Method for evaluating immunotoxicity of pesticide based on three-dimensional cell electrochemical sensor

A cell sensor, three-dimensional cell technology, applied in the field of pesticide immunotoxicity evaluation based on three-dimensional cell electrochemical sensor, can solve problems such as inability to accurately reflect cell responses, and achieve the effects of convenient equipment, fast response, and accelerated detection speed.

Active Publication Date: 2022-03-18
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these sensors are based on two-dimensional cell culture, which cannot accurately reflect the response of cells when they are grown under natural conditions.

Method used

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  • Method for evaluating immunotoxicity of pesticide based on three-dimensional cell electrochemical sensor
  • Method for evaluating immunotoxicity of pesticide based on three-dimensional cell electrochemical sensor
  • Method for evaluating immunotoxicity of pesticide based on three-dimensional cell electrochemical sensor

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Embodiment 1

[0059] (1) Preparation of FeTCP@RGO;

[0060] Add 4 mg of graphene oxide to 16.5 mL of ultrapure water, and ultrasonically dissolve it at room temperature for 20 min. Subsequently, 17.6 mg of FeTCP was dispersed in 3.5 mL of DMF / water mixture (4:1, v / v) to obtain mixed solution A; it was added dropwise into the GO suspension. The resulting mixture was further sonicated for 30 minutes and then stirred overnight at 70 °C to obtain a reddish-brown solution. During this process, FeTCP self-assembles onto graphene oxide via hydrophobic interactions and strong π–π stacking. Then, 3.2 μL of hydrazine solution (volume concentration 85%) and 64 μL of ammonia water (25 wt%) were added to the above solution, and stored at 95° C. for 1 h under vigorous stirring. The obtained FeTCP@RGO was centrifuged at 10000 g for 20 min and washed 3 times with ultrapure water. Finally, 15 mg of the obtained FeTCP@RGO was dispersed in 4 mL of ultrapure water by ultrasonic waves to obtain a FeTCP@RGO s...

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Abstract

The invention belongs to the technical field of biosensing, and particularly relates to a method for evaluating pesticide immunotoxicity by using a three-dimensional cell electrochemical sensor. The preparation method comprises the following steps: firstly, preparing a ferriporphyrin-modified reduced graphene oxide compound, and then modifying the ferriporphyrin-modified reduced graphene oxide compound onto a working electrode of a screen-printed carbon electrode; meanwhile, synthesizing a GelMA precursor; secondly, packaging the mouse macrophage RAW264.7 in GelMA hydrogel, and carrying out three-dimensional culture in a culture pool on the electrode; finally, the cells are pretreated through pesticide atrazine or metabolite of the pesticide atrazine, then lipopolysaccharide is used for inducing the cells to release NO, differential pulse voltammetry is adopted for determining an electrical signal of NO, and evaluation of pesticide immunotoxicity is achieved by calculating the inhibition rate of pesticide treatment on NO release of the cells. The electrochemical cell sensor prepared by the invention is rapid, sensitive and reliable, and the cells are more close to in-vivo physiological conditions after being subjected to three-dimensional culture, so that the electrochemical cell sensor has a good application prospect in the aspect of evaluating the immunotoxicity of pesticides.

Description

technical field [0001] The invention relates to the technical field of biosensing, in particular to a method for evaluating the immunotoxicity of pesticides based on a three-dimensional cell electrochemical sensor. Background technique [0002] Pesticides are various chemicals used to control pests and disease vectors. The widespread use of pesticides helps increase agricultural productivity but has resulted in major public health concerns worldwide due to their harmful effects on animals and humans. Adverse effects of exposure to pesticides, such as mutagenicity, teratogenicity, and carcinogenicity, have been widely reported. However, immunotoxicity, a toxic effect that usually does not directly lead to death, has received little public attention. Studies have reported that triazines, organophosphorus and carbamate pesticides can affect immunity and exert immunotoxic effects. Atrazine (AT), a member of the triazine herbicide family, is considered an endocrine disruptor, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48
CPCG01N27/48
Inventor 邹小波魏晓鸥张迪石吉勇刘超孙悦翟晓东
Owner JIANGSU UNIV
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