In vitro and in vivo systems toxicity analysis

Biosafety Level 2 laboratory is equipped with laminar airflow biological safety cabinet apparatus, incubator systems, thermoregulated water bath and inverted microscope.

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The cell lines in use are: alveolar type II human cells (A549), human bronchial cells (BEAS-2B), human epithelial cells of the lung (NCI-H441), endothelial cell lines (HPMEC-ST1.6R), monocytic cells (THP-1), human continuous epithelial cells (A431, A30), glioma cells (C6), primary cultures of nervous cells (granular and ippocampus rat cells).

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The biological parameters analysed are: cell viability (MTT and Alamar Blue Assay, LDH test), ROS production (DCFH), inflammatory proteins release (ELISA), DNA damage, cell cycle alterations, specific protein expressions (Western blot).

At present the research is oriented to the evaluation of the biological effects produced by: particulate matter (PM10, PM2.5; PM1 and Ultrafine Particles), emissions from materials of tyre productions, nanomaterials of industrial origin and metal oxides nanoparticles.

In vivo systems (mice) are analysed in an animal breeding lab equipped with appropriate instruments for experiments on wild type and genetically modified mice.

The facilities are: a MicroSprayer® Aerosolizer (Model IA-1C FMJ-250) High Pressure Syringe (Penn Century, USA); ImageQuant LAS4000 (GE); Ultracentrifuge Optima L90K (Beckman Coulter); Spettrofotometer UV-visible (JASCO V-530 UV-VIS); Beta-Imager 2000Z (Biospace).

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Mice are used to study ambient particle toxicity induced by different PM fractions.

At present the effects produced on the respiratory, cardiovascular and nervous systems are under analysis through the evaluation of: inflammatory proteins release (cytokines and chemokines, Hsp70, MPO); citotoxicity induction (Alcalin phosphatase, LDH, Caspasis), oxidative stress markers (SOD, GSH/GSS, HO-1, iNOS); endothelial dysfunction activation (PAI-1, fibrinogen, ET-1, sP-selectin, angiotensin II).