MIP-1α/CCL3 and SDF-1α/CXCL12 Production by Pulp Fibroblasts Stimulated by Materials

Objectives: The aim of this in vitro study was to quantify MIP-1α/CCL3 and SDF-1α/CXCL12 production in the supernatants of cultured human dental pulp fibroblasts challenged by different dental materials. Methods: Dental pulp of one lower third molar was collected and cultured in DMEM. Cells (50,000 cells/cm2) were plated on 24-well plates and incubated in a humidified incubator at 37oC with 5% CO2 and 95% air until reach confluence. Solid materials [Group 1 (G1) - Single Bond(3M ESPE), G2 – Vitrebond(3M ESPE), G3 - Ketac Molar(3M ESPE), G4 – Dycal(Dentsply)] were applied as round shaped specimens (2mm thick and 5mm in diameter) prepared according to the respective manufacturer's instructions into transwell membranes. Fluid materials [G5 - HEMA 10nM, G6 - HEMA 100nM, G7 - HEMA 1000nM, G8 - Single Bond unpolymerized in the concentrations of 1:100; G9 - 1:1,000 and G10 - 1:10,000] were added to the culture medium. G11 - control group (medium alone) and G12 - control group (transwell alone) were also evaluated. Chemokines production analysis was performed using ELISA at times of 6 and 24 hours. Data were compared by one way ANOVA (p<0.05) followed by Tukey's post hoc test (p<0.05). Results: Results were statistically significant for group G3 when comparing MIP-1α/CCL3 production at 6h and 24h (0±0 and 87.65±41.05 pg/mL, respectively, p<0.05). A time-dependent difference was also observed for SDF-1α/CXCL12 production in G12 at 6h and 24h (14.21±10.83 pg/mL and 77.39±15.94 pg/mL, respectively, p<0.05). Interestingly, G4 suppressed SDF-1α/CXCL12 production at 24h as compared to transwell alone (control group-G12) (77.39±15.94 pg/mL and 0±0, respectively, p<0.05). Conclusion: There was a time-dependent increase in MIP-1α/CCL3 production for G3, and for the control group (G12) regarding SDF-1α/CXCL12 production. There was a suppression in SDF-1α/CXCL12 production for G4 compared to the control group (transwell alone-G12). Financial support: FAPESP 2005/60167-0.