BioSInt Unit Superfici ed Interfacce biofunzionali Cecilia Pederzolli

1. BioSInt UnitSuperficiedInterfaccebiofunzionaliCecilia Pederzolli Istituto "Marie Curie" Pergine Eleonora Busana ChiaraCondler Stage estivi studenti 2009-2010

2. LaboratorioScienzeBiomolecularied Interfacce Centro Materiali e Microsistemi CNR Centro Nazionale Ricerche IBF Istituto di Biofisica M. Dalla Serra • Studio dei fenomeni che avvengono all’interfaccia - biosuperfici, biomembrane- • con duplice obiettivo: • Aumentare la comprensione dei meccanismi molecolari alla base delle malattie • Sviluppare sistemi innovativi di diagnosi e terapia (biomedicina) Settore di integrazione di diverse discipline: fisica, biologia, chimica

3. Biointerfaces The problem Materials for medical devices have been selected on the basis of mechanical and physical properties, with less consideration given to their interactions with the biological environment Poor interfacial biocompatibility Protein-protein interaction Cell Cell-protein layer interaction Protein layer BIOMATERIAL Surface properties play a crucial role protein adsorption, cell adhesion etc. are all surface-induce phenomena and depend on a large extent on the properties of the outer few atomic layers of materials

4. DNA surface immobilisation surface passivation + + + + + + + + ligand oligonucleotide blocking agent ligand spacer arm Surface modification processes to exercise a control over the way in which the biological fluids or individual biomolecules respond to the material surface 1) Reductionof the aspecificadhesion 2) Specificimmobilisationofbiomolecules Goal: Methods for covalent immobilization of biomolecules resulting in better biomolecule activity, reduced nonspecific adsorption, and greater stability.

5. Drug delivery systems no treatment RFMP treatment 3x3 m Lipid-based nano-structured materials cell membrane Partnership CNR-IBF Trento, MIT Boston Using micro nano-capsules (liposomes, particles) the transport and release of drugs and active molecules can be possible in a controlled way System - in solution - on implants surface Active principles - PFT bacterial toxins inhibitors - traditional antibacterial molecules - peptides Inhibition of bacterial grown Active multilayers deposited from liposomes

6. diagnose Sviluppo di materiali per la realizzazione di microsistemi per l’analisi del DNA in diagnostica molecolare time scale (minutes) <120 0 blood inheriteddiseases Infections by pathogens ... Eluted DNA Mix, Incubation & purification PCR & detection biological sample Mixer lysis buffer PCR mix surface

7. Molecular diagnostics Surface analysis and chemical modification of silicon-based materials for the development of a lab-on-chip capable to perform a genetic analysis starting from the extraction of genomic DNA directly from blood to the genetic identification of specific mutations and/or SNPs (single nucleotide polymorphism). DNA IN 1st channel Channels 2-9 DNA OUT 10th channel PicoGreen stained genomic DNA The system is a silicon/pyrex microchip (1x1cm, 6 µL total volume), where whole blood is injected (0.5 µL), DNA is extracted and used as a template for on-chip PCR. Products are revealed by fluorescence detection directly on chip. Partnership Olivetti Jet, Biodiversity, Telethon, Politecnico di Torino

8. Research overview Molecular diagnostics Carbon nanotube-based sensors for DNA detection SEM and TEM analysis • CNT – fluorescein π stacking interaction • ECL detection by means of Ru(bpy)32+labelled DNA 1. Realization of electric contact on CNT cylinder 2. CNT embedding in PDMS matrix 3. Thermal treatment at 100 °C 4. Electrode detaching from silicon support 5. Electric conductivity test Electrodes As expected, the maximum ECL intensity falls in correspondence of Ru(bpy)32+ electrochemical oxidation (+1.2 V vs. Ag) The first of three voltammetric cycles is indicated by solid line, while the subsequent two by dashed lines. The curve modifications from first to third cycle are indicated by the arrow directions Partnership Politecnico di Torino, University of Bologna

9. Il gruppo di ricerca Valeria Antonini Ramona Dallapiccola Lorenzo Lunelli Marta Marchioretto Lorenza Marocchi Laura Pasquardini Federica Piras Cristina Potrich Mayra Tejuca Laura Tosatto Michele Vinante Manuela Zanetti