In vitro screening of topical formulation excipients for epithelial toxicity in cancerous and non-cancerous cell lines

Authors

  • Farzaneh Forouz Therapeutics Research Group, Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia; School of Pharmacy, The University of Queensland, Woolloongabba, QLD 4102, Australia https://orcid.org/0000-0001-8290-3817
  • Yousuf Mohammed Therapeutics Research Group, Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia; E-mail: y.mohammed@uq.edu.au https://orcid.org/0000-0002-7825-7557
  • Hamid S. A. Shobeiri Nejad School of Mathematics, Physics and Computing, University of Southern Queensland, QLD 4350, Australia https://orcid.org/0009-0009-0250-840X
  • Michael S. Roberts Therapeutics Research Group, Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia; School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA 5000, Australia; Therapeutics Research Centre, Basil Hetzel Institute for Translational Medical Research, The Queen Elizabeth Hospital, Woodville, SA 5011, Adelaide, Australia https://orcid.org/0000-0003-3894-5301
  • Jeffrey E. Grice Therapeutics Research Group, Frazer Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia, Tel.: +61-7-344-38032; E-mail: jeff.grice@uq.edu.au https://orcid.org/0000-0002-8862-4461

DOI:

https://doi.org/10.17179/excli2023-6072

Keywords:

cytotoxicity, cell sensitivity, MTT, crystal violet, ROS, cell cycle analysis

Abstract

Chemical excipients used in topical formulations may be toxic to living skin cells. Here, we compared the in vitro toxicity of some common solubilizing excipients against human melanoma cells, human keratinocytes (HaCaT) and primary skin fibroblasts (FB) as examples of cancerous, immortalized and primary human skin cells, often used as experimental models representative of in vivo conditions. Two distinct endpoint assays (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV)) were used. The mechanism of cell death after excipient exposure was assessed through Reactive Oxygen Species (ROS) production, cell membrane integrity and cell cycle progression. Results showed that the surfactants, Labrasol®, Labrafil® and Transcutol®, were less toxic than Triton X-100 (a model irritant) in all cell types whereas the oil, Labrafac®, was non-toxic. The human melanoma WM164 cell line showed the greatest sensitivity toward cytotoxicity after chemical exposure, while the other cell lines were more resistant. The relative excipient cytotoxicity responses observed in the MTT and CV assays were comparable and similar trends were seen in their estimated 50 % inhibitory concentration (IC50) values. DNA fragmentation by flow cytometry after exposing the cells to IC50 concentrations of the excipients showed negligible apoptotic populations. ROS production was increased in all cell types after toxic exposure; however, ROS elevation did not lead to apoptosis. The toxicity profiles of each excipient are not only relevant to their use in formulating safe topical products but also in the potential synergistic efficacy in the topical treatment of melanoma.

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Published

2023-11-16

How to Cite

Forouz, F., Mohammed, Y., Shobeiri Nejad, H. S. A., Roberts, M. S., & Grice, J. E. (2023). In vitro screening of topical formulation excipients for epithelial toxicity in cancerous and non-cancerous cell lines. EXCLI Journal, 22, 1173–1199. https://doi.org/10.17179/excli2023-6072

Issue

Vol. 22 (2023)

Section

Original articles

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Copyright (c) 2023 Farzaneh Forouz , Yousuf Mohammed, Hamid S. A. Shobeiri Nejad , Michael S. Roberts, Jeffrey E. Grice

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