title: Co-delivery of SOX9 genes and anti-Cbfa-1 siRNA coated onto PLGA nanoparticles for chondrogenesis of human MSCs.
abstract: Some genes expressed in stem cells interrupt and/or enhance differentiation. Therefore, the aim of this study was to inhibit the expression of unnecessary genes and enhance the expression of specific genes involved in stem cell differentiation by using small interfering RNA (siRNA) and plasmid DNA (pDNA) incorporated into cationic polymers as co-delivery factors. To achieve co-delivery of siRNA and pDNA to human mesenchymal stem cells (hMSCs), two different genes were complexed with poly(ethyleneimine) (PEI) and then coated onto poly(lactide-co-glycolic acid) (PLGA) nanoparticles (NP). To evaluate co-delivery of siRNA and pDNA into hMSCs, cells were transfected with green fluorescence protein (GFP) pDNA (GFP pDNA) and GFP siRNA (GFP siRNA). The percentage of GFP-expressing hMSCs decreased from 25.35 to 3.7% after transfection with GFP-DNA/PLGA NP (NPs) or GFP siRNA/PLGA NPs, whereas GFP-DNA/PLGA NPs and scramble siRNA (MOCK)/PLGA NPs had no effect on GFP expression. hMSCs cotransfected with coSOX9-pDNA/NPs and Cbfa-1-siRNA/NPs were tested both in vitro and in vivo using gel retardation, dynamic light scattering (DLS), and scanning electron microscope (SEM). The expression of genes and proteins associated with chondrogenesis was evaluated by FACS, RT-PCR, real time-qPCR, Western blotting, immunohistochemistry, and immunofluorescence imaging.
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, 3F, Yatap Acecore, 502 Yatap-dong Bundang-gu, Seongnam-si, Republic of Korea.
authors.forename: Su Yeon
authors.initials: SY
authors.lastname: Jeon
authors.forename: Ji Sun
authors.initials: JS
authors.lastname: Park
authors.forename: Han Na
authors.initials: HN
authors.lastname: Yang
authors.forename: Dae Gyun
authors.initials: DG
authors.lastname: Woo
authors.forename: Keun-Hong
authors.initials: KH
authors.lastname: Park
mesh_terms: D000818:Animals; D001673:Biodegradation, Environmental; D015153:Blotting, Western; D016923:Cell Death; D020219:Chondrogenesis; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry*; D008027:Light; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D034741:RNA, Small Interfering / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D012542:Scattering, Radiation; D014162:Transfection
chemical_list: D020099:Coated Materials, Biocompatible; D050659:Core Binding Factor Alpha 1 Subunit; D034741:RNA, Small Interfering; D055758:SOX9 Transcription Factor; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid