In Vitro Evaluation of the Protective and Immunomodulatory Potential of Annexin A1 During the Inflammatory Process: A Study in Primary Gingival Fibroblasts

• Grant number: 25/20352-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2025
• End date: October 31, 2026
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Principal Investigator: Mabelle de Freitas Monteiro
• Grantee: Melina Zambrotti Machado Donatoni
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil

Abstract

Periodontitis is a chronic inflammatory disease resulting from an imbalance between the host's immunoinflammatory response and the dental biofilm. Therefore, it is essential to investigate approaches that enable a better understanding of this pathology. In this context, Annexin A1 (ANXA1) stands out as a protein that modulates the inflammatory response, predominantly expressed in immune system cells, and is responsible for maintaining tissue homeostasis and promoting inflammatory resolution. Although its role is well established in systemic pathologies, its impact in the dental context remains poorly explored, making it crucial to deepen knowledge in this area to develop innovative diagnostic and therapeutic strategies for periodontitis. Therefore, the aim of this study will be to evaluate the effect of ANXA1-modulating molecules on gingival fibroblasts exposed or not to an inflammatory stimulus. For this purpose, primary cells collected from periodontally healthy patients in previous studies will be used. The obtained cells will undergo in vitro culture, followed by cell viability analysis to ensure the quality of the biological material. Subsequently, the cells will be exposed to a universal inflammatory inducer (PMA), along with the use of ANXA1-modulating molecules: an ANXA1 blocker (neutralizing monoclonal anti-ANXA1 antibody) and an inducer of this protein (AC2-26 peptide). These will be organized into six experimental groups to assess the cellular modulations that occur in the presence or absence of inflammation. Tissue RNA isolation and RT-PCR assays will be performed to evaluate gene expression, complemented by immunoenzymatic analyses. Finally, data will be subjected to statistical analysis, initially using the Shapiro-Wilk test to assess data normality. Parametric (ANOVA) or non-parametric (Kruskal-Wallis and Mann-Whitney) tests will then be applied, with the significance level set at 5%.