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Functional self-assembled nano-peptide hydrogels

A self-assembly, multi-functional technology, applied in medical science, prostheses, etc., can solve the problems of vascular endothelial cells that do not simulate VEGF fragments, low efficiency, and limited cell functions, and achieve the effect of promoting cell adhesion

Inactive Publication Date: 2016-08-31
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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  • Functional self-assembled nano-peptide hydrogels
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Examples

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 Patents(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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