In-vitro transfection potential of fluorinated G5 PAMAM dendrimers for miRNA delivery to MRC-5 cells

Abstract

Objectives. MicroRNAs (miRNAs) are involved in the regulation of most biological processes and alsocontribute to many types of disease. Fibroblast cells, such as MRC-5, are often used in biological researchesutilizing cell transfection methods due to their difficult to transfect nature. Cells can be genetically engineeredby using viral and non-viral methods. Poly(amidoamine) (PAMAM) dendrimers are very promising alternativeas a delivery vehicle due to their well-defined characteristics. In this study, in vitro transfection potential ofcystamine core generation five (G5) PAMAM dendrimers fluorinated with 2,3,4,5,6-pentafluorobenzoic acid(PFB) and pentafluoropropionic acid (PFP) for miRNA delivery to MRC-5 cells was examined. Methods.Spectroscopic techniques were used in the characterization of the prepared dendrimers. miRNA binding andcondensation capability of dendrimers was examined by gel retardation assay. Characterization of dendriplexeswas made by zeta potential, particle size measurements and transmission electron microscopy. Transfectionefficiencies of the dendriplexes were determined by flow cytometry and intracytoplasmic distribution of thedendriplexes was shown by laser scanning confocal microscopy. Also, quantitative structure-activityrelationship and molecular docking calculations were used to be able to discuss transfection efficiencies of thedendriplexes into the cell. Results. While high level of viability on MRC-5 cells was observed for dendriplexesprepared with PFB and PFP, transfection efficiency with PFP was higher than PFB. Transfection efficiencydifference between these two compounds was attributed to their molecular structures. Conclusions. Obtainedresults hold promise for the usage of these compounds as a transfection reagent at MRC-5 cells. Further studiesare needed to support these findings