In vitro evaluation of adhesion/proliferation of human gingival fibroblasts on demineralized root surfaces by toluidine blue O in antimicrobial photodynamic therapy

Abstract

BackgroundAntimicrobial photodynamic therapy (aPDT) in Dentistry has important effects as bacterial destruction in areas with periodontal disease. Some dyes applied in aPDT could present low pH and, consequently, result in tooth demineralization. This study evaluated demineralization produced by aPDT with toluidine blue O (TBO) at low pH and analyzed adhesion/proliferation of human gingival fibroblasts (HGF). MethodsIn the 1st phase, bovine enamel and root dentin fragments received 2 treatments: PDT4 group (TBO–100μg/ml—pH 4–60s) plus laser (660nm, 45J/cm2, 1.08J, 30mW, 30s, spot 0.024cm2, 1.25W/cm2, sweeping, non-contact) and CA group (citric acid plus tetracycline—pH 1–180s). Surface hardness loss and tooth wear were statistically analyzed (Student’s t test, ANOVA/Tukey, p<0.05). In the 2nd phase, human dentin fragments were divided in C (control group—scaling and root planing), PDT4 and CA. HGF (104, 5th passage) were cultured on these fragments for 24, 48 and 72h and counted in scanning electron microscopy photographs. Number of HGF was analyzed using repeated-measures ANOVA and Tukey (p<0.05). ResultsPercentage of surface hardness loss was similar in dentin for PDT4 (71.5%) and CA (76.1%) (p>0.05) and higher in enamel for CA (68.0%) compared to PDT4 (34.1%) (p<0.05). In respect to wear, no difference was found between PDT4 (dentin: 12.58μm, enamel: 12.19μm respectively) and CA (dentin: 11.74μm and enamel: 11.03μm) (p>0.05). Number of HGF was higher after 72h in CA group (2.66, p<0.05) compared to PDT4 (2.2) and C (1.33). ConclusionPDT4 is not as aggressive as CA for enamel. However, dentin demineralized promoted by PDT4 does not stimulate HGF adhesion and proliferation as CA.