ORMOPLEXEs for gene therapy: In vitro and in vivo assays

• NH2-ORMOSIL NPs were eco-friendly synthesized.
• pDNA:NH2-ORMOSIL NPs conjugation was confirmed by agarose gel electrophoretic mobility.
• ORMOPLEXEs exhibited cellular uptake and transfection properties in in vitro.
• ORMOPLEXEs biodistribution was studied in D. rerio embryos.
• NH2-ORMOSIL NPs showed no ecotoxicity in D. rerio eggs.

Gene therapy stays on the cutting edge of biomedical research, being the design of the optimal gene delivery vector one of the key requests. Silica-based nanoparticles (NPs) have emerged as promising non-viral gene delivery vector, due to their high biocompatibility, nontoxicity, non-immunogenicity, biodegradability and enormous bioconjugation versatility. In this work a sol-gel methodology for the synthesis of amino-functionalized silica NPs (NH2-ORMOSIL NPs) was optimized, and NPs were characterized by TEM and FTIR. In a first step NH2-ORMOSIL NPs were bioconjugated with a plasmid DNA, pVAX1-GFP, assembling an ORMOPLEXE, confirmed by agarose gel electrophoresis. In a second step, in vitro studies have been performed with cultured CHO cells, where ORMOPLEXEs transfection was proved by CLSM. In vivo transfection efficiency and bio-distribution were performed in Zebrafish (Danio rerio) embryos, assessed by FM. Finally, NPs ecotoxicity was studied in zebrafish embryos by following the mortality and developmental endpoints.

Figure optionsDownload as PowerPoint slide