Cellular responses induced by multi-walled carbon nanotubes: in vivo and in vitro studies on the medicinal leech macrophages

The core characteristics of multi-wall carbon nanotubes (MWCNTs) are impressive and attractive for technology however, since their production and use is steadily increasing, their environmental dispersion could be potentially hazardous to animal and human health. For this reason, the identification of new methods and of reliable models to better understand MWCNT effects is essential. Here we propose the medicinal leech as an alternative model to assess the effects of MWCNTs on immune system. Our previous studies have already demonstrated that in vivo MWCNT treatment induces the activation of leech’s macrophages. Here we will focus on the direct effects of MWCNTs on these cells by isolating and culturing leech's macrophages by means of the consolidated Matrigel technique, followed by MWCNT in vitro treatment. Our results indicate that MWCNT administration causes both the decrease of cell proliferation rate and the increase of the apoptotic rate. Furthermore, since oxidative stress is linked with inflammation, reactive oxygen species has been evaluated confirming that their production rate increases after MWCNT treatment. Our experimental approaches demonstrate the ability of MWCNTs inducing a powerful inflammatory response and confirm that the medicinal leech is a good alternative model to study the possible harmful effects of any nanomaterial.