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Ultrastructural Features of Gold Nanoparticles Interaction with HepG2 and HEK293 Cells in Monolayer and Spheroids.

ABSTRACT: Use of multicellular spheroids in studies of nanoparticles (NPs) has increased in the last decade, however details of NPs interaction with spheroids are poorly known. We synthesized AuNPs (12.0 ± 0.1 nm in diameter, transmission electron microscopy (TEM data) and covered them with bovine serum albumin (BSA) and polyethyleneimine (PEI). Values of hydrodynamic diameter were 17.4 ± 0.4; 35.9 ± 0.5 and ±125.9 ± 2.8 nm for AuNPs, AuBSA-NPs and AuPEI-NPs, and Z-potential (net charge) values were -33.6 ± 2.0; -35.7 ± 1.8 and 39.9 ± 1.3 mV, respectively. Spheroids of human hepatocarcinoma (HepG2) and human embryo kidney (HEK293) cells (Corning ® spheroid microplates CLS4515-5EA), and monolayers of these cell lines were incubated with all NPs for 15 min-4 h, and fixed in 4% paraformaldehyde solution. Samples were examined using transmission and scanning electron microscopy. HepG2 and HEK2893 spheroids showed tissue-specific features and contacted with culture medium by basal plasma membrane of the cells. HepG2 cells both in monolayer and spheroids did not uptake of the AuNPs, while AuBSA-NPs and AuPEI-NPs readily penetrated these cells. All studied NPs penetrated HEK293 cells in both monolayer and spheroids. Thus, two different cell cultures maintained a type of the interaction with NPs in monolayer and spheroid forms, which not depended on NPs Z-potential and size.

SUBMITTER: Chelobanov B

PROVIDER: S-EPMC7650816 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Ultrastructural Features of Gold Nanoparticles Interaction with HepG2 and HEK293 Cells in Monolayer and Spheroids.

Chelobanov Boris B Poletaeva Julia J Epanchintseva Anna A Tupitsyna Anastasiya A Pyshnaya Inna I Ryabchikova Elena E

Nanomaterials (Basel, Switzerland) 20201016 10

Use of multicellular spheroids in studies of nanoparticles (NPs) has increased in the last decade, however details of NPs interaction with spheroids are poorly known. We synthesized AuNPs (12.0 ± 0.1 nm in diameter, transmission electron microscopy (TEM data) and covered them with bovine serum albumin (BSA) and polyethyleneimine (PEI). Values of hydrodynamic diameter were 17.4 ± 0.4; 35.9 ± 0.5 and ±125.9 ± 2.8 nm for AuNPs, AuBSA-NPs and AuPEI-NPs, and Z-potential (net charge) values were -33.6 ...[more]

PMID: 33081137

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Project description:In order to determine how functionalized gold nanoparticles (AuNPs) interact in a near-physiological environment, we performed all-atom molecular dynamics simulations on the icosahedral Au144 nanoparticles each coated with a homogeneous set of 60 thiolates selected from one of these five (5) types: 11-mercapto-1-undecanesulfonate -SC11H22(SO3(-)), 5-mercapto-1-pentanesulfonate -SC5H10(SO3(-)), 5-mercapto-1-pentaneamine -SC5H10(NH3(+)), 4-mercapto-benzoate -SPh(COO(-)), or 4-mercapto-benzamide -SPh(CONH3(+)). These thiolates were selected to elucidate how the aggregation behavior of AuNPs depends on ligand parameters, including the charge of the terminal group (anionic vs. cationic), and its length and conformational flexibility. For this purpose, each functionalized AuNP was paired with a copy of itself, placed in an aqueous cell, neutralized by 120 Na(+)/Cl(-) counter-ions and salinated with a 150 mM concentration of NaCl, to form five (5) systems of like-charged AuNPs pairs in a saline. We computed the potential of mean force (the reversible work of separation) as a function of the intra-pair distance and, based on which, the aggregation affinities. We found that the AuNPs coated with negatively charged, short ligands have very high affinities. Structurally, a significant number of Na(+) counter-ions reside on a plane between the AuNPs, mediating the interaction. Each such ion forms a "salt bridge" (or "ionic bonds") to both of the AuNPs when they are separated by its diameter plus 0.2-0.3 nm. The positively charged AuNPs have much weaker affinities, as Cl(-) counter-ions form fewer and weaker salt bridges between the AuNPs. In the case of Au144(SC11H22(SO3(-)))60 pair, the flexible ligands fluctuate much more than the other four cases. The large fluctuations disfavor the forming of salt bridges between two AuNPs, but enable hydrophobic contact between the exposed hydrocarbon chains of the two AuNPs, which are subject to an effective attraction at a separation much greater than the AuNP diameter and involve a higher concentration of counter ions in the inter-pair space.

Dataset Information

Ultrastructural Features of Gold Nanoparticles Interaction with HepG2 and HEK293 Cells in Monolayer and Spheroids.

Publications

Ultrastructural Features of Gold Nanoparticles Interaction with HepG2 and HEK293 Cells in Monolayer and Spheroids.

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