In this study, graphene oxide (GO) has been applied as a matrix for enzyme immobilization. The protein adsorption capacity of GO is much higher than of other large surface area carbonaceous materials. Its structure and physicochemical properties are reported beneficial also for enzymatic activity modifications. The experimental proof was done here that GO-based biocatalytic systems with immobilized catalase are modifiable in terms of catalyzed reaction kinetic constants. It was found that activity and stability of catalase, considered here as model enzyme, closely depend on enzyme/GO ratio. The changes in kinetic parameters can be related to secondary structure alterations. The correlation between enzyme/GO ratio and kinetic and structure parameters is reported for the first time and enables the conscious control of biocatalytic processes and their extended applications. The biological activity of obtained biocatalytic systems was confirmed in vitro by the use of functional test. The addition of immobilized catalase improved the cells’ viability after they were exposed to hydrogen peroxide and tert-butyl-hydroperoxide used as source of reactive oxygen species.
1 January 2017 in Advances in Dermatology and Allergology
The link between air pollution, UV irradiation and skin carcinogenesis has been demonstrated within a large number of epidemiological studies. Many have shown the detrimental effect that UV irradiation can have on human health as well as the long-term damage which can result from air pollution,...
The cytotoxic influence of two different carbonaceous nanomaterials on human mesenchymal stem cells (MSCs) cultured in vitro was compared in the short (1–3 days) and long term (up to 60 days). Amorphous carbon and single-walled carbon nanotubes were chosen and evaluated due to their contrasting physicochemical properties. Both materials, though supposed similarly low-toxic in basic short-term cytotoxicity assays, demonstrated dramatically different properties in the long-term study. The surface chemistry and biomolecule-adsorption capacity turned out to be crucial factors influencing cytotoxicity. We proved that amorphous carbon is able to weakly bind a low-affinity protein coat (so-called soft corona), while carbon nanotubes behaved oppositely. Obtained results from zeta-potential and adsorption measurements for both nanomaterials confirmed that a hard protein corona was present on the single-walled carbon-nanotube surface that aggravated their cytotoxic influence. The long-term exposure of the mesenchymal stem cells to carbon nanotubes, coated by the strongly bound proteins, showed a significant decrease in cell-growth rate, followed by cell senescence and death. These results are of great importance in the light of increasing nanomaterial applications in biomedicine and cell-based therapies. Our better understanding of the puzzling cytotoxicity of carbonaceous nanomaterials, reflecting their surface chemistry and interactions, is helpful in adjusting their properties when tailored for specific applications.
26 May 2018 in The Journal of Physical Chemistry Letters
Carbon-based quantum dots are widely suggested as fluorescent carriers of drugs, genes or other bioactive molecules. In this work, we thoroughly examine the easy-to-obtain, biocompatible, nitrogen-containing carbonaceous quantum dots (N-CQDs) with stable fluorescent properties that are resistant to wide-range pH changes. Moreover, we explain the mechanism of fluorescence quenching at extreme pH conditions. Our in vitro results indicate that N-CQDs penetrate the cell membrane; however, fluorescence intensity measured inside the cells was lower than expected from carbonaceous dots extracellular concentration decrease. We studied the mechanism of quenching and identified reduced form of β-nicotinamide adenine dinucleotide (NADH) as one of the intracellular quenchers. We proved it experimentally that the elucidated redox process triggers the efficient reduction of amide functionalities to non-fluorescent amines on carbonaceous dots surface. We determined the 5 nm–wide reactive redox zone around the N-CQD surface. The better understanding of fluorescence quenching will help to accurately quantify and dose the internalized carbonaceous quantum dots for biomedical applications.
1 July 2018 in Materials Science and Engineering: C