The activities of the Department of Biomedical and Biotechnological Sciences of the University of Catania

The Laboratory of Cellular and Molecular Physiology and the Pharmacology Laboratory of the Dipt BIOMETEC of the University of Catania (Unict) are both involved in preclinical studies, in vitro and in vivo, useful for evaluating the effectiveness of new generation LSD systems in comparison with the already patented LSD system.

In detail, the BIOMETEC research team is working on: 

• screening of different LSD systems with and without drugs in different immortalized and primary hepatocarcinoma cell lines;
• selection of the most effective LSD system; 
• analysis of the cellular and molecular mechanisms underlying the effectiveness of the new selected LSD system;
• generation of the mouse model of hepato-carcinoma;
• evaluation of the effects of the innovative LSD system compared to the previously tested system;
• analysis of the molecular mechanisms underlying the effectiveness of the new system;
• In vivo analyses of pharmacokinetics and toxicology, with particular emphasis to the evaluation of the minimum dose able to produce toxic effects and the maximum dose lacking of any effect.

Specifically, following the physicochemical characterization of the drug-delivery system conjugated with the anticancer drug Sorafenib, hepatocarcinoma cell cultures were set up in order to evaluate the biological effects of the selected best-performing LSD system.
For this purpose, three different hepato-carcinoma cell lines were set up to perform the various in vitro assays. The selected cell lines are two immortalized lines and one primary line obtained from human hepatocarcinoma. The latter will be isolated, cultured and characterized from histological pieces of hepato-carcinoma at the "G.F. Ingrassia" Department. 

The immortalized cell lines were treated with LSD systems conjugated or not with the drug Sorafenib. Following treatment, assays of: MTT cell viability, Caspase3/7 and Caspase9 assay were performed.

Pharmacological assays are also being planned to evaluate the effects of the next-generation LSD systems compared with the one already in use.After evaluating the efficacy of the LSD system in vitro, a mouse model of hepato-carcinoma is planned to be developed with one of the cell lines (both immoralized and primary) previously tested in vitro. 

Specifically, 5x106 hepatocarcinoma cells will be inoculated subcutaneously into immunodeficient nude mice of the NU(NCr)-Foxn1nu strain. As soon as the tumor mass has formed (5 mm), the animals will be divided into experimental groups and treated with the LSD system used in vitro. Following the treatments with the LSD systems, the animals will be monitored to assess the biological efficacy of the individual treatments. For this purpose, the presence and growth of tumor masses will be assessed through both palpation of the implantation zone and measurement with a digital micrometer caliper. 

In vivo studies will make use of PET/CT imaging analysis in order to study molecular mechanisms of response to treatment. In addition, animals will also be analyzed at defined intervals with additional in vivo preclinical imaging methods (Vevo ultrasound, Bruker in-vivo xtreme 2) in order to define the growth rate of the tumor mass, possible tumor spread, and lavascularization over time. Pharmacological and toxicological assays will be performed to evaluate the efficacy of individual treatments with the chosen next-generation LSD system.

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