Comparison of nanoparticle-mediated transfection methods for DNA expression plasmids: Efficiency and cytotoxicity
Creators: name: Durán, Maria Carolina
name: Willenbrock, Saskia
name: Barchanski, Annette
name: Müller, Jessika-M.V.
name: Maiolini, Arianna
name: Soller, Jan T.
name: Barcikowski, Stephan
name: Nolte, Ingo
name: Feige, Karsten
name: Murua Escobar, Hugo
Publisher: name: London : Biomed Central
Year: 2011
Subjects: subject: Conventional approach
subject: Conventional methods
subject: Cytotoxic
subject: Cytotoxic effects
subject: Different sizes
subject: DNA expressions
subject: Ligand-free
subject: Ligand-stabilized
subject: Mammalian cell lines
subject: Nanoparticle formulation
subject: Propidium iodide
subject: Recombinant protein
subject: Side effect
subject: Stabilizer-free
subject: Transfection efficiency
subject: Transfection methods
subject: Transfection protocol
subject: Transfection reagents
subject: Cell culture
subject: Cell proliferation
subject: Cells
subject: Cytotoxicity
subject: DNA
subject: Efficiency
subject: Flow cytometry
subject: Fluorescence microscopy
subject: Ligands
subject: Magnets
subject: Mammals
subject: Nanoparticles
subject: Proteins
subject: Toxicity
subject: Molecular biology
subject: nanoparticle
subject: plasmid DNA
subject: propidium iodide
subject: gold
subject: green fluorescent protein
subject: metal nanoparticle
subject: animal cell
subject: article
subject: cell proliferation
subject: comparative study
subject: cytotoxicity
subject: DNA transfection
subject: flow cytometry
subject: fluorescence microscopy
subject: gene expression
subject: immunofluorescence
subject: mammal cell
subject: nonhuman
subject: protein expression
subject: cell line
subject: cell survival
subject: chemistry
subject: drug effect
subject: genetic transfection
subject: genetics
subject: human
subject: laser
subject: metabolism
subject: plasmid
subject: Eukaryota
subject: Mammalia
subject: Cell Line
subject: Cell Proliferation
subject: Cell Survival
subject: Flow Cytometry
subject: Gold
subject: Green Fluorescent Proteins
subject: Humans
subject: Lasers
subject: Metal Nanoparticles
subject: Microscopy, Fluorescence
subject: Plasmids
subject: Transfection
subject: Dewey Decimal Classification::500 | Naturwissenschaften::540 | Chemie
subject: Dewey Decimal Classification::600 | Technik::610 | Medizin, Gesundheit

description: Background: Reproducibly high transfection rates with low methodology-induced cytotoxic side effects are essential to attain the required effect on targeted cells when exogenous DNA is transfected. Different approaches and modifications such as the use of nanoparticles (NPs) are being evaluated to increase transfection efficiencies. Several studies have focused on the attained transfection efficiency after NP-mediated approaches. However, data comparing toxicity of these novel approaches with conventional methods is still rare.Transfection efficiency and methodology-induced cytotoxicity were analysed after transfection with different NP-mediated and conventional approaches. Two eukaryotic DNA-expression-plasmids were used to transfect the mammalian cell line MTH53A applying six different transfection protocols: conventional transfection reagent (FuGENE HD, FHD), FHD in combination with two different sizes of stabilizer-free laser-generated AuNPs (PLAL-AuNPs_S1,_S2), FHD and commercially available AuNPs (Plano-AuNP), and two magnetic transfection protocols. 24 h post transfection efficiency of each protocol was analysed using fluorescence microscopy and GFP-based flow cytometry. Toxicity was assessed measuring cell proliferation and percentage of propidium iodide (PI%) positive cells. Expression of the respective recombinant proteins was evaluated by immunofluorescence.Results: The addition of AuNPs to the transfection protocols significantly increased transfection efficiency in the pIRES-hrGFPII-eIL-12 transfections (FHD: 16%; AuNPs mean: 28%), whereas the magnet-assisted protocols did not increase efficiency. Ligand-free PLAL-AuNPs had no significant cytotoxic effect, while the ligand-stabilized Plano-AuNPs induced a significant increase in the PI% and lower cell proliferation. For pIRES-hrGFPII-rHMGB1 transfections significantly higher transfection efficiency was observed with PLAL-AuNPs (FHD: 31%; PLAL-AuNPs_S1: 46%; PLAL-AuNPs_S2: 50%), while the magnet-assisted transfection led to significantly lower efficiencies than the FHD protocol. With PLAL-AuNPs_S1 and _S2 the PI% was significantly higher, yet no consistent effect of these NPs on cell proliferation was observed. The magnet-assisted protocols were least effective, but did result in the lowest cytotoxic effect.Conclusions: This study demonstrated that transfection efficiency of DNA-expression-plasmids was significantly improved by the addition of AuNPs. In some combinations the respective cytotoxicity was increased depending on the type of the applied AuNPs and the transfected DNA construct. Consequently, our results indicate that for routine use of these AuNPs the specific nanoparticle formulation and DNA construct combination has to be considered.