Oxygen gradient induced organization of skeletal tissue engineering constructs.

The spatial distribution and orientation of cells within the growth plate result in a highly organized tissue structure that provides a beneficial and robust environment for new bone formation. The bone forming potential has inspired researchers to re-create this structured environment and explore the use of growth plate-like intermediates in skeletal tissue engineering. Clear clues are available that oxygen plays an essential role in shaping the growth plate, however the exact mechanisms and contribution to new bone formation are largely unknown. This study aims to use oxygen as an instructive guide for the production of spatially organized spheroid-containing constructs that can be applied as skeletal tissue intermediates. It envisions the application of biosensors for measurement of oxygen and oxygen-related pathway activation, and their combination with advanced light sheet microscopy. Instructive oxygen gradients will be generated by integrating a microfluidics gradient generator in the microscopy setup. The pathway activation patterns and induced cellular responses will provide crucial information that helps unraveling the mechanisms by which spatial organization of constructs can contribute to new bone formation, and that is crucial for the rational design of new tissue engineering strategies.