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. 2020 Aug 15;9(8):1905. doi: 10.3390/cells9081905

Figure 2.

Figure 2

In vitro performance of ManNP and NonNP. (A) Cell viability of THP-1 macrophages and SVEC4-10 liver endothelial cells exposed to escalating doses of scrambled siRNA (scsiRNA)-loaded ManNP/NonNP showing that both scsiRNA loaded carriers were well tolerated by the cell-lines, as determined by the MTT assay (10:1 weight-to-weight ratio w/w of (Non-)ManNP: siRNA, incubation time 72 h, n = 2, *, **, **** p < 0.05, 0.001, 0.00001—NonNP vs. ManNP). (B) Cellular uptake of Cy5-scsiRNA-loaded and Oregon Green 488 labeled (Non-)ManNP (488-(Non-)ManNP) in human hepatocytes (HepG2), murine and human macrophages (RAW and THP-1 macrophages), and liver endothelial cells (SVEC4-10) at 25, 50, and 100 nM siRNA concentrations after 1 h incubation, followed by flow cytometry assessment (exemplary dot plots shown in Figure S2, n = 3, * p < 0.05) (C). Cellular uptake of scsiRNA-loaded 488-ManNP in native M0 and M2-, M1-polarized primary macrophages at 25 and 50 nM siRNA concentrations after 1 h incubation and analyzed by flow cytometry. Cellular uptake of 488-ManNP was significantly higher in M2 polarized macrophages compared to M0- and M1-polarized macrophages (MFI: mean fluorescence intensity of 488-(Non-)ManNP loaded with (Cy5-)scsiRNA, n = 3, ** p < 0.001, *** p < 0.0001).