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Combined application of low-temperature plasma and ascorbic acid

A technology of low-temperature plasma and low-temperature plasma, which is applied in the field of biomedicine to achieve the effect of inducing cell death

Inactive Publication Date: 2020-08-07
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can only penetrate the surface layer of the skin or act on the surface of tumor cells, which has certain limitations

Method used

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  • Combined application of low-temperature plasma and ascorbic acid
  • Combined application of low-temperature plasma and ascorbic acid
  • Combined application of low-temperature plasma and ascorbic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Determination of the inhibitory effect of low-temperature plasma-irradiated medium and ascorbic acid alone or in combination on U251 / U87 glioma cells.

[0023] In order to judge whether the low-temperature plasma irradiated medium and ascorbic acid alone have an effect on the cell viability of glioma cell lines, the effects of low-temperature plasma-irradiated medium and ascorbic acid on the viability of glioma cell lines were tested separately. First, the U251 cell line and the U87 cell line were divided into 10*10 4 The density of cells / well was seeded in 96-well plate, and after culturing for 24 hours:

[0024] (1) Ascorbic acid acts alone: ​​configure the culture medium with ascorbic acid concentrations of 0, 1, 2, 4, 6, and 8 mmol / L to treat the cells, and after 1h, change the ordinary medium to continue to cultivate for 2h;

[0025] (2) The culture medium irradiated by low-temperature plasma alone acts: the culture medium (volume of 10 ml) after being ...

Embodiment 2

[0033] Example 2 Determination of the effect of low temperature plasma combined with ascorbic acid in inducing the death of U251 / U87 glioma cells

[0034] After culturing U251 and U87 glioma cells in 96-well plates for 24 hours, they were further cultured for 1 hour with the medium with ascorbic acid concentration of 4 mmol / L, and then incubated with the medium after low temperature plasma irradiation for 6 minutes for 2 hours.

[0035] refer to Figure 2A , indicating that the combination of low-temperature plasma irradiated medium and ascorbic acid (indicated by combined in the figure) rounded the cells, showing a death-like morphology. refer to Figure 2B , detected by the cell death kit, compared with the single treatment, the combination can indeed significantly induce cell death, the scale bar in the figure = 100 μm, the lower part of the figure is the corresponding statistical data: the relative ratio of dead cells to live cells. P values ​​were calculated by t-test f...

Embodiment 3

[0036] Example 3 An analysis experiment on the mechanism of the synergistic inhibitory effect of low temperature plasma and ascorbic acid in combination was carried out.

[0037]1) After irradiating 10 ml of culture medium with low temperature plasma for 6 minutes, use a hydrogen peroxide kit to detect the hydrogen peroxide level in the culture medium; as Figure 3A shown, use H 2 O 2 Detection kit for the detection of H in common medium (High DMEM) and 6min CAM 2 O 2 The results in the figure show that the low temperature plasma significantly increases the hydrogen peroxide level in the cell culture medium. Among them, the t-test calculates the P value compared to the High DMEME group: ****p<0.001 has a significant difference relative to the control group.

[0038] 2) After culturing U251 and U87 glioma cells in a 96-well plate for 24 hours, they were cultured for 1 hour with media with ascorbic acid concentrations of 0, 4 and 8 mmol / L, and then 300 U of catalase was adde...

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Abstract

The invention discloses combined application of low-temperature plasma and ascorbic acid, and particularly relates to combined application of a culture medium irradiated by the low-temperature plasmaand the ascorbic acid in in-vitro non-therapeutic induction of glioma cell death. It is shown that the culture medium irradiated by the low-temperature plasma and the ascorbic acid are combined to have an obvious synergistic inhibition effect on glioma cell lines U251 and U87 cultured in vitro, and cell death can be remarkably induced. Through the scheme provided by the invention, a new strategy can be provided for researching the combined application of low-temperature plasma and ascorbic acid in glioma treatment, and a new thought is expected to be provided for designing a new product or a new scheme for treating glioma.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the combined application of low-temperature plasma and ascorbic acid. Background technique [0002] Brain gliomas are the most common primary intracranial tumors arising from carcinogenesis of glial cells in the brain and spinal cord, including astrocytic tumors, oligodendroglial tumors, mixed glial tumors, and ependymal tumors Tumors, of which astrocytomas are the most common among gliomas, accounting for approximately 65% ​​of gliomas. The World Health Organization (WHO) classifies gliomas into grades I to IV according to their malignancy. Among them, grades I and II are well-differentiated low-grade gliomas, which are benign tumors; grades III and IV are poorly differentiated high-grade gliomas, which are malignant tumors, and GBM accounts for 50% to 80%. Glioma is the most common malignant tumor in the nervous system. In the past 10 years, even though surgery, radiation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N13/00
CPCC12N5/0693C12N13/00C12N2500/38
Inventor 孙敏轩周元帅杨姣俞惠丹赵容川孟星君撒小涵欧阳楠黄煜伦李炎炎乔治·塞吉亚迪斯
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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