Spontaneous Differentiation of Human Neural Stem Cells on Nanodiamonds.

The potential use of stem cells in regenerative medicine requires the ability to be able to control stem cell fate as cellular networks are developed. Here, nanodiamonds (≈10 nm) are supported on glass and shown to be an excellent host for the attachment and proliferation of human neural stem cells. Moreover, it is shown that spontaneous differentiation into neurons occurs on nanodiamonds. The use of variously oxygen terminated and hydrogen terminated nanodiamonds has been explored. It is shown that O-ND monolayers promote the differentiation of human neural stem cells into neurons with increased total neurite length, degree of branching, and density of neurites when compared with H-NDs or the glass control. The total number of neurites and total neurite length expressing MAP2, a protein enriched in dendrites, is over five times higher for spontaneously differentiated neurones on the O-NDs compared to the control. The fact that inexpensive nanodiamonds can be attached through simple sonication from water on 2D and 3D shapes indicates significant promise for their potential as biomaterials in which neuro-regenerative diseases can be studied.

Surface functionalisation of nanodiamonds for human neural stem cell adhesion and proliferation.

Biological systems interact with nanostructured materials on a sub-cellular level. These interactions may govern cell behaviour and the precise control of a nanomaterial's structure and surface chemistry allow for a high degree of tunability to be achieved. Cells are surrounded by an extra-cellular matrix with nano-topographical properties. Diamond based materials, and specifically nanostructured diamond has attracted much attention due to its extreme electrical and mechanical properties, chemical inertness and biocompatibility. Here the interaction of nanodiamond monolayers with human Neural Stem Cells (hNSCs) has been investigated. The effect of altering surface functionalisation of nanodiamonds on hNSC adhesion and proliferation has shown that confluent cellular attachment occurs on oxygen terminated nanodiamonds (O-NDs), but not on hydrogen terminated nanodiamonds (H-NDs). Analysis of H and O-NDs by Atomic Force Microscopy, contact angle measurements and protein adsorption suggests that differences in topography, wettability, surface charge and protein adsorption of these surfaces may underlie the difference in cellular adhesion of hNSCs reported here.

Patterning of nanodiamond tracks and nanocrystalline diamond films using a micropipette for additive direct-write processing.

The ability to pattern the seeding of nanodiamonds (NDs), and thus selectively control areas of diamond growth, is a useful capability for many applications, including photonics, microelectromechanical systems (MEMS) prototyping, and biomaterial design. A microprinting technique using a computer-driven micropipette has been developed to deposit patterns of ND monolayers from an unreactive water/glycerol ND ink to 5-µm resolution. The concentration and composition of the ND solution were optimized to realize high-density monolayers of NDs and consistent ND printing. Subsequent nanocrystalline diamond (NCD) patterns grown using chemical vapor deposition showed a high level of compliance with the printed ND pattern. This "direct-write", bottom-up, and additive process offers a versatile and simple alternative to pattern diamond. The process has the particular advantage that it does not require lithography or destructive processing such as reactive-ion etching (RIE) and, pertinently, does not involve reactive chemicals that could alter the surface chemistry of NDs. Furthermore, given that this process obviates the use of conventional lithography, substrates that are not suitable for lithographic processing (e.g., excessively small or three-dimensional structured substrates) can be inscribed with ND patterns. The technique also allows for the growth of discrete, localized, single-crystal nanodiamonds with applications in quantum technology.

Biocompatibility of nanostructured boron doped diamond for the attachment and proliferation of human neural stem cells.

OBJECTIVE: We quantitatively investigate the biocompatibility of chemical vapour deposited (CVD) nanocrystalline diamond (NCD) after the inclusion of boron, with and without nanostructuring. The nanostructuring method involves a novel approach of growing NCD over carbon nanotubes (CNTs) that act as a 3D scaffold. This nanostructuring of BNCD leads to a material with increased capacitance, and this along with wide electrochemical window makes BNCD an ideal material for neural interface applications, and thus it is essential that their biocompatibility is investigated. APPROACH: Biocompatibility was assessed by observing the interaction of human neural stem cells (hNSCs) with a variety of NCD substrates including un-doped ones, and NCD doped with boron, which are both planar, and nanostructured. hNSCs were chosen due to their sensitivity, and various methods including cell population and confluency were used to quantify biocompatibility. MAIN RESULTS: Boron inclusion into NCD film was shown to have no observable effect on hNSC attachment, proliferation and viability. Furthermore, the biocompatibility of nanostructured boron-doped NCD is increased upon nanostructuring, potentially due to the increased surface area. SIGNIFICANCE: Diamond is an attractive material for supporting the attachment and development of cells as it can show exceptional biocompatibility. When boron is used as a dopant within diamond it becomes a p-type semiconductor, and at high concentrations the diamond becomes quasi-metallic, offering the prospect of a direct electrical device-cell interfacing system.