Sánchez Arribas, NataliaVelasco Rodríguez, BrendaAicart Sospedra, EmilioGuerrero Martínez, AndrésJunquera González, María ElenaTaboada, Pablo2024-11-142024-11-142024Sánchez-Arribas, Natalia, et al. «Lipid Nanoparticles as Nano-Trojan-Horses for siRNA Delivery and Gene-Knockdown». Journal of Colloid and Interface Science, vol. 679, febrero de 2025, pp. 975-87. DOI.org (Crossref), https://doi.org/10.1016/j.jcis.2024.10.1151095-71030021-979710.1016/j.jcis.2024.10.115https://hdl.handle.net/20.500.14352/110601Received 20 June 2024; Received in revised form 17 October 2024; Accepted 19 October 2024 Available online 22 October 2024 0021-9797/© 2024 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/bync-nd/4.0/ ). NOTA IMPORTANTE: Esta publicación ha sido aceptada en Octubre de 2024 (https://doi.org/10.1016/j.jcis.2024.10.115), aunque la editorial la ha encajado en el Volumen 679, Part B, February 2925, con referencia Journal of Colloid and Interface Science 679 (2025) 975–987The therapeutic messenger RNA strategies, such as those using small interfering RNAs, take several advantages (versatility, efficiency and selectivity) over plasmid DNA-based strategies. However, the challenge remains to find nanovectors capable of properly loading the genetic material, transporting it through troublesome environments, like a tumoral site, and delivering it into the cytoplasm of target cells. Here, lipid nanoparticles, consisting of a gemini cationic/neutral helper lipid mixture, are proposed as siRNA nanovector. Cells from cervical and brain cancer overexpressing the green fluorescent protein (GFP) were chosen to analyse the biological response as well as the efficiency and safety of the siRNA-loaded nanovector according to the cell phenotype. Flow cytometry and epifluorescence or confocal microscopy were used to follow the gene knockdown in these overexpressed cells. The effect of the nanovector on cellular proliferation was evaluated with cytotoxicity assays while their potential oxidative stress generation was determined by quantifying the generation of reactive oxygen species. To explore the mechanism of cellular uptake, different inhibitors of endocytic pathways were used during incubation with cells. Finally, nanovectors were incubated in 3D-grown cells (spheroids) to see whether they can penetrate the complex tumoral microenvironments, their efficiency to knockdown GFP expression being monitored by confocal microscopy.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal articlehttps://doi.org/10.1016/j.jcis.2024.10.115https://www.sciencedirect.com/science/article/pii/S0021979724024688open access544Lipid-based nanovectorLipid nanoparticlesGene knockdownsiRNAInternalization routeReactive oxygen species3D cell spheroidsCytotoxicityLamellar phaseQuímica física (Química)Bioquímica (Química)2307 Química Física