Evaluation of the remineralizing potential of poly-aspartic acid (pAsp) and poly-glutamic acid (pGLU) in a 3D bioprinted dentin model using stem cells from the apical papilla (SCAPs) and its application in dentin affected by caries.

• Grant number: 23/18285-3
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: July 01, 2024
• End date: May 04, 2025
• Field of knowledge: Health Sciences - Dentistry - Dental Materials
• Principal Investigator: Fábio Dupart Nascimento
• Grantee: Marina Damasceno e Souza de Carvalho Chiari
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

This project aims to: 1) evaluate and characterize and cellular interactions with pAsp e/ou pGlu 2) bioprint a mineralizable 3D dentin matrix (scaffold) and 3) evaluate the mechanisms by which pAsp and/or pGlu can modulate the dentin neoformation process. Therefore, this project was divided in phases. Phase I: evaluation and characterization of cellular interactions with the pAsp and/or pGlu peptides. MDPC-23 and SCAPs cells will be cultivated in media containing or not the isolated and/or associated peptides and subsequently analyzed for: cell proliferation; expression of genes related to osteogenic differentiation (Osteogenesis RT2 ProfilerTM PCR array); protein immunodetection (Western Blotting); alkaline phosphatase assay and intracellular mapping of proteins modulated by peptides. Furthermore, the interaction of pAsp and/or pGlu with type I collagen will be evaluated by molecular docking and circular dichroism (CD). Phase II: scaffold bioprinting using a bioink loaded with MDPC-23 or SCAPs. The bioprinted dentin matrix will or will not receive the injection of pAsp and/or pGlu and will be characterized regarding: mechanical properties (compression test); degradation; rheology; microstructural analysis (scanning electron microscopy/SEM); total RNA (RNAt) extraction and mineral deposition assay (alizarin assay). Phase III: evaluation of the remineralizing potential of the bioprinted scaffold associated or not with peptides when applied to human dentin affected by caries. Remineralization will be quantified through the nanoindentation test (nanohardness/ elastic modulus) carried out on the transverse surface of the dentin after 30 days of storage in contact with the bioprinted material. Mineral recovery will also be assessed qualitatively through scanning electron microscopy/ energy dispersive x-ray spectroscopy (SEM/EDX). Quantifiable data will be subjected to ANOVA (analysis of variance). Tukey test will be used for pair-wise comparisons. The global significance level (±) will be set at 5%.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CHIARI, M. DAMASCENO E SOUZA; VICHI, F. MARON; SANTOS, E. IRAE ALMEIDA HUMMEL PIMENTA; DE OLIVEIRA, F. RODRIGUES; HONORIO, H. MARQUES; NASCIMENTO, F. DUPART; BRAGA, R. RUGGIERO. Calcium Silicate Particules: Biological and Mechanical Role in the Hybrid Layer. JOURNAL OF DENTAL RESEARCH, v. N/A, p. 9-pg., 2025-06-25. (20/05986-5, 23/10291-4, 23/18285-3)