Proteomic analysis in cells treated with pristine carbon nano-onions and its subcellular localization

Abstract

Nanoparticles toxicity and cellular uptake are substantial factors that regulate the utility of nanoparticles for biomedical applications in diagnostic imaging and therapy. In this work, the cellular uptake of pristine carbon nano-onions (CNOs) produced by submerged arc discharge was confirmed by transmission electron microscopy (TEM) analysis. Carbon nano-onions were localized within early and late endosomes of human keratinocyte cells. TEM images suggest the possible degradation of these structures, in late endosomes of keratinocyte cells after 24 h. Proteomic shotgun analysis revealed that carbon nano-onions cause statistically significant protein abundance changes in a total of 168 proteins. Most of the differentially expressed proteins are involved in biological processes such as metabolic, cellular and immune system processes. Treated cells showed increased expression of clathrin heavy chain 1, calnexin and isoforms β/α, ε and σ 14-3-3 protein, which are related to clathrin-mediated endocytosis. Thus changes in protein abundance concerning clathrin-mediated endocytosis may suggest this as a plausible way by which the carbon nano-onions intracellular traffic occurs.

Fluorescent Carbon Nano-onion as Bioimaging Probe

“…The different types of CNO-based nanoprobes have been developed for cell imaging and some of the representative images are shown in Figure . Cellular uptake of hydrophobic pristine CNO (p-CNO) has been explored by synthesizing the CNO via the arc discharge method and their internalization was confirmed by TEM analysis, Figure a . TEM images show the aggregation tendency of p-CNO because of its hydrophobicity and its accumulation in endosomes of human keratinocyte cells.…”

“…p-CNO is non-toxic in nature up to 30 μg/mL, which is confirmed by trypan blue assay. Generally, hydrophobic CNO was transformed into hydrophilic to overcome the aggregation tendency and was used as a cell imaging probe …”

“…Abbreviations: cm, cell membrane; is, intercellular space; g, Golgi complex; e, endosome. Reproduced with permission from ref . Copyright 2019 Vietnam Academy of Science & Technology.…”

“…Cellular uptake of hydrophobic pristine CNO (p-CNO) has been explored by synthesizing the CNO via the arc discharge method and their internalization was confirmed by TEM analysis, Figure 4a. 98 TEM images show the aggregation tendency of p-CNO because of its hydrophobicity and its accumulation in endosomes of human keratinocyte cells. p-CNO may enter into the cells via clathrin-mediated endocytosis pathway as the clathrin-mediated endocytosisrelated protein clathrin heavy chain 1, calnexin, and isoforms β/ α, ε, and σ 14−3−3 expression increases in the treated cells.…”

“…Generally, hydrophobic CNO was transformed into hydrophilic to overcome the aggregation tendency and was used as a cell imaging probe. 98 Fluoresceinamine-terminated CNO was developed for imaging of cancerous C57BL/6 μ B DCs cells and their cell uptake ability compared with similarly functionalized CNT. Efficient cell uptake was confirmed from the image in Figure 4b.…”