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. 2019 Nov 12;11(11):600. doi: 10.3390/pharmaceutics11110600

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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(A,D) Absence of early cytotoxic effects, as determined in lactate dehydrogenase (LDH) release assays for P10Y/siCtrl and P5Y/siCtrl complexes in PC3 (A) and G55T2 (D) cells. LDH release was quantitated 24 h after transfection of 30 pmol siCtrl/polymer per 24 well. (B,E) Knockdown of the Enhanced Green Fluorescent Protein (EGFP) reporter gene in stably expressing PC3-EGFP/Luc (B) and G55T2-EGFP/Luc (E) cells. Cells were transfected with P10Y and P5Y siRNA complexes and analyzed after 72 h by flow cytometry. (C,F) Cell viabilities of PC3 cells (C) and G55T2 cells (F) 48 h after transfection of P10Y/siCtrl and P5Y/siCtrl complexes at mass ratio 2.5 with increasing siRNA-concentrations. (G) Hemolysis assay for P5Y/siRNA complexes, demonstrating the absence of erythrocyte damage over a wider range of different concentrations. (H) Erythrocyte aggregation assay, further confirming the biocompatibility of P5Y/siRNA complexes as shown for 3 µg complexed siRNA (upper left; all lens magnifications: 4×). A total of 750 kDa bPEI complexes served as positive control, which led to a dose-dependent, profound aggregation of the red blood cells.