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Bifunctional exosome, preparation method thereof and application in cerebral apoplexy repair

An exosome and dual-function technology, applied in biochemical equipment and methods, cell dissociation methods, pharmaceutical formulations, etc., can solve the problems of cell toxicity, interference with protein activity, etc., to increase neurogenesis, promote neural differentiation, Improve the effect of accumulation

Active Publication Date: 2021-09-03
上海市伤骨科研究所
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Problems solved by technology

However, the use of Cu(I) ions is toxic to cells and interferes with protein activity

Method used

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  • Bifunctional exosome, preparation method thereof and application in cerebral apoplexy repair
  • Bifunctional exosome, preparation method thereof and application in cerebral apoplexy repair
  • Bifunctional exosome, preparation method thereof and application in cerebral apoplexy repair

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Embodiment Construction

[0039] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be described in detail below in conjunction with the examples. It must be pointed out that the following examples are only used to explain and illustrate the present invention, and are not intended to limit the present invention . Some non-essential improvements and adjustments made by those skilled in the art based on the above content of the invention still belong to the protection scope of the present invention.

[0040] Example of implementation

[0041] 1. BV2 microglia culture, polarization and identification

[0042] Cells were cultured in a DMEM medium containing 10% FBS (fetal bovine serum) and 1% penicillin-streptomycin in a 37°C incubator. The method of polarizing BV2 to M2 was as follows: BV2 cells were treated with 20ng / mL IL-4 (interleukin) for 48h, and Iba-1, CD206 and ARG antibodies were detected by immunofluorescence. Weste...

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Abstract

The invention provides a bifunctional exosome, a preparation method thereof and application in cerebral apoplexy repair. The preparation method comprises the following steps of firstly, through amidation reaction, carrying out azidation group modification on DA7R polypeptide and SDF-1 factor separately; secondly, introducing a DBCO group on the surface of the exosome through an addition reaction of succinimide and an amino group to construct a DBCO-exosome; and finally, constructing the DA7R-SDF-1 dual-modified functionalized exosome through a copper-free click chemistry method. The bifunctional exosome obtained by the method shows targeting to injured blood vessels of a stroke part and has the characteristic of recruitment of neural stem cells, so that the capability of inducing the neural stem cells to directionally differentiate to neurons is enhanced, and finally neural restoration is realized through the 'integrated multifunctional' exosome. The invention paves a way for treatment of central nervous system injury based on the exosome and reasonable design of functionalized exosome.

Description

technical field [0001] The invention belongs to the technical field of biological carrier targeted drug delivery, and specifically relates to a bifunctional exosome, a preparation method thereof, and an application in stroke repair. Background technique [0002] Injuries to the central nervous system (such as stroke, traumatic brain injury, spinal cord injury, etc.) are often difficult to heal, can easily lead to long-term disability, and seriously affect the quality of life of patients. This is mainly due to the two major difficulties in nerve repair: the limited ability of neurons to regenerate and the formation of astrocyte scars. Numerous scientific researchers have carried out a lot of research on the repair of central nervous system injury, and have made some progress. [0003] In vitro neural stem cell transplantation is widely used in nerve repair therapy. However, due to the influence of the damaged microenvironment, the survival rate of neural stem cells is low, a...

Claims

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Application Information

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IPC IPC(8): C12N5/079A61K35/30A61K47/64A61P9/10A61P25/00A61P25/28
CPCC12N5/0622C12N5/0006A61K35/30A61K47/64A61P9/10A61P25/00A61P25/28C12N2501/2304C12N2509/00
Inventor 崔文国阮慧瞳汤耀辉李勇芳杨国源
Owner 上海市伤骨科研究所
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