Effect of sugarcane cystatin CaneCPI-5 on osteogenic differentiation of human dental pulp cells

Abstract

For pulp and periapical repair, or endodontic regeneration to occur, it is necessary for mesenchymal stem cells from the apical papilla and/or dental pulp to proliferate, migrate to the site of injury, and differentiate into cells that produce mineralized tissue. Therefore, materials used in endodontic therapy should stimulate those events. Phytocystatins are plant-derived cystatins capable of inhibiting cathepsins. Some of them are produced recombinantly, such as CaneCPI-5 (derived from sugar cane). Considering their pro-osteogenic potential, this study aimed to assess the cytocompatibility and effect of CaneCPI-5 on the proliferation, migration, and osteogenic differentiation of human dental pulp stem cells (hDPSCs). The hDPSCs exposed to CaneCPI-5 and unexposed (control) were evaluated as follows: cell viability, by alamarBlue assay; proliferation, by bromodeoxyuridine (BrdU) incorporation assay; migration, by transwell assay; deposition of mineralized nodules, by alizarin red staining; and activity of tissue-nonspecific alkaline phosphatase (TNAP). Data were evaluated by one- or two-way ANOVA, followed by Tukey or Kruskal-Wallis post-test and Dunn or Mann Whitney post-test. CaneCPI-5 was cytocompatible and induced higher migration, proliferation, formation of mineralized nodules, and TNAP activity in hDPSCs compared to control. In conclusion, CaneCPI-5 represents a molecule with promising application in endodontic therapy to stimulate events necessary for pulp and periapical repair/regeneration.