Biological Analysis of Simvastatin-releasing Chitosan Scaffold as a Cell-free System for Pulp-dentin Regeneration

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Soares, Diana G.
Anovazzi, Giovanna [UNESP]
Bordini, Ester Alves F. [UNESP]
Zuta, Uxua O. [UNESP]
Silva Leite, Maria Luísa A. [UNESP]
Basso, Fernanda G. [UNESP]
Hebling, Josimeri [UNESP]
de Souza Costa, Carlos A. [UNESP]

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Introduction: The improvement of biomaterials capable of driving the regeneration of the pulp-dentin complex mediated by resident cells is the goal of regenerative dentistry. In the present investigation, a chitosan scaffold (CHSC) that released bioactive concentrations of simvastatin (SIM) was tested, aimed at the development of a cell-free tissue engineering system. Methods: First, we performed a dose-response assay to select the bioactive dose of SIM capable of inducing an odontoblastic phenotype in dental pulp cells (DPCs); after which we evaluated the synergistic effect of this dosage with the CHSC/DPC construct. SIM at 1.0 μmol/L (CHSC-SIM1.0) and 0.5 μmol/L were incorporated into the CHSC, and cell viability, adhesion, and calcium deposition were evaluated. Finally, we assessed the biomaterials in an artificial pulp chamber/3-dimensional culture model to simulate the cell-free approach in vitro. Results: SIM at 0.1 μmol/L was selected as the bioactive dose. This drug was capable of strongly inducing an odontoblastic phenotype on the DPC/CHSC construct. The incorporation of SIM into CHSC had no deleterious effect on cell viability and adhesion to the scaffold structure. CHSC-SIM1.0 led to significantly higher calcium-rich matrix deposition on scaffold/dentin disc assay compared with the control (CHSC). This biomaterial induced the migration of DPCs from a 3-dimensional culture to its surface as well as stimulated significantly higher expressions of alkaline phosphatase, collagen type 1 alpha 1, dentin matrix acidic phosphoprotein 1, and dentin sialophosphoprotein on 3-dimensional–cultured DPCs than on those in contact with CHSC. Conclusions: CHSC-SIM1.0 scaffold was capable of increasing the chemotaxis and regenerative potential of DPCs.



Cell differentiation, dental pulp, scaffolds, tissue engineering

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Journal of Endodontics, v. 44, n. 6, p. 971-976.e1, 2018.