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Brain-targeted exosome and preparation method and applications thereof

An exosome and brain technology, which is applied to brain-targeted exosomes and its application in the treatment of brain diseases, can solve the problems of incomplete and single detection after traumatic brain injury, and overcome the problems of Difficulties in brain targeting, overcoming toxic side effects, overcoming inefficient effects

Active Publication Date: 2019-05-03
SHANGHAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the current biomedical imaging technology is relatively single, and the post-healing detection of traumatic brain injury is not comprehensive, and now the in vivo imaging technology has attracted attention. Therefore, it is necessary to develop a natural drug delivery system that can be used for precise treatment, and at the same time can realize the visualization of real-time monitoring treatment has become an urgent problem to be solved

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  • Brain-targeted exosome and preparation method and applications thereof
  • Brain-targeted exosome and preparation method and applications thereof
  • Brain-targeted exosome and preparation method and applications thereof

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preparation example Construction

[0029] Another aspect of the embodiments of the present invention also provides a method for preparing brain-targeted exosomes, including:

[0030] Make brain-targeted neural stem cells secrete exosomes with brain-targeted peptides;

[0031] collecting and purifying the exosomes;

[0032] The fluorescent material is allowed to enter the inner cavity of the exosome, thereby obtaining the brain-targeted exosome.

[0033] Among them, exosomes are derived from genetically engineered neural stem cells. The neural stem cells can be modified first by genetic engineering, so that the surface of the neural stem cell cell membrane carries a targeting peptide targeting the brain. Exosomes with brain-targeted peptides secreted by brain-targeted modified neural stem cells have the function of crossing the blood-brain barrier and can treat traumatic brain injury. Such a natural nanovesicle can achieve low Toxic side effects, efficient and precise treatment.

[0034] Fluorescent materials...

Embodiment

[0067] (1) Extraction of neural stem cells

[0068] Step 1: Take 4-week-old Balb / C mice, kill them by cervical dislocation, cut off the head, take out the intact brain, and put it on ice-precooled DMEM-F12 medium (containing 1% double antibody), Carefully peel off the hypothalamus (SVZ) area, place it on ice, and use forceps to peel off the meninges and blood vessels; mince the hypothalamus (SVZ) with a scalpel, put it into a 15 mL centrifuge tube containing 5 mL of DMEM-F12 serum-free medium, add 0.5mL of neural stem cells were digested with trypsin Accutase at 37°C for 20min, shaking every 5min. Centrifuge at 200 g for 5 min to remove the supernatant; add 1 mL of DMEM-F12, resuspend the cells in the medium, add 20 μl of DNase, and gently pipette the tissue 10 times with a pipette, without generating air bubbles during pipetting. Add 1 mL of fresh medium and a small amount of DNase, slowly pipet 15 times on ice, let stand for 1-2 min, suck the supernatant into a new 15 mL ce...

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Abstract

The present invention discloses a brain-targeted exosome which comprises an exosome capable of passing through blood brain barrier and a fluorescent material with an imaging function, wherein the fluorescent material is distributed in the entocoele of the exosome. The invention also discloses a preparation method of the brain-targeted exosome, and the method comprises: enabling brain-targeted neural stem cells to secret an exosome with a brain-targeted peptide; collecting and purifying the exosome; and enabling the fluorescent material to enter the entocoele of the exosome so as to obtain thebrain-targeted exosome. The brain-targeted exosome provided by the present invention is mainly formed by natural materials, has a function of passing through blood brain barrier, can overcome the problem that traditional nano-particles have low efficiency of passing through blood brain barrier and have toxic or side effect, overcomes the difficulty of brain targeting of traditional particles, andprovides a visible and accurate treatment strategy for treating brain-related diseases such as traumatic brain injury and brain tumors.

Description

technical field [0001] The invention relates to the field of nanobiological materials, in particular to a brain-targeted exosome and its application in the treatment of brain diseases. Background technique [0002] In recent years, Traumatic Brain Injury (Traumatic Brain Injury, TBI) has become the neurological disease second only to stroke, and has become one of the causes of death and disability worldwide. TBI is initially a primary trauma, and then over time Over time, TBI becomes a secondary brain injury, which leads to a cascade of pathophysiological responses leading to circulatory disturbance, blood-brain barrier disruption, neuronal death and subsequent neurological dysfunction after the primary injury. [0003] Studies have shown that in addition to the traditional liposome drug-loaded treatment, exosomes are now a kind of cell vesicles with a size of 30-100nm formed by the invagination of the cell membrane, which has attracted everyone's attention. Exosomes contain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61P25/00A61P35/00
Inventor 王强斌王仟伍董津辰陈光村
Owner SHANGHAI UNIV
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