Ricerc@Sapienza

Gene therapy represents a new technique for the treatment of cancers and diseases due to genomic defects or alterations. Although different approaches have been investigated, successful transport of foreign genetic materials through the cell membrane remains a major hurdle. Viral vehicles are the most commonly used carriers for DNA delivery but they need a precise controlled concentration to reduce toxicity and pathogenic effects. The micro-scaled environment of in vitro models could significantly improve gene transfection by eliminating diffusion issues associated with standard cell culture system but, due to long time experimental setting, it could result in cell nutrient deprivation potentially damaging cells. The development of a rapid and safe delivery technique maximizing efficiency, cell viability and reducing time, is crucial. Ultrasound and micro bubbles (USMB) together with microfluidic and carrier-based techniques can be combined to obtain the desired effect in shorter time. The main goal of this project is to investigate MBs stable cavitation effects on transfection efficiency, evoked by low-intensity US exposure, in a microfluidic platform. The insect baculovirus modified to vehicle actin-Green Fluorescent Protein (actin-GFP) is used in live cells for real time fluorescence imaging to visualize cellular cytoskeleton rearrangements. A flow containing growth media and high concentration of baculovirus is injected in the device seeded with Human Umbilical Vein Endothelial Cells (HUVECs). 2-hours break every 10 minutes is needed to let the virus explain its effect. The change in GFP expression over time is evaluated with a confocal microscope through a dedicated set-up in presence or not of US-activated MBs driven by 1MHz unfocused transducer. An increase of GFP transfection is expected with reduced viral-exposure time.