Serum inflammatory markers
Serum inflammatory status parameters (e.g. IL-18, TNF-alpha, TGF- beta 1, TGF-beta 2, PCR) were measured at Labospace s.r.l. (via Virgilio Ranzato, 12, 20128, Milano, Italy) by a bead-based multiplex assay (Luminex) on Bio-Plex® 3D Reader (Luminex FLEXMAP 3D, Bio-Rad Laboratories, Inc, Hercules, CA, United States) with the relevant commercial reagent kits.The unknown sample cytokine concentrations were then calculated by Bio-Plex Manager software v6.0. using a standard curve derived from the known reference cytokine concentrations supplied by the manufacturer.