Synthesis and characterization of polycaprolactone membranes incorporated with 45S5, 58S and S53P4 bioglass: in vitro osteogenesis and in vivo bone neoformation comparative analysis

• Grant number: 21/09544-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: May 01, 2022
• End date: September 01, 2023
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Luana Marotta Reis de Vasconcellos
• Grantee: Marina Santos Fernandes
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
• Associated scholarship(s): 22/09045-6 - Evaluation of inflammatory response and immunomodulation in polycaprolactone membranes incorporated with bioactive glasses 45S5, 58S and S53P4, BE.EP.MS

Abstract

The advancement of regenerative medicine in tissue repair and biomaterials leads to improvement on existing treatments and technologies, aiming to optimize the resolution of injuries related to bone diseases, and allows increasingly efficient treatments for the population. Thus, the aim of this study will be to evaluate the influence of polycaprolactone (PCL) membranes incorporated with three different types of bioglass: 45S5 bioglass (PB45), 58S bioglass (PB58) and S53P4 bioglass (PB53), on the activity and differentiation of osteoblasts in vitro, as well as in bone neoformation in vivo. The synthesis of all bioglasses will be carried out by the Sol-Gel route, which will be incorporated into the PCL solution prior to the production of the membranes through the electrospinning process, under three fixed parameters: 2 mLh-1 flow rate, 10 kV of voltage and 12 cm of distance. The membranes will be characterized morphologically (MEV, fiber diameter analysis and wettability analysis) and chemically (EDS, DRX, FTIR and bioglass zeta potential values). After characterization, the membranes will be submitted to in vitro biological tests of cell viability, evaluation of total protein content, alkaline phosphatase activity, analysis of cell morphology by direct fluorescence, genotoxicity and formation of mineralization nodules and calcium quantification. At this stage, mesenchymal cells obtained from rat femurs will be isolated and plated with the membranes, in order to assess their influence on the activity and cell differentiation of osteoblasts. In the antimicrobial analysis, the membranes will be cultivated with strains of Staphylococcus aureus and Streptococcus mutans and Pseudomonas aeruginosa in order to evaluate the formation of monotypic biofilms, which will be quantified by the MTT method. For the biological tests performed in the in vivo step, 30 rats will undergo surgery for critical bone defect in the right and left tibias, which will be filled with the same material, PCL, PB45, PB58, PB53. There will also be a control group, in which the filling of defects will be with a clot. The animals will be euthanized 4 weeks after the surgical procedure. For all groups, the right femurs will be submitted to histological, histomorphometric and immunohistochemical analysis, while the left femurs will be submitted to the three-point flexion test. 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)