DIACEREIN EFFECT ON BONE CELLS ACTIVITY MARKERS IN INDUCED PERIODONTITIS

• Grant number: 23/15699-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2024
• End date: February 28, 2025
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Paulo Sergio Cerri
• Grantee: Lays Cristina Gouvea
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Periodontitis (P) is a multifactorial and immunoinflammatory disease that culminates with the degradation of periodontal tissues (soft and hard tissues). P progression is under a complex control of several mediators including interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-±), matrix metalloproteinases (MMPs). These mediators promote the osteoclast formation, differentiation and activity leading to bone resorption. Diacerein, an anti-inflammatory drug which has an effect inhibitory on IL-1 and TNF-±, is used for the treatment of rheumatoid arthritis, disease similar to P. The aim of the study will be to evaluate whether diacerein reduces the immunoexpression of MMP-9 and ATVase, enzymes involved with osteoclast activity, in osteoclasts of alveolar bone in molars with induced P. Moreover, will be evaluated whether diacerein interferes with osterix and alkaline phosphatase, factors associated with differentiation and activity of osteoblasts. Immunoexpression of IL-10 which participates in the tissue repair will also be evaluated in induced P. 54 Holtzman rats will be distributed into PDG (periodontitis diacerein-treated group), PSG (periodontitis sham group) and CG (Control group; healthy periodontium). P will be induced with the insertion of a cotton thread in the cervical colon of the upper first molars. After 7 days, the ligature will be removed and the rats with P will receive 100 mg/kg body weight of diacerein (PDG) or physiological solution (PSG) by gavage for 7, 15 and 30 days. Thus, in each group will be allocated 6 animals per period. After paraffin-embedded, the non-serial sections will be stained with hematoxylin and eosin (HE) for selection of choice region (interdental gingiva, located between first and second molars, and alveolar process around the roots of the first molar). Sections will be submitted to immunofluorescence/immunohistochemistry reactions for detection of V-ATPase, MMP-9, osterix, alkaline phosphatase and IL-10. The quantification of V-ATPase and MMP-9 immunoexpression in osteoclasts will be performed using a fluorescent microscope and a analysis software (Leica Application Suite-LAS4.3). Total area of osteoclast will be measured and, subsequently, V-ATPase and MMP-9 immunofluorescent area will be also measured. Thus, will be obtained the percentual of immunofluorescent area for V-ATPase and MMP-9 in osteoclast. Number of immunolabelled cells for alkaline phosphatase, osterix and IL-10 will be computed in a standardized area using a light microscopy Olympus and an image analysis software. The quantitative data will be submitted to two-way ANOVA and Tukey post-test (p < 0.05).