Immunological monitoring
STERILE
The supragingival biofilm of the selected dental elements was removed and the sites were carefully dried with air jets and then isolated with sterile cotton rolls. GCF samples were obtained with Periopaper® strips (Oralflow Inc., Amityville, NY, USA). The strips were carefully inserted close to the margin of the gingival sulcus, remaining for a period of 30 s. The amount of total protein in each sample was determined by conventional enzyme immunoassays (ELISA) using specific kits (DCTM Protein Assay; Bio-Rad Laboratories, Inc. Berkeley, CA, USA). Cytokine levels (IL-1α, IL-1β, monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1β (MIP-1β), and IL-8) were determined (pg/μl) using commercially available kits (HCYTOMAG-60 K–Milliplex