Graphene 2D platform is safe and cytocompatibile for HaCaT cells growing under static and dynamic conditions

Abstract

The study concerns the influence of graphene monolayer, as a 2 D platform, on cell viability, cytoskeleton, adhesions sites andmorphology of mitochondria of keratinocytes (HaCaT) under static conditions. Based on quantitative and immunofluorescent analysis, it could be stated that graphene substrate does not cause any damage to membrane or disruption of other monitored parameters. Spindle poles and cytokinesis bridges indicating proliferation of cells on this graphene substrate were detected. Moreover, the keratinocyte migration rate on the graphene substrate was comparable to control glass substrate when the created wound was completely closed after 38 hours. HaCaT morphology and viability were also assessed under dynamic conditions (lab on a chip – micro scale). For this purpose, microfluidic graphene system was designed and constructed. No differences as well as no anomalies were observed during cultivation of these cells on the graphene or glass substrates in relation to cultivation conditions: static (macro scale) and dynamic (micro scale). Only natural percentage of dead cells was determined using different methods, which proved that the graphene as the 2 D platform is cytocompatible with keratinocytes. The obtained results encourage the use of the designed lab on a chip system in toxicity testing of graphene also on other cells and further research on the use of graphene monolayers to produce bio-bandages for skin wounds in animal tests.

Keywords:

• Graphene
• keratinocytes
• lab on a chip
• cytoskeleton
• mitochondria

Acknowledgments

JS thank Dr. Michał Młotek and Prof. Wioletta Raróg-Pilecka for the use of their contact angle measurement setup.

Author contributions

IL was the research leader, design the study project, produced and analyzed the data for all experiments as well as generating the figures for all data and drafted the manuscript itself. EJ generated the data for the micro analysis and wrote that section of the manuscript. ZA generated the data for the micro analysis ES participated in the data analysis MS has intellectually accompanied all experimental work, LS-D contributing to the interpretation of the data and has been involved in critically revising the manuscript, IP provided characterization data for the graphene monolayer. JS analyzed graphene monolayer, MCHK was involved in planning the design of the study, has intellectually accompanied all experimental work, contributing to the analysis and interpretation of the data and has been involved in critically revising the manuscript. All authors reviewed and approved the final manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).