Evaluation of inflammatory response and immunomodulation in polycaprolactone membranes incorporated with bioactive glasses 45S5, 58S and S53P4

• Grant number: 22/09045-6
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: September 30, 2022
• End date: February 28, 2023
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Luana Marotta Reis de Vasconcellos
• Grantee: Marina Santos Fernandes
• Supervisor: Rene Olivares-Navarrete
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
• Institution abroad: Virginia Commonwealth University (VCU), United States
• Associated to the scholarship: 21/09544-0 - Synthesis and characterization of polycaprolactone membranes incorporated with 45S5, 58S and S53P4 bioglass: in vitro osteogenesis and in vivo bone neoformation comparative analysis, BP.MS

Abstract

Associated with the process 2021/09544-0, in which the synthesis and characterization of membranes of polycaprolactone (PCL) incorporated with bioactive glasses 45S5, 58S and S53P4 are being evaluated, as well as in vitro osteogenesis and in vivo bone neoformation, the present project applies for a five-month research internship in the Virginia Commonwealth University (VCU). Currently, one of the biggest challenges in the field of biomaterials and regenerative medicine is to understand the inflammatory response and immunomodulation of the host after implantation. It is known immune cells are directly interacting with each other and the biomaterial, generating a cascade of events that can lead to the success of integration or complete failure. Therefore, the aim of this project is to complement the biological study of these biomaterials through the analysis of the inflammatory response and immunomodulation, for a better understanding of the host response, in addition to the investigation of cellular and molecular events resulting from the implantation. For this, in vitro and in vivo tests will be performed. For in vitro tests, bone marrow cells will be isolated from the femur of C57Bl6 mice and cultured for seven days, and naive macrophages will be generated in a culture medium supplemented with a macrophage colony-stimulating factor. After 24 hours the media will be collected and, the analysis of the inflammatory cytokines of IL-1², IL-4, IL-6, IL-10, Il-17, and TNF± will be performed by enzyme immunosorbent assays (ELISA), as so analysis of the expression of the inflammatory genes Arg1, l1b, Nos2, Tnf, Il1rn, Tgfb1. For the in vivo tests, C57BL/6 10 weeks male mice (Jackson Strain #000664) will be submitted to the same surgery performed in the student's master's research project to create a bone defect and fill it with the biomaterials, after 24 hours, 3 and 7 days postoperatively, blood will be collected by cardiac puncture for the Multiplex ELISA assay. Femurs will be removed for flow cytometry multicolor analysis, which it will characterize the percentage of immune cells present at the site of the injury. The data will be analyzed by software programs suitable for testing, then the data will be statistically analyzed, initially through the normality test aiming at the application of the appropriate statistical test. The significance level adopted will be the conventional value of 5%. (AU)