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 (NOx) levels using embedded sensors is of high importance particularly in the context of cellular biocompatibility testing. However, NOx 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 NOx 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 NOx detection method can be an integral part of cell response to biomaterials.

Automated summary

Nitric oxide (NO) and its by-products are important biological signals in human physiology and pathology, particularly in the vascular and immune systems. In situ determination of NO-related molecule (NOx) levels using embedded sensors is important for cellular biocompatibility testing, but a dedicated NOx analytical reference method is lacking. A novel PAPA-NONOate-based reference method has been demonstrated for NOx sensor calibration, showing potential for detection of oxidative stress-related NO secretion in vascular endothelial cells in 3D tissue from 3D printing.