Effect of conditioned medium from stem cells of the apical papilla on peripheral monocytes surveillance in vitro

• Grant number: 22/05303-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2022
• End date: August 31, 2023
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Carla Renata Sipert
• Grantee: Rafael Victor Albuquerque de Abreu
• Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Bone and root resorption are among the main responsible causes of tooth loss. Regenerative endodontic procedures have been recently proposed as an alternative for the treatment of external root resorption. Monocytes are the osteoclast precursor and they are considered key cells in this context. Stem cells of the apical papilla (SCAP) are in close contact with bone along root formation and they may display some role in bone metabolism. This study aims to investigate the potential of conditioned media from SCAP on in vitro surveillance of monocytes. Primary cultures of SCAP will be established for conditioned media obtaining (CM). Human peripheral blood monocytes will be isolated and cultured. These cells will be cultured with CM or medium only being both added by M-CSF and RANKL for 7, 14, and 21 days. At each time point, cytotoxicity will be evaluated by using MTT [(3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)] assay. Next, monocytes will be subjected to dUTP nick end labeling (TUNEL) assay at determined time points. Finally, SCAP supernatants will be quantified for osteoprotegerin, interleukin-33, and TNF-related apoptosis-inducing ligand (TRAIL). Statistical analysis will be performed by ANOVA or Kruskal Wallis test (p<0.05). (AU)