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Introdution: The aim of this study was to compare levels of neurokinin A (NKA), substance P (SP), interleukin (IL)-8, and matrix metalloproteinase-8 (MMP-8) in pulp tissue and gingival crevicular fluid (GCF) samples of healthy and symptomatic irreversible pulpitis teeth., Methods: Forty patients diagnosed with healthy and symptomatic irreversible pulpitis teeth were included in this study. NKA, SP, IL-8, and MMP-8 levels were measured using the enzyme-linked immunosorbent assay test after pulp and GCF samples were obtained from healthy (n = 20) and symptomatic irreversible pulpitis teeth (n = 20). GCF sampling of 40 teeth was repeated 1 week later. Routine root canal treatment procedures of the teeth were performed, and the treatment process was completed. As a control group, GCF samples were taken from the contralateral teeth in both groups. Statistical analysis was performed using dependent and independent t tests, analysis of variance, Kruskal-Wallis, Mann-Whitney U tests, and Pearson correlation analysis., Results: Comparing the groups, all mediator levels were significantly higher in the pulp samples in the pulpitis group compared with the healthy group (NKA: P < .001, SP: P = .005, IL-8: P < .001, and MMP-8: P < .001). Likewise, in the pulpitis group, all mediator levels were significantly higher in the first GCF samples compared with the healthy group (NKA: P = .01, SP: P < .001, IL-8: P = .001, and MMP-8: P < .001)., Conclusions: It was observed that NKA, SP, IL-8, and MMP-8 increased significantly in pulp tissue and GCF specimens of symptomatic irreversible pulpitis teeth compared with pulp tissue and GCF specimens of healthy teeth. Second, it was determined that NKA, SP, IL-8, and MMP-8 levels decreased significantly in GCF samples in teeth diagnosed with symptomatic irreversible pulpitis 1 week after the removal of inflamed pulp. Finally, SP, IL-8, and MMP-8 levels were found to be higher in pulp tissue samples of the patients with symptomatic irreversible pulpitis with higher pain scores than those with low pain scores., (Copyright © 2020 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)