Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells

Abstract

Background: Mesenchymal stem cell transplantation is a promising method in regenerative medicine. Gene-modified mesenchymal stem cells possess superior characteristics of specific tissue differentiation, resistance to apoptosis, and directional migration. Viral vectors have the disadvantages of potential immunogenicity, carcinogenicity, and complicated synthetic procedures. Polyethylene glycol-grafted polyethylenimine (PEG-PEI) holds promise in gene delivery because of easy preparation and potentially targeting modification. Methods: A PEG8k-PEI25k graft copolymer was synthesized. Agarose gel retardation assay and dynamic light scattering were used to determine the properties of the nanoparticles. MTT reduction, wound and healing, and differentiation assays were used to test the cytobiological characteristics of rat mesenchymal stem cells, fluorescence microscopy and flow cytometry were used to determine transfection efficiency, and atomic force microscopy was used to evaluate the interaction between PEG-PEI/plasmid nanoparticles and mesenchymal stem cells. Results: After incubation with the copolymer, the bionomics of mesenchymal stem cells showed no significant change. The mesenchymal stem cells still maintained high viability, resettled the wound area, and differentiated into adipocytes and osteoblasts. The PEG-PEI completely packed plasmid and condensed plasmid into stable nanoparticles of 100–150 nm diameter. After optimizing the N/P ratio, the PEG-PEI/plasmid microcapsules delivered plasmid into mesenchymal stem cells and obtained an optimum transfection efficiency of 15%–21%, which was higher than for cationic liposomes. Conclusion: These data indicate that PEG-PEI is a valid gene delivery agent and has better transfection efficiency than cationic liposomes in mesenchymal stem cells.

Efficient delivery of C/EBP beta gene into human mesenchymal stem cells via polyethylenimine-coated gold nanoparticles enhances adipogenic differentiation

“…In the present study, we modified the surface of AuNPs with 25 kDa branched polyethylenimine (PEI), a commercially available cationic polymer, to enhance their transfection efficiency in difficult-to-transfect cells, such as hMSCs. PEI is a well-studied cationic polymer, which on its own has been used as a non-viral gene delivery vector or more often to modify the surfaces of nanovectors due to their controllable synthesis, highly abundant surface amino groups, and their ability to compact large amounts of nucleic acids 13 30 37 38 39 40 41 .…”

Mesenchymal Stem Cell-Based Therapy

“…85 Nonviral vectors, which require the use of cationic polymers and liposomes for condensation are plagued with the problem of poor transfection efficiency. 86 Although Adv transduction of MSCs has been demonstrated, transduction efficiency is poor because of low expression of coxsackievirus and adenovirus receptor (CAR) by MSCs. 87 Adv vectors have been modified to target integrin overexpressing on MSCs.…”

Adenovirus-mediated RAR-β over-expression enhances ATRA-induced neuronal differentiation of rat mesenchymal stem cells

“…Because of stem cell derivation, a general eukaryotic expression vector is hardly transfected into MSCs by lipofection or calcium phosphate co-precipitation [25]. In this study, we successfully constructed an adenovirus vector carrying the RAR-β gene by using the replication-defective AdEasy adenovirus system.…”