The influence of simvastatin on osteoblast functionality in the presence of titanium dioxide particles In-vitro

Abstract

Objective

Leaching of particles from dental titanium implant surfaces into preimplant microenvironment causes detrimental effects on bone cells. The current study investigated influence of simvastatin in mitigating adverse pro-inflammatory effects of titanium dioxide (TiO₂) micro (MP) and nano (NP) particles on hFOB 1.19 cells in vitro.

Design

Viability of hFOB 1.19 cells following exposure to varying concentrations of TiO₂ MPs and NPs and simvastatin were measured by XTT assay. hFOB 1.19 cells were treated with 100 µg/mL of TiO₂ MPs, 100 µg/mL of TiO₂ NPs, 0.1 µM simvastatin, 100 µg/mL of TiO₂ MPs+ 0.1 µM simvastatin and 100 µg/mL of TiO₂ NPs+ 0.1 µM simvastatin. After 24 h, ROS was measured by flow cytometry. On day 14, real-time PCR analysis for pro-inflammatory cytokines and bone formation markers was done for TNFα, IL1β, osteocalcin, ALP, and Col1 markers; while ALP and RANKL/OPG ratio were determined by colorimetric and ELISA assays respectively. Further, mineralization study using Alizarin Red S staining (ARS) and calcium quantification were performed.

Results

Exposure of hFOB to TiO₂ MPs and NPs generated ROS and reduced cell viability significantly, with upregulation of pro-inflammatory markers TNFα and IL1β and downregulation of bone formation markers OC and increased RANKL/OPG ratio and lowered degree of mineralization. Treatment with 0.1 µM of simvastatin treatment reversed the effects by mitigating oxidative stress, dampening pro-inflammatory markers, upregulation of bone formation markers, lowering RANKL/OPG ratio and increasing degree of mineralization.

Conclusion

Simvastatin possesses antioxidant, anti-inflammatory, and pro-osteogenic properties that may support bone healing around titanium implants.