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Sugarcane-derived cysteine protease inhibitor: effect on osteoblastic and odontoblastic differentiation of human dental pulp cells

Grant number: 22/07823-1
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): April 01, 2023
Effective date (End): February 28, 2027
Field of knowledge:Health Sciences - Dentistry - Endodontics
Principal Investigator:Gisele Faria
Grantee:Luana Raphael da Silva
Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil


Cathepsins, a group of cysteine proteases, play an important role in some pathological processes. It is known that an increase in their activity, especially cathepsin K, is related to the development of diseases such as osteoporosis and apical periodontitis, as they participate in the bone resorption process. Cystatins are natural and reversible inhibitors of cysteine proteases. The cystatins that inhibit cathepsin K constitute an emerging class of drugs that are potent antagonists of osteoclastic activity, reducing bone loss. In addition to the inhibition of cathepsin K, some studies have shown that cystatins may have a pro-osteogenic effect. Phytocystatins are plant cystatins, and some of them have already been produced recombinantly, such as CaneCPI-5 derived from sugarcane. The CaneCPI-5 anti-inflammatory effect and ability to inhibit human cathepsins have already been demonstrated. Furthermore, a preliminary study by our research group showed, in pre-osteoblasts (MC3T3), that it has pro-osteogenic potential. The aim of this study will be evaluate the effect of CaneCPI-5, in contact with dentin blocks, on human dental pulp cells (hDPCs). Different methodological approaches will be used, such as viability, adhesion and spread assays, as well as differentiation in mineralizing phenotype of hDPCs. Additionally, assays will be carried out seeking to know the involvement of the BMP/SMAD pathway in the osteogenic and/or odontogenic differentiation of hDPCs stimulated by CaneCPI-5 using quantitative real-time polymerase chain reaction (RT-qPCR) and Western Blot tests. If the results are promising, CaneCPI-5 could constitute a molecule with potential for biotechnological applications, such as in treatment techniques that aim to repair the dentin-pulpal complex and/or periapical region, including endodontic regeneration techniques.

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