Author: Abdullah Alnuqaydan
Alnuqaydan, Abdullah, 2017 Determination of Cancer Induction and Cell-Killing Potential of Beauty Products and Personal Care Products Using Human Skin Cells, Flinders University, School of Medicine
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The in vitro toxicity and genotoxicity of four cosmetic products were determined using a human keratinocyte (HaCaT) and human fibroblast CCD-1064SK cells lines. The products were: Nivea Visage Q10plus Anti-Wrinkle cream including synthetic chemicals with TiO2 (NVAW+ TiO2); Nivea Visage Q10plus Anti-Wrinkle cream including synthetic chemicals without TiO2 (NVAW); Facial Moisturizer: Camellia & Geranium Blossom (FMCGB); with glycerol as the negative control. Two human skin cell lines (human keratinocytes HaCaT and human fibroblast CCD1064SK) were employed. Four popular products for head lice treatment were also examined for both types of toxicity: Lice breaker (Permethrin 1% w/w); KP24 Medicated lotion (Maldison0.5% w/w); Organix Pyrethrum treatment (4g/L Pyrethrins, 16g/LPiperonyl Butoxide); Tea Tree Oil (100% pure); and Lavender oil (100% pure) on a human skin cell line (HaCaT). Toxicity was measured by Crystal Violet assay and Methyl tetrazolium (MTT) assay and the proportion of apoptosis or necrosis was monitored by Flow Cytometry. Genotoxicity was detected via the cytokinesis block micronucleus (CBMN) assay. NVAW+ TiO2 induced the highest toxicity and genotoxicity levels of all the tested beauty products. No toxicity or genotoxicity observed with FMCGB, but there was a significant necrosis. Glycerol did not induce any toxicity or genotoxicity. Populations of cells treated with diluted (NVAW+ TiO2) and (NVAW) products showed increased proportions of apoptosis and necrosis. The decrease in NDI by NVAW+ TiO2 and NVAW (1.4 (P < 0.05)) was observed at the 0.3% w/v dose and by FMCGB at the 0.05% w/v dose. (NVAW+ TiO2) and (NVAW) products showed increased frequency of micronuclei (MNi). (NVAW+ TiO2) product proved to induce significantly more micronuclei (MNi) than the product without TiO2. (NVAW+ TiO2) product induced genetic damage manifested as chromosomal damage determined by CBMN assay at a frequency significantly higher (43 MNi/1000 binucleated cells, n=3) than the background frequency (media alone control; MNi range= 9 MNi /1000 binucleated cells, n=3). Head lice treatments Tea Tree Oil (TTO), Pure Lavender oil and Pyrethrum did induce significant cytotoxicity. Also, they enhanced both early apoptosis and late apoptosis or necrosis. However, two head lice treatments, Permethrin (Lice Breaker) and Maldison (Malathion) (KP24) did not induce cytotoxicity. Early apoptosis and necrosis were observed in Permethrin treatment, and late apoptosis and early necrosis were measured in Maldison (Malathion) (KP24). Moreover, Permethrin (Lice Breaker) and Maldison (Malathion) (KP24) induced micronuclei (MNi) at a frequency significantly higher (range= 15-25 MNi/1000 binucleated cells, n=3) than the background frequency (media alone control; MNi range= 6 MNi /1000 binucleated cells, n=3). Moreover, Calendula officinalis extracts A (C5), B (C6), D and C were examined for their ability to protect against hydrogen peroxide (H2O2) induce cell killing and chromosomal damage to HaCaT skin cells. Using the MTT cytotoxicity assay and CBMN genotoxicity assay, it was observed that extracts of Calendula officinalis gave time-dependent and concentration-dependent protection against H2O2-induced oxidative stress in vitro using human skin cells. Pre-incubation with the Calendula extracts for 24 and 48 hours increased survival relative to the population without extract by 20% and 40% respectively following oxidative challenge. The frequency of MNi in the presence of extracts was reduced by an average increment of 20 MNi/1000 BN cells to 2-9 MNi/1000 BN cells at all doses tested. Finally, Calendula extract was examined for its protective effects against the cytotoxicity of selected beauty products and head lice treatments and titanium dioxide (TiO2) on HaCaT cells. Pre-incubation with the Calendula extract for 48 hours significantly increased survival relative to the population without the extract by 30% and 50%, following treatment with personal care products.
Keywords: Toxicity, Genotoxicity, in vitro toxicity, Chromosomal damage, Beauty Products, Personal care Products, Cell culture, Skin cells, MTT Assay, Crystal Violet Assay, Flow cytometry Assay, CBMN Assay.
Subject: Medicine thesis
Thesis type: Doctor of Philosophy
Completed: 2017
School: School of Medicine
Supervisor: Associate Professor Barbara Sanderson