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

Author(s)

Salesa, Beatriz ^[1] ; Tunon-Molina, Alberto ^[1] ; Cano-Vicent, Alba ^[1] ; Assis, Marcelo ^[2] ; Andres, Juan ^[2] ; Serrano-Aroca, Angel ^[1]

Affiliation

^[1] Univ Catolica Valencia San Vicente Martir, Ctr Invest Traslac San Alberto Magno, Biomat & Bioengn Lab, C-Guillem Castro 94, Valencia 46001 - Spain

^[2] Univ Jaume, Dept Phys & Analyt Chem, Castellon de La Plana 12071 - Spain

• Document type: Journal article
• Source: APPLIED SCIENCES-BASEL; v. 12, n. 2 JAN 2022.
• Web of Science Citations: 1

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 mu g/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 mu g/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 mu g/mL for 24 h, and at 12.5 mu g/mL for 72 h. Moreover, GNPs present time-dependent cytotoxicity (EC50 of 1.142 mu g/mL and 0.760 mu g/mL at 12 h and 24 h, respectively) and significant proliferative activity at the non-toxic concentrations of 0.005 and 0.01 mu g/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. (AU)

• FAPESP's process: 13/07296-2 - CDMF - Center for the Development of Functional Materials
• Grantee: Elson Longo da Silva
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC