Dts1060 capillary cells

Chitosan-TPP nanoparticles complexed with pCpG-siRNA-Scramble or pCpG-sh2GDH were prepared following the ionic gelation method as described (González et al., 2016) . Three-hundred µg of pCpG-siRNA-Scramble or pCpG-sh2GDH were added to 1.2 ml of 0.84 mg/ml TPP.
Thereafter, TPP-DNA solutions were added dropwise to 3 ml of 2 mg/ml low molecular weight chitosan-acetate buffer (1:0.4 chitosan/TPP ratio). Chitosan-TPP-DNA nanoparticles were pelleted, rinsed twice with ultrapure water and resuspended in 2 ml of 2 % w/v mannitol as cryoprotector during lyophilisation. After a freeze-dry cycle at -47 °C, an additional drying step was performed at 25 °C to remove residual water. Chitosan-TPP-DNA nanoparticles were characterised by atomic force microscopy using peak force tapping mode (Multimode 8 AFM attached to a Nanoscope III Electronics, Bruker, USA). Z potential was determined using laser Doppler microelectrophoresis in a Zetasizer NanoZ equipped with DTS1060 capillary cells (Malvern Instruments, Malvern, UK).
Chitosan-TPP-DNA nanoparticles were resuspended in 0.9 % NaCl previous administration to S. aurata.

Morphological characterisation of the nanoparticles was performed by atomic force microscopy using peak force tapping mode (Multimode 8 AFM attached to a Nanoscope III Electronics, Bruker, USA). The Z potential values, as a measure of surface charge, were determined by means of laser Doppler microelectrophoresis using a Zetasizer NanoZ (Malvern Instruments, Malvern, UK) equipped with DTS1060 capillary cells (Malvern Instruments).