title: ORMOPLEXEs for gene therapy: In vitro and in vivo assays.
abstract: 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.
authors.affiliation: Departamento de Engenharia Química, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal; CQE, Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Portugal.
authors.forename: J C
authors.initials: JC
authors.lastname: Matos
authors.affiliation: Departamento de Biologia & CESAM, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
authors.forename: A R
authors.initials: AR
authors.lastname: Soares
authors.affiliation: Departamento de Biologia & CESAM, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
authors.forename: I
authors.initials: I
authors.lastname: Domingues
authors.affiliation: Departamento de Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisboa, Portugal; IBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Portugal.
authors.forename: G A
authors.initials: GA
authors.lastname: Monteiro
authors.affiliation: Departamento de Engenharia Química, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal; CQE, Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Portugal. Electronic address: clara.goncalves@ist.utl.pt.
authors.forename: M C
authors.initials: MC
authors.lastname: Gonçalves
mesh_terms: D000588:Amines / Q000737:chemistry; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D024202:Electrophoretic Mobility Shift Assay; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism; D018014:Gene Transfer Techniques; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure; D010316:Particle Size; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D018675:Toxicity Tests; D015027:Zebrafish / Q000254:growth & development
chemical_list: D000588:Amines; D012822:Silicon Dioxide
keywords: Gene therapy; Nanoparticles; Non-viral vector; ORMOPLEXE; ORMOSIL; Sol-gel; Transfection