TRACKING TRANSCRIPTION FACTOR ACTIVITY IN 3D NANOSCAFFOLD

Mesenchymal stem cells (MSCs) are multipotent cells present in the stromal fraction of many tissues. Their immunomodulatory properties and differentiation potential have been proven beneficial in number of autoimmune disease therapy models. However a reliable in vitro approach to study MSCs differentiation in inflamed tissue and a clear insight into the involved signalling pathways may accelerate their application in vivo. Fibrin nanoscaffolds have been developed for cartilage tissue engineering and provide a natural matrix that can mimic synovium microenvironment. Moreover fibrin hydrogels allow recreating in vitro the same stiffness chondrocytes face in vivo, leading to improved chondrogenesis differentiation. Besides all these properties, fibrin can be completely degraded by nattokinase, allowing an easy single cell analysis. The aim of this work is to investigate the role of NF-kB inflammatory pathway in MSCs chondrogenesis performed within fibrin nanoscaffolds. Thus we established NF-kB luciferase reporter line to monitor transcription factor activation. Combining degradable fibrin nanoscaffold technology to reporter line for monitoring transcription factors activity will provide a powerful platform to understand the role of inflammation in different stages of chondrogenesis.