title: Ultrapure chitosan oligomers as carriers for corneal gene transfer.
abstract: NOVAFECT chitosans are ultrapure chitosan oligomers that were recently marketed as carriers for non-viral gene therapy. There are no reports on systematic design and improvement of formulations based on NOVAFECT chitosans for gene delivery. Therefore, we have designed and characterized chitosan-DNA nanoparticles based on NOVAFECT. We found that the size of oligomeric chitosan-DNA nanoparticles is small, <or=98.2+/-4.4 nm. Zeta potential measurements of oligomeric chitosan-DNA nanoparticles exhibited a strong positive charge >or=44.1+/-3.5 millivolt. In vitro transfection studies demonstrated the ability of oligomeric chitosan-DNA nanoparticles to effectively transfect COS-7 cells. In rat corneas, injection of a select formulation of oligomeric chitosan-DNA nanoparticles into the stroma showed that (a) luciferase gene expression was 5.4 times greater than following administration of polyethylenimine-DNA nanoparticles; and (b) the cells that express the transgene, green fluorescent protein, were keratocytes (corneal fibroblasts). This study lays the foundation for evaluating oligomeric chitosan-DNA nanoparticles as pharmaceuticals for corneal gene therapy, a promising approach for the treatment of acquired and inherited corneal diseases that otherwise lead to blindness. Oligomeric chitosan-DNA nanoparticles can also be evaluated for the treatment of ocular diseases outside of the cornea, and for various additional gene therapy applications.
authors.affiliation: Department of Pharmaceutical Sciences, Midwestern University Chicago College of Pharmacy, Downers Grove, IL 60515, USA. eklausner@midwestern.edu
authors.forename: Eytan A
authors.initials: EA
authors.lastname: Klausner
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zhang
authors.forename: Robert L
authors.initials: RL
authors.lastname: Chapman
authors.forename: Richard F
authors.initials: RF
authors.lastname: Multack
authors.forename: Michael V
authors.initials: MV
authors.lastname: Volin
mesh_terms: D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D002522:Chlorocebus aethiops; D003315:Cornea / Q000378:metabolism*; D004247:DNA / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry; D010316:Particle Size; D051381:Rats; D014162:Transfection
chemical_list: D004337:Drug Carriers; D049452:Green Fluorescent Proteins; D004247:DNA; D048271:Chitosan; D008156:Luciferases