Cytotoxicity of M-MSNs and its functionalized counterpart.

<p>(a) The curves show the MTT cell viability results (reported as a % of the medium-treated (control) cells) after 8 h treatment with different particles doses. (b) Cell images: cytotoxicity comparison between the two particles at a concentration of 500 μg/ml. Bar = 100 μm.</p

Confocal laser scanning microscopic images of RAW264.7 cells incubated with M-MSN-P-CpG nanoparticles for 3 h.

<p>(1) The particle is labeled with fluorescein isothiocyanate (FITC, emission of green fluorescence), (2) the endolysosomes in cells are stained with LysoTrackerRed (emission of red fluorescence), (3) the nuclei are stained with 4,6-diamino-2-phenyl indole (DAPI, emission of blue fluorescence), bar = 10 ìm, (4) the overlapped image of 1–3.</p

Cell uptake of CpG loaded in M-MSN-P.

<p>The cell uptake efficiency was examined by labeling the CpG with Cy3. The free oligo CpG-Cy3 and Lipofectamine-CpG-Cy3 were utilized as controls. M-MSNs are unable to carry CpG-Cy3 thus no Cy3 signal is observed. M-MSN-P with CpG-Cy3 shows a positive signal. Bar = 50 μm. The doses of free CpG or CpG in loaded carriers were equivalent to 15μg/ml.</p

TNF-αsecretion at 8 h after the addition of CpG loaded complexes to RAW264.7 cells.

<p>Results are expressed as mean±SD of three independent measurements.</p

Amine functionalized nanoparticles are denoted as M-MSN-A.

<p>Following further PEGylation, they are denoted as M-MSN-P. (a) TEM micrograph for M-MSN-A. The bar represents 50 nm. The inset shows the pore size distribution plot for M-MSN-A. (b) TG and DTG curves, (c) Zeta potential and (d) particle size distributions of M-MSN-A and M-MSN-P.</p

Inhibition of Hela cells proliferation by CpG loaded particles and doxorubicin hydrochloride (DOX).

<p>RAW264.7 and Hela were placed in the upper and lower chambers of Transwell plates, respectively. CpG loaded complexes and DOX were added to the upper side at a final concentration of 10 μg CpG/ml and 1 μg DOX/ml. The proliferative activity of Hela cells was measured at 48 h with the MTT assay and was converted to a percentage of the medium-treated (control) cells. Results are expressed as mean±SD of three independent measurements. CpG (and CpG loaded particle) group displayed significant differences, compared to non-CpG controls. CpG (and CpG loaded particles) also present different cell viabilities compared to the DOX treatment group (**<i>P</i><0.01).</p

IL-12 concentration in the serum after intravenous injection of free CpG or the M-MSN-P/CpG complex.

<p>At 6 h after injection, serum samples were collected and the concentration of IL-12 was measured by ELISA. Results are expressed as mean±SD of three mice (*<i>P</i><0.05, ***<i>P</i><0.001).</p

Schematic illustration of the steps for the amino-modification (M-MSN-A) and PEG (MW 2000) grafting (M-MSN-P) as well as CpG loading into the particles.

<p>Schematic illustration of the steps for the amino-modification (M-MSN-A) and PEG (MW 2000) grafting (M-MSN-P) as well as CpG loading into the particles.</p

CpG adsorption against M-MSN-P and M-MSNs.

<p>CpG adsorption against M-MSN-P and M-MSNs.</p

GC/MS of MEO.

<p>GC/MS of MEO.</p