Simvastatin-Loaded Chitosan Microspheres as a Biomaterial for Dentin Tissue Engineering

In the present study, chitosan microspheres (MSCH) loaded with different concentrations of simvastatin (2%, 5%, and 10%) were synthesized as a biomaterial for dentin tissue engineering. The microspheres were prepared by emulsion crosslinking method, and simvastatin was incorporated during the process. The microspheres were then physicochemically and morphologically characterized. Scanning electron microscopy and infrared spectroscopy confirmed the spherical morphology of synthesized microspheres and the chemical incorporation of simvastatin into MSCH, respectively. UV-visible absorption confirmed the controlled and continuous release pattern of the drug. To mimic the clinical application in vitro, the microspheres were applied onto three-dimensional (3D) cultures of human dental pulp cells (HDPCs). Cell viability, proliferation, and in situ-mineralized matrix deposition were evaluated. The results indicated no cytotoxic effects for all 3D cultures for all tested biomaterials, with cells being able to proliferate significantly over time. HDPCs showed a significant increase in the deposition of mineralization nodules when 3D cultures were in direct contact with chitosan microspheres in comparison to control; nevertheless, the highest expression was observed for MSCH encapsulated with 5% and 10% simvastatin, which was significantly higher than plain MSCH. Therefore, chitosan microsphere systems loaded with 5%-10% simvastatin provided the development of a controlled release system in bioactive dosages for dentin tissue engineering.