SERRS multiplexing with multivalent nanostructures for the identification and enumeration of epithelial and mesenchymal cells.

Liquid biopsy represents a new frontier of cancer diagnosis and prognosis, which allows the isolation of tumor cells released in the blood stream. The extremely low abundance of these cells needs appropriate methodologies for their identification and enumeration. Herein we present a new protocol based on surface enhanced resonance Raman scattering (SERRS) gold multivalent nanostructures to identify and enumerate tumor cells with epithelial and mesenchimal markers. The validation of the protocol is obtained with spiked samples of peripheral blood mononuclear cells (PBMC). Gold nanostructures are functionalized with SERRS labels and with antibodies to link the tumor cells. Three types of such nanosystems were simultaneously used and the protocol allows obtaining the identification of all individual tumor cells with the help of a Random Forest ensemble learning method.

Targeted killing of prostate cancer cells using antibody-drug conjugated carbon nanohorns.

The ability of carbon nanohorns (CNHs) to cross biological barriers makes them potential carriers for delivery purposes. In this work, we report the design of a new selective antibody-drug nanosystem based on CNHs for the treatment of prostate cancer (PCa). In particular, cisplatin in a prodrug form and the monoclonal antibody (Ab) D2B, selective for PSMA+ cancer cells, have been attached to CNHs due to the current application of this antigen in PCa therapy. The hybrids Ab-CNHs, cisplatin-CNHs and functionalised-CNHs have also been synthesized to be used as control systems. The efficacy and specificity of the D2B-cisplatin-CNH conjugate to selectively target and kill PSMA+ prostate cancer cells have been demonstrated in comparison with other derivatives. The developed strategy to functionalise CNHs is fascinating because it can allow the fine tuning of both drug and Ab molecules attached to the nanostructure in order to modulate the activity of the nanosystem. Finally, the herein described methodology can be used for the incorporation of almost any drugs or Abs in the platforms in order to create new targeted drugs for the treatment of different diseases.

Evidencing the mask effect of graphene oxide: a comparative study on primary human and murine phagocytic cells.

Graphene oxide (GO) is attracting an ever-growing interest in different fields and applications. Not much is known about the possible impact of GO sheet lateral dimensions on their effects in vitro, especially on human primary cells. In an attempt to address this issue, we present a study to evaluate, how highly soluble 2-dimensional GO constituted of large or small flakes affects human monocyte derived macrophages (hMDM). For this purpose, the lateral size of GO was tuned using sonication and three samples were obtained. The non sonicated one presented large flakes (~1.32 µm) while sonication for 2 and 26 hours generated small (~0.27 µm) and very small (~0.13 µm) sheets of GO, respectively. Cell studies were then conducted to evaluate the cytotoxicity, the oxidative stress induction, the activation potential and the pro-inflammatory effects of these different types of GO at increasing concentrations. In comparison, the same experiments were run on murine intraperitoneal macrophages (mIPM). The interaction between GO and cells was further examined by TEM and Raman spectroscopy. Our data revealed that the GO sheet size had a significant impact on different cellular parameters (i.e. cellular viability, ROS generation, and cellular activation). Indeed, the more the lateral dimensions of GO were reduced, the higher were the cellular internalization and the effects on cellular functionality. Our data also revealed a particular interaction of GO flakes with the cellular membrane. In fact, a GO mask due to the parallel arrangement of the graphene sheets on the cellular surface was observed. Considering the mask effect, we have hypothesized that this particular contact between GO sheets and the cell membrane could either promote their internalization or isolate cells from their environment, thus possibly accounting for the following impact on cellular parameters.