Optimization of human skin keratinocyte culture protocols using bioactive molecules derived from olive oil Ortiz Arrabal, Olimpia Bermejo Casares, Fabiola Garzón Bello, Ingrid Johanna Mesa García, María Dolores Gómez Llorente, Carolina Alaminos Mingorance, Miguel Human keratinocytes Tissue engineering Skin Cell culture Cell proliferation Olive oil The author (s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by projects from the Consejeria de Salud y Familias, Junta de Andalucia, Spain (PE-0395-2019) , and proyectos de I+D+i en el marco del Programa Operativo FEDER Andalucia 2014-2020, Universidad de Granada and Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (B-CTS-450-UGR20) . Supported by the Spanish Plan Nacional de Investigacion Cientifica, Desarrollo e Innovacion Tecnologica (I+D+i) of the Spanish Ministry of Science and Innovation, Instituto de Salud Carlos III (grants FIS PI20/0317 and FIS PI21/0980) , and from Instituto de Salud Carlos III (ICI19-00024) (BIOCLEFT) , co-financed by the European Regional Development Fund (ERDF) through the "Una manera de hacer Europa" program, European Union. Skin damage due to severe burns can compromise patient life. Current tissue engineering methods allow the generation of human skin substitutes for clinical use. However, this process is time-consuming, as the kerati-nocytes required to generate artificial skin have a low proliferation rate in culture. In this study, we evaluated the pro-proliferative effects of three natural biomolecules isolated from olive oil: phenolic extract (PE), DL-3,4-dihydroxyphenyl glycol (DHFG), and oleuropein (OLP), on cultured human skin keratinocytes. The results showed that PE and OLP increased the proliferation of immortalized human skin keratinocytes, especially at concentrations of 10 and 5 & mu;g/mL, respectively, without altering cell viability. In contrast, DHFG did not produce a significant improvement in keratinocyte proliferation. In normal human skin keratinocytes obtained from skin biopsies, we found that PE, but not OLP, could increase the number of keratinocyte colonies and the area occupied by these cells. Furthermore, this effect was associated with increased KI-67 and Proliferating cell nu-clear antigen (PCNA) gene expression. Thus, we propose that PE positively affects keratinocyte proliferation and could be used in culture protocols to improve bioartificial skin generation by tissue engineering. 2023-09-18T07:41:28Z 2023-09-18T07:41:28Z 2023-08 journal article O. Ortiz-Arrabal et al. Optimization of human skin keratinocyte culture protocols using bioactive molecules derived from olive oil. Biomedicine & Pharmacotherapy 164 (2023) 115000. [https://doi.org/10.1016/j.biopha.2023.115000] https://hdl.handle.net/10481/84466 10.1016/j.biopha.2023.115000 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 Internacional Elsevier