In vitro cytocompatibility assessment of amorphous carbon structures using neuroblastoma and Schwann cells

The development of scaffolds for neural tissue engineering application requires an understanding of cell adhesion, proliferation, and migration of neuronal cells. Considering the potential application of carbon as scaffold materials and the lack of understanding of compatibility of amorphous carbon with neuronal cells, the carbon-based materials in the forms of carbon films and continuous electrospun carbon nanofibers having average diameter of approximate to 200 nm are being investigated with or without ultraviolet (UV) and oxy-plasma (OP) treatments for cytocompatibility property using mouse Neuroblastoma (N2a) and rat Schwann cells (RT4-D6P2T). The use of Raman spectroscopy in combination with Fourier transform infrared (FTIR) and X-ray diffraction establishes the amorphous nature and surface-bonding characteristics of the studied carbon materials. Although both UV and OP treatments make carbon surfaces more hydrophilic, the cell viability of N2a cells is statistically more significant on OP treated fibers/films compared to UV fiber/film substrates after 4 days in culture. The electrospun carbon fibrous substrate provides the physical guidance to the cultured Schwann cells. Overall, the experimental results of this study demonstrate that the electrospun amorphous carbon nanofibrous scaffolds can be used as a suitable biomaterial substrate for supporting cell adhesion and proliferation of neuronal cells in the context of their applications as artificial nerve implants. (c) 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2013.

Vertical electric field stimulated neural cell functionality on porous amorphous carbon electrodes

We demonstrate the efficacy of amorphous macroporous carbon substrates as electrodes to support neuronal cell proliferation and differentiation in electric field mediated culture conditions. The electric field was applied perpendicular to carbon substrate electrode, while growing mouse neuroblastoma (N2a) cells in vitro. The placement of the second electrode outside of the cell culture medium allows the investigation of cell response to electric field without the concurrent complexities of submerged electrodes such as potentially toxic electrode reactions, electro-kinetic flows and charge transfer (electrical current) in the cell medium. The macroporous carbon electrodes are uniquely characterized by a higher specific charge storage capacity (0.2 mC/cm(2)) and low impedance (3.3 k Omega at 1 kHz). The optimal window of electric field stimulation for better cell viability and neurite outgrowth is established. When a uniform or a gradient electric field was applied perpendicular to the amorphous carbon substrate, it was found that the N2a cell viability and neurite length were higher at low electric field strengths (<= 2.5 V/cm) compared to that measured without an applied field (0 V/cm). While the cell viability was assessed by two complementary biochemical assays (MTT and LDH), the differentiation was studied by indirect immunostaining. Overall, the results of the present study unambiguously establish the uniform/gradient vertical electric field based culture protocol to either enhance or to restrict neurite outgrowth respectively at lower or higher field strengths, when neuroblastoma cells are cultured on porous glassy carbon electrodes having a desired combination of electrochemical properties. (C) 2013 Elsevier Ltd. All rights reserved.

Vertical electric field induced bacterial growth inactivation on amorphous carbon electrodes

The objective of the present work is to understand the vertical electric field stimulation of the bacterial cells, when grown on amorphous carbon substrates in vitro. In particular, the antibacterial activity against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli are studied using MTTassay, live/dead assay and inner membrane permeabilization assays. In our experiments, the carbon substrate acts as one electrode and the counter electrode is positioned outside the culture medium, thus suppressing the current, electrokinetic motions and chemical reactions. Guided by similar experiments conducted in our group on neuroblastoma cells, the present experimental results further establish the interdependence of field strength and exposure duration towards bacterial growth inactivation in vitro. Importantly, significant reduction in bacterial viability was recorded at the 2.5 V/cm electric field stimulation conditions, which does not reduce the neural cell viability to any significant extent on an identical substrate. Following electrical stimulation, the bacterial growth is significantly inhibited for S. aureus bacterial strain in an exposure time dependent manner. In summary, our experiments establish the effectiveness of the vertical electric field towards bacterial growth inactivation on amorphous carbon substrates, which is a cell type dependent phenomenon (Gram-positive vs. Gram-negative). (C) 2014 Elsevier Ltd. All rights reserved.

Air Stable Iron/Iron Carbide Magnetic Nanoparticles Embedded in Amorphous Carbon Globules

We have synthesized Fe/Fe3C magnetic nanoparticles embedded in an amorphous carbon globule by pyrolysing of benzene, ferrocene and hydroboric acid. The diameter of the globules is similar to 1 mu m and that of Fe/Fe3C magnetic nanoparticles is similar to 40 nm. The globules exhibit ferromagnetic like behavior and the magnetization as well as the coercivity is found to increases with decreasing temperature.

Influence of ion energy and substrate temperature on the optical and electronic properties of tetrahedral amorphous carbon (ta-C) films

The properties of amorphous carbon (a-C) deposited using a filtered cathodic vacuum arc as a function of the ion energy and substrate temperature are reported. The sp3 fraction was found to strongly depend on the ion energy, giving a highly sp3 bonded a-C denoted as tetrahedral amorphous carbon (ta-C) at ion energies around 100 eV. The optical band gap was found to follow similar trends to other diamondlike carbon films, varying almost linearly with sp2 fraction. The dependence of the electronic properties are discussed in terms of models of the electronic structure of a-C. The structure of ta-C was also strongly dependent on the deposition temperature, changing sharply to sp2 above a transition temperature, T1, of ≈200°C. Furthermore, T1 was found to decrease with increasing ion energy. Most film properties, such as compressive stress and plasmon energy, were correlated to the sp3 fraction. However, the optical and electrical properties were found to undergo a more gradual transition with the deposition temperature which we attribute to the medium range order of sp2 sites. We attribute the variation in film properties with the deposition temperature to diffusion of interstitials to the surface above T1 due to thermal activation, leading to the relaxation of density in context of a growth model. © 1997 American Institute of Physics.

Nature of disorder and localization in amorphous carbon

The electronic structure of amorphous diamond-like carbon is studied. Analysis of the participation ratio shows that π states within the σ-σ* gap are localized. The localization arises from dihedral angle disorder. The localization of π states causes the mobility gap to exceed the optical gap, which accounts for the low carrier mobility and the flat photoluminesence excitation spectrum. © 1998 Elsevier Science B.V. All rights reserved.

Doping mechanism in tetrahedral amorphous carbon

Doping in hydrogenated amorphous silicon occurs by a process of an ionised donor atom partially compensated by a charged dangling bond. The total energies of various dopant and dopant/bonding combinations are calculated for tetrahedral amorphous carbon. It is found that charged dangling bonds are less favoured because of the stronger Coulombic repulsion in ta-C. Instead the dopants can be compensated by weak bond states in the lower gap associated with odd-membered π-rings or odd-numbered π-chains. The effect is that the doping efficiency is low but there are not charged midgap recombination centres, to reduce photoconductivity or photoluminescence with doping, as occurs in a-Si:H.

Properties of tetrahedral amorphous carbon deposited by a filtered cathodic vacuum arc

The properties of a highly sp3 bonded form of amorphous carbon denoted ta-C deposited from a filtered cathodic vacuum arc (FCVA) are described as a function of ion energy and deposition temperature. The sp3 fraction depends strongly on ion energy and reaches 85% at an ion energy of 100 eV. Other properties such as density and band gap vary in a similar fashion, with the optical gap reaching a maximum of 2.3 eV. These films are very smooth with area roughness of order 1 nm. The sp3 fraction falls suddenly to almost zero for deposition above about 200 °C.

Doping mechanism in tetrahedral amorphous carbon

Doping in hydrogenated amorphous silicon occurs by a process of an ionized donor atom partially compensated by a charged dangling bond. The total energies of various dopant and dopant/bonding combinations are calculated for tetrahedral amorphous carbon. It is found that charged dangling bonds are less favored because of the stronger Coulombic repulsion in ta-C. Instead the dopants can be compensated by weak bond states in the lower gap associated with odd-membered π-rings or odd-numbered π-chains. The effect is that the doping efficiency is low but there are not charged midgap recombination centres, to reduce photoconductivity or photoluminescence with doping, as occurs in a-Si:H.

Surface atomic properties of tetrahedral amorphous carbon

The surface energy and surface atomic structure of tetrahedral amorphous carbon has been calculated by an ab-initio method. The surface atoms are found to reconstruct into sp2 sites often bonded in graphitic rings. Placing the dangling bonds on adjacent surface atoms lower their energy by π-bonding and this is the source of the low surface energy. The even lower surface energy of hydrogenated amorphous carbon (a-C:H) is due to the hydrogenation of all broken surface bonds. © 2005 Elsevier B.V. All rights reserved.

Cross-sectional structure of tetrahedral amorphous carbon thin films

A cross-sectional transmission electron microscope study of the low density layers at the surface and at the substrate-film interface of tetrahedral amorphous carbon (ta-C) films grown on (001) silicon substrates is presented. Spatially resolved electron energy loss spectroscopy is used to determine the bonding and composition of a tetrahedral amorphous carbon film with nanometre spatial resolution. For a ta-C film grown with a substrate bias of -300 V, an interfacial region approximately 5 nm wide is present in which the carbon is sp2 bonded and is mixed with silicon and oxygen from the substrate. An sp2 bonded layer observed at the surface of the film is 1.3 ± 0.3 nm thick and contains no detectable impurities. It is argued that the sp2 bonded surface layer is intrinsic to the growth process, but that the sp2 bonding in the interfacial layer at the substrate may be related to the presence of oxygen from the substrate.