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A kind of polypeptide for preparing hydrogel and hydrogel prepared therefrom

A technology of hydrogel and buffer solution, applied in the field of polypeptide and its prepared hydrogel, can solve the problems of difficult to achieve cell regulation, limited wide application of chemical residues, limited gel strength, etc., so as to avoid potential toxicity and immunogens. Effects of Sexual Risk

Active Publication Date: 2019-02-26
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The synthesis of small molecules is relatively easy, but due to the artificial design of its structure and the use of secondary bond forces to form gels, it is difficult to degrade the gel and the strength of the formed gel is limited.
Polymer gels have the characteristics of simple preparation, easy structure control, and considerable strength, but it is difficult to achieve biodegradation; at the same time, it is difficult to achieve the regulation of specific cell functions
Currently used polymer materials such as chitin, sodium alginate, collagen or chemically modified synthetic polymers such as polyethylene glycol, etc., these polymer materials have good support, but the chemical residues in the synthesis , free radical initiators, ultraviolet rays and other issues limit its wide application

Method used

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  • A kind of polypeptide for preparing hydrogel and hydrogel prepared therefrom
  • A kind of polypeptide for preparing hydrogel and hydrogel prepared therefrom
  • A kind of polypeptide for preparing hydrogel and hydrogel prepared therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation method of the present embodiment hydrogel comprises the following steps:

[0043] Peptide Ac-I 3 SLKGQ-NH 2 Under the action of the buffer solution Hepes buffer solution, dilute to a concentration of 8mM, vortex and sonicate, heat in a water bath at 85°C and cool to room temperature, add transglutaminase (TGase) to make the final concentration of TGase The concentration was 0.9U / mL, and incubated at 37°C. After 24 hours and 15 days, hydrogels were able to form. Rheologically tested, such as Figure 4 As shown, Ac-I 3 SLKGQ-NH 2 The storage modulus G' of the polypeptide hydrogel is about 800 Pa, and the loss modulus G" is about 1 / 10 of the storage modulus G', indicating that the system forms a typical hydrogel.

[0044] Hydrogel degradation test: at 37°C, matrix metalloproteinase II (MMP-2) was added to the self-assembled hydrogel to make the final concentration 100ng / mL, and the gel was placed in a 37°C water bath After incubation for 15 days, the...

Embodiment 2

[0046] Ac-I 3 SLKGQ-NH 2 Gel preparation method:

[0047] Select the short peptide Ac-I prepared by the aforementioned solid-phase synthesis method 3 SLKGQ-NH 2Under the action of Hepes buffer, dilute to a concentration of 8mM, after oscillating and sonicating, heat in a water bath at 85°C for 2 hours, naturally cool to room temperature, add dissolved in Hepes buffer containing 5mM CaCl 2 and 2mM dithiothreitol (DTT) transglutaminase, so that the final concentration of the enzyme is 0.9U / mL, and the final concentration of the short peptide is 7.27mM. The resulting solution was incubated in a water bath at 37°C for 24 hours to obtain an inverted self-supporting hydrogel.

[0048] The gel in embodiment 2 is assayed:

[0049] Pipette 20 μL of Ac-I 3 SLKGQ-NH 2 The gel was added dropwise on the cleaned silicon wafer, and then 300 μL of ultrapure water was added quickly, and after standing for 10 seconds for adsorption, the sample was blown dry with nitrogen gas, and scanned...

Embodiment 3

[0052] Characterization of Ac-I by Haake rheometer 3 SLKGQ-NH 2 For the mechanical properties (viscoelasticity) of hydrogel samples, a cone-plate with a diameter of 35mm and a taper of 2° was used as the rotor, and a flat plate with a diameter of 35mm was used as the stator. The sample volume used for each measurement was 400 μL, and the experimental temperature was constant at 25°C. Stress scanning is carried out at a frequency of 1Hz, and the scanning range is from 0.01% to 100%. The linear viscoelastic region of the gel is measured, and the appropriate stress is selected from the linear viscoelastic region for dynamic frequency scanning, and the scanning range is from 0.01Hz to 100Hz. The relationship between the energy modulus G' and the energy loss modulus G".

[0053] Ac-I 3 SLKGQ-NH 2 Dilute to a concentration of 8mM under the action of Hepes buffer, add transglutaminase according to the method of Example 2, and place it in a water bath at 37°C for 24 hours, then tak...

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Abstract

The present invention provides a polypeptide for preparing a hydrogel, the amino acid sequence of which is SEQ ID NO: 1; the polypeptide of the present invention is used for preparing a hydrogel; the prepared hydrogel can be used for preparing a cell culture scaffold. Under physiological conditions, the polypeptide of the present invention is catalyzed by the enzyme of the organism itself to form a hydrogel, which avoids the damage to the tissue caused by external chemical agents, ultraviolet light and the like. And it can be degraded by biological enzymes, realizing the degradation of the gel by endogenous substances in the organism, avoiding the potential toxicity and immunogenicity risks caused by the introduction of exogenous chemical agents; it is an ideal tissue engineering material. Human life and health are of great significance.

Description

technical field [0001] The invention relates to the technical field of preparation of hydrogel, in particular to a polypeptide for preparing hydrogel and the prepared hydrogel. Background technique [0002] The preparation of hydrogels in the prior art can be divided into two categories: macromolecular crosslinking and small molecule self-assembly. The synthesis of small molecules is relatively easy, but because its structure is artificially designed and the use of secondary bond forces to form the gel makes it difficult to degrade the gel and the strength of the formed gel is limited. Polymer gels have the characteristics of simple preparation, easy structure control, and considerable strength, but it is difficult to achieve biodegradation; at the same time, it is difficult to achieve the regulation of specific cell functions. Currently used polymer materials such as chitin, sodium alginate, collagen or chemically modified synthetic polymers such as polyethylene glycol, et...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12N5/00
CPCC07K7/06C12N5/0068C12N2533/50
Inventor 陈翠霞徐海张宇
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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