To evaluate siRNA-mediated knockdown following systemic nanoparticle administration, the bioluminescence signal from the B16F10 tumors in the lungs was monitored by IVIS. Prior to imaging, 3.75 mg D-luciferin (Cayman Chemical Company) in 150 μL volume was injected intraperitoneally (i.p.) in each mouse. Image analysis was carried out using Living Image software to quantify the total bioluminescence of the colonized tumors. The nanoparticles were loaded with either siRNA targeting firefly luciferase (siLuc), siRNA targeting Bcl-2 (siBcl-2), or negative control siRNA (scRNA), and bioluminescence was used to determine whether successful delivery was achieved.
Any potential hepatotoxicity of the systemically delivered photocrosslinked nanoparticles was examined. Animals received four repeated i.v. injections of nanoparticles and untreated animals were used as controls. Blood was collected from each animal after 8 days after initiation of treatment, and the serum was collected by centrifugation at 1500 rcf for 15 min at 4 °C. The serum in treated and untreated animals was analyzed for AST activity and ALT (Sigma-Aldrich), key biomarkers for liver health. The AST and ALT activity assays were performed according to manufacturer’s instructions.
Any potential hepatotoxicity of the systemically delivered photocrosslinked nanoparticles was examined. Animals received four repeated i.v. injections of nanoparticles and untreated animals were used as controls. Blood was collected from each animal after 8 days after initiation of treatment, and the serum was collected by centrifugation at 1500 rcf for 15 min at 4 °C. The serum in treated and untreated animals was analyzed for AST activity and ALT (Sigma-Aldrich), key biomarkers for liver health. The AST and ALT activity assays were performed according to manufacturer’s instructions.