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Method for treating cerebral ischemia reperfusion injury by using dental pulp stem cell exosome

A cerebral ischemia-reperfusion and dental pulp stem cell technology, applied in the field of dental pulp stem cell therapy, can solve the problems of limited clinical application and damage to the body, and achieve the effect of reducing the area of ​​cerebral infarction

Inactive Publication Date: 2021-09-21
优牙生物科技(上海)有限公司
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Problems solved by technology

[0003] Exosomes derived from bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells can play a role in resisting cerebral ischemia-reperfusion injury, but the acquisition of these two kinds of mesenchymal stem cells is a harmful to the body Therefore, it is necessary to use dental pulp stem cell exosomes to treat cerebral ischemia-reperfusion injury to improve the above problems

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  • Method for treating cerebral ischemia reperfusion injury by using dental pulp stem cell exosome
  • Method for treating cerebral ischemia reperfusion injury by using dental pulp stem cell exosome
  • Method for treating cerebral ischemia reperfusion injury by using dental pulp stem cell exosome

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Embodiment 1

[0028] Example 1, the method of using dental pulp stem cell exosomes to treat cerebral ischemia-reperfusion injury, the specific steps are

[0029] S1: Exosomes derived from dental pulp stem cells were extracted from healthy volunteers aged 18 to 30 years without caries and periodontal lesions from third molars - 75% alcohol disinfection of the tooth surface - dental drill to remove the pulp - Cut into small pieces of 1 mm3 - Rinse 3 times with PBS containing 2.5% antibiotics - Put the pulp tissue in a 1.5ml EP tube and digest it with 3mg / ml type I collagenase - 4mg / mL dispase at 37°C for 30 minutes - Cells were resuspended - cultured in a 37°C 5% CO2 incubator (medium: DMEM + 10% fetal bovine serum + 100U / mL penicillin and 100ug / mL streptomycin) - change the medium after 5 days, and then every three days Once; when the cells are fused to about 80%, digest and centrifuge, and transfer to the third generation; when the third generation cells are fused to about 80%, remove the u...

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Abstract

The invention relates to the technical field of dental pulp stem and fine treatment, in particular to a method for treating cerebral ischemia reperfusion injury by using a dental pulp stem cell exosome. The method comprises the steps: extraction of exosomes from dental pulp stem cells, extraction of third molars without dental caries and periodontal lesions from healthy volunteers at age of 18 to 30 years-disinfection of tooth surface with 75% alcohol-removal of dental pulp from dental drill-cutting into pieces of 1 mm3-flushing three times with PBS containing 2.5% antibiotic-placement of pulp tissue in 1.5 ml EP tube with digestion of 3 mg / ml type I collagenase-placement in dispase of 4 mg / mL for 30 minutes at 37 DEG C-cell resuspension- and culturing in a 5% CO2 incubator at 37 DEG C; brain tissue is an organ most sensitive to ischemia and hypoxia of the body, irreversible brain injury occurs after blood flow supply is interrupted for several minutes, animal experiments find that the exosome from the dental pulp stem cells can reduce the cerebral infarction area after cerebral ischemia-reperfusion injury of a mouse, and a key technology is provided for clinical application of the dental pulp stem cell exosome in treatment of cerebral ischemia-reperfusion injury.

Description

technical field [0001] The invention relates to the technical field of dental pulp stem cell therapy, in particular to a method for using dental pulp stem cell exosomes to treat cerebral ischemia-reperfusion injury. Background technique [0002] At present, a variety of exosomes derived from mesenchymal stem cells can alleviate cerebral ischemia-reperfusion injury, such as exosomes derived from adipose-derived stem cells and exosomes derived from bone marrow stem cells. However, whether exosomes derived from another important type of mesenchymal stem cells (dental pulp stem cells) can alleviate cerebral ischemia-reperfusion injury is unknown. The acquisition of dental pulp stem cells comes from exfoliated wisdom teeth, so compared with other types of mesenchymal stem cells, the acquisition of dental pulp stem cells is less damaging and easier to obtain, so exosomes from dental pulp stem cells are used to alleviate cerebral ischemia regeneration. Perfusion injury has a bette...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A01K67/027A61K35/28A61P9/10
CPCC12N5/0664A01K67/027A61K35/28A61P9/10C12N2509/00C12N2509/10A01K2207/30A01K2227/105A01K2267/0375
Inventor 李松高亮王伦云叶青松
Owner 优牙生物科技(上海)有限公司
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