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

Year 2018, Volume: 4 Issue: 2, 92 - 100, 04.04.2018

Ali Öztuna Hasan Nazır

https://doi.org/10.18621/eurj.346612

Cited By: 2

Abstract

Objectives. MicroRNAs (miRNAs) are involved in the regulation of
most biological processes and also contribute to many types of disease.
Fibroblast cells, such as MRC-5, are often used in biological researches
utilizing cell transfection methods due to their difficult to transfect nature.
Cells can be genetically engineered by using viral and non-viral methods.
Poly(amidoamine) (PAMAM) dendrimers are very promising alternative as a
delivery vehicle due to their well-defined
characteristics. In this study, in vitro
transfection potential of cystamine 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 and condensation capability of dendrimers was examined by gel
retardation assay. Characterization of dendriplexes was made by zeta potential,
particle size measurements and transmission electron microscopy. Transfection
efficiencies of the dendriplexes were determined by flow cytometry and
intracytoplasmic distribution of the dendriplexes was shown by laser scanning
confocal microscopy. Also, quantitative structure-activity relationship and molecular
docking calculations were used to be able to discuss transfection efficiencies
of the dendriplexes into the cell. Results. While high level of viability
on MRC-5 cells was observed for dendriplexes prepared with PFB and PFP,
transfection efficiency with PFP was higher than PFB. Transfection efficiency
difference between these two compounds was attributed to their molecular
structures. Conclusions. Obtained results hold promise for the usage of
these compounds as a transfection reagent at MRC-5 cells. Further studies are
needed to support these findings. 

Keywords

Cystamine-G5-PAMAM, pentafluorobenzoic acid, pentafluoropropionic acid, miRNA, MRC-5

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