R.U.2 TUSCIA UNIVERSITY - VITERBO

The activities of RU2 will focus on the biological characterization of extracts prepared with NADES or with conventional solvents and procedures. This will help define the functional properties of the extracts and their potential applications in various fields. 

Cytotoxic and antiproliferative activities will be carried out on human cell lines, which represent the best in vitro study models for various types of diseases, such as breast adenocarcinoma cells (MCF-7 and MDA), neuroblastoma (SH-SY5Y), colorectal adenocarcinoma (Caco-2), and acute leukemia (CCRF-CEM and HL60). A normal cell line (MCF 10A) will be used as a control. The first approach will evaluate the effects of extracts and fractions on cell viability using the MTT assay.

The anti-inflammatory effects of the extracts will also be studied in the Caco-2 cell line using flow cytometry focused on cytokine secretion and the synthesis of Interleukin-8 (IL-8), Interleukin-1ß (IL-1ß), Interleukin-6 (IL-6), Interleukin-10 (IL-10), tumour necrosis factor (TNF), and interleukin-12p70 (IL-12p70). In addition, morphological alterations induced in cell models will be evaluated by optical microscopy, fluorescence microscopy, and transmission and scanning electron microscopy. Antibodies against specific proteins and fluorescent dyes for cellular targets will be used in immunofluorescence methodologies, and electron microscopy techniques will define cell-cell interactions and morphological and ultrastructural alterations of cellular components.

Antibacterial activity against certain environmental and pathogenic bacteria will also be studied, using five different strains of environmental bacteria and Erwinia amylovora as a phytopathogenic bacterium, defining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for each of them, according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI).