Role of Common Cell Culture Media Supplements on Citrate-Stabilized Gold Nanoparticle Protein Corona Formation, Aggregation State, and the Consequent Impact on Cellular Uptake

Abstract

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.

A Review for Uncovering the “Protein-Nanoparticle Alliance”: Implications of the Protein Corona for Biomedical Applications

“…Investigations of the internalization mechanism responsible for AuNP uptake suggested that this process was not significantly influenced by either clathrin receptors or lipid rafts. These findings indicate that under cell culture conditions, the primary factor influencing cellular uptake is NP aggregation [66].…”

“…CCMs are aqueous ionic solutions containing growth substances, often combined with various additives such as antioxidants, dyes, and antibiotics. Barbero et al conducted a study to examine the influence of additives, namely phenol red, penicillin-streptomycin, l-glutamine, and β-mercaptoethanol, on the development of the PC in cell culture media [66]. PCs with comparable characteristics were acquired, with the exception of the inclusion of antibiotics.…”

Neuroinflammation-Related Proteins NOD2 and Spp1 Are Abnormally Upregulated in Amyotrophic Lateral Sclerosis

“…In-gel digestion protocol and liquid chromatography–mass spectrometry (LC-MS) analysis protocol were both performed as previously reported. 16 …”

“…In-gel digestion protocol and liquid chromatography-mass spectrometry (LC-MS) analysis protocol were both performed as previously reported. 16 Protein Identification and Quantitative Differential Analysis Protein identification and quantitative differential analysis protocols were adapted from previous reports. 17,18 Progenesis QI for proteomics software v3.0 (Nonlinear dynamics, UK) was used for MS data analysis using default settings.…”

Biomolecular Corona of Gold Nanoparticles: The Urgent Need for Strong Roots to Grow Strong Branches

“…Another key aspect is the type of biological fluid. Most studies investigate the BMC forming in serum or cell culture medium, [61,64,127,152,155,[204][205][206] but others use plasma (or directly in vivo) [64,65,70,207,208] or individual proteins. [193,205,209] Notably, different cell culture media (e.g., RPMI, DMEM) can lead to the formation of different GNP-BMCs.…”