Collagen – hyaluronan biomimetic hybrid networks with embedded fluorescent force sensors
Vinod Subramaniam a, Galja Pletikapic a, Gijsje Koenderink a
a Center for Nanophotonics, AMOLF, The Netherlands, Science Park, 104, Amsterdam, Netherlands
Proceedings of New Advances in Probing Cell-ECM Interactions (CellMatrix)
Berlin, Germany, 2016 October 20th - 21st
Organizers: Ovijit Chaudhuri, Allen Liu and Sapun Parekh
Oral, Galja Pletikapic, presentation 009
Publication date: 25th July 2016

Collagen and proteoglycans are the main components of the extracellular matrix and are thus frequently used in tissue engineering and medical applications. Here we combine collagen type II and hyaluronan, two key constituents of articular cartilage, in a composite network. While collagen forms stiff fibers and serves as a structural backbone providing tensile strength to the cartilage, hyaluronic acid binds water and facilitates swelling. Our goal is to relate the unique macroscopic mechanical properties of such networks (for example high water content and swelling pressure without syneresis) to the balance of local forces between the molecular components. We measured the macroscopic mechanical properties by rheology in combination with various structural techniques (EM, AFM, confocal microscopy, FRAP, and turbidity) to characterize fibril self-assembly, bundling, and network formation across nano- to micrometer scales. To measure the local forces at the molecular level, we are developing a fluorescent force sensor whose FRET signal changes in response to mechanical stimuli such as gel swelling or stretching. We will present first results obtained with this FRET based force sensor, which is based on a DNA hairpin and a Cy3/Cy5 FRET pair. Further, we will employ embedded fluorescent viscosity sensor to probe changes in the local viscosity upon perturbations (i.e. swelling) 



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