Host-pathogen interaction between a three-dimensional tissue model with a binary cell system and dual species biofilm Under Normoglycemic and Diabetic Conditions

• Grant number: 25/13858-0
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: October 01, 2025
• End date: March 31, 2026
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Flaviana Bombarda de Andrade
• Grantee: Mirela Cesar de Barros
• Supervisor: Anil Kishen
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
• Institution abroad: University of Toronto (U of T), Canada
• Associated to the scholarship: 24/17193-0 - Impact of endodontic treatment of teeth with pulp necrosis and periapical lesion in patients with type II diabetes mellitus: Microbiological and tomographic evaluation, BP.DR

Abstract

Apical periodontitis is characterized by inflammation and bone destruction in the periapical tissues in response to bacteria and their virulence factors within the root canal system. Studies have shown that diabetes mellitus (DM) influences both the pathogenesis and prognosis of pulpal and periapical diseases, favoring the proliferation and colonization of microorganisms, particularly strict anaerobes. Therefore, understanding the interactions between the microbiota, the immune response, and host tissue cells is essential for optimizing treatment strategies that promote periradicular healing. This study aims to evaluate the in vitro response of a three-dimensional tissue model composed of periodontal ligament fibroblasts (PdLFs) and macrophages (MQs) under normoglycemic and diabetes-simulated conditions, in the presence of a dual-species biofilm and proinflammatory cytokines. PdLFs and MQs will be seeded into 3D-printed dumbbell-shaped molds to generate tissue constructs with a binary cell population. Fusobacterium nucleatum and Parvimonas micra biofilms will be grown in 24-well plates under anaerobic conditions for 7 days. Three experimental conditions-biofilm supernatant, interleukin-1 beta, and tumor necrosis factor-alpha-as well as a control (no stimulation), will be applied to the tissue constructs to assess tissue characteristics, viable cell volume, and morphology using confocal laser scanning microscopy over a 0- to 7-day period. Experiments will be conducted in triplicate, and data will be tested for normality and subsequently analyzed using appropriate parametric or non-parametric statistical tests, with a significance level set at 5%. (AU)