A polypeptide nanofiber hydrogel for slow release of exosomes and its preparation method and application

A technology of nanofibers and hydrogels, which is applied in the field of medicine, can solve the problems that polypeptide hydrogels cannot provide heart protection functions, and achieve the effects of increasing residence time, improving curative effect, and improving heart function

Active Publication Date: 2021-12-24
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the self-assembling peptide hydrogel does not provide cardioprotection

Method used

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  • A polypeptide nanofiber hydrogel for slow release of exosomes and its preparation method and application
  • A polypeptide nanofiber hydrogel for slow release of exosomes and its preparation method and application
  • A polypeptide nanofiber hydrogel for slow release of exosomes and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Separation of exosomes derived from umbilical cord mesenchymal stem cells

[0035] (1) Human umbilical cord mesenchymal stem cells UMSC (UMSC) grown adherently were obtained by resuscitation. When the degree of cell aggregation in the 10cm cell culture dish reaches more than 90%, the cells are subcultured, digested with 0.125% trypsin, and then terminated with complete medium. After centrifugation, the obtained cells are resuspended and divided equally among Three 10cm cell culture dishes were cultured.

[0036] (2) Normal fetal bovine serum was ultracentrifuged at 110,000 g for 8 hours at 4°C to remove exosomes, and the centrifuged serum was filtered with a 0.22 μm disposable sterile filter to obtain exosome-free serum.

[0037] (3) Collect exosomes from passage 6-8 cells, digest and centrifuge the cells, resuspend them in α-MEM medium with 10% fetal bovine serum free of exosomes, and culture them for 48 h after passage according to the normal ratio. super...

Embodiment 2

[0040] Example 2, Synthesis and Identification of Self-Assembled Polypeptides

[0041] 1. Synthetic peptides PA-GG-Hexarelin-S and NapFF.

[0042] The sequence of the polypeptide PA-GG-Hexarelin-S is: Palmitoyl-Gly-Thr-Ala-Gly-Leu-Ile-Gly-Gln-Gly-Gly-His-D-2-methyl-Trp-Ala-Trp-D- Phe-Lys-Ser. The peptide was synthesized in Nanjing GenScript Biotechnology Co., Ltd. The specific synthesis steps are as follows: a seventeen amino acid peptide containing MMP-2 sensitive sequence (GTAGLIQQE) and cardioprotective peptide (Hexarelin) was synthesized by standard FMOC chemical synthesis method , followed by alkylation of the peptide at the N-terminus, the specific steps are as follows: The polypeptide was alkylated at the N-terminus by two 12-hour reactions with palmitic acid in the presence of diethylethylamine (DIPEA) dissolved in dimethylformamide (DMF). Then, the polypeptide was cleaved from the resin for 2 h with a cleavage solution composed of trifluoroacetic acid (TFA), deioni...

Embodiment 3

[0054] Example 3, self-assembled polypeptide PA-GG-Hexarelin-S protective effect on cardiomyocytes

[0055] Different concentrations of PA-GG-Hexarelin-S and cardiomyocytes were pre-incubated for 4 hours, followed by 100 μM H 2 o 2 Treated for 4h, CCK-8 experiment was used to detect cell activity, the result showed that 10 -6 -10 -8 mol / L concentration of PA-GG-Hexarelin-S can improve the viability of cardiomyocytes ( Figure 4 ), indicating that the self-assembling polypeptide PA-GG-Hexarelin-S of the present invention has the protective function of cardiomyocytes.

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Abstract

The invention belongs to the technical field of medicine, and discloses a self-assembled polypeptide, a polypeptide nanofiber hydrogel made from the self-assembled polypeptide NapFF and exosomes, a preparation method and an application. The amino acid sequence of the self-assembling polypeptide of the present invention is shown in SEQ ID No.1. The self-assembled polypeptide of the present invention has cardioprotective function. The polypeptide nanofiber hydrogel of the present invention is made of the self-assembling polypeptide, NapFF and exosomes, the self-assembling polypeptide and NapFF form a hydrogel, and the exosomes are encapsulated in the hydrogel middle. The polypeptide nanofiber hydrogel of the present invention can not only improve the residence time of exosomes with cardioprotective function in tissues, improve the curative effect of exosomes on improving cardiac function, but also directly provide cardioprotective function, further improving cardioprotective function. Cardiac function in the infarct area, can be used for the treatment of cardiovascular diseases such as myocardial infarction.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a polypeptide nanofiber hydrogel and its preparation method and application, in particular to a polypeptide nanofiber hydrogel for slow-release exosomes and its preparation method and application. Background technique [0002] Myocardial infarction is pathologically defined as myocardial cell death due to prolonged ischemia. Ischemic heart disease, generally due to persistent coronary ischemia that results in myocardial cell damage and death, oxygen deprivation and myocardial ischemia trigger an inflammatory response, myocardial cell necrosis and apoptosis, resulting in myocardial remodeling and irreversible cardiac function Decline, even cardiac insufficiency, sudden death, etc., seriously threaten human health. Myocardial damage due to ischemia and hypoxia is also called acute coronary syndrome. According to statistics, about 700,000 patients die of acute coronar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/02A61K35/51A61K35/545A61K35/34A61K9/06A61K38/08A61K47/42A61P9/10
CPCA61K38/08A61K47/42A61P9/10A61K9/0024A61K9/06A61K35/34A61K35/51A61K35/545A61K2300/00
Inventor 李杨欣韩超珊
Owner SUZHOU UNIV
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