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) levels using embedded sensors is of high importance particularly in the context of cellular biocompatibility testing. However, NO 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 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 detection method can be an integral part of cell response to biomaterials. Graphical abstract.