Purpose: To evaluate clinical periodontal findings and GCF levels of tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6) and interleukin 8 (IL-8) in patients with overhang amalgam restorations before and after overhang restoration removal. Methods: 22 volunteer subjects (age range: 22-42 years old) with 22 overhang Class II amalgam restorations were selected. GCF samples were obtained from adjacent and contralateral teeth for IL-6, IL-8 and TNF-alpha, measurements and analyzed using enzyme-linked immunosorbent assay (ELISA) kits. The amalgam overhangs were removed and the readings were repeated at the end of 1 week. Clinical periodontal assessments, including gingival index (GI) and plaque index (PI) were performed at baseline and after 1 week. Data were analyzed using the paired t test and independent sample t-test at a significance level of 0.05. Correlations were investigated using Pearson correlation analysis. Results: A significant reduction in GI and PI was observed after removal of the overhanging restoration on Day 7. (P< 0.05) However, there was no significant difference between IL-6 (P= 0.857), IL-8 (P= 0.579) and TNF-alpha (P= 0.958) levels before and after overhang restoration removal. There were no significant correlations between laboratory findings and clinical parameters.