Graphene Nanoplatelets: In Vivo and In Vitro Toxicity, Cell Proliferative Activity, and Cell Gene Expression

Beatriz Salesa,Marcelo Assis,Ángel Serrano-Aroca,Alba Cano-Vicent,Juan Andrés,Alberto Tuñón-Molina

doi:10.3390/app12020720

Beatriz Salesa, Marcelo Assis + Show 4 more

Open Access

https://doi.org/10.3390/app12020720

Copy DOI

Abstract

Multi-layer graphene (2–10 layers), also called graphene nanoplatelets (GNPs), is a carbon-based nanomaterial (CBN) type with excellent properties desirable for many biomedical applications. Despite the promising advantages reported of GNPs, nanoscale materials may also present a potential hazard to humans. Therefore, in this study, the in vivo toxicity of these nanomaterials at a wide range of concentrations from 12.5 to 500 µg/mL was evaluated in the Caenorhabditis elegans model for 24 h (acute toxicity) and 72 h (chronic toxicity). Furthermore, their in vitro toxicity (from 0 to 10 µg/mL for 12 and 24 h), proliferative activity at 72 and 96 h, and their effect on the expression of thirteen genes in human keratinocytes HaCaT cells were studied. The physico-chemical and morphological aspects of the GNPs used in this study were analyzed by Raman scattering spectroscopy, electron microscopy, zeta potential as a function of pH, and particle size measurements by dynamic light scattering. The results of this study showed that GNPs showed in vivo non-toxic concentrations of 25 and 12.5 µg/mL for 24 h, and at 12.5 µg/mL for 72 h. Moreover, GNPs present time-dependent cytotoxicity (EC50 of 1.142 µg/mL and 0.760 µg/mL at 12 h and 24 h, respectively) and significant proliferative activity at the non-toxic concentrations of 0.005 and 0.01 μg/mL in the HaCaT cell line. The gene expression study showed that this multi-layer-graphene is capable of up-regulating six of the thirteen genes of human keratinocytes (SOD1, CAT, TGFB1, FN1, CDH1, and FBN), two more genes than other CBNs in their oxidized form such as multi-layer graphene oxide. Therefore, all these results reinforce the promising use of these CBNs in biomedical fields such as wound healing and skin tissue engineering.

Highlights

Introduction published maps and institutional affilCarbon-based nanomaterials (CBNs) are very promising functional materials with many advanced biomedical applications, including medical imaging and nanotherapeutics [1,2,3]
CBNs have been recently proposed as next-generation antimicrobials to combat infectious diseases such as the current Coronavirus disease 2019 (COVID-19)
The antimicrobial mechanism of CBNs is usually attributed to a combination of physical and chemical processes such as membrane structure disruption, microorganism entrapment, electron transfer, and/or oxidative stress by the action of reactive oxygen species (ROS) [5]

Summary

Introduction

Introduction published maps and institutional affilCarbon-based nanomaterials (CBNs) are very promising functional materials with many advanced biomedical applications, including medical imaging and nanotherapeutics [1,2,3]. Because they have unique biological properties such as antimicrobial activity against a broad range of microorganisms, the capacity of inducing tissue regeneration, and low risk of antimicrobial resistance [4]. The antimicrobial mechanism of CBNs is usually attributed to a combination of physical and chemical processes such as membrane structure disruption, microorganism entrapment, electron transfer, and/or oxidative stress by the action of reactive oxygen species (ROS) [5]. They possess many other excellent physical and biological properties such as high thermal and electrical conductivity, excellent mechanical performance, immunomodulatory potential and can be combined with stem cells iations.

Methods

Results

Conclusion