title: Cellular uptake and fate of fibroin microspheres loaded with randomly fragmented DNA in 3T3 cells.
abstract: Purified fibroin protein can be obtained in large quantities from silk fibers and processed to form microscopic particles as delivery vehicles for therapeutic agents. In this study, we demonstrated that fibroin microspheres were taken up by 3T3 cells, localized in the nonlysosomal compartment, and secreted from the cytoplasm after medium replenishment. DNA-loaded microspheres were taken up by >95% of 3T3 cells. DNA cargo had no influence on the intracellular trafficking of microspheres, while fluorescently labeled cargo DNA was observed in the lysosomal compartment and in the microspheres. These results indicate that fibroin microspheres can travel through 3T3 cells without making any contact with the lysosomal compartments. The amount of DNA loaded in the microspheres taken up by 3T3 cells was estimated up to 831.0 pg/cell. Thus, fibroin microspheres can deliver a large amount of randomly fragmented DNA (<10 kb) into the cytoplasmic compartment of 3T3 cells.
authors.affiliation: Department of Bioengineering and Technology, Kangwon National University, Chuncheon, South Korea.
authors.forename: Jin Sil
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Department of Bioengineering and Technology, Kangwon National University, Chuncheon, South Korea.
authors.forename: Won
authors.initials: W
authors.lastname: Hur
mesh_terms: D016475:3T3 Cells; D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D005349:Fibroins / Q000737:chemistry / Q000493:pharmacokinetics*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry / Q000493:pharmacokinetics; D008247:Lysosomes; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D011897:Random Allocation; D013347:Subcellular Fractions
chemical_list: D004247:DNA; D005349:Fibroins; D016650:Fluorescein-5-isothiocyanate
keywords: DNA delivery; fibroin; microspheres; transcellular transport