Ferritin-Based Systems as a Platform for DNA Construct Delivery: Preparation, Characterization, and In Vitro Evaluation

• Published in: BioNanoScience 2025, Vol.15 (1)
• Main Authors: Paiva, Natália F., Rios, Wendy M., Antonio, Gabriel Pocci, Umino, Gisele M., Ferreira, Maíra Peres, Brandão, Izaíra Tincani, Del Ciampo, José Orestes, Bentley, Maria Vitória Lopes Badra, Casagrande, Rúbia, Verri, Waldiceu A., Fonseca, Maria José Vieira, de Freitas, Osvaldo, Silva, Célio L., de Carvalho Vicentini, Fabiana Testa Moura
• Format: Article
• Language: English
• Subjects: Apoferritin; Biological and Medical Physics; Biological properties; Biomaterials; Biomedical materials; Biophysics; Cell activation; Cell membranes; Cell size; Cellular structure; Chitosan; Circuits and Systems; Combined vaccines; DNA damage; DNA structure; Drug delivery; Drug delivery systems; Electrostatic properties; Engineering; Ferritin; Growth factors; Immune response; Immune system; Immunotherapy; Internalization; Melanoma; Nanoparticles; Nanotechnology; Nucleic acids; pH effects; Plasmids; Polysaccharides; Proteins; Transfection; Vaccines; Zeta potential
• ISSN: 2191-1630; 2191-1649
• DOI: 10.1007/s12668-024-01676-7

Strategies based on nucleic acids combined with immunotherapy are promising alternatives to improve the therapeutic outcome of various tumors, even those at an advanced stage. The use of nanoparticles as delivery systems for nucleic acid vaccines is advantageous because it allows the protection of the nucleic acid against degradation, overcomes the barriers of cellular internalization, and increases stability and there is also the possibility of delivery of several vaccine nucleic acids to the same target cell to improve the immune response activation. Ferritin nanoparticles, which are protein-based supramolecular complexes that have a highly symmetrical and uniformly sized spherical hollow architecture, have emerged as one of the promising biomaterials for use as a drug delivery system due to its unique structural characteristics and biological properties, being identified as a new platform for use in the bio nanotechnology. In this context, the structure of apoferritin was disorganized based on two conditions of pH (pH ≤ 2 and pH ≥ 11) to break the electrostatic interactions that maintain its spherical hollow structure and consequent incorporation of the plasmid DNA that expresses the variant III of the epidermal growth factor receptor (EGFrvIII). Then, these obtained product systems were characterized according to the physical–chemical characteristics of size, shape, and zeta potential; pDNA encapsulation efficiency and evaluation of HEK cell transfection; and consequent production of the protein of interest. The early results of the present study showed that the systems containing ferritin:pDNA were not able to transfect HEK cells under the initial tested conditions, possibly due to the electrostatic repulsion between the negative residual charges of ferritin nanoparticles and cell membranes that can overcome the natural absorption capacity of ferritin. So, a promising alternative to enhance transfection into cells was the physical or chemical binding of ferritin with natural charged polysaccharides, such as chitosan. Thus, in the second part of this study, we proposed to obtain and characterize a ferritin associated with chitosan delivery system as a platform to carry DNA construct that expresses EGFrvIII for further evaluation and use in the treatment of melanoma. The obtained results, in view of the cell transfection assessments and the characterizations performed, provided a positive scenario for the use of the ferritin as a plasmid DNA delivery sy