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Functional self-assembling nano peptide hydrogel

A Self-Assembling, Versatile Technology

Inactive Publication Date: 2015-02-18
孙念峰 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although RADA16-I / KLT has the function of simulating VEGF fragments, as a tissue engineering material, its function in cell adhesion is relatively limited, which makes its efficiency low in practical applications; although RADA16-I / RGD can Promotes cell adhesion, but does not mimic the specific function of VEGF fragments for vascular endothelial cells

Method used

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Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1, design and synthesis of polypeptide

[0034] 1.1. The sequence of ion-complementary self-assembled nanopolypeptide RADA16-I: Ac-(RADA) 4 -NH 2 , which can ensure the self-assembly of the peptide solution after mixing.

[0035] 1.2. The RGD short peptide sequence Arg-Gly-Asp widely exists in various extracellular matrix proteins such as cohesin, and specifically binds to integrin on the cell membrane, which is a key binding site for cell attachment. RADA16-I The carboxy-terminal synthetic functional fragment RGD short peptide sequence is the RGD functional polypeptide used in the present invention, and its sequence is Ac-RGD-(RADA) 4 -NH 2 .

[0036] 1.3. The VEGF mimic polypeptide fragment GGGGGKLTWQELYQLKYKGI, as an analogue of VEGF, mimics the helical region of 17-25 interval of VEGF. This region is capable of activating VEGF receptors, thereby activating VEGF-related cell signaling pathways including vascular endothelial cell proliferation. The ca...

Embodiment 2

[0038] Example 2, Preparation of Polypeptide Solution and Atomic Force Microscope Detection

[0039] The three synthetic peptide powders of RADA16-I, KLT and RGD were respectively dissolved in biological grade deionized ultrapure water to obtain three peptide solutions with a mass concentration of 10g / L (1%). Sonicate for 15 minutes to dissolve the peptide completely. After designing a large number of experimental groups to study the self-assembly gel effect of various ratio mixing methods, it was determined to mix according to the molar ratio of RADA16-I:KLT:RGD=2:1:1. Mix the aqueous solutions of RADA16-I polypeptide, KLT functional polypeptide and RGD functional polypeptide at a molar ratio of 2:1:1 and ultrasonically mix to obtain a mixed polypeptide solution (RADA16-I / KLT / RGD), which is ready to be used to prepare functional Self-assembling peptide hydrogels. The mixed polypeptide solution was quickly filtered and sterilized with a 0.22 μm sterile filter for use. After...

Embodiment 3

[0040] Example 3. Preparation of functional self-assembled polypeptide hydrogel and observation of gel microstructure by scanning electron microscope

[0041] 3.1. In order to assemble into a complete hydrogel with a certain shape, this study uses a transwell chamber to complete the self-assembly induced by saline solution. Put the required number of transwell chambers into a 24-well culture plate with 400uL of 0.01M PBS buffer per well, and put the culture plate in a 37°C incubator overnight to permeabilize the basement membrane of the transwell chambers. Then suck out the permeated liquid in the transwell chamber, add 100 μL of mixed polypeptide solution (RADA16-I / KLT / RGD) into the chamber, and then incubate at 37°C (this temperature can accelerate gel formation) for at least 1 hour to form a gel. After the gel was formed, take out a small chamber, carefully cut off the basement membrane and take out the gel, fixed with 2.5% glutaraldehyde for 30 minutes, dehydrated with gra...

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Abstract

The invention relates to functional self-assembling nano peptide hydrogel (RADA16-I / KLT / RGD) which can promote cell adhesion and can promote vascular endothelial cell proliferation and migration. The functional self-assembling nano peptide hydrogel comprises ion complementary type self-assembling nano peptide RADA16-I, KLT function peptide containing VEGF (vascular endothelial growth factor) mimetic peptide fragments and RGD function peptide containing RGD short peptide sequences. The functional self-assembling nano peptide hydrogel can promote cell adhesion, can promote vascular endothelial cell proliferation and migration, is a good tissue engineering framework material, and can be used for scientific research of tissue engineering technology or cytobiology and the like. The invention further provides physical characteristics, a preparation method and an application of the functional self-assembling nano peptide hydrogel.

Description

technical field [0001] The invention relates to a functional self-assembled nanometer polypeptide hydrogel and a preparation method thereof. The functional self-assembled nanopolypeptide hydrogel of the present invention can not only promote cell adhesion, but also promote the proliferation and migration of vascular endothelial cells, and can be used as a tissue engineering framework material in the field of vascular tissue engineering, and can also be used in tissue engineering technology or Scientific research such as cell biology. Background technique [0002] In 1993, Professor Zhang Shuguang and others discovered an ion-complementary polypeptide that could self-assemble, and used it to synthesize a hydrogel. Self-assembled polypeptide hydrogels are stable aggregates formed spontaneously between polypeptide molecules through non-covalent bonds such as hydrogen bonds, electrostatic interactions, and hydrophobic interactions, from which materials with different structures...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08J3/28C08J3/24C08L89/00A61L27/38A61L27/52A61L27/22A61L27/54
Inventor 孙念峰刘占鳌黄文柏周冠洲范鲁峰邵闻冲
Owner 孙念峰
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