Abstract

The increasing biological use of graphene-based materials has prompted research inquiries on their effects on microorganisms. The work herein reported different types of microbiological activity of reduced graphene oxide (RGO). At relatively high concentrations (200 and 400 μg/mL), RGO exhibited antibacterial activity on the model bacterium Escherichia coli, while at lower concentrations (10 and 50 μg/mL), interestingly, no antibacterial effect was observed. Instead, an increase in the viable population after exposure at lower concentrations was observed, verified by colony counting and fluorescence microscopy. Further investigation ruled out the possibility of nutrient release from RGO being responsible for this growth-enhancing effect, whereby a comparable number of viable cells were found in the particle-free RGO leachate systems relative to the control. A before and after exposure X-ray photoelectron spectroscopy (XPS) analysis of the RGO detected less presence of C-C bond on the particle surface, suggesting the ability of the bacterium for the use of the carbon-based materials for growth. This potential RGO-cell interaction is further supported by the observed emergence of C-N bond on the particle surface, the nitrogen moieties most likely of bacterial (cell envelope) origins. Although still an early evidence, such RGO-cell interactions could explain the viable cell increase observed at the lower concentration RGO systems. The present study highlights the concentration-dependent microbiological effects of RGO, clarifying the contradicting reports on the growth enhancing versus antibacterial effect of graphene-based materials. The knowledge is important not only for the antibacterial formulation of carbon-based materials but also when assessing their environmental impact.

Highlights

  • The indiscriminate use of antibiotics has led to the emergence of many drug-resistant bacteria, urging the need to explore for alternative antimicrobial agents

  • The acquired reduced graphene oxide (RGO) was characterized by Xray diffraction (XRD) and transmission electron microscopy (TEM)

  • RGO exhibited cellkilling activities on E. coli at relatively high particle concentrations (200 and 400 μg/mL), while cell proliferation was observed at lower particle concentrations (10 and 50 μg/mL), being validated by both agar colony counting and fluorescence viability staining methods

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Summary

Introduction

The indiscriminate use of antibiotics has led to the emergence of many drug-resistant bacteria, urging the need to explore for alternative antimicrobial agents. Studies have observed the antibacterial effects of carbon-based materials, including the GO and RGO; yet, interestingly, more recent work have shown the possibility of these materials, for example, with GO, to promote cell proliferation, that is, as a potential carbon source. Using the model bacterium Escherichia coli, the present work studied the viability of cells when exposed to a relatively wide concentration range of RGO, including treatments with the particle-free leachate systems. The latter is to validate the potential release of carbon source from RGO for cell growth. The work used spectroscopy analysis to examine RGO-cell interactions, characterizing physical contact that could lead to the antibacterial versus growth enhancer effects in question

Experimental Section
Results and Discussion
Microbiological Effects of RGO
Evaluation method
Conclusions
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