Persistent apical periodontitis after root canal treatment: clinical-laboratory evaluation associated with bioinformatics analysis applied to endodontics

• Grant number: 23/16757-5
• Support Opportunities: Regular Research Grants
• Start date: February 01, 2025
• End date: January 31, 2027
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Manoel Damiao de Sousa Neto
• Grantee: Manoel Damiao de Sousa Neto
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Precision medicine is part of the evolution of contemporary medicine based on scientific evidence and proof, that seeks to reduce errors and optimize results when making health decisions and recommendations, as well as being a potentially practical and economically viable alternative to current practices in disease prevention, diagnosis, treatment and prognosis determination. In parallel, through advances in Bioinformatics analysis, the search for the "golden biomarker" could be simplified, especially in more complex disease states, and the investigation of biosignatures could be the future, as it could lead to the development of simplified diagnostic tests that benefit clinicians and patients, leading to the development of Precision Dentistry. In endodontics, in addition to microbial and mechanical factors related to root canal treatment itself, intrinsic host factors can interfere with treatment success and the need for reintervention. In view of this, the aim of this study is to investigate the involvement of molecular aspects of the host related to the development of persistent periapical lesions after root canal treatment, by means of PCR allelic discrimination of TP53, CCL5 and CCR5 gene polymorphisms, PCR gene expression analysis of TP53, EP300, CCL5, CCR1, CCR3 and CCR5, histopathological and immunohistochemical evaluation of P53, P300, CCL5, CCR1, CCR3 and CCR5, as well as total RNA sequencing (RNA-Seq) of these lesions. In addition, the clinical and imaging aspects observed in the development, progression and repair of apical periodontitis, by means of semiologic examinations and cone beam computed tomography (CBCT), will be used as phenotypic parameters for understanding the molecular aspects involved in the etiology of persistent endodontic infection. Patients diagnosed with pulp necrosis who, at least one year after endodontic treatment, still have a persistent periapical lesion and patients with complete repair of the lesion (control group) will be selected for the study. Saliva samples will be collected as a source of genomic DNA and, in cases requiring surgical intervention, the periapical lesion will be collected. After processing the samples, TP53, CCL5 and CCR5 genotyping will be carried out, as well as TP53, EP300, CCL5, CCR1, CCR3 and CCR5 gene expression analysis, histopathological classification and immunohistochemical analysis of P53, P300, CCL5, CCR1, CCR3 and CCR5, in addition to total RNA sequencing associated with Bioinformatics analysis. In this way, the project aims to contribute to a change in the scientific paradigm in relation to the diagnosis, prevention and treatment of periapical lesions persistent to root canal treatment. Thus, being able to play a fundamental and decisive role in the planning and execution of clinical procedures and their prognosis through precision dentistry. (AU)