Delivery system of biomimetic modification of cerium oxide nanoparticles by exosomes and its application in hair cells

A delivery system and nanoparticle technology, applied in the field of medicine and biology, can solve the problems of inability to efficiently remove cell ROS, achieve hearing protection, restore auditory function, and inhibit cell apoptosis

Active Publication Date: 2021-11-09
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] This application provides a biomimetic modification of cerium oxide nanoparticles by exosomes and its application in the hair cells of the outer ear and inner ear to solve the technical problem of inability to efficiently remove cell ROS in the prior art

Method used

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  • Delivery system of biomimetic modification of cerium oxide nanoparticles by exosomes and its application in hair cells
  • Delivery system of biomimetic modification of cerium oxide nanoparticles by exosomes and its application in hair cells
  • Delivery system of biomimetic modification of cerium oxide nanoparticles by exosomes and its application in hair cells

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preparation example Construction

[0034] A preparation method of a delivery system for exosome biomimetic modification of cerium oxide nanoparticles, the method comprising the following steps:

[0035] obtaining the neural stem cell exosomes;

[0036] mixing and incubating the neural stem cell exosomes with the cerium ion solution, and purifying to obtain a purified solution;

[0037] The purified solution is contacted and reacted with a reducing agent to obtain exosomes modified with cerium oxide nanoparticles, that is, the delivery system, that is, engineered neural stem cell exosomes (ANEN) chimeric with antioxidant nanomaterials.

[0038] In the examples of the present application, the neural stem cell exosomes and cerium ion solution were mixed and incubated, and the purification included: in PBS (phosphate) aqueous solution, the exosomes were mixed with Ce(NO 3 ) 3 ·6H 2 O (0.1 mM) was placed at 37°C and incubated for 16 h to make Ce 3+ Fully adsorbed on the surface of exosomes, the mixture was centr...

Embodiment 1

[0052] Example 1. Construction of engineered neural stem cell stably transformed strains and isolation and extraction of engineered exosomes

[0053] First, use the sequence design of the Palm membrane localization peptide to connect the primer to mCherry, and the A666 targeting peptide (CLEPRWGFGWWLH) is connected to mCherry by enzyme digestion. The expression vector is the PLVX vector for packaging lentivirus, and the promoter is CMV (Cytomegalovirus) , using lentivirus infection to construct a stable cell line ( figure 1 a-b in ). After the above-mentioned stable transduction lines were constructed, A666-modified neural stem cell exosomes (ANE) with a size of about 100 nm were separated and screened by ultracentrifugation through expanded culture, and analyzed by TEM, NTA, flow cytometry and WB. The morphology, particle size, and composition were analyzed. The TEM results showed that it presented an obvious cup holder-like structure. The particle size was about 100 nm as d...

Embodiment 2、A666

[0054] Example 2, A666-NSC-Exo in situ modification of cerium oxide nanoparticles

[0055] Cerium oxide nanoparticles were synthesized in situ on the exosomes of neural stem cells. After optimization of the synthesis conditions, cerium oxide nanoparticles with uniform particle size and moderate size were preliminarily synthesized, and ANEN was constructed. The TEM results were as follows: figure 2 As shown in a, obvious cerium oxide nanoparticles were seen on the exosomes. At the same time, WB analysis found that exosomes modified with cerium oxide nanoparticles contained markers of neural stem cells (Sox2, Bmil, Nestin), indicating that the modification did not affect the physiological activity of exosomes ( figure 2 b), and the modified exosomes have little effect on particle size ( figure 2 in c), has good stability in aqueous solution ( figure 2 d), where A666-NSC-Exo represents neural stem cell exosomes with surface-modified hair cell targeting molecules, and AMNE r...

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Abstract

The present invention relates to the field of medical biotechnology, in particular to a delivery system of exosome biomimetic modification of cerium oxide nanoparticles and its application in hair cells. The components of the delivery system include: cerium oxide nanoparticles, neural stem cell exosomes, the neural stem cell exosomes include surface proteins for inhibiting cell apoptosis, and the cerium oxide nanoparticles have peroxidase activity ; To solve the technical problem of being unable to efficiently remove cellular ROS in the prior art.

Description

technical field [0001] The present invention relates to the field of medical biotechnology, and in particular relates to a delivery system of exosome biomimetic modification of cerium oxide nanoparticles and its application in hair cells. Background technique [0002] Protecting and repairing hair cells is critical to preventing and treating hearing loss. A large number of studies have shown that hearing loss is closely related to the increase of reactive oxygen species (ROS), and ROS is involved in the apoptosis process of hair cells. It has been analyzed that ROS interacts with hair cell membrane phospholipids to generate lipid peroxides, some of which act as mediators of auditory nerve and hair cell apoptosis to trigger hair cell apoptosis. When exposed to noise, local ischemia, or ototoxic drugs, the production of ROS increases and exceeds the scavenging capacity of the cochlear antioxidant system, which will trigger a series of oxidative stress damage such as apoptosis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/69A61K47/64A61K33/24A61K35/30A61P39/06A61P27/16B82Y5/00B82Y30/00B82Y40/00C12N5/10
CPCA61K47/6923A61K47/64A61K33/24A61K35/30A61P39/06A61P27/16B82Y5/00B82Y30/00B82Y40/00C12N5/0623A61K47/6901C12N2510/02
Inventor 丁辉王晓君王大吉李子威黄海冰刘洋聂国辉
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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