title: Efficient gene delivery to primary human retinal pigment epithelial cells: The innate and acquired properties of vectors.
abstract: The purpose of this study is designing non-viral gene delivery vectors for transfection of the primary human retinal pigment epithelial cells (RPE). In the design process of gene delivery vectors, considering physicochemical properties of vectors alone does not seem to be enough since they interact with constituents of the surrounding environment and hence gain new characteristics. Moreover, due to these interactions, their cargo can be released untimely or undergo degradation before reaching to the target cells. Further, the characteristics of cells itself can also influence the transfection efficacy. For example, the non-dividing property of RPE cells can impede the transfection efficiency which in most studies was ignored by using immortal cell lines. In this study, vectors with different characteristics differing in mixing orders of pDNA, PEI polymer, and PLGA/PEI or PLGA nanoparticles were prepared and characterized. Then, their characteristics and efficacy in gene delivery to RPE cells in the presence of vitreous or fetal bovine serum (FBS) were evaluated. All formulations showed no cytotoxicity and were able to protect pDNA from premature release and degradation in extracellular media. Also, the adsorption of vitreous or serum proteins onto the surface of vectors changed their properties and hence cellular uptake and transfection efficacy.
authors.affiliation: Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran. Electronic address: n_tasharrofi@yahoo.com.
authors.forename: Nooshin
authors.initials: N
authors.lastname: Tasharrofi
authors.affiliation: Stem Cell Technology Research Center, Tehran, Iran. Electronic address: f.kouhkan@stemcellstech.com.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Kouhkan
authors.affiliation: Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran. Electronic address: soleim_m@modares.ac.ir.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Institute of Medical Biotechnology, Faculty of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Electronic address: soheili@nigeb.ac.ir.
authors.forename: Zahra-Soheila
authors.initials: ZS
authors.lastname: Soheili
authors.affiliation: Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran; Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Atyabi
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran. Electronic address: akbarijo@tums.ac.ir.
authors.forename: Hamid
authors.initials: H
authors.lastname: Akbari Javar
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran; Medical Biomaterial Research Center (MBRC), Faculty of Pharmacy, Tehran University of Medical Science, Tehran, Iran. Electronic address: dorkoosh@tums.ac.ir.
authors.forename: Farid
authors.initials: F
authors.lastname: Abedin Dorkoosh
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D004847:Epithelial Cells / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D007223:Infant; D019344:Lactic Acid / Q000008:administration & dosage / Q000737:chemistry; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D011094:Polyethyleneimine / Q000008:administration & dosage / Q000737:chemistry; D011100:Polyglycolic Acid / Q000008:administration & dosage / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D055213:Retinal Pigment Epithelium / Q000166:cytology*
chemical_list: D003470:Culture Media; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011094:Polyethyleneimine; D004247:DNA
keywords: PLGA/PEI nanoparticles; Primary human RPE cells; Protein adsorption; Transfection; Vitreous