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Development and characterization of titanium surfaces coated with bioactive molecules

Grant number: 19/20783-6
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): October 01, 2020
Effective date (End): September 30, 2023
Field of knowledge:Health Sciences - Dentistry - Dental Materials
Principal Investigator:Carlos Alberto de Souza Costa
Grantee:Taisa Nogueira Pansani
Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil


Modifications on implant surface aims to obtain a bioactive substrate capable of improving the interaction between implants and oral tissues specially in challenging and difficult osseointegration conditions. One of the most common surface modification method actually used is the chemical/thermic treatments, which produces a porous bioactive layer on Titanium (Ti) surface allowing the direct binding of molecules on it. Extracellular Vesicles (EVs) are specialized messengers present in biological fluids that deliver their components to other cells, promoting physiological changes and inducing tissue regeneration. Similarly, Hydroxyapatite (HA) is an osteoconductive and bioactive material used for bone neoformation and mineralization. Therefore, the objectives of this project are: to isolate and characterize extracellular vesicles (EVs) from mature osteoblasts (study 1); to determine the cytocompatible concentration of EVs and poly(e-caprolactone) nanofibers (PCL) containing hydroxyapatite nanoparticles (nHA) in pre-osteoblast culture (study 2); to develop and characterize Ti surfaces coated with EVs and PCL-nanofibers containing nHA (study 3); and to evaluate in vivo new Ti surfaces coated with bioactive molecules (study 4). EVs will be isolated from mature osteoblasts (MC3T3-E1) and characterized regarding their concentration, size and composition. The nHA will be incorporated into the PCL nanofibers at different concentrations and then, the best cytocompatible concentration of EVs and PCL nanofibers containing nHA will be determined in pre-osteblasts obtained from mouse calvaria. For these purposes, the surface of Ti discs will be treated with 5M of NaOH alkaline solution and EVs applied on them or the Ti surface will be covered with nHA-containing PCL nanofibers. Cell viability, total proteins and alkaline phosphatase synthesis, formation of mineralization nodules, gene expression and protein synthesis of some osteogenic markers will be assessed, as well as the morphology of the cells seeded on the bioactive Ti surfaces. For in vivo analysis (Study 4), histological and histomorphometric evaluations of Ti surfaces implanted into calvaria of rats will be performed. Quantitative data will be evaluated for normality and homoscedasticity and appropriate statistical tests will be selected considering a significance level of 5%. (AU)

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