Multi-Layer Graphene Oxide in Human Keratinocytes: Time-Dependent Cytotoxicity, Proliferation, and Gene Expression

Beatriz Salesa,Ángel Serrano-Aroca

doi:10.3390/coatings11040414

Beatriz Salesa, Ángel Serrano-Aroca

Open Access

https://doi.org/10.3390/coatings11040414

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Journal: Coatings	Publication Date: Apr 2, 2021
Citations: 12	License type: CC BY 4.0

Affiliation: Valencia Catholic University Saint Vincent Martyr

Abstract

Few-layer graphene oxide (GO) has shown no or very weak cytotoxicity and anti-proliferative effects in a wide range of cell lines, such as glioma cells and human skin HaCaT cells at concentrations up to 100 µg/mL. However, as multi-layer GO has hardly been explored in the biomedical field, in this study, this other type of GO was examined in human keratinocyte HaCaT cells treated with different concentrations, ranging from 0.01 to 150 µg/mL, for different periods of time (3, 12, and 24 h). The results revealed a time–concentration dependence with two non-cytotoxic concentrations (0.01 and 0.05 µg/mL) and a median effective concentration value of 4.087 µg/mL at 24 h GO exposure. Contrary to what has previously been reported for few-layer GO, cell proliferation of the HaCaT cells in contact with the multi-layer GO at 0.01 μg/mL showed identical proliferative activity to an epidermal growth factor (1.6-fold greater than the control group) after 96 h. The effects of the multi-layer GO on the expression of 13 genes (SOD1, CAT, MMP1, TGFB1, GPX1, FN1, HAS2, LAMB1, LUM, CDH1, COL4A1, FBN, and VCAN) at non-cytotoxic concentrations of GO in the HaCaT cells were analyzed after 24 h. The lowest non-cytotoxic GO concentration was able to upregulate the CAT, TGFB1, FN1, and CDH1 genes, which confirms multi-layer GO’s great potential in the biomedical field.

Highlights

Graphene oxide (GO) is a carbon nanomaterial with great potential in the biomedical field due to its excellent physical, and unique biological properties such antimicrobial activity that render them very promising for a wide range of potential industrial applications [1,2]
The time-dependent cytotoxicity, proliferation and gene expression results determined for multi-layer GO in human keratinocyte HaCaT cells is presented in the following subsections
The results of this study show that the exposure of GO for 24 hours did not produced any effect in human keratinocyte HaCaT cells at the highest concentration (0.05 μg/mL)

Summary

Introduction

Graphene oxide (GO) is a carbon nanomaterial with great potential in the biomedical field due to its excellent physical, and unique biological properties such antimicrobial activity that render them very promising for a wide range of potential industrial applications [1,2]. GO possess unique properties such as broad-spectrum antimicrobial properties capable of inactivating enveloped RNA viruses such as SARS-CoV-2 and bacteria such as Streptococcus pneumoniae, it presents low risk of inducing microbial resistance and it is able to induce tissue regeneration. Multiple variants of this material have been synthesized and studied such as fewlayer nanosheets, multi-layer nanosheets, dots, nanocaps, flakes, nanoribbons or nanotubes, among many others [4,5]. Few-layer GO has shown no cytotoxicity in human A549 lung epithelial cells up to 48 h[6]. Few-layer GO dose less than 20μg/mL did not exhibit toxicity to human fibroblast cells either[10]

Methods

Results

Conclusion