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Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/221527
Applicability of the comet assay in hacat cells for the evaluation of photogenotoxicity
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The establishment of protocols to ensure the safety of pharmaceuticals and cosmetics is
becoming increasingly important. Regulatory authorities require the evaluation of their
phototoxic potential and, consequently, their photogenotoxicity.
This study aimed to validate the comet assay as a reliable method for assessing
photogenotoxicity of chemical compounds in human keratinocytes (HaCaT). It focused on
analyzing sodium dodecyl sulfate (SDS) and benzophenone-3 (BZ) as negative and positive
controls, respectively. The cells were incubated with the test chemicals for 1 hour, followed
by irradiation with 4 J/cm² UVA for the designated time. Cell viability was measured using LDH
assays, and a comet assay was performed immediately after irradiation at non-cytotoxic
concentrations of the compounds. Methyl methanesulfonate (MMS) was used as a positive
control for the comet assay.
SDS was classified as non-phototoxic and non-photogenotoxic. In contrast, BZ was classified
as phototoxic in the 24-hour MTT assay and photogenotoxic in the comet assay. The MTT
assay proved to be more sensitive than the LDH assay in assessing photocytotoxicity.
The comet assay conducted immediately after irradiation was determined to be the more
effective for accurately evaluating DNA damage. It was concluded that a 24-hour time interval
post-irradiation is not optimal, as it allows the activation of cellular repair mechanisms.
Overall, the comet assay shows promise as a reliable tool for detecting photogenotoxic
substances. However, to reduce variability and improve robustness, further assays are
needed. This study highlights the importance of integrating additional assays to fully assess
the significance of photosafety.
Key words: Phototoxicity, photogenotoxocity, in vitro, HaCat, comet assay
Descripció
Treballs Finals de Grau de Farmàcia, Facultat de Farmàcia i Ciències de l'Alimentació, Universitat de Barcelona, 2025. Tutora: Elisabet Teixidó
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POUS VINYALS, Laura. Applicability of the comet assay in hacat cells for the evaluation of photogenotoxicity. [consulta: 28 de novembre de 2025]. [Disponible a: https://hdl.handle.net/2445/221527]