Fig. 6.
Cp-LPP/shRunx2 lipopolyplexes reduce the ALP activity and block the formation of calcium deposits. (A) Quantitative assay of ALP activity performed at 7 and 14 days after a single transfection of HGOM-activated VIC on day 5 or two transfections (on days 5 and 12) with different nanoparticle formulations. (B) Quantitative assay of calcium concentration at 7 and 14 days after a single transfection of HGOM-activated VIC on day 5 or two transfections (on days 5 and 12) with different nanoparticle formulations. For both assays, data are means ± S.D. of three experiments in triplicate and presented as fold change relative to HGOM condition (considered 1). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 when compared with HGOM at 7 days; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 when compared with HGOM at 14 days. (C) Alizarin Red-stained calcium deposits in 3D-cultured VIC after 14 days of HGOM exposure and two transfections performed on days 5 and 12 with different nanoparticles. The graph displays the quantitative measurement of the dye released from the hydrogel. Data are means ± S.D. of two experiments made in triplicates and were represented as fold change relative to HGOM condition (considered 1). ∗∗p < 0.01, ∗∗∗p < 0.001 when compared with HGOM; #p < 0.05, ##p < 0.01 when compared with NM.