The use of biodegradable PLGA nanoparticles to mediate SOX9 gene delivery in human mesenchymal stem cells (hMSCs) and induce chondrogenesis.
In stem cell therapy, transfection of specific genes into stem cells is an important technique to induce cell differentiation. To perform gene transfection in human mesenchymal stem cells (hMSCs), we designed and fabricated a non-viral vector system for specific stem cell differentiation. Several kinds of gene carriers were evaluated with regard to their transfection efficiency and their ability to enhance hMSCs differentiation. Of these delivery vehicles, biodegradable poly (DL-lactic-co-glycolic acid) (PLGA) nanoparticles yielded the best results, as they complexed with high levels of plasmid DNA (pDNA), allowed robust gene expression in hMSCs, and induced chondrogenesis. Polyplexing with polyethylenimine (PEI) enhanced the cellular uptake of SOX9 DNA complexed with PLGA nanoparticles both in vitro and in vivo. The expression of enhanced green fluorescent protein (EGFP) and SOX9 increased up to 75% in hMSCs transfected with PEI/SOX9 complexed PLGA nanoparticles 2 days after transfection. SOX9 gene expression was evaluated by RT-PCR, real time-qPCR, glycosaminoglycan (GAG)/DNA levels, immunoblotting, histology, and immunofluorescence.
Kim
Jae-Hwan
JH
Department of Biomedical Science, College of Life Science, CHA University 606-16, Yeoksam 1-Dong, Kangnam-gu, Seoul 135-081, Republic of Korea.
Park
Ji Sun
JS
Yang
Han Na
HN
Woo
Dae Gyun
DG
Jeon
Su Yeon
SY
Do
Hyun-Jin
HJ
Lim
Hye-Young
HY
Kim
Jung Mo
JM
Park
Keun-Hong
KH
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002470:Cell Survival; D020219:Chondrogenesis; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry*; D008156:Luciferases / Q000378:metabolism; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / 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; D055758:SOX9 Transcription Factor / Q000235:genetics*; D055672:Static Electricity; D014162:Transfection; D055815:Young Adult
D001672:Biocompatible Materials; D055758:SOX9 Transcription Factor; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D008156:Luciferases