Reference method for off-line analysis of nitrogen oxides in cell culture media by an ozone-based chemiluminescence detector

Nitric oxide (NO) and its by-products are important biological signals in human physiology and pathology particularly in the vascular and immune systems. Thus, in situ determination of the NO-related molecule (NO x ) levels using embedded sensors is of high importance particularly in the context of cellular biocompatibility testing. However, NO x analytical reference method dedicated to the evaluation of biomaterial biocompatibility testing is lacking. Herein, we demonstrate a PAPA-NONOate-based reference method for the calibration of NO x sensors. After, the validation of this reference method and its potentialities were demonstrated for the detection of the oxidative stress-related NO secretion of vascular endothelial cells in a 3D tissue issued from 3D printing. Such NO x detection method can be an integral part of cell response to biomaterials. Graphical abstract