title: Chondroitin sulfate-polyethylenimine copolymer-coated superparamagnetic iron oxide nanoparticles as an efficient magneto-gene carrier for microRNA-encoding plasmid DNA delivery.
abstract: MicroRNA-128 (miR-128) is an attractive therapeutic molecule with powerful glioblastoma regulation properties. However, miR-128 lacks biological stability and leads to poor delivery efficacy in clinical applications. In our previous study, we demonstrated two effective transgene carriers, including polyethylenimine (PEI)-decorated superparamagnetic iron oxide nanoparticles (SPIONs) as well as chemically-conjugated chondroitin sulfate-PEI copolymers (CPs). In this contribution, we report optimized conditions for coating CPs onto the surfaces of SPIONs, forming CPIOs, for magneto-gene delivery systems. The optimized weight ratio of the CPs and SPIONs is 2 : 1, which resulted in the formation of a stable particle as a good transgene carrier. The hydrodynamic diameter of the CPIOs is ∼136 nm. The gel electrophoresis results demonstrate that the weight ratio of CPIO/DNA required to completely encapsulate pDNA is ≥3. The in vitro tests of CPIO/DNA were done in 293 T, CRL5802, and U87-MG cells in the presence and absence of an external magnetic field. The magnetofection efficiency of CPIO/DNA was measured in the three cell lines with or without fetal bovine serum (FBS). CPIO/DNA exhibited remarkably improved gene expression in the presence of the magnetic field and 10% FBS as compared with a gold non-viral standard, PEI/DNA, and a commercial magnetofection reagent, PolyMag/DNA. In addition, CPIO/DNA showed less cytotoxicity than PEI/DNA and PolyMag/DNA against the three cell lines. The transfection efficiency of the magnetoplex improved significantly with an assisted magnetic field. In miR-128 delivery, a microRNA plate array and fluorescence in situ hybridization were used to demonstrate that CPIO/pMIRNA-128 indeed expresses more miR-128 with the assisted magnetic field than without. In a biodistribution test, CPIO/Cy5-DNA showed higher accumulation at the tumor site where an external magnet is placed nearby.
authors.affiliation: Department of Medicinal & Applied Chemistry, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan. lfwang@kmu.edu.tw.
authors.forename: Yu-Lun
authors.initials: YL
authors.lastname: Lo
authors.forename: Han-Lin
authors.initials: HL
authors.lastname: Chou
authors.forename: Zi-Xian
authors.initials: ZX
authors.lastname: Liao
authors.forename: Shih-Jer
authors.initials: SJ
authors.lastname: Huang
authors.forename: Jyun-Han
authors.initials: JH
authors.lastname: Ke
authors.forename: Yu-Sheng
authors.initials: YS
authors.lastname: Liu
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Chiu
authors.forename: Li-Fang
authors.initials: LF
authors.lastname: Wang
mesh_terms: D002809:Chondroitin Sulfates / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis; D003911:Dextrans / Q000737:chemistry* / Q000648:ultrastructure; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008422:Materials Testing; D035683:MicroRNAs / Q000235:genetics*; D053769:Nanocapsules / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D011094:Polyethyleneimine / Q000737:chemistry; D014162:Transfection / Q000379:methods*
chemical_list: D020099:Coated Materials, Biocompatible; D003911:Dextrans; D058185:Magnetite Nanoparticles; D035683:MicroRNAs; D053769:Nanocapsules; C097921:ferumoxtran-10; D011094:Polyethyleneimine; D002809:Chondroitin Sulfates