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The rapid uptake of nanomaterials in life sciences calls for the development of universal, high-yield techniques for their production and interfacing with biomolecules. Top-down methods take advantage of the existing variety of bulk and thin-film solid-state materials for improved prediction and control of the resultant nanomaterial properties. We demonstrate the power of this approach using high-energy ball milling (HEBM) of alumina (Al2O3). Nanoalumina particles with a mean size of 25 nm in their most stable α-crystallographic phase were produced in gram quantities, suitable for biological and biomedical applications. Nanomaterial contamination from zirconia balls used in HEBM was reduced from 19 to 2% using a selective acid etching procedure. The biocompatibility of the milled nanomaterial was demonstrated by forming stable colloids in water and physiological buffers, corroborated by zeta potentials of +40 mV and -40 mV and characterized by in vitro cytotoxicity assays. Finally, the feasibility of a milled nanoalumina surface in anchoring a host of functional groups and biomolecules was demonstrated by the functionalization of their surface using facile silane chemistry, resulting in the decoration of the nanoparticle surface with amino groups suitable for further conjugation of biomolecules.