title: Muscle gene electrotransfer is increased by the antioxidant tempol in mice.
abstract: Electropermeabilization (EP) is an effective method of gene transfer into different tissues. During EP, reactive oxygen species (ROS) are formed, which could affect transfection efficiency. The role of generated ROS and the role of antioxidants in electrotransfer in myoblasts in vitro and in Musculus tibialis cranialis in mice were, therefore, investigated. We demonstrate in the study that during EP of C2C12 myoblasts, ROS are generated on the surface of the cells, which do not induce long-term genomic DNA damage. Plasmid DNA for transfection (pEGFP-N1), which is present outside the cells during EP, neutralizes the generated ROS. The ROS generation is proportional to the amplitude of the electric pulses and can be scavenged by antioxidants, such as vitamin C or tempol. When antioxidants were used during gene electrotransfer, the transfection efficiency of C2C12 myoblasts was statistically significantly increased 1.6-fold with tempol. Also in vivo, the transfection efficiency of M. tibialis cranialis in mice was statistically significantly increased 1.4-fold by tempol. The study indicates that ROS are generated on cells during EP and can be scavenged by antioxidants. Specifically, tempol can be used to improve gene electrotransfer into the muscle and possibly also to other tissues.
authors.affiliation: Department of Experimental Oncology, Institute of Oncology Ljubljana, Ljubljana, Slovenia.
authors.forename: B
authors.initials: B
authors.lastname: Markelc
authors.forename: G
authors.initials: G
authors.lastname: Tevz
authors.forename: M
authors.initials: M
authors.lastname: Cemazar
authors.forename: S
authors.initials: S
authors.lastname: Kranjc
authors.forename: J
authors.initials: J
authors.lastname: Lavrencak
authors.forename: B
authors.initials: B
authors.lastname: Zegura
authors.forename: J
authors.initials: J
authors.lastname: Teissie
authors.forename: G
authors.initials: G
authors.lastname: Sersa
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology* / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival; D003497:Cyclic N-Oxides / Q000494:pharmacology* / Q000633:toxicity; D018274:Electroporation / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000378:metabolism*; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013113:Spin Labels
chemical_list: D000975:Antioxidants; D003497:Cyclic N-Oxides; D017382:Reactive Oxygen Species; D013113:Spin Labels; C001803:tempol