Study of cytotoxic and therapeutic effects of stable and purified silver nanoparticles on tumor cells.

We have synthesized and purified silver nanoparticles (Ag NPs) (11.3+/-2.3 nm) that are stable (non-aggregated) in cell culture medium and inside single living cells. We have developed new imaging methods to characterize sizes and number of single NPs in the medium and in single living cells in real-time and determine their stability (non-aggregation) in the medium and in single living cells at single NP resolution. These new approaches allow us to study toxic and therapeutic effects of single Ag NPs on tumor cells (L929, mouse fibroblast cells) with determined sizes and concentrations (doses) of NPs over time at single NP and single cell resolution. We found that Ag NPs inhibited the growth and division of tumor cells and their nuclei, in a dose and time dependent manner, showing significant inhibitory effects and abnormal cells with giant undivided nuclei or multiple nuclei beyond 12 h incubation. The results show that Ag NPs inhibited the segregation of chromosomes, but not their replication. Intracellular Ag NPs were well distributed in the cell population, and located in the nuclei and cytoplasm with higher numbers in the cytoplasm. This study demonstrates the possibility of using Ag NPs to inhibit the growth and division of tumor cells and using their cytotoxicity for potential therapeutic treatments. This study offers a new method to count the number of single NPs in the medium for characterization of their concentration and stability at single NP resolution over time.