The present contribution is focused on feasibility of using comb-like copolymers of polyethylenimine with poly(2-ethyl-2-oxazoline) (LPEI-comb-PEtOx) with varying grafting densities and degrees of polymerization of PEI and PEtOx to deliver DNA molecules into cells. The copolymers form small and well-defined particles at elevated temperatures, which are used as platforms for binding and condensing DNA. The electrostatic interactions between particles and DNA result in formation of sub-100 nm polyplex particles of narrow size distribution and different morphology and structure. The investigated gene delivery systems exhibit transfection efficiency dependent on the copolymer chain topology, shape of the polyplex particles, and internalization pathway. Flow cytometry shows enhanced transfection efficiency of the polyplexes with elongated and ellipsoidal morphology. The preliminary biocompatibility study on a panel of human cell lines shows that pure copolymers and polyplexes thereof are practically devoid of cytotoxicity.

Chlorogenic (5-CQA), 1,5-, 3,5-, 4,5-and 3,4-dicaffeoylquinic (DCQA) acids were identified and quantified in the methanol extracts of Inula oculus-christi L., I. bifrons L., I. aschersoniana JANKA var. aschersoniana, I. ensifolia L., I. conyza (GRIESS.) DC. and I. germanica L. by HPLC analysis. The amount of 5-CQA varied from 5.48 to 28.44 mg/g DE and the highest content was detected in I. ensifolia. 1,5-DCQA (4.05-55.25 mg/g DE) was the most abundant dicaffeoyl ester of quinic acid followed by 3,5-DCQA, 4,5-DCQA and 3,4-DCQA. The extract of I. ensifolia showed the highest total phenolic content (119.92 � 0.95 mg GAE/g DE) and exhibited the strongest DPPH radical scavenging activity (69.41 � 0.55 %). I. bifrons extract was found to be the most active sample against ABTS * + (TEAC 0.257 � 0.012 mg/mL) and the best tyrosinase inhibitor. The studied extracts demonstrated a low inhibitory effect towards acetylcholinesterase and possessed low cytotoxicity in concentration range from 10 to 300 μg/mL toward non-cancer (MDCK II) and cancer (A 549) cells. Figure 2. HPLC chromatograms of the studied extracts and standard mixture.[a] Results are presented as mean � SD from three replicates (n = 3). Means in the columns with the same letter are not significantly different from each other (p > 0.05) (t-test).

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Comblike polyethylenimines with varying degrees of polymerization of both the main and side chains as well as different grafting densities were evaluated as gene delivery vectors. They were able to condense linear and plasmid DNA into nanosized polyplex particles with dimensions and surface potentials in the 130-330 nm and -30 to +15 mV ranges, respectively, depending on the amine/phosphate (N/P) ratio. The polyplexes remained stable in aqueous and buffer solutions from several hours up to several days. The moderate colloidal stability was also manifested in a relatively broad size distribution (PDI typically above 0.2) and structural polymorphism observed by transmission electron microscopy. Both the neat polymers and polyplexes displayed low cytotoxicity in WISH cells as the relative cell viability was more than 60%. Experiments with lysosomal fluorescence staining revealed that the internalization pathways and, in turn, transfection efficiency of the polyplex nanoparticles depended on the polymer chain topology. The vector systems based on the polymers of denser structure can be considered to be promising systems for gene transfection in eukaryotic cells.

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