A responsive nanoprobe for ratiometric florescence detection of hydroxyl radicals in macrophage polarization†

Abstract

Quantification of hydroxyl radicals (˙OH), one form of reactive oxygen species (ROS), plays critical roles in early diagnosis and treatment monitoring of various diseases. In this work, we report the development of a responsive nanoprobe for ratiometric fluorescence detection and imaging of ˙OH in macrophage polarization. The nanoprobe, BSA-CCA@LDH-SRB, was designed and prepared using coumarin 3-carboxylic acid (CCA) as the sensing unit for ˙OH, and sulforhodamine B (SRB) loaded on layered double hydroxide (LDH) served as the fluorescent reference component. The coupling of CCA to bovine serum albumin (BSA) and the loading of BSA–CCA on the surface of LDH enabled the nanoprobe for fluorescence detection of ˙OH with high sensitivity and minimal interference from other biomolecules, ions, and ROS. The emission of the prepared BSA-CCA@LDH-SRB at 444 nm emerged and the intensity was increased according to the concentration of ˙OH, while the emission at 580 nm was maintained, allowing the nanoprobe for ratiometric fluorescence (F_444/580) detection of ˙OH. Loading of the BSA protein on the LDH surface and the biocompatibility and colloidal stability of the LDH-based fluorescent nanoprobe were further improved, facilitating the detection of ˙OH generation in macrophage polarization stimulated by both biomolecules and physical ultrasound irradiation. This study thus offers a new nanoprobe as the tool for investigating ˙OH evolutions, advancing the biomedical investigations of macrophage polarization associated inflammation.