We show that the cytotoxicity of water-soluble fullerene species is a sensitive function of surface derivatization; in two different human cell lines, the lethal dose of fullerene changed over 7 orders of magnitude with relatively minor alterations in fullerene structure. In particular, an aggregated form of C60, the least derivatized of the four materials, was substantially more toxic than highly soluble derivatives such as C3, Na+ 2 - 3[C60O7 - 9(OH)12 - 15](2-3)-, and C60(OH)24. Oxidative damage to the cell membranes was observed in all cases where fullerene exposure led to cell death. We show that under ambient conditions in water fullerenes can generate superoxide anions and postulate that these oxygen radicals are responsible for membrane damage and subsequent cell death. This work demonstrates both a strategy for enhancing the toxicity of fullerenes for certain applications such as cancer therapeutics or bactericides, as well as a remediation for the possible unwarranted biological effects of pristine fullerenes.

Upon contact with water, under a variety of conditions, C60 spontaneously forms a stable aggregate with nanoscale dimensions (d = 25−500 nm), termed here “nano-C60”. The color, hydrophobicity, and reactivity of individual C60 are substantially altered in this aggregate form. Herein, we provide conclusive lines of evidence demonstrating that in solution these aggregates are crystalline in order and remain as underivatized C60 throughout the formation/stabilization process that can later be chemically reversed. Particle size can be affected by formation parameters such as rates and the pH of the water addition. Once formed, nano-C60 remains stable in solution at or below ionic strengths of 0.05 I for months. In addition to demonstrating aggregate formation and stability over a wide range of conditions, results suggest that prokaryotic exposure to nano-C60 at relatively low concentrations is inhibitory, indicated by lack of growth (≥0.4 ppm) and decreased aerobic respiration rates (4 ppm). This work demonstrates the fact that the environmental fate, distribution, and biological risk associated with this important class of engineered nanomaterials will require a model that addresses not only the properties of bulk C60 but also that of the aggregate form generated in aqueous media.

While the reactivity of C60 has been described in a variety of organic solvents, little information is available regarding aqueous-based reactions due to solubility limitations. In this study, a reaction between C60, as a nanoscale suspension, and dissolved ozone in the aqueous phase was investigated. Findings indicate a facile reaction occurs, resulting in aggregate dissolution concurrent with formation of water-soluble fullerene oxide species. Product analyses, including 13C NMR, MS (LDI), FTIR, UV-Vis, and XPS, indicate highly oxidized fullerene with an average of approximately 29 oxygen additions per molecule, arranged in repeating hydroxyl and hemiketal functionalities. These findings are significant in that they (1) demonstrate the feasibility of other aqueous-based fullerene chemistries, including those for alternative synthesis routes, which might otherwise be considered prohibitive on the basis of solubility limitations, and (2) imply that the aqueous reactivity of fullerene-based materials must be considered appropriately for accurate assessment of their transport, fate, and potential risk(s) in environmental systems.