Applications of the Neutral Red Cytotoxicity Assay to Risk Assessment of Aquatic Contaminants: An Overview

2009 ◽  
pp. 215-215-15 ◽  
Author(s):  
B Babich ◽  
E Borenfreund
Keyword(s):  
Risk Assessment ◽  
Neutral Red ◽  
Cytotoxicity Assay ◽  
Aquatic Contaminants

Use of the Rainbow Trout Hepatoma Cell Line, RTH-149, in a Cytotoxicity Assay

1989 ◽  
Vol 17 (2) ◽  
pp. 67-71
Author(s):  
Harvey Babich ◽  
Nieves Martin-Alguacil ◽  
Ellen Borenfreund
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
Rainbow Trout ◽  
Cell Line ◽  
Aromatic Hydrocarbons ◽  
Hepatoma Cell ◽  
Neutral Red ◽  
Cytotoxicity Assay ◽  
Hepatoma Cell Line ◽  
Polycyclic Aromatic ◽  
Direct Acting

The rainbow trout hepatoma cell line, RTH-149, was evaluated for use as a bioindicator cell type in the neutral red cytotoxicity assay. The cells were exposed for six days to various polycyclic aromatic hydrocarbons, including chemicals that are direct-acting toxicants and chemicals that require enzymatic biotransformation to cytotoxic metabolites. Whereas benzo[a]pyrene was only slightly cytotoxic, its metabolites — (±)trans-7,8-diol-benzo[a]pyrene and 3-hydroxy-benzo[a]pyrene — were highly cytotoxic. 7,12-Dimethylbenz[a]anthracene was cytotoxic, but cytotoxicity did not occur with benzo[a]anthracene, benzo[b]fluoranthene and benzo[k]fluoranthene. This cell line appears to lack sufficient xenobiotic metabolising capacity to biotransform many of these polycyclic aromatic hydrocarbons to activated cytotoxic metabolites.

Validation Cytotoxicity Assay for Lipophilic Substances

2018 ◽  
Vol 18 (4) ◽  
pp. 275-286 ◽  
Author(s):  
Natalia Mencacci Esteves-Pedro ◽  
Kenji Sugibayashi ◽  
Elissa A. Ostrosky ◽  
Marcio Ferrari ◽  
Bianca da Silva Sufi ◽  
...  
Keyword(s):  
Essential Oils ◽  
Culture Medium ◽  
False Negative ◽  
Neutral Red ◽  
Cytotoxicity Assay ◽  
Equivalence Test ◽  
Polysorbate 20 ◽  
Negative Results ◽  
False Negative Results ◽  
Lipophilic Substances

It is challenging to disperse lipophilic substances in a validated cytotoxicity assay, especially for compounds with log Kow greater than or equal to 5 that may show false negative results. The purpose of this study was to explain the challenges in conducting a cytotoxicity validated test of lipophilic substances: Minthostachys setosa, Pimenta pseudocaryophyllus, and Drimysbrasiliensis essential oils. Additionally, we compared the equivalence of Neutral Red (NR) and 3- (4,5-dimethylthiazol-2-yl) -5- (3- carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H -tetrazolium, inner salt (MTS) in detecting cell viability. The Hydrophile-Lipophile Balance (HLB) technique was used to evaluate the dispersion of essential oils and cytotoxicity in accordance to the guidelines of the OECD / GD 129 validated cytotoxicity assay. We compared the equivalence of vital dyes by TOST equivalence test. According to the results, we demonstrated the possibility of using other ways to disperse the lipophilic substances. Based on the HLB theory, we selected polysorbate 20 as the best solubilizing agent of the essential oils studied in D10 culture medium.

Applications of the Neutral Red Cytotoxicity Assay to In Vitro Toxicology

1990 ◽  
Vol 18 (1_part_1) ◽  
pp. 129-144 ◽  
Author(s):  
Harvey Babich ◽  
Ellen Borenfreund
Keyword(s):  
Mammalian Cells ◽  
Toxicity Testing ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Cytotoxicity Assay ◽  
Chemical Test ◽  
Neutral Red Assay ◽  
Cells In Culture ◽  
Viable Cells

A concerted effort is currently in progress to develop alternatives to the use of live animals for the acute toxicity testing of xenobiotics. To this end, the neutral red in vitro cytotoxicity assay was developed which, although initially based on the use of mammalian cells in culture, has also been adapted for ecotoxicity studies using fish cells in culture. The neutral red assay is based on the binding of neutral red, a weakly cationic supravital dye, to the lysosomal matrix of viable cells after their incubation with toxic agents. Spectrophotometric quantitation of the extracted dye with a scanning microtitre well reader at 540nm was found to be linear with the number of surviving, viable cells. The assay has been used to determine the relative acute cytotoxicities of a broad spectrum of chemical test agents, to establish structure-toxicity relationships for series of related chemicals, to study metabolism-mediated cytotoxicity, to evaluate interactions between combinations of test agents, to evaluate differential and selective toxicities of cancer chemotherapeutics and other pharmaceuticals, and to study temperature-toxicity interactions.

Interleukin-1α and Interleukin-8 Release by Human Keratinocyte Cell Culture Treated with Surfactants

1997 ◽  
Vol 25 (2) ◽  
pp. 161-171
Author(s):  
Michio Shibata ◽  
Takanari Tsuda ◽  
Hiroshi Itagaki ◽  
Shinobu Kato ◽  
Toshiaki Kobayashi ◽  
...  
Keyword(s):  
Culture Medium ◽  
Membrane Damage ◽  
Human Keratinocytes ◽  
Neutral Red ◽  
Cytotoxicity Assay ◽  
Cytotoxic Effects ◽  
Human Keratinocyte ◽  
Keratinocyte Cell ◽  
Interleukin 1Α ◽  
Cocamidopropyl Betaine

The effects of four cosmetic surfactants on interleukin (IL)-1α and IL-8 release from human keratinocytes were studied to investigate the feasibility of using these effects for the prediction of the irritation potential of chemicals. After exposure of cells to surfactants, the amounts of IL-1α and IL-8 released into culture medium were measured by ELISA. Cytotoxicity was evaluated by using the neutral red uptake (NRU) cytotoxicity assay. Cytokine release was increased 7–15 times by sodium lauryl sulphate (SLS), laurtrimonium chloride, cocamidopropyl betaine (CPB) and Oleth-5 at cytotoxic concentrations. IL-8 release was increased 3–4 times by SLS, CPB and Oleth-5 at subcytotoxic concentrations. After exposure to SLS, IL-1α was released within 1 hour, suggesting that IL-1α release is associated with membrane damage, whereas IL-8 release continued for 24 hours, suggesting that IL-8 was produced within the cells. Cytotoxicity tests and IL-8 release assays were also performed on seven other surfactants. The results show that moderate irritants CPB and PEG-4 dioleate, which have weak cytotoxic effects, significantly increased IL-8 release from human keratinocytes. It is suggested that measurement of IL-8 release is useful for predicting the irritation potential of chemicals which cannot be detected by using the NRU cytotoxicity assay.

Risk assessment in dentistry

1998 ◽  
Vol 62 (10) ◽  
pp. 756-761 ◽  
Author(s):  
CW Douglass
Keyword(s):  
Risk Assessment
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