title: Construction of PLGA Nanoparticles Coated with Polycistronic SOX5, SOX6, and SOX9 Genes for Chondrogenesis of Human Mesenchymal Stem Cells.
abstract: Transfection of a cocktail of genes into cells has recently attracted attraction in stem cell differentiation. However, it is not easy to control the transfection rate of each gene. To control and regulate gene delivery into human mesenchymal stem cells (hMSCs), we employed multicistronic genes coupled with a nonviral gene carrier system for stem cell differentiation. Three genes, SOX5, SOX6, and SOX9, were successfully fabricated in a single plasmid. This multicistronic plasmid was complexed with the polycationic polymer polyethylenimine, and poly(lactic-co-glycolic) acid (PLGA) nanoparticles were coated with this complex. The uptake of PLGA nanoparticles complexed with the multicistronic plasmid was tested first. Thereafter, transfection of SOX5, SOX6, and SOX9 was evaluated, which increased the potential for chondrogenesis of hMSCs. The expression of specific genes triggered by transfection of SOX5, SOX6, and SOX9 was tested by RT-PCR and real-time qPCR. Furthermore, specific proteins related to chondrocytes were investigated by a glycosaminoglycan/DNA assay, Western blotting, histological analyses, and immunofluorescence staining. These methods demonstrated that chondrogenesis of hMSCs treated with PLGA nanoparticles carrying this multicistronic genes was better than that of hMSCs treated with other carriers. Furthermore, the multicistronic genes complexed with PLGA nanoparticles were more simple than that of each single gene complexation with PLGA nanoparticles. Multicistronic genes showed more chondrogenic differentiation than each single gene transfection methods.
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Ji Sun
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Se Won
authors.initials: SW
authors.lastname: Yi
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Seong Min
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Jae-Hwan
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University , 6F, CHA Bio-complex, 689 Sampyeong-dong Bundang-gu, Seongnam-si 134-88, Korea.
authors.forename: Keun-Hong
authors.identifier: http://orcid.org/0000-0002-3489-2890
authors.initials: KH
authors.lastname: Park
mesh_terms: D002454:Cell Differentiation; D019902:Chondrocytes; D020219:Chondrogenesis; D006801:Humans; D059630:Mesenchymal Stem Cells; D053758:Nanoparticles; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D055758:SOX9 Transcription Factor; D055755:SOXD Transcription Factors
chemical_list: C490301:SOX5 protein, human; C486270:SOX6 protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D055755:SOXD Transcription Factors; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
keywords: PEI; SOX9; hMSC; nanoparticle; polycistronic vector