Osteogenic differentiation of pulp cells induced by cystatin derived from sweet orange (CsinCPI-2): evaluation of the involvement of the signaling pathway BMP2-Smad1/5/8

Abstract

Results published by our research group showed that cystatin CsinCPI-2, derived from citrus sinensis (sweet orange), was able to inhibit the gene expression and enzymatic activity of human cysteine cathepsins, showed anti-inflammatory activity and, had pro-osteogenic effect on human dental pulp cells. This pro-osteogenic effect was demonstrated by the increase in the alkaline phosphatase (ALP) activity, deposition of mineralized nodules and the gene expression of osteogenic markers such as bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX2), ALP, osteocalcin and bone sialoprotein (BSP). Considering that CsinCPI-2 induced the differentiation of human dental pulp cells into osteogenic phenotype, it is important to know the signaling pathways involved in this differentiation, for a better understanding of the effect of the protein on undifferentiated mesenchymal cells and, consequently, to indicate it or not for reparative therapy in endodontics and other areas. Thus, the objective of the scientific initiation project is to study the possible role of the BMP2-Smad1/5/8 signaling pathway in differentiating of human pulp cells in osteogenic phenotype induced by CsinCPI-2. Specifically, in the scientific initiation project, it will be evaluate, in the human dental pulp cells exposed to CsinCPI-2, treated or not with BMP inhibitor, the gene expression of BMP-2, SMAD 1, SMAD 5 and RUNX2 by means of reverse transcription polymerase chain reaction quantitative real time (qRT-PCR).