For the analysis of cytokines levels in vitro, J774 cells were seeded at 5×104 cells well−1 into a tissue-culture treated 96-well plate and incubated overnight. Following incubation, cells were treated with oxidized and APTES-modified MSV, mLLV, hLLV, and zymosan (100 μg mL−1, positive control) in complete media. Untreated cells represented a negative control. After 3 h and 24 h of treatment, media from each sample was collected and centrifuged at 10 000 × g. The supernatant was then collected and analyzed for IL-1β, IL-6, IL-10, IL-12 p40, IL-12 p70, and TNF-α using the MILLIPLEX MAP Mouse Cytokine/Chemokine premixed Immunoassay plate (Millipore).
Cytokines, hepatic, and renal function for in vivo experiments were evaluated by collecting blood from treated BALB/c mice (n=3) and evaluated for cytokines as mentioned using a MILLIPLEX MAP Mouse Cytokine/Chemokine premixed Immunoassay plate (Millipore). Hepatic and renal function was evaluated by evaluating blood serum for AST/ALT using an ALT Colorimetric Activity Assay and AST Colorimetric Activity Assay (Sigma). Creatinine levels were evaluated using a Creatinine Assay Kit (Abcam).
Cytokines, hepatic, and renal function for in vivo experiments were evaluated by collecting blood from treated BALB/c mice (n=3) and evaluated for cytokines as mentioned using a MILLIPLEX MAP Mouse Cytokine/Chemokine premixed Immunoassay plate (Millipore). Hepatic and renal function was evaluated by evaluating blood serum for AST/ALT using an ALT Colorimetric Activity Assay and AST Colorimetric Activity Assay (Sigma). Creatinine levels were evaluated using a Creatinine Assay Kit (Abcam).