Peroxidase-induced degradation of single-walled carbon nanotubes: hypochlorite is a major oxidant capable of in vivo degradation of carbon nanotubes

Abstract

Due to their extraordinary properties, single-walled carbon nanotubes (SWNTs) have a tremendous potential for medical applications such as clinical diagnostics, targeted drug (or gene) delivery and cancer therapy. Hence, effects of SWNTs on living systems as well as mechanisms for biodegradation of SWTNs are of great importance and must be studied before starting to explore SWNTs for medical use. This study was undertaken to compare the potential of different peroxidases in degrading carboxylated SWNT (c-SWNT) and to elucidate the role of peroxidase-generated reactive products in this process. A detailed study showed that neither reactive intermediate products nor free radicals generated via peroxidase cycle can considerably oxidize c-SWNT. Biodegradation of c-SWNT in model system can be induced by free radicals generated as a result of heme degradation. The latter explains why hemoglobin, which is a pseudo-peroxidase possessing low peroxidase activity, is able to oxidize carbon nanotubes with a higher efficiency than horseradish peroxidase. However, c-SWNT in the presence of blood plasma (15 vol %) demonstrated no degradation even at high concentrations of hemoglobin and H2O2. The comparison of the ability of various peroxidases to degrade SWNTs in vitro revealed that MPO, due to its ability to produce hypochlorite, and lactoperoxidase, due to its ability to produce hypobromite, are extremely efficient in degrading carbon nanotubes. Since neutrophils are a main source of human MPO, we tested the effect of SWNTs on these cells. SWNTs were unable to stimulate neutrophils. On the other hand, they dose-dependently enhanced opsonized zymosan-induced cell stimulation as detected by measuring the amount of hypochlorite produced. This finding may be relevant to the in vivo situation, for example, at inflammatory sites. In order to imitate conditions characteristic of phagosomes and inflammatory sites, we titrated the suspension of c-SWNT in the presence of diluted blood plasma at pH 5.8 with high concentrations of (MPO + H2O2) or hypochlorite and found significant degradation of nanotubes. Collectively, our results indicate that hypochlorite is a main candidate for oxidative degradation of carbon nanotubes in vivo.