Method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum

Abstract

The invention discloses a method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum. The method includes the steps of: 1) freezing to-be-tested tobacco chewing gum in liquid nitrogen; 2) instantly crushing the tobacco chewing gum by means of a crusher; 3) freezing the crushed tobacco chewing gum in the liquid nitrogen; 4) soaking the crushed tobacco chewing gum in artificial saliva; 5) centrifugally separating an extract liquid; 6) detecting a supernatant liquid by means of an osmotic pressure measuring instrument and regulating the osmotic pressure to be consistent with that of a cell culture medium; 7) measuring the content of nicotine in the extract liquid; 8) culturing human oral mucosa epithelial cells as a detection object; 9) adding the extract liquid of the tobacco chewing gum to the cultured human oral mucosa epithelial cells; and 10) culturing the cells for 4 hours, and detecting the change situation of the [gamma] H2AX of the cells by means of a high-content imaging instrument.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to the determination of the effect of gum-based tobacco products on rH2AX of human oral cavity cells. Background technique [0002] Gum-based smokeless tobacco product (Tobacco chewing gum) is an oral smokeless tobacco product made by adding a certain amount of tobacco extract, spices and other edible additives to the gum base. Gum-based tobacco products are used by putting them in the mouth and chewing them. The chewing extracts will merge with saliva and gather in the mouth, and be spit out or swallowed according to different consumption habits. Since the product mainly acts on the oral cavity and directly contacts oral cells, it is necessary to establish a method for detecting its effect on the γH2AX level of human oral cells to evaluate its safety. [0003] The existing safety evaluation methods of tobacco products are mainly aimed at traditional cigarettes, usually by ...

AI Technical Summary

Problems solved by technology

However, gum-based tobacco products do not produce smoke and directly affect the oral cavity, so the safety assessment methods of traditional cigarettes are not applicable to this product

Snus products in pouches are generally used by sticking them between the gums and lips, and their contents can be dissolved by ordinary soaking methods. Due to the existence of gum base, most of the contents of gum-based tobacco products can be dissolved by direct soaking methods. Contents are difficult to dissolve, making accurate biosafety testing impossible

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