overnight under UV irradiation.

[0061] Cytocompatibility analysis of hydrogel: Digest cultured L-929 cells and HUVEC cells respectively, and under sterile conditions after ce...

Embodiment 3

[0067] Characterization of antibacterial and anti-biofilm properties of hydrogel

[0068] Prepare polypeptide hydrogels with mass fractions of 0.15wt.%, 0.2wt.%, and 0.3wt.% according to the hydrogel preparation method in Example 1, that is, weigh 1.5mg, 2mg, and 3mg of polypeptide freeze-dried powders respectively (Fmoc-LFKFFK-NH 2 ) into the EP tube, add 3 μL of dimethyl sulfoxide (DMSO) to dissolve the polypeptide completely, then add 1 mL of PBS buffer solution with pH=6, and then use a pipette to fully mix the polypeptide solution to make the polypeptide in the Evenly distributed in the solution, sterilized and stable polypeptide hydrogels with mass fractions of 0.15wt.%, 0.2wt.%, and 0.3wt.% can be obtained respectively. Finally, the amount of 250 μL per well was respectively added into a 24-well plate, and finally the 24-well plate was sterilized overnight under UV irradiation.

[0069] Plate count characterization of hydrogel antibacterial properties: First, glycerol...

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Abstract

The invention relates to a self-assembled polypeptide. The sequence structure of the polypeptide is as follows: Fmoc-LFKFFK-NH2. The invention further relates to polypeptide hydrogel and a preparation method and application thereof. The polypeptide hydrogel disclosed by the invention has the beneficial effects that the polypeptide hydrogel prepared by the preparation method has relatively good cell compatibility and blood compatibility, and can promote proliferation, adhesion and extension of L-929 cells; migration of HUVEC cells can be effectively promoted, and formation of new blood vessels is promoted through VEGFA and HIF-1alpha signal channels; gram-positive bacteria (MRSA) and gram-negative bacteria (pseudomonas aeruginosa) can be killed in a mode of cell membrane destruction; after the MRSA-infected wound of a diabetic mouse is injected, the MRSA-infected wound can be effectively killed, the healing of the wound and the formation of new blood vessels at the wound are remarkably promoted, and the toxicity to other tissues of the mouse is avoided.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a preparation method and application of a polypeptide hydrogel, in particular to a preparation method and application of a polypeptide hydrogel for promoting the healing of diabetic chronic wounds. Background technique [0002] Chronic wounds have become a serious threat to public healthcare, imposing a huge medical and financial burden worldwide. Chronic wounds, especially diabetic ulcers, are often accompanied by persistent microbial infection, and about 41% of patients with diabetic foot ulcers eventually have to amputate. What's more frightening is that biofilms are more likely to form on the surface of chronic wounds, thereby protecting the internal microorganisms from the penetration and invasion of antibiotics, so as to produce more drug-resistant bacteria. It turns out that this biofilm is present in more than 90 percent of chronic wounds. In addition, extracellular matrix (ECM)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61K9/06A61P17/02A61P3/10A61P31/04A61L26/00
CPCC07K7/06A61K9/06A61K9/0014A61K47/20A61P17/02A61P3/10A61P31/04A61L26/008A61L26/0066A61L26/0047A61L2300/404A61L2300/412A61L2300/252A61K38/00C08L89/00
Inventor 陈超玄起泽王平张琦章苇
Owner EAST CHINA UNIV OF SCI & TECH