The transfection efficiency of photosensitizer-induced gene delivery to human MSCs and internalization rates of EGFP and Runx2 genes

• Published in: Biomaterials Vol. 33; no. 27; pp. 6485 - 6494
• Main Authors: Park, Sin-jung, Na, Kun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.09.2012
• Subjects: Advanced Basic Science; Blotting, Western; Cell Death - drug effects; Cell Death - radiation effects; Cell Survival - drug effects; Cell Survival - radiation effects; Core Binding Factor Alpha 1 Subunit - genetics; Core Binding Factor Alpha 1 Subunit - metabolism; Dentistry; DNA - metabolism; Endocytosis - drug effects; Endocytosis - radiation effects; Gene delivery; Gene Expression Regulation - drug effects; Gene Expression Regulation - radiation effects; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Human mesenchymal stem cell; Humans; Light; Mesenchymal Stromal Cells - drug effects; Mesenchymal Stromal Cells - metabolism; Mesenchymal Stromal Cells - radiation effects; Microscopy, Confocal; Osteogenesis; Osteogenesis - drug effects; Osteogenesis - radiation effects; Pheophorbide-a; Photosensitizing Agents - pharmacology; Plasmids - metabolism; Polyethyleneimine - chemistry; Polymerase Chain Reaction; Runx2; Time Factors; Transfection - methods; Human mesenchymal stem cell; Gene delivery; Pheophorbide-a; Osteogenesis; Runx2
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2012.05.040

Abstract To improve the transfection efficiency of non-viral gene vectors to human mesenchymal stem cells (hMSCs), a photosensitizer (PS)-induced gene delivery system was designed by using pheophorbide-a (pheo-a) as a PS. In FACS results, this system showed excellent gene transfection efficiency depending on irradiation power. The result was strongly supported by western blot and real-time quantitative PCR (RT-qPCR) assays. The protein and mRNA expression of enhanced green fluorescent protein (EGFP) in hMSCs treated with 0.9 J/cm2 irradiation increased 9.8- and 8.7-fold compared with non-irradiated hMSCs, respectively. Furthermore, the internalization of PEI/pDNA complexes in hMSCs was enhanced by light irradiation even under conditions that inhibited endocytosis. The hemolytic activity of PS with irradiation (0.9 J/cm2 ) significantly increased to 55%. Thus, PS with light irradiation facilitated both the internalization and endosomal escape of gene complexes. For osteogenic induction, the Runt-related transcription factor 2 (Runx2) gene was transferred to hMSCs via PS-induced transfection. Von Kossa staining indicated that Runx2 overexpression significantly enhanced the osteogenesis of hMSCs. Therefore, this PS-induced gene delivery method has potential value for stem cell therapy via gene delivery.