Pro-osteogenic effect of recombinant sugarcane cystatin, CaneCPI-5, on human dental pulp cells

Author(s)

Ana Flávia Balestrero Cassiano

• Document type: Master's Dissertation
• Press: Araraquara. 2022-08-25.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Odontologia. Araraquara
• Defense date: 2022-06-28
• Advisor: Gisele Faria

Abstract

The increase in cysteine cathepsins activity (a group of cysteine proteases) is related to the development of osteolytic diseases, such as osteoporosis and apical periodontitis, as they participate in bone resorption through the proteolytic degradation of organic matrix components. Cystatins are natural and reversible inhibitors of cysteine proteases. Phytocystatins are plant cystatins, some of them have already been produced recombinantly, such as CaneCPI-5 (derived from sugarcane), which is the focus of this study because it showed pro-osteogenic potential observed in a preliminary study of our research group. The aim of this study is to evaluate the cytocompatibility and effect of CaneCPI-5 on proliferation, migration, osteogenic and odontogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs exposed to CaneCPI-5 and unexposed (control) were evaluated for viability using the alamarBlue assay, proliferation by bromodeoxyuridine incorporation (BrdU) assay, migration by transwell assay, deposition of inorganic precipitates by alizarin red staining and tissue-nonspecific alkaline phosphatase (TNAP) enzyme activity. Data were evaluated by one-way or two-way ANOVA, followed by Tukey or Kruskal-Wallis post-test and Dunn or Mann Whitney post-test, with α = 0.05. CaneCPI-5 was cytocompatible, induced higher migration, proliferation, formation of mineralized nodules and TNAP activity in hDPSCs compared to control. Thus, it can be concluded that CaneCPI-5 constitutes a molecule with promising potential to be used in therapies that aim the pulp and periapical repair/regeneration. (AU)

• FAPESP's process: 20/09576-6 - Evaluation of the pro-osteo/odontogenic effect of recombinant sugarcane cystatin, CaneCPI-5, on human pulp cells
• Grantee: Ana Flávia Balestrero Cassiano
• Support Opportunities: Scholarships in Brazil - Master