Optimisation and Characterisation of Anti-Fouling Ternary SAM Layers for Impedance-Based Aptasensors.

An aptasensor with enhanced anti-fouling properties has been developed. As a case study, the aptasensor was designed with specificity for human thrombin. The sensing platform was developed on screen printed electrodes and is composed of a self-assembled monolayer made from a ternary mixture of 15-base thiolated DNA aptamers specific for human thrombin co-immobilised with 1,6-hexanedithiol (HDT) and further passivated with 1-mercapto-6-hexanol (MCH). HDT binds to the surface by two of its thiol groups forming alkyl chain bridges and this architecture protects from non-specific attachment of molecules to the electrode surface. Using Electrochemical Impedance Spectroscopy (EIS), the aptasensor is able to detect human thrombin as variations in charge transfer resistance (Rct) upon protein binding. After exposure to a high concentration of non-specific Bovine Serum Albumin (BSA) solution, no changes in the Rct value were observed, highlighting the bio-fouling resistance of the surface generated. In this paper, we present the optimisation and characterisation of the aptasensor based on the ternary self-assembled monolayer (SAM) layer. We show that anti-fouling properties depend on the type of gold surface used for biosensor construction, which was also confirmed by contact angle measurements. We further studied the ratio between aptamers and HDT, which can determine the specificity and selectivity of the sensing layer. We also report the influence of buffer pH and temperature used for incubation of electrodes with proteins on detection and anti-fouling properties. Finally, the stability of the aptasensor was studied by storage of modified electrodes for up to 28 days in different buffers and atmospheric conditions. Aptasensors based on ternary SAM layers are highly promising for clinical applications for detection of a range of proteins in real biological samples.

Differential patterning of neuronal, glial and neural progenitor cells on phosphorus-doped and UV irradiated diamond-like carbon.

Diamond-like carbon (DLC) is an attractive biomaterial for coating human implantable devices. Our particular research interest is in developing DLC as a coating material for implants and electrical devices for the nervous system. We previously reported that DLC is not toxic to N2a neuroblastoma cells or primary cortical neurons and showed that phosphorus-doped DLC (P:DLC) could be used to produce patterned neuron networks. In the present study we complement and extend these findings by exploring patterning of dorsal root ganglion (DRG) explants, human neural progenitor cells (hNPC) and U-87 astroglioma cells on P:DLC. Further P:DLC data is provided to highlight that P:DLC can be used as an effective coating material for in vitro multi-electrode arrays (MEAs) with potential for patterning groups of neurons on selected electrodes. We also introduce ultraviolet (UV) irradiation as a simple treatment to render DLC neurocompatible. We show that UV:DLC can be used to support patterned and unpatterned cortical neuron growth. These findings strongly support the use of DLC as tailorable and tuneable substrate to study neural cell biology in vitro and in vivo. We conclude that DLC is a well-suited candidate material for coating implantable devices in the human nervous system.

Patterned growth of neuronal cells on modified diamond-like carbon substrates.

Diamond-like carbon (DLC) has been explored as a biomaterial with potential use for coating implantable devices and surgical instruments. In this study the interaction of DLC with mammalian neuronal cells has been studied along with its modifications to improve its function as a biomaterial. We describe the use of DLC, oxidised DLC and phosphorus-doped DLC to support the growth and survival of primary central nervous system neurones and neuroblastoma cells. None of these substrates were cytotoxic and primary neurones adhered better to phosphorus-doped DLC than unmodified DLC. This property was used to culture cortical neurones in a predetermined micropattern. This raises the potential of DLC as a biomaterial for central nervous system (CNS) implantation. Furthermore, patterned DLC and phosphorus-doped DLC can direct neuronal growth, generating a powerful tool to study neuronal networks in a spatially distinct way. This study reports the generation of nerve cell patterns via patterned deposition of DLC.