Complex assessment of a plasmid DNA mechanism of action in development of gene-activated materials

Abstract

The number of studies related with gene-activated matrices is increasing annually; the first-in-class product has been already implemented into clinical practice for bone grafting indications. Considering specificity of the gene-activated matrices mechanism of action determined by gene constructs, there is a demand to standardize the methods allowing to characterize all the stages of biological action in vivo. Here, using on the example of a gene-activated hydrogel consisting of type I collagen and plasmid DNA with the vascular endothelial growth factor gene (VEGF165), the main steps of the plasmid DNA mechanism of action were confirmed by various methods. For this, a fluorescent Cy3, reporter plasmid DNA with the firefly luciferase gene (Luc), RT-PCR and ELISA, immunohistochemical study with antibodies to CD31 were used. The results were compared with the other scientific papers, some recommendations were formulated to determine a minimally required list of studies for the development of gene-activated materials.

Keywords

VEGF / gene-activated matrix / hydrogel / plasmid DNA / Cy3 / luciferase / VEGF

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