Evaluation of Phytotoxicity and Mutagenicity of Novel DMAEMA-Containing Gene Carriers
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uation of Phytotoxicity and Mutagenicity of Novel DMAEMA-Containing Gene Carriers N. Finiuka, b, N. Romanyukb, N. Mitinac, O. Lobachevskad, A. Zaichenkoc, O. Terekb, and R. Stoikaa, b, * aInstitute
of Cell Biology, National Academy of Sciences of Ukraine, Lviv, 79005 Ukraine b Ivan Franko National University of Lviv, Lviv, 79005 Ukraine c Lviv Polytechnic National University, Lviv, 79013 Ukraine d Institute of Ecology of the Carpathians, National Academy of Sciences of Ukraine, Lviv, 79026 Ukraine *e-mail: [email protected] Received October 16, 2019; revised January 16, 2020; accepted September 18, 2020
Abstract—The use of novel carriers for gene delivery has been rapidly growing; thus, investigation of potential phytotoxic and mutagenic action of gene delivery carriers is important for preventing their negative side effects. We found that poly-DMAEMA carriers used in a 0.0025% dose exhibited a weak cytotoxic effect towards Allium cepa plants. In a higher dose (0.025%), they slightly (by 26–55%) increased the level of catalase activity but did not affect the level of superoxide dismutase activity and malondialdehyde content in roots of A. cepa. Results of the ana-telophase test in A. cepa demonstrated no genotoxic activity of the polymeric carriers used in a 0.0025% concentration and the higher dose (0.025%). Slight genotoxic activity was detected only for BGP24 and BGP26, PEG-containing poly-DMAEMA carriers, used in 0.025% concentration. The Ames test (–S9 and +S9) revealed no mutagenic potential of the DMAEMA-based polymers. Thus, low phytotoxicity and no mutagenicity of novel polymeric carriers suggest their potential as promising nanocarriers for gene delivery into plant cells. Keywords: poly(2-dimethylamino)ethyl methacrylate, polymeric carrier, ana-telophase test, Ames test, catalase, superoxide dismutase, malondialdehyde DOI: 10.3103/S0095452720050096
INTRODUCTION Nanomaterials are widely used to deliver genetic material to target cells. Cationic polymers are promising systems for delivering genetic material to cells for biotechnological and biomedical needs [1–4]. Representatives of polycationic carriers include poly(L-lysine) (PLL), poly(amidoamine) (PAMAM), poly(ethyleneimine) (PEI), and polymethacrylates (for example, poly(2-dimethylamino) ethyl methacrylate (DMAEMA)), which have been proposed for delivery of target genes to cells [4–7]. DMAEMA-based polymers were effective in transferring target genes to cells in mammals [8–11], yeast [12], and plants [13, 14]. However, the toxicity of polymeric carriers can impede their effective use in gene therapy and biotechnology [8, 15, 16]. It has been found that nanomaterials can be stressors that disrupt the oxidative status and enhance lipid peroxidation in target cells [17, 18]. Therefore, the development of safe and effective carriers of genetic material is important for the advancement of genetic engineering and gene therapy. Evaluation of the cytotoxicity of polymeric carriers is necessary to identify potential risks of their use. It has been sho
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