Multilineage differentiation of human-derived dermal fibroblasts transfected with genes coated on PLGA nanoparticles plus growth factors

Wounded tissues and cells may be treated with growth factors and specific genes for the purpose of tissue repair and regeneration. To deliver specific genes into tissues and cells, this study presents the use of fabricated poly (dl-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) complexed with the cationic polymer poly (ethleneimine) (PEI). Through complexation with PEI, several types of genes (SOX9, Cbfa1, and C/EBP-α) were coated into PLGA NPs, which enhanced gene uptake into normal human-derived dermal fibroblast cells (NFDHCs) in vitro and in vivo. Several cell types (293T, HeLa, and fibroblast cells) were transfected with fluorescence-tagged PEI/SOX9, PEI/Cbfa1, and PEI/C/EBP-α gene-complexed PLGA NPs. The gene and protein expression levels in the cells were evaluated by RT-PCR, real-time quantitative PCR, Western blotting, and confocal laser microscopy. Fibroblast cells encapsulated in fibrin gels were transfected with the gene-complexed NPs plus specific growth factors (TGF-β3, BMP-2, or IGF/bFGF), which induced chondrogenesis, osteogenesis, or adipogenesis both in vitro and after transplantation into nude mouse.