Mariana P. Arce

The rise of antimicrobial resistance (AMR) has become a critical health challenge. This, plus the antimicrobial discovery void, had led scientists to search for an effective alternative to antimicrobials. In this context, nanomaterials, such as graphene oxide (GO), a two-dimensional (2D) carbon molecule with oxidized functional groups, have been shown to interact physically and chemically with bacteria. Moreover, the addition of polyethylene glycol (PEG) to its surface enhances GO’s biocompatibility and water solubility, making it a promising candidate for biomedical applications. This study evaluates the antimicrobial efficacy of GO and its polyethylene glycol-modified form (GO-PEG) against Staphylococcus aureus, a bacterium responsible for numerous hospital-acquired and multidrug-resistant infections. After their production, both nanomaterials were characterized using various techniques to provide insight into their morphology, stability, and functional group composition. Then, the antimicrobial activity of GO and GO-PEG was assessed using the Müeller–Hinton broth microdilution method, determining the minimum inhibitory concentration (MIC) for S. aureus among ten different concentrations of both nanomaterials (from 0.0625 to 32 mg/mL). The results demonstrate the potential of GO as an effective antimicrobial agent at 16 and 32 mg/mL, offering new strategies in the fight against AMR. Further research could establish its role in future therapeutic applications.

In this work, a stability study of dispersions of graphene oxide and graphene oxide functionalized with polyethylene glycol (PEG) in the presence of bovine serum albumin is carried out. First, a structural characterization of these nanomaterials is performed by scanning electron microscopy, atomic force microscopy, and ultraviolet visible spectroscopy, comparing the starting nanomaterials with the nanomaterials in contact with the biological material, i.e., bovine fetal serum. The different experiments were performed at different concentrations of nanomaterial (0.125–0.5 mg/mL) and BSA (0.01–0.04 mg/mL), at different incubation times (5–360 min), with and without PEG, and at different temperatures (25–40 °C). The SEM results show that BSA is adsorbed on the surface of the graphene oxide nanomaterial. Using UV-Vis spectrophotometry, the characteristic absorption peaks of BSA are observed at 210 and 280 nm, corroborating that the protein has been adsorbed. When the time increases, the BSA protein can be detached from the nanomaterial due to a desorption process. The stability of the dispersions is reached at a pH between 7 and 9. The dispersions behave like a Newtonian fluid with viscosity values between 1.1 and 1.5 mPa·s at a temperature range of 25 to 40 °C. The viscosity values decrease as the temperature increases.

Carbon nanomaterials have received increased attention in the last few years due to their potential applications in several areas. In medicine, for example, these nanomaterials could be used as contrast agents, drug transporters, and tissue regenerators or in gene therapy. This makes it necessary to know the behavior of carbon nanomaterials in biological media to assure good fluidity and the absence of deleterious effects on human health. In this work, the rheological characterization of different graphene nanomaterials in fetal bovine serum and other fluids, such as bovine serum albumin and water, is studied using rotational and microfluidic chip rheometry. Graphene oxide, graphene nanoplatelets, and expanded graphene oxide at concentrations between 1 and 3 mg/mL and temperatures in the 25–40 °C range were used. The suspensions were also characterized by transmission and scanning electron microscopy and atomic force microscopy, and the results show a high tendency to aggregation and reveals that there is a protein–nanomaterial interaction. Although rotational rheometry is customarily used, it cannot provide reliable measurements in low viscosity samples, showing an apparent shear thickening, whereas capillary viscometers need transparent samples; therefore, microfluidic technology appears to be a suitable method to measure low viscosity, non-transparent Newtonian fluids, as it is able to determine small variations in viscosity. No significant changes in viscosity are found within the solid concentration range studied but it decreases between 1.1 and 0.6 mPa·s when the temperature raises from 25 to 40 °C.

The coupling reactions of polyethylene glycol (PEG) with two different nano-carbonaceous materials, graphene oxide (GO) and expanded graphene oxide (EGO), were achieved by amide bond formations. These reactions yielded PEGylated graphene oxides, GO-PEG and EGO-PEG. Whilst presence of the newly formed amide links (NH-CO) were confirmed by FTIR stretches observed at 1732 cm⁻¹ and 1712 cm⁻¹, the associated Raman D- and G-bands resonated at 1311/1318 cm⁻¹ and 1584/1595 cm⁻¹ had shown the carbonaceous structures in both PEGylated products remain unchanged. Whilst SEM images revealed the nano-sheet structures in all the GO derivatives (GO/EGO and GO-PEG/EGO-PEG), TEM images clearly showed the nano-structures of both GO-PEG and EGO-PEG had undergone significant morphological changes from their starting materials after the PEGylated processes. The successful PEGylations were also indicated by the change of pH values measured in the starting GO/EGO (pH 2.6–3.3) and the PEGylated GO-PEG/EGO-PEG (pH 6.6–6.9) products. Initial antifungal activities of selective metallic nanomaterials (ZnO and Cu) and the four GO derivatives were screened against Candida albicans using the in vitro cut-well method. Whilst the haemocytometer count indicated GO-PEG and copper nanoparticles (CuNPs) exhibited the best antifungal effects, the corresponding SEM images showed C. albicans had, respectively, undergone extensive shrinkage and porosity deformations. Synergistic antifungal effects all GO derivatives in various ratio of CuNPs combinations were determined by assessing C. albicans viabilities using broth dilution assays. The best synergistic effects were observed when a 30:70 ratio of GO/GO-PEG combined with CuNPs, where MIC₅₀ 185–225 μm/mL were recorded. Moreover, the decreased antifungal activities observed in EGO and EGO-PEG may be explained by their poor colloidal stability with increasing nanoparticle concentrations.