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Method for predicting eye irritation by using various corneal structure cell combinations

A combination of forecasting and corneal structure technology, applied in measuring devices, biological testing, instruments, etc., can solve the problems of long time-consuming experiments, high costs, and certain difficulties in operation

Active Publication Date: 2019-12-13
GUANGZHOU HUADAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the current combined evaluation of eye stimulation methods in vitro has problems such as long experimental time, high cost, difficult operation, and high inter-laboratory variability, which also need to be optimized to further improve the detection throughput of the method

Method used

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  • Method for predicting eye irritation by using various corneal structure cell combinations
  • Method for predicting eye irritation by using various corneal structure cell combinations
  • Method for predicting eye irritation by using various corneal structure cell combinations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Evaluation of Eye Irritation of a Certain Brand of Amino Acid Foaming Cleanser Based on Multiple Corneal Structural Cytotoxicity Methods

[0062] 1. Corneal Epithelial Irritation Prediction

[0063] 1.1 Routine culture of corneal epithelial cells: the cells used are rabbit corneal epithelial SIRC cells.

[0064] 1.2 Cytotoxicity test

[0065] (1) Determination and preparation of the concentration of the test substance: Weigh 200 mg facial cleanser with an analytical balance, prepare a stock solution with a serum-free medium as a solvent, and prepare a 100 mg / mL mother solution for use.

[0066] (2) Plating: When the density of corneal epithelial cells in the cell bottle reaches 80%, add 0.25% trypsin / 0.02% EDTA to digest and separate the cells from the cell bottle, centrifuge at 1200rpm / min for 5 minutes to remove trypsin, Add new medium to resuspend, take 1mL of cell suspension and dilute to 4mL, take 10μL and drop it on a cell counting plate, count under a...

Embodiment 2

[0118] Example 2 Evaluating the eye irritation of a certain brand of eye drops based on a variety of corneal structural cytotoxicity methods

[0119] 1. Corneal Epithelial Irritation Prediction

[0120] 1.1 Corneal epithelial cells: the cells used are human corneal cell line THCE cells.

[0121] 1.2 Cytotoxicity test

[0122] (1) Determination and preparation of the concentration of the test substance: Weigh 200 mg of eye drops with an analytical balance, prepare a stock solution with a serum-free medium as a solvent, and prepare a 100 mg / mL mother solution for use.

[0123] (2) Plating: When the density of corneal epithelial cells in the cell bottle reaches 80%, add 0.25% trypsin / 0.02% EDTA to digest and separate the cells from the cell bottle, centrifuge at 1200rpm / min for 5 minutes to remove trypsin, Add new medium to resuspend, take 1mL of cell suspension and dilute to 4mL, take 10μL and drop it on a cell counting plate, count under a 40× microscope, adjust the concentra...

Embodiment 3

[0188] Example 3 Evaluating the Eye Irritation of a Certain Brand of Shampoo Based on Multiple Corneal Structure Cytotoxicity Methods

[0189] 1. Corneal Epithelial Irritation Prediction

[0190] 1.1 Routine culture of corneal epithelial cells: primary human corneal epithelial cells.

[0191] 1.2 Cytotoxicity test

[0192] (1) Determination and preparation of the concentration of the test substance: Weigh 200 mg of shampoo with an analytical balance, prepare a stock solution with a serum-free medium as a solvent, and prepare a 100 mg / mL mother solution for use.

[0193] (2) Plating: When the density of corneal epithelial cells in the cell bottle reaches 80%, add 0.25% trypsin / 0.02% EDTA to digest and separate the cells from the cell bottle, centrifuge at 1200rpm / min for 5 minutes to remove trypsin, Add new medium to resuspend, take 1mL of cell suspension and dilute to 4mL, take 10μL and drop it on a cell counting plate, count under a 40× microscope, adjust the concentration ...

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Abstract

The invention discloses a method for predicting eye irritation by using various corneal structure cell combinations. The method comprises the following steps of (1) predicting the irritation of corneal epithelial cells; (2) predicting the irritation of corneal stromal fibroblasts, wherein (2.1) neutral red release (NRR) testing is carried out and (2.2) inflammatory factor testing is carried out; (3) predicting the irritation of corneal endothelial cells; and (4) predicting the establishment of an eye irritation model based on various corneal structure cell combinations. The method can comprehensively reflect the toxic effect of a tested substance (such as a stimulant or a corrosive substance) on corneal structure cells; non-animal testing is realized; the irritation degree of eye irritation substances can be predicted; data information is rich; and the method can be widely applied.

Description

technical field [0001] The invention belongs to the technical field of non-animal testing methods, and in particular relates to a method for predicting eye irritation by using a combination of various corneal structural cells. Background technique [0002] In daily life, people's eyes may be actively exposed to or accidentally exposed to exogenous substances such as medicines, daily chemical products, pesticides, etc., which enter the eyes in the original or diluted form, causing local irritation and even corrosion, which may affect human health. . Therefore, it is of great significance to evaluate the possible eye irritation of foreign substances. [0003] Eye irritation belongs to the category of health toxicology. The traditional method is the rabbit eye test proposed by Draize et al. in 1944. The simple process is to directly drop the test sample into the eyes of rabbits and observe the damage reaction caused. This method is still widely used internationally. But the ...

Claims

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Application Information

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IPC IPC(8): G01N33/50
CPCG01N33/5044
Inventor 罗婷婷程树军黄健聪
Owner GUANGZHOU HUADAI BIOLOGICAL TECH CO LTD
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