These results suggest that the membranes of HeLa cells were more resistant to PEI. Interestingly, the PHMBG-M/SiO2 particles were more cytotoxic than PHMBG in HeLa cells (Figure 3(d)). Although the external magnetic field helped reduce its cytotoxicity, the particles still remained more cytotoxic than PHMBG (Figure 3(d)). Figure 3 Effect of nanoparticle/siRNA (N/P) ratio on membrane permeability of CHO-K1 ((a) and (b)) and HeLa ((c) and (d)) cell lines. Comparing the transfection efficiency,

cell viability, and cell membrane integrity of all materials at the optimum N/P ratios (from 34 to 43) for siRNA transfection Inhibitors,research,lifescience,medical shows that, although PEI is an efficient transfecting vehicle for CHO-K1 cells, it is highly cytotoxic (100% LDH released). Our Inhibitors,research,lifescience,medical results show that the PEI-modified PEI-M/SiO2 particles possessed higher transfecting potential and substantially reduced

cytotoxicity than PEI. Application of the external magnetic field (PEI-M/SiO2-magnetofection) did not alter the cell viability or cytotoxicity of the particles, but it did significantly increase the transfection efficiency of PEI-M/SiO2 in CHO-K1 cells (Figure 4(a)). The siRNA transfection efficiency of PEI and PEI-M/SiO2 in HeLa cells was similar, Inhibitors,research,lifescience,medical and PEI-M/SiO2-magnetofection did not improve the siRNA uptake at this particular N/P ratio. No decrease in cell Inhibitors,research,lifescience,medical viability and or increase in cytotoxicity were observed with PEI-M/SiO2 and PEI-M/SiO2-magnetofection

in HeLa cells (Figure 4(b)). Both in CHO-K1 and HeLa cells, PHMBG’s NPs were less efficient transfecting vehicles than PEI’s modified NPs, but in CHO-K1, they were less cytotoxic than PEI, whereas in HeLa they were more cytotoxic. Surprisingly, PHMBG-M/SiO2-magnetofection caused significant membrane disruption to CHO-K1 cells (Figure 4(a)). Surprisingly, in HeLa cells, PHMBG-M/SiO2-magnetofection was a less efficient transfecting vehicle than PHMBG-M/SiO2 Inhibitors,research,lifescience,medical (Figure 4(b)). Figure 4 Panel (a) CHO-K1; (b) HeLa. N/P ratios: 39 for PEI, 34 for PEI-M/SiO2, 43 for PHMBG, and 42 for PHMBG-M/SiO2. The last column in Figures 4(a) and 4(b) demonstrates that siRNA cannot cross cell membranes by itself, as demonstrated by the transfection of siRNA without any of the NP materials. As previously discussed, an efficient delivery vehicle carrying siRNA across a cell membrane Histone demethylase to downregulate the expression of the target gene requires the successful completion of several key steps [57, 58], the first one being the ability of the NPs to bind siRNA. Towards this end, we employed the ethidium bromide displacement assay to CDK inhibitor assess the relative degree of binding between the respective polyelectrolyte and siRNA. Our results show that increasing the N/P ratios also increase the binding between the delivery vehicle and siRNA (the relative fluorescence intensity decreases, Figure 5(a)).