BUSCADOR RECOLECTA

Sodium citrate-stabilized gold nanoparticles (AuNPs) are destabilized when dispersed in cell culture media (CCMs). This may promote their aggregation and subsequent sedimentation, or under the proper conditions, their interaction with dispersed proteins can lead to the formation of a NP-stabilizing protein corona. CCMs are ionic solutions that contain growth substances which are typically supplemented, in addition to serum, with different substances such as dyes, antioxidants, and antibiotics. In this study, the impact of phenol red, penicillin-streptomycin, l-glutamine, and β-mercaptoethanol on the formation of the NP-protein corona in CCMs was investigated. Similar protein coronas were obtained except in the presence of antibiotics. Under these conditions, the protein corona took more time to be formed, and its density and composition were altered, as indicated by UV-vis spectroscopy, Z potential, dynamic light scattering, and liquid chromatography-mass spectrometry analyses. As a consequence of these modifications, a significantly different AuNP cellular uptake was measured, showing that NP uptake increased as did the NP aggregate formation. AuNP uptake studies performed in the presence of clathrin- and caveolin-mediated endocytosis inhibitors showed that neither clathrin receptors nor lipid rafts were significantly involved in the internalization mechanism. These results suggest that in these conditions, NP aggregation is the main mechanism responsible for their cellular uptake.

Título del postprint: The Role of Common Cell Culture Media Supplements on the Nanoparticle Protein Corona Formation, Aggregation State, and the Consequent Impact on Cellular Uptake, Sodium citrate-stabilized gold nanoparticles (AuNPs) are destabilized when dispersed in cell culture media (CCMs). This may promote their aggregation and subsequent sedimentation, or under the proper conditions, their interaction with dispersed proteins can lead to the formation of a NP-stabilizing protein corona. CCMs are ionic solutions that contain growth substances which are typically supplemented, in addition to serum, with different substances such as dyes, antioxidants, and antibiotics. In this study, the impact of phenol red, penicillin–streptomycin, l-glutamine, and β-mercaptoethanol on the formation of the NP–protein corona in CCMs was investigated. Similar protein coronas were obtained except in the presence of antibiotics. Under these conditions, the protein corona took more time to be formed, and its density and composition were altered, as indicated by UV–vis spectroscopy, Z potential, dynamic light scattering, and liquid chromatography–mass spectrometry analyses. As a consequence of these modifications, a significantly different AuNP cellular uptake was measured, showing that NP uptake increased as did the NP aggregate formation. AuNP uptake studies performed in the presence of clathrin- and caveolin-mediated endocytosis inhibitors showed that neither clathrin receptors nor lipid rafts were significantly involved in the internalization mechanism. These results suggest that in these conditions, NP aggregation is the main mechanism responsible for their cellular uptake., We acknowledge financial support from the Pandora European Training Network (GA-671881) funded in the framework of H2020 Marie Skłodowska-Curie ITN programme, Spanish Ministerio de Ciencia, Innovación y Universidades (MCIU) (RTI2018-099965-B-I00, AEI/FEDER, UE), and Catalan Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) (2017-SGR-1431). L.M. thanks the financial support from AGAUR by means of the grants for the incorporation of post-doctoral research staff into the Catalan science and technology system within the Beatriu de Pinós program (grant no. 2016 BP 00350). N.G.B. acknowledges financial support by MINECO through the Ramon y Cajal program (RYC-2012-10991). ICN2 is supported by the Severo Ochoa program from Spanish MINECO (grant no. SEV-2017-0706) and is funded by the CERCA Programme/Generalitat de Catalunya. The present work has been performed in the framework of the doctorate in Chemistry and Materials Science at the Autonomous University of Barcelona (UAB) and has been supported by the Priority program ACBN, University of Salzburg. O.H.M. acknowledges financial support from the Spanish Ministry of Science and Innovation (MICINN) through a FPI fellowship, resolved by the Agencia Estatal de Investigación (AEI) with reference (BES-2017-083043). The Proteomics Unit at VHIO is a member of Proteored, PRB3, supported by grant PT17/0019 and funded by ISCIII and ERDF., Peer reviewed