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Effects of the phytocystatin CsinCPI-2 in the experimental periodontal disease induced in mice

Grant number: 17/19863-0
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): December 01, 2017
Effective date (End): November 30, 2019
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Joni Augusto Cirelli
Grantee:Carlos Eduardo de Azevedo Mariano
Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Connective tissue destruction in periodontal diseases can potentially involve the action of several types of host-derived proteinases. Among them, the cysteine proteinases, especially cathepsins, have the ability to degrade many extracellular matrix components, mainly collagen. The cysteine proteinase is also known to play an important role in the beginning and progression of immune-inflammatory processes. In this context, the cystatins are reversible inhibitors of cysteine proteinases whose action mechanism is based on competitive inhibition by blocking the proteolytic activity. A new cystatin derived from orange was characterized and named CsinCPI-2. This inhibitor produced in recombinant form was demonstrated to be efficient in inhibiting human cathepsins, as well as other cysteine proteinases. The understanding of the role of cysteine proteinases, especially cathepsins, in the progression of periodontal disease may lead to new therapies for the periodontal disease control. Thus, the aim of this study is to evaluate the effect of CsinCPI-2 inhibitory activity on proinflammatory cathepsins and cytokines in the immune-inflammatory response and progression of periodontal destruction in mice submitted to experimental induction of periodontal disease. For this, male Swiss mice will be distributed in 4 experimental groups (n = 8 animals / group): Group 1 - negative control group; Group 2 - periodontal disease will be induced by placing ligatures on the maxillary first molar bilaterally and animals will receive intraperitoneal PBS injection daily; Groups 3 and 4 - periodontal disease will be induced and animals will receive intraperitoneal injection of CsinCPI-2 (1 ¼l / g of weight) daily at concentrations of 0.8 ¼g / ¼l and 1.6 ¼g / ¼l, respectively. The immuno-inflammatory process present in the gingival tissue will be evaluated through histological analysis and real-time RT-PCR. The evaluation of alveolar bone loss will be evaluated by histomorphometry (AU)