title: Insonation facilitates plasmid DNA transfection into the central nervous system and microbubbles enhance the effect.
abstract: Many of the diseases which affect the central nervous system are intractable to conventional therapies and therefore require alternative treatments such as gene therapy. Therapy requires safety, since the central nervous system is a critical organ. Choice of nonviral vectors such as naked plasmid DNA may have merit. However, transfection efficiencies of these vectors are low. We have investigated the use of 210.4 kHz ultrasound and found that 5.0 W/cm(2) of insonation for 5 s most effectively transfected a plasmid DNA into culture slices of mouse brain (147.68-fold increase compared with 0 W/cm(2) of insonation for 5 s). The effect was reinforced by combination with echo contrast agent, Levovist. One hundred fifty mg/mL of Levovist significantly increased gene transfection by ultrasound (5.23-fold when insonated at 5.0 W/cm(2) for 5 s). When DNA was intracranially injected, Levovist also enhanced gene transfection in newborn mice (4.49-fold increase when insonated at 5.0 W/cm(2) for 5 s). Since ultrasound successfully transfected naked plasmid DNA into the neural tissue and Levovist enhanced the effect, this approach may have a significant role in gene transfer to the central nervous system.
authors.affiliation: Department of Molecular Cell Biology, Institute of DNA Medicine, Research Center for Medical Science, Jikei University School of medicine, 3-25-8 Nishishinbashi, Minato-ku, Tokyo, Japan 105-8461. manome@jikei.ac.jp
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Manome
authors.forename: Naoto
authors.initials: N
authors.lastname: Nakayama
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nakayama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Furuhata
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000648:ultrastructure; D016923:Cell Death; D003287:Contrast Media / Q000008:administration & dosage; D004247:DNA / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D051379:Mice; D008813:Mice, Inbred ICR; D045423:Microbubbles; D008855:Microscopy, Electron, Scanning / Q000379:methods; D009457:Neuroglia; D009474:Neurons; D010957:Plasmids; D011134:Polysaccharides / Q000008:administration & dosage; D046509:Tissue Culture Techniques / Q000379:methods; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics; D014465:Ultrasonics
chemical_list: D003287:Contrast Media; D011134:Polysaccharides; C059891:SHU 508; D004247:DNA