Irrigation with Ambroxol Hydrochloride and Disodium EDTA in Regenerative Procedures: Effect on Growth Factor Expression

• Grant number: 25/06358-1
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: August 01, 2025
• End date: January 31, 2026
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Marco Antonio Hungaro Duarte
• Grantee: Letycia Accioly Simões Coelho
• Supervisor: Leticia Chaves de Souza
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
• Institution abroad: University of Pittsburgh (Pitt), United States
• Associated to the scholarship: 24/18863-0 - IN VITRO EVALUATION OF AMBROXOL HYDROCHLORIDE AS AN ALTERNATIVE IRRIGANT IN REGENERATIVE ENDODONTIC PROCEDURES: ANTIMICROBIAL, ANTI-BIOFILM, AND DENTINAL ULTRASTRUCTURAL EFFECTS ANALYSIS, BP.DR

Abstract

Regenerative endodontic procedures are indicated for the treatment of immature necrotic teeth, as they allow for root development, resulting in increased root length, thickness, and apical closure. Effective disinfection of the environment is a prerequisite for these procedures, and the elimination of the instrumentation step complicates cleaning and decontamination, which are carried out only by chemical solutions. The combined use of low concentrations of sodium hypochlorite (NaOCl) and 17% ethylenediaminetetraacetic acid (EDTA) has been recommended for the cleaning and disinfection of canals in necrotic immature teeth. However, studies have shown that NaOCl has deleterious effects on growth factors in the dentin matrix, and 17% EDTA has low antimicrobial efficacy. Therefore, the objectives of this research are to evaluate the use of Ambroxol Hydrochloride and 15% disodium EDTA, alone or in combination with different irrigating solutions, on the expression of VEGF-A, IGF-1, TGF-¿1, BMP-7, and BMP-2 from the dentin matrix and SCAPs culture (in vitro). For all tests, specimens will be divided into 9 groups (G1-G9) according to the irrigation protocol and further subdivided according to the irrigation method (n=3): conventional syringe and needle irrigation (SG1), ultrasound agitation (SG2), and Thera Lase Surgery laser agitation (SG3). To apply the irrigation protocols, bovine root dentin cylinders will be prepared and placed under a three-dimensional (3D) SCAPs culture simulating the apical papilla. Subsequently, the final concentrations of VEGF-A, IGF-1, TGF-¿1, BMP-7, and BMP-2 and their association with the dentin matrix and 3D cultures will be measured by enzyme-linked immunosorbent assay (ELISA). For all tests, the Shapiro-Wilk normality test will be applied, followed by parametric or non-parametric tests, with a 5% significance level adopted. (AU)