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Evaluation of biocompatibility and efficiency of plasmid DNA delivery by gene-activated hydrogels in vitro

Genes & Cells: Vol XIV, №2, 2019, pp: 40-46 DOI: 10.23868/201906017


I.Y. Bozo, A.A. Titova, M.N. Zhuravleva, A.I. Bilyalov, M.O. Mavlikeev, I.A. Yakovlev, I.I. Eremin, A.A. Pulin, V.S. Komlev, R.V. Deev

Injectable forms of biomaterials have significant prospects for development of minimally invasive medical technologies in regenerative medicine. In this study, we evaluated the opportunities of using synthetic and natural hydrogels, including those containing calcium phosphates and hydroxyapatite, to deliver gene constructs, plasmid DNA molecules carrying some genes (encoding vascular endothelial growth factor, green fluorescent protein, or luciferase), to cells in vitro. The study of plasmid DNA delivery was performed in the line of mouse embryonic fibroblasts 3T3 using fluorescence and luminescent methods. Hydrogel biocompatibility was characterized in cultures of multipotent mesenchymal stromal cells.
We found that the extracts of investigated hydrogels did not exert cytotoxic effects, however, with direct contact of materials containing either calcium phosphate particles or hydroxyapatite, a cytotoxic effect was observed. In the case of hydrogels based on collagen and hyaluronic acid that showed optimal biocompatibility, the transfection efficiency turned out to be low without using of a transfection agent and when used, it was 24 and 31%, respectively.
Other variants of hydrogels did not provide plasmid DNA delivery, possibly due to cytotoxic effects. Further in vivo studies will clarify the features of plasmid DNA delivery with hydrogels, as well as evaluate the effectiveness of gene-activated hydrogels in tissue regeneration of the musculoskeletal system.

Keywords: gene-activated hydrogels, plasmid DNA, biocompatibility,luciferase.

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