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


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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.

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About the authors

I. Y Bozo

Histograft, LLC; Research Institute of General Pathology and Pathophysiology; A.I. Burnazyana Federal Medical Biophysical Center, FMBA of Russia

Email: bozo.ilya@gmail.com

M. O Mavlikeev

Research Institute of General Pathology and Pathophysiology; I.I. Mechnikov North-Western State Medical University

A. A Titova

Research Institute of General Pathology and Pathophysiology; Kazan (Volga Region) Federal University

A. I Bilyalov

Kazan (Volga Region) Federal University

F. A Indeykin

Kazan Federal Medical University

A. A Pulin

Research Institute of General Pathology and Pathophysiology

I. I Eremin

Research Institute of General Pathology and Pathophysiology

V. S Komlev

AA. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences

A. A Isaev

Institute of Human Stem Cells, PJSC

R. V Deev

I.I. Mechnikov North-Western State Medical University; Institute of Human Stem Cells, PJSC

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