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Effect of phytocystatin Citrus CPI-2 on the proliferation, migration and differentiation of cells from human apical papilla and dental pulp

Abstract

Cathepsins are enzymes that participate in the process of bone resorption by proteolytic degradation of the matrix components, and that, in disorder, lead to the development of some pathological conditions as osteoporosis, periodontitis, apical periodontitis (periapical lesion), among others. In this context, it is known that the use of cathepsin inhibitors, known as cystatins, can control bone resorption. Phytocystatins are class of specific cystatins obtained from plants, as is the case of Citrus CPI-2 derived from citrus. Previous results from our research group showed that Citrus CPI-2 was able to inhibit the enzymatic activity of human cathepsins and had anti-inflammatory potential. Furthermore, Citrus CPI-2 showed potential to induce differentiation of undifferentiated cells into osteo/odontogenic phenotype, indicating its potential use in the modulation of cellular metabolism. Based on the above considerations, the aim of the project is to study the effect of phytocystatin Citrus CPI-2 on the proliferation, migration, differentiation and mineralization of primary culture cells from human apical papilla and dental pulp. If it is confirmed that Citrus CPI-2 stimulates osteo/odontogenic differentiation of pulp and apical papilla cells, additional tests will be performed, seeking to understand the involvement of the BMP / SMAD pathway in this process. Different methodological approaches, such as proliferation and cell migration assays, gene expression analysis by qPCR, protein expression analysis by immunobloting, alkaline phosphatase activity and alizarin red staining for the detection of inorganic precipitates will be performed. If the hypothesis that Citrus CPI-2 induces osteo/odontogenic differentiation of cells from the dental pulp and apical papilla is confirmed, it may be suggested as a promising molecule for biotechnological applications, especially for direct pulp capping and endodontic regeneration procedures, or as a component of intracanal medication to promote pulp and periapical repair/regeneration. In addition, other potential applications occur at systemic levels, as in the case of bone pathologies that affect the whole skeleton. (AU)

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VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
LEGUIZAMON, NATALIA DA PONTE; RODRIGUES, ELISANDRA MARCIA; DE CAMPOS, MICHEL LEANDRO; BOAS NOGUEIRA, ANDRESSA VILAS; VIOLA, KENNIA SCAPIN; SCHNEIDER, VANESSA KARINE; NEO-JUSTINO, DANIELA MORILHA; TANOMARU-FILHO, MARIO; ZAMBUZZI, WILLIAN FERNANDO; HENRIQUE-SILVA, FLAVIO; SOARES-COSTA, ANDREA; FARIA, GISELE; CIRELLI, JONI AUGUSTO. In vivo and in vitro anti-inflammatory and pro-osteogenic effects of citrus cystatin CsinCPI-2. CYTOKINE, v. 123, NOV 2019. Web of Science Citations: 0.

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