Dipartimento di Scienza e Tecnologia del Farmaco

Descrizione del progetto

Membri:

Dott. Gloria Berlier  (Dip. Chimica) 
Dott. Ivana Fenoglio   (Dip. Chimica) 

Partners:
Department of Chemistry, University of Turin, Italy 

Description

In recent years a large number of new nanomaterials for a wide range of application have been introduced in the market. Concerns exist about possible adverse health effects following human exposure to nanomaterials. Moreover, most of the nanoparticles are able to generate Reactive Oxygen Species (ROS), both as particle-derived and as cell-derived radicals, and thus oxidative stress constitutes one of the principal injury mechanisms caused by engineered nanoparticles.

The aim of this project is to study the potential toxicity of different nanomaterials (i.e. titanium dioxide, carbon soot and amorphous silica nanoparticles) on biological system. The damage to cells, DNA, proteins and lipids is evaluated by:

1)      in vitro cytotoxicity screening

2)      plasmid DNA strand breaks. The potential induction of DNA damage is checked by the conversion of supercoiled plasmid double-stranded DNA to open circular and linear form

3)      the 8-hydroxy-2’deoxyguanosine (8-OHdG) formation, a biomarker of the oxidative stress of DNA

4)      the denaturation, aggregation, fragmentation and oxidation of proteins

5)      the effect on the expression and activity of proteins and enzymes belonging to stress-activated signalling cascade

6)      the formation of 4-hydroxy-nonenal, final product of advanced lipoxidation

 Techiques and facilities:

• Human cell culture
• Colorimetric assays
• Cytotoxicity assays in cell cultures (sulforhodamine B or MTT)
• DNA agarose gel electrophoresis
• Protein electrophoresis  
• Immunoassays (western Blot, ELISA)
• UV-Vis spectroscopy

Keywords:  Human cell culture, Nanoparticles, Interaction cell-nanoparticles, Cytotoxicity, ROS, Stress oxidative, DNA damage, Protein damage, Lipid damage