Quantitative Analysis of MMPs and Cytokines

MMPs and cytokines analysis was performed by using xMAP technology (Luminex 200, Luminex, Austin, TX, USA) to measure MMP-1, MMP-2, MMP-8, MMP-9, IL-1β, and IL-8. The Milliplex MAP multiplex assay Human MMP Panel 2 Magnetic Bead HMMP2MAG-55 K and HSP2MAG-63 K (Millipore, Billerica, MA, USA) kits were conducted in 96-well microplate format according to the manufacturer’s recommendations. Concentrations of MMPs were determined on the Bio-Plex Protein Array System (Bio-Rad, Hercules, CA, USA). Briefly, each of the bead solutions was transferred into a mixing vial and adjusted to a volume of 3 ml with bead diluents. Internal controls and standards were included in every assay. Following the addition of sample supernatants and beads, the resulting mixture was incubated overnight at 4 °C. The plate was read by Bio-Plex array reader with the Luminex 200 software system [20 (link)].