At baseline, all subjects underwent a full-mouth clinical periodontal examination by a calibrated periodontist, including PD and AL, using a William’s periodontal probe at six sites for each tooth. In addition, the bleeding index (BI) of each tooth was recorded. Periodontal parameters were re-evaluated by the same examiner at 3 months after non-surgical periodontal therapy.
Fasting blood samples were collected from each subject into ethylenediaminetetraacetic acid (EDTA)-containing tubes and analyzed by a technician who was blinded to the case status using a calibrated automated hematology analyzer (Sysmex, Corporation, Kobe, Japan) and a biochemistry Automatic Analyzer (model 7180; Hitachi High-Technologies Corporation, Tokyo, Japan). Serum was separated and stored immediately at −80°C. Serum concentrations of serum high-sensitivity C-reactive protein (hs-CRP), iron, ferritin and transferrin, in addition to transferrin saturation (TS), and total iron binding capacity (TIBC) were measured with an Automatic Analyzer (model 7170A; Hitachi High-Technologies Corporation). Enzyme-linked immunosorbent assay kits were used to measure serum concentrations of IL-6 (#KE1368; ImmunoWay Biotechnology Company, Plano, TX, U.S.A.) and hepcidin (#DHP250; R&D Systems, Inc., Minneapolis, MN, U.S.A.) in accordance with the manufacturers’ instructions.
Fasting blood samples were collected from each subject into ethylenediaminetetraacetic acid (EDTA)-containing tubes and analyzed by a technician who was blinded to the case status using a calibrated automated hematology analyzer (Sysmex, Corporation, Kobe, Japan) and a biochemistry Automatic Analyzer (model 7180; Hitachi High-Technologies Corporation, Tokyo, Japan). Serum was separated and stored immediately at −80°C. Serum concentrations of serum high-sensitivity C-reactive protein (hs-CRP), iron, ferritin and transferrin, in addition to transferrin saturation (TS), and total iron binding capacity (TIBC) were measured with an Automatic Analyzer (model 7170A; Hitachi High-Technologies Corporation). Enzyme-linked immunosorbent assay kits were used to measure serum concentrations of IL-6 (#KE1368; ImmunoWay Biotechnology Company, Plano, TX, U.S.A.) and hepcidin (#DHP250; R&D Systems, Inc., Minneapolis, MN, U.S.A.) in accordance with the manufacturers’ instructions.