Marina Massaro1, Giampaolo Barone1, Giuseppa Biddeci2, Giuseppe Cavallaro3, Francesco Di Blasi4, Giuseppe Lazzara3, Giuseppe Nicotra5, Corrado Spinella5, Gaetano Spinelli4, Serena Riela6. 1. Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy. 2. Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy; Institute of Biomedicine and Molecular Immunology, CNR, IBIM, Via Ugo La Malfa, 153, 90146 Palermo, Italy. 3. Dipartimento di Fisica e Chimica (DiFC), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy. 4. Institute of Biomedicine and Molecular Immunology, CNR, IBIM, Via Ugo La Malfa, 153, 90146 Palermo, Italy. 5. CNR-IMM, Zona Industriale Strada VIII, 5, 95121 Catania, Italy. 6. Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy. Electronic address: serena.riela@unipa.it.
Abstract
HYPOTHESIS: The use of non-viral vectors for gene therapy is hindered by their lower transfection efficiency and their lacking of self-track ability. EXPERIMENTS: This study aims to investigate the biological properties of halloysite nanotubes-carbon dots hybrid and its potential use as non-viral vector for oral gene therapy. The morphology and the chemical composition of the halloysite hybrid were investigated by means of high angle annular dark field scanning TEM and electron energy loss spectroscopy techniques, respectively. The cytotoxicity and the antioxidant activity were investigated by standard methods (MTS, DPPH and H2O2, respectively) using human cervical cancer HeLa cells as model. Studies of cellular uptake were carried out by fluorescence microscopy. Finally, we investigated the loading and release ability of the hybrid versus calf thymus DNA by fluorescence microscopy, circular dichroism, dynamic light scattering and ζ-potential measurements. FINDINGS: All investigations performed confirmed the existence of strong electrostatic interactions between the DNA and the halloysite hybrid, so it shows promise as a multi-functional cationic non-viral vector that has also possesses intracellular tracking capability and promising in vitro antioxidant potential.
HYPOTHESIS: The use of non-viral vectors for gene therapy is hindered by their lower transfection efficiency and their lacking of self-track ability. EXPERIMENTS: This study aims to investigate the biological properties of halloysite nanotubes-carbon dots hybrid and its potential use as non-viral vector for oral gene therapy. The morphology and the chemical composition of the halloysite hybrid were investigated by means of high angle annular dark field scanning TEM and electron energy loss spectroscopy techniques, respectively. The cytotoxicity and the antioxidant activity were investigated by standard methods (MTS, DPPH and H2O2, respectively) using human cervical cancerHeLa cells as model. Studies of cellular uptake were carried out by fluorescence microscopy. Finally, we investigated the loading and release ability of the hybrid versus calf thymus DNA by fluorescence microscopy, circular dichroism, dynamic light scattering and ζ-potential measurements. FINDINGS: All investigations performed confirmed the existence of strong electrostatic interactions between the DNA and the halloysite hybrid, so it shows promise as a multi-functional cationic non-viral vector that has also possesses intracellular tracking capability and promising in vitro antioxidant potential.