Differential effects of graphene materials on the metabolism and function of human skin cells

Abstract

Graphene-related materials (GRMs) such as graphene oxide (GO) and few-layer graphene (FLG) are used in multiple biomedical applications; however, there is still insufficient information available regarding their interactions with the main biological barriers such as skin. In this study, we explored the effects of GO and FLG on HaCaTs human skin keratinocytes, using NMR-based metabolomics and fluorescence microscopy to evaluate the global impact of each GRM on cell fate and damage. GO and FLG at low concentrations (5 μg mL-1) induced a differential remodeling of the metabolome, preceded by an increase in the level of radical oxygen species (ROS) and free cytosolic Ca2+. These changes are linked to a concentration-dependent increase in cell death by triggering apoptosis and necrosis, the latter being predominant at higher concentrations of the nanostructures. In addition, both compounds reduce the ability of HaCaT cells to heal wounds. Our results demonstrate that the GO and FLG used in this study, which mainly differ in their oxidation state, slightly trigger differential effects on HaCaTs cells, but with evident outcomes at the cellular and molecular levels. Their behavior as pro-apoptotic/necrotic substances and their ability to inhibit cell migration, even at low doses, should be considered in the development of future applications.

Skin irritation potential of graphene-based materials using a non-animal test

“…In addition, the inability of GBMs to reach the stratum basale may explain the apparent discrepancy between these results and those previously obtained on HaCaT skin keratinocytes, at least for FLG and GO, that induced significant cytotoxicity at high concentrations. [11][12][13] Indeed, HaCaT cells are a simplified model of proliferating keratinocytes more compliant with those typical of the stratum basale.…”

“…8 However, despite the importance of skin exposure and the dermotoxic potential of GBMs, very limited data on the cutaneous toxicity of these materials are currently available. Only few in vitro data regarding the effects on skin fibroblasts 9 and keratinocytes [10][11][12][13] are available, so far. In HaCaT skin keratinocytes, we previously demonstrated that FLG and GO reduced cell viability after interaction with plasma membrane.…”

Graphenic Materials for Biomedical Applications

“…Previous studies have shown that the information obtained on the in vitro and in vivo toxicity of GBMs due to their dimensions and variation in oxidation states is far from complete [60]. The interactions of living cells with GBMs depends on their hydrophilicity, surface chemistry, purity, lateral dimensions, layer number, and dose [61][62][63][64]. These properties vary greatly according to their synthesis eventually to methods of functionalization.…”

“…It will probably react first when it comes into direct contact with GBMs. Therefore Frontinan-Rubio et al studied the effects of GO and GR on human HaCaT keratinocytes at the molecular level on cell, metabolomic, Ca 2+ , and ROS processes, as well as cell motility and death [62]. They observed different effects of GBMs on HaCaT cells and varying amounts of metabolites occurring in biochemical processes, for example, in butanoate metabolism, glycolysis, and the citric acid cycle.…”

Experimental, Numerical, and Analytical Study on The Effect of Graphene Oxide in The Mechanical Properties of a Solvent-Free Reinforced Epoxy Resin

“…This reinforcement is obtained by the application of a modified Hummers method to carbon nano-fibres, following the procedure detailed in [35]. The physicochemical characterization of GO is detailed in [36]. Some preliminary tests of nanocomposite preparation with the GO, as supplied, revealed that achieving even a minimal level of dispersion in the absence of solvents is impossible, despite the application of harsh ultrasounds.…”