Abstract
BACKGROUND: The aim of this study was to evaluate the indirect effects of residual silver nanoparticles (AgNPs) gel on human dental pulp stromal cells (DPSCs). METHODS: Ninety-five dentin discs (4x4x1 mm) were prepared from freshly extracted human single-rooted teeth following institutional ethical approval and informed consent. Samples were cleaned, autoclaved, and treated with: 1.5%NaOCl, Saline and 17% EDTA then randomly assigned to 5 groups that received 50 μl of one of the following treatments: 0.01%AgNPs, 0.015%AgNPs, 0.02%AgNPs, Calcium hydroxide (Ca (OH)(2)) or no treatment for 1 week. Discs were washed with Saline and 17%EDTA then seeded with DPSCs and incubated for 3 and 7 days. At 24 hours unattached cells were collected and counted. At each time point cytotoxicity (LDH assay), cell viability (live/dead staining and confocal microscopy) and cell proliferation (WST1 assay) were assessed. All experiments were repeated a minimum of 3 times using DPSCs isolated from 3 different donors for each time point assessed (n = 9/group). Statistical analysis was done using One-Way ANOVA followed by Tukey's test and Kruskal Wallis followed by post-hoc comparisons with significance set at p ≤ 0.05. RESULTS: After 24 hours, the percentage of DPSCs attachment ranged between 92.66% ±4.54 and 95.08% ±1.44 with no significant difference between groups (P = 0.126). Cell viability was ≥92% at 24 hours for all groups. However this percentage dropped to less than 60% at 3 days then started to rise again at 7 days. There was no significant difference in cytotoxicity between different groups at all time points except for 0.01%AgNPs group which had the highest cytotoxicity. DPSCs proliferation increased significantly from 3 to 7 days in all groups except for Ca (OH)(2) which showed lower proliferation rates at both 3 (45.89%) and 7 days (79.25%). CONCLUSION: Dentin discs treated for 7 days with concentrations of AgNPs gel (0.01-0.02%) allowed more than 90% DPSCs cell attachment after 24 hours. The cytotoxicity and proliferation of DPSCs in response to AgNPs gel were comparable to those with calcium hydroxide. This suggests that AgNPs gel may represent a promising future candidate for clinical use in regenerative endodontics. However, its effects may be concentration-dependent warranting further investigation.