In vitro evaluation of hesperitin nanofibers on inflammatory cells and bone metabolism

Abstract

Nanofibers is a new class of biomaterials with a wide range of possible applications to the area of nanobiotechnology and tissue engineering. Due to their different characteristics, nanofibers can promote greater cell/matrix-cell interaction, as well as the incorporation of bioactive molecules, reinforcing their biological properties. Hesperitin is a hesperidin metabolite with effective therapeutic properties such as antioxidant, anti-inflammatory, anticancer, analgesic and with beneficial effects on bone metabolism, thus can be consider as a new therapeutic agent for the treatment of periodontal disease. Therefore, our hypothesis is that hesperitin nanofibers reduce the production of inflammatory mediators, promotes osteogenesis and blocks osteoclastogenesis. To test our hypothesis, we propose the following aims: Objective 1: To evaluate the action of hesperitin nanofibers on the expression of inflammatory mediators and cytokine gene expression. RAW 264.7 macrophage-like cell cultures will be seeded and stimulated with Porphyromonas gingivalis and Escherichia coli LPSs, and evaluated by Bioplex 200 and RT-PCR. Objective 2: To evaluate the influence of hesperitin nanofibers on the differentiation and metabolic activity of osteoblasts and osteoclasts. MC3T3-E1 pre-osteoblastic cells will be used to evaluate the osteoblast-scaffold interactions by RT2 Profiler PCR microarray and formation of mineralized nodules. To evaluate the osteoclast-scaffold interactions will be used Human osteoclast precursor cells derived from hematopoietic stem cells isolated from human bone marrow stimulated with RANKL and analyzed the cell viability and osteoclasts activity by immunohistochemistry of TRAP. The numerical data obtained will be submitted to the specific statistical analysis using the software GraphPad Prism 6, and all the tests of this study will be applied with significance level of 5% (p <0.05). (AU)