Development of a versatile monitoring technique for real-time protein activity tracking within cellular environment and biomimetic tissue engineering scaffold systems

Abstract

Crowded cellular environments with complex and intricate molecular interactions underpin biological processes. High biological noise is intrinsic within these biological systems, and this poses critical challenges to the in situ detection and measurement of biomolecular and protein activities important to advancing approaches to disease and injury treatment. Currently the techniques available to characterize protein behaviours in living biological systems are highly elaborate and are generally greatly hindered by the high background noise of the cellular environments. Here we present a versatile and straight forward technique for monitoring proteins and protein interactions within cells, based on a novel nano-bio-technology method based on nanoparticles Local Surface Plasmon Resonance (LSPR). High sensitive gold edge coated triangular silver nanostructures (AuTSNP), which exhibit a highly sensitive spectral response to the molecular interactions on their surfaces, are used to probe protein behaviours within complex cellular and tissue regeneration environments. In this work, monitoring of the dynamic behaviour of a critical extracellular protein, Fibronectin (Fn) in its active form, within the presence of bone tissue regeneration scaffolds and living cells (C2C12 myoblasts, MC3T3-E1 pre-osteoblasts) is presented. The excellent sensitivity and straight forward application within cellular environments, demonstrates AuTSNP as powerful new tools to signature protein conformational transitions and monitor essential protein activity.