Temporal and mechanistic tracking of cellular uptake dynamics with novel surface fluorophore-bound nanodiamonds.

Nanoparticles (NPs) offer promise for a multitude of biological applications including cellular probes at the bio-interface for targeted delivery of anticancer substances, Raman and fluorescent-based imaging and directed cell growth. Nanodiamonds (NDs), in particular, have several advantages compared to other carbon-based nanomaterials - including a rich surface chemistry useful for chemical conjugation, high biocompatibility with little reactive oxygen species (ROS) generation, physical and chemical stability that affords sterilization, high surface area to volume ratio, transparency and a high index of refraction. The visualization of ND internalization into cells is possible via photoluminescence, which is produced by direct dye conjugation or high energy irradiation that creates nitrogen vacancy centers. Here, we explore the kinetics and mechanisms involved in the intracellular uptake and localization of novel, highly-stable, fluorophore-conjugated NDs. Examination in a neuronal cell line (N2A) shows ND localization to early endosomes and lysosomes with eventual release into the cytoplasm. The addition of endocytosis and exocytosis inhibitors allows for diminished uptake and increased accumulation, respectively, which further corroborates cellular behavior in response to NDs. Ultimately, the ability of the NDs to travel throughout cellular compartments of varying pH without degradation of the surface-conjugated fluorophore or alteration of cell viability over extended periods of time is promising for their use in biomedical applications as stable, biocompatible, fluorescent probes.

Effect of proton irradiation on photoluminescent properties of PDMS-nanodiamond composites.

Pure poly(dimethylsiloxane) (PDMS) films, PDMS-nanodiamond (ND) and pure nanodiamond powder were irradiated with 2 MeV protons under a variety of fluence and current conditions. Upon proton irradiation, these samples acquire a fluence-dependent photoluminescence (PL). The emission and excitation spectra, photostability and emission lifetime of the induced photoluminescence of PDMS and PDMS-ND samples are reported. Pure PDMS exhibits a noticeable stable blue PL, while the PDMS-ND composites exhibit a pronounced stable green PL under 425 nm excitation. The PL of PDMS-ND composites is much more prominent than that of pure PDMS or pure ND powder even when irradiated at higher doses. The origin of the significantly enhanced PL intensity for the proton-irradiated PDMS-ND composite is explained by the combination of enhanced intrinsic PL within ND particles due to ion-implantation-generated defects and by PL originating from structural transformations produced by protons at the nanodiamond/matrix interface.

Guidance of actin filament elongation on filament-binding tracks.

Biomolecular motors, which convert chemical energy into mechanical work in intracellular processes, have high potential in bionanotechnology in vitro as molecular shuttles or nanoscale actuators. In this context, guided elongation of actin filaments in vitro could be used to lay tracks for myosin motor-based shuttles or to direct nanoscale actuators based on actin filament end-tracking motors. To guide the direction of filament polymerization on surfaces, microcontact printing was used to create tracks of chemically modified myosin, which binds to, but cannot exert force on, filaments. These filament-binding tracks captured nascent filaments from solution and guided the direction of their subsequent elongation. The effect of track width and protein surface density on filament alignment and elongation rate was quantified. These results indicate that microcontact printing is a useful method for guiding actin filament polymerization in vitro for biomolecular motor-based applications.