Assessment of gold nanoparticles on human peripheral blood cells by metabolic profiling with 1H-NMR spectroscopy, a novel translational approach on a patient-specific basis

Abstract

Human peripheral blood cells are relevant ex vivo models for characterizing diseases and evaluating the pharmacological effects of therapeutic interventions, as they provide a close reflection of an individual pathophysiological state. In this work, a new approach to evaluate the impact of nanoparticles on the three main fractions of human peripheral blood cells by nuclear magnetic resonance spectroscopy is shown. Thus, a comprehensive protocol has been set-up including the separation of blood cells, their in vitro treatment with nanoparticles and the extraction and characterization of metabolites by nuclear magnetic resonance. This method was applied to assess the effect of gold nanoparticles, either coated with chitosan or supported on ceria, on peripheral blood cells from healthy individuals. A clear antioxidant effect was observed for chitosan-coated gold nanoparticles by a significant increase in reduced glutathione, that was much less pronounced for gold-cerium nanoparticles. In addition, the analysis revealed significant alterations of several other pathways, which were stronger for gold-cerium nanoparticles. These results are in accordance with the toxicological data previously reported for these materials, confirming the value of the current methodology.

A Translational In Vivo and In Vitro Metabolomic Study Reveals Altered Metabolic Pathways in Red Blood Cells of Type 2 Diabetes

“…In this context, the study of the metabolic profile of RBCs is a suitable approach for personalized medicine in T2D. We recently set up a new protocol [ 32 ] that allows the quantification of more than 80 different metabolites in RBCs by 1 H nuclear magnetic resonance (NMR) that can be linked to different metabolic pathways. Although several studies about the metabolomic profile of serum/plasma samples have been published [ 33 , 34 , 35 ], little information exist regarding NMR-based studies about RBCs, and to the best of our knowledge there are no studies focused on the metabolomic assessment of RBCs in T2D.…”

“…This suggests that the RBC is especially sensitive to T2D and, therefore, thus could eventually provide additional biomarkers [ 36 ]. In addition, the metabolites identified in RBCs are different to the metabolites than can be quantified from plasma and several metabolites that can only be found at low concentrations in plasma are much more concentrated in RBCs (i.e., nicotinamide adenine dinucleotide derivatives, glutathione) [ 32 ]. Moreover, cells may be less sensitive to on-off changes than plasma/serum samples, reducing false positive/negative results [ 37 ].…”

Metabolic fingerprint of insulin resistance in human polymorphonuclear leucocytes

“…PMN samples of the two cohorts were analysed following a recent approach described by our group [ 21 ], represented in S4 Fig . Briefly, PMNs were first isolated from peripheral blood by a double Ficoll-Paque gradient, with purity higher than 90% tested by flow cytometry being obtained.…”

“…Metabolites were extracted following the methanol-chloroform-water protocol that was recently optimized for PMN cells [ 20 , 21 ]. Briefly, frozen samples were placed on ice and allowed to thaw for 5 min.…”

A translational approach to assess the metabolomic impact of stabilized gold nanoparticles by NMR spectroscopy

“…In this context, a series of metabolomics studies, focused on different model systems, have been recently published describing alterations in different metabolic pathways due to the presence of specific NPs. 19,20,22 In particular, several metabolomics studies, based on the effects of AuNPs, have provided insights into their mechanism of action 3,23 revealing that they can bind to intracellular metabolites, 24 influence different metabolic processes (energy and choline metabolism, HBP pathway, oxidative stress 25 ), lead to cell apoptosis 26 and affect cancer cells differently from healthy cells 27 .…”