Evaluation and comparison of cytotoxicity, genotoxicity, and apoptotic effects of poly-l-lysine/plasmid DNA micro- and nanoparticles

Abstract

<jats:p> The principal impediment to gene therapy is the development of efficient, nontoxic gene carriers that can handle and deliver foreign genetic materials into various cell types, including healthy and cancerous cells. Poly-l-lysine (PLL) polymers are one of the most favorable gene carriers among nonviral vectors, and PLL had low transfection and safety issues. The purpose of this study was to measure cellular toxicity, DNA damage, and apoptotic effects of PLL nanoparticles. Neuro2A mammalian cells were cultured and exposed to PLL/DNA complexes at different polymer/DNA ratios ( C/ P ratio 2 and 6) for 24 h. To evaluate metabolic activity, genotoxicity, and apoptotic influences of PLL nanoparticle, the following experimental methods were employed, in order: 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), DNA damage (COMET analysis) assay, and sub-G1 peak apoptosis assay. Our data indicate that toxicity is concentration dependent and a high concentration of polymer declined the metabolic activity. In addition, largest complexes ( C/ P 6 in HEPES buffered saline buffer) have slighter negative impact on metabolic activity. In agreement with our cytotoxicity data, apoptotic assay result represented that increase in size of PLL/DNA complexes decrease the number of apoptotic cells. Also, there was a remarkable increase in percent tail DNA of Neuro2A cells treated with higher concentration of PLL and its polyplexes. The present study demonstrated that PLL/DNA complexes caused cytotoxic, apoptotic, and genotoxic effects in a dose-dependent and weight ratio-dependent manner, which also affected the size of polyplexes. </jats:p>

Study of lead-induced neurotoxicity in cholinergic cells differentiated from bone marrow-derived mesenchymal stem cells

“…The present study showed that PDL inhibits the growth and proliferation of BMSCs; This may be due to the cytotoxic effects on HeLa cells and Neuro2A mammalian cells mentioned in several studies as a result (Alinejad-Mofrad et al, 2019;Arnold et al, 1979). In our study, plates coated with gelatin 0.1% were used for culture and differentiation and cytotoxicity assessment.…”

Multilayered siRNA nanoparticles as a delivery system for extended gene silencing effect

“…NPs system even at day 21. Whereas, there was a complete release of siRNA from the PLL-coated 7 layered NPs over just a period of 5 days as reported by Lee et al The PLR coated particles have also been observed to possess a higher transfection efficiency as compared to PLL [95] . With a higher transfection efficiency, PLR would be able to protect and provide sustained and prolonged release of siRNA into the cytoplasm, as well as to silence the gene.…”

Advances in Nanomaterials Used in Co-Delivery of siRNA and Small Molecule Drugs for Cancer Treatment

“…However, this results in increased cytotoxicity. With the use of higher molecular weight PLL, there is a trade-off between greater transfection efficiency and the accompanying higher cytotoxicity [52]. It has been shown that grafting of PEG to PLL can reduce cytotoxicity without reduction of transfection efficiency [53].…”