Kevin M. Metz

Gastrointestinal biodurability of engineered nanoparticles: Development of an in vitro assay

The toxicity of engineered nanoparticles is expected to depend in part on their stability in biological systems. To assess the biodurability of engineered nanomaterials in the human digestive system, we adapted an in vitro assay previously used to evaluate the bioaccessibility of metals in contaminated soils. The compositions of the simulated gastric and intestinal fluids, temperature and residence times were designed to closely mimic conditions in the stomach and duodenum of the small intestine. We demonstrated the utility of the assay using CdSecore/ZnSshell quantum dots functionalized with polyethylene glycol (PEG) thiol of two different molecular masses (PEG350 and PEG5000). Under gastric conditions, removal of the PEG ligand diminished the stability of PEG350-quantum dot suspensions, while PEG5000-quantum dots were severely degraded. Inclusion of the glycoprotein mucin, but not the digestive protein pepsin, in simulated gastric fluids provided both PEG350- and PEG5000-coated quantum dots partial protection from transformations induced by gastric conditions.

Toxicity of Oxidatively Degraded Quantum Dots to Developing Zebrafish (Danio rerio)

Once released into the environment, engineered nanoparticles (eNPs) are subjected to processes that may alter their physical or chemical properties, potentially altering their toxicity vis-à-vis the as-synthesized materials. We examined the toxicity to zebrafish ( Danio rerio ) embryos of CdSecore/ZnSshell quantum dots (QDs) before and after exposure to an in vitro chemical model designed to simulate oxidative weathering in soil environments based on a reductant-driven Fentons reaction. Exposure to these oxidative conditions resulted in severe degradation of the QDs: the Zn shell eroded, Cd(2+) and selenium were released, and amorphous Se-containing aggregates were formed. Products of QD weathering exhibited higher potency than did as-synthesized QDs. Morphological endpoints of toxicity included pericardial, ocular and yolk sac edema, nondepleted yolk, spinal curvature, tail malformations, and craniofacial malformations. To better understand the selenium-like toxicity observed in QD exposures, we examined the toxicity of selenite, selenate, and amorphous selenium nanoparticles (SeNPs). Selenite exposures resulted in high mortality to embryos/larvae while selenate and SeNPs were nontoxic. Co-exposures to SeNPs + CdCl2 resulted in dramatic increase in mortality and recapitulated the morphological endpoints of toxicity observed with exposure to products of QD weathering. Cadmium body burden was increased in larvae exposed to weathered QDs or SeNP + CdCl2 suggesting the increased potency of products of QD weathering was due to selenium modulation of cadmium toxicity. Our findings highlight the need to examine the toxicity of eNPs after they have undergone environmental weathering processes.

Electrical characterization of nanowire bridges incorporating biomolecular recognition elements

We have investigated the formation and electrical properties of nanowire bridges formed when nanowires modified with the biomolecule biotin span across a gap between gold microelectrodes functionalized with the complementary biomolecule, avidin. Dielectrophoretic manipulation with a 1 MHz AC voltage is used to manipulate biotin-modified nanowires into the inter-electrode gap. Biomolecular recognition between the biotin-modified nanowires and the avidin-modified gold microelectrodes then holds the nanowires securely in place. By simultaneously applying a second, lower-frequency AC voltage and using lock-in detection, we are able to monitor individual bridging events in real time and to characterize the change in electrical response associated with individual nanowire bridges. The combined use of physical manipulation with biomolecular recognition can be used for selective assembly of nanoscale materials, as well as analytical application as a biologically activated switch in which an electrical contact is controlled by a biomolecular recognition process.