CCL2‐Driven Inflammation Links Periodontitis to Anxiety

Abstract

AimThis study investigates the association between periodontitis and anxiety, focusing on the role of the chemokine CCL2 in mediating this relationship.Materials and MethodsThis study comprises an analytical cross‐sectional study and a preclinical in vivo study. In the analytical cross‐sectional study, anxiety levels were assessed in individuals with periodontitis and healthy controls using the Hamilton Anxiety Rating Scale (HAMA). Blood and periodontal tissue samples were analysed for CCL2 levels via ELISA and monocyte counts via flow cytometry. In the preclinical in vivo study, a mouse model of ligature‐induced periodontitis was established. Anxiety‐like behaviours were evaluated using the Open Field Test and Elevated Plus Maze. Blood as well as periodontal and brain tissues were collected to measure CCL2 levels and monocyte/macrophage infiltration through ELISA and flow cytometry. Tight junction proteins in periodontal tissues and brain microvessels were analysed via Western blotting and immunofluorescence. Blood–brain barrier (BBB) permeability was assessed using Evans Blue extravasation. A CCL2‐neutralising antibody was administered to assess its effects on anxiety and periodontal pathology.ResultsIn the analytical cross‐sectional study (50 individuals with periodontitis and 50 healthy controls), individuals with periodontitis showed higher anxiety levels, correlating with elevated CCL2 levels in blood (56.84 [15.87] pg/mL vs. 19.28 [7.47] pg/mL in controls, p < 0.0001) and periodontal tissues (67.37 [23.10] pg/mL vs. 22.77 [10.21] pg/mL in controls, p < 0.0001), as well as increased monocyte counts (CD14+ monocytes in blood: 8.08 [3.01]% vs. 5.28 [1.84]% in controls, p < 0.01). In the preclinical in vivo study (total n = 80 mice, with n = 8 per group in analyses), periodontal inflammation increased CCL2 expression in periodontal tissues (125.80 [55.10] pg/mL vs. 25.13 [4.89] pg/mL in controls, p < 0.001) and blood (111.10 [47.80] pg/mL vs. 22.21 [5.39] pg/mL in controls, p < 0.001), enhanced monocyte/macrophage infiltration into periodontal tissues (7.75 [1.96]% vs. 4.82 [0.82]% in controls, p < 0.01) and brain (2.78 [0.91]% vs. 1.38 [0.47]% in controls, p < 0.01), disrupted the integrity of periodontal tight junction and blood–brain barrier and increased anxiety‐like behaviours. Administration of CCL2‐neutralising antibody reduced anxiety‐like behaviours, attenuated alveolar bone loss and local inflammation and decreased monocyte/macrophage infiltration and barrier integrity disruption.ConclusionThe study identifies the association between periodontitis and anxiety, with the CCL2‐mediated inflammatory pathogenesis as an underlying mechanism.