Tomato Seed Coat Permeability to Selected Carbon Nanomaterials and Enhancement of Germination and Seedling Growth.

Seed coat permeability was examined using a model that tested the effects of soaking tomato (Solanum lycopersicon) seeds in combination with carbon-based nanomaterials (CBNMs) and ultrasonic irradiation (US). Penetration of seed coats to the embryo by CBNMs, as well as CBNMs effects on seed germination and seedling growth, was examined. Two CBNMs, C60(OH)20 (fullerol) and multiwalled nanotubes (MWNTs), were applied at 50 mg/L, and treatment exposure ranged from 0 to 60 minutes. Bright field, fluorescence, and electron microscopy and micro-Raman spectroscopy provided corroborating evidence that neither CBNM was able to penetrate the seed coat. The restriction of nanomaterial (NM) uptake was attributed to the semipermeable layer located at the innermost layer of the seed coat adjacent to the endosperm. Seed treatments using US at 30 or 60 minutes in the presence of MWNTs physically disrupted the seed coat; however, the integrity of the semipermeable layer was not impaired. The germination percentage and seedling length and weight were enhanced in the presence of MWNTs but were not altered by C60(OH)20. The combined exposure of seeds to NMs and US provided insight into the nanoparticle-seed interaction and may serve as a delivery system for enhancing seed germination and early seedling growth.

Biophysical methods for assessing plant responses to nanoparticle exposure.

As nanotechnology rapidly emerges into a new industry-driven by its enormous potential to revolutionize electronics, materials, and medicine-exposure of living species to discharged nanoparticles has become inevitable. Despite the increased effort on elucidating the environmental impact of nanotechnology, literature on higher plants exposure to nanoparticles remains scarce and often contradictory. Here we present our biophysical methodologies for the study of carbon nanoparticle uptake by Allium cepa cells and rice plants. We address the three essential aspects for such studies: identification of carbon nanoparticles in the plant species, quantification of nanotransport and aggregation in the plant compartments, and evaluation of plant responses to nanoparticle exposure on the cellular and organism level. Considering the close connection between plant and mammalian species in ecological systems especially in the food chain, we draw a direct comparison on the uptake of carbon nanoparticles in plant and mammalian cells. In addition to the above studies, we present methods for assessing the effects of quantum dot adsorption on algal photosynthesis.

Cytoprotective properties of a fullerene derivative against copper.

To delineate the complexity of the response of cells to nanoparticles we have performed a study on HT-29 human colon carcinoma cells exposed first to a fullerene derivative C(60)(OH)(20) and then to physiological copper ions. Our cell viability, proliferation, and intracellular reactive oxygen species (ROS) production assays clearly indicated that C(60)(OH)(20) suppressed cell damage as well as ROS production induced by copper, probably through neutralization of the metal ions by C(60)(OH)(20) in the extracellular space, as well as by adsorption and uptake of the nanoparticles surface-modified by the biomolecular species in the cell medium. This double-exposure study provides new data on the effects of nanoparticles on cell metabolism and may aid the treatment of oxidant-mediated diseases using nanomedicine.

In vitro polymerization of microtubules with a fullerene derivative.

Fullerene derivative C(60)(OH)(20) inhibited microtubule polymerization at low micromolar concentrations. The inhibition was mainly attributed to the formation of hydrogen bonding between the nanoparticle and the tubulin heterodimer, the building block of the microtubule, as evidenced by docking and molecular dynamics simulations. Our circular dichroism spectroscopy measurement indicated changes in the tubulin secondary structures, while our guanosine-5'-triphosphate hydrolysis assay showed hindered release of inorganic phosphate by the nanoparticle. Isothermal titration calorimetry revealed that C(60)(OH)(20) binds to tubulin at a molar ratio of 9:1 and with a binding constant of 1.3 ± 0.16 × 10(6) M(-1), which was substantiated by the binding site and binding energy analysis using docking and molecular dynamics simulations. Our simulations further suggested that occupancy by the nanoparticles at the longitudinal contacts between tubulin dimers within a protofilament or at the lateral contacts of the M-loop and H5 and H12 helices of neighboring tubulins could also influence the polymerization process. This study offered a new molecular-level insight on how nanoparticles may reshape the assembly of cytoskeletal proteins, a topic of essential importance for illuminating cell response to engineered nanoparticles and for the advancement of nanomedicine.

Experimental and simulation studies of a real-time polymerase chain reaction in the presence of a fullerene derivative.

The real-time polymerase chain reaction of the plant gene heat shock transcription factor was fully inhibited in the presence of a fullerene derivative C(60)(OH)(20) at a concentration of 4 x 10(-4) mM. This inhibition was attributed to the interaction between the nanoparticle and Taq DNA polymerase in the reaction. Atomistic molecular dynamics simulations showed a clear tendency for hydrogen bonding between C(60)(OH)(20) and both the dNTPs and ssDNA components of the polymerase chain reaction. These studies facilitate our understanding of the fate of nanoparticles in biomolecular systems, a topic of tremendous importance for addressing the biological and environmental implications of nanomaterials.