Validation Cytotoxicity Assay for Lipophilic Substances

Author(s): Natalia Mencacci Esteves-Pedro, Kenji Sugibayashi, Elissa A. Ostrosky, Marcio Ferrari, Bianca da Silva Sufi, Monica Beatriz Mathor, Paulo Roberto H. Moreno, Felipe R. Lourenco, Vladi Olga Consiglieri, Andre R. Baby, Telma Mary Kaneko*.

Journal Name: Current Topics in Medicinal Chemistry

Volume 18 , Issue 4 , 2018

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Abstract:

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.

Keywords: Cytotoxicity, essential oil, Minthostachys setosa, Pimenta pseudocaryophyllus, Drimysbrasiliensis, neutral red, thiazolyl blue tetrazolium bromide.

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Article Details

VOLUME: 18
ISSUE: 4
Year: 2018
Page: [275 - 286]
Pages: 12
DOI: 10.2174/1568026618666180410142829
Price: $58

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