Chitosan-Based Nanoparticles for Intracellular Delivery of ISAV Fusion Protein cDNA into Melanoma Cells: A Path to Develop Oncolytic Anticancer Therapies

Figure 1

NP characterization and evaluation. (a) Complex ensemble was evaluated at various ratios between chitosan and pGFP by electrophoretic migration of pDNA in the presence of chitosan on an agarose gel. DNA ladder (L) is shown in lane 1, naked vector (NV) was used as a negative control in lane 2, and the different N/P complexes are shown in lanes 3-6. (b) Complex formation efficiency was evaluated as described in Materials and Methods, and it corresponds to insoluble pDNA with respect to total pDNA and represented as . Representative histograms from physical parameters such as (c) size and (d) zeta potential for N/P 4, 20, 28, and 40 are shown in red, green, blue, and black, respectively. (e) NP morphology was determined by atomic force microscopy for N/P 20 and 28. (f) The effect of the naked vector (NV) and NP (N/P from 4 to 40) on cell viability was evaluated by MTT assays at 24, 48, and 120 h after challenge and was normalized against untreated cells (control). (g) Transfection efficiency was evaluated using GFP as a reporter with NPs or Lipofectamine and evaluated by flow cytometry. A minimum of three independent experiments was performed. Bars correspond to , and statistical analyses were performed using the Mann–Whitney test ().

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