Ratiometric Fluorescent Microgels for Sensing Extracellular Microenvironment pH during Biomaterial Degradation

[Image: see text] Bone regeneration has attracted extensive attention in the field of regenerative medicine. The influence of biomaterial on the extracellular environment is important for regulating the biological functions of cells for tissue regeneration. Among the various influencing factors, we had previously demonstrated that the extracellular pH value in the local microenvironment during biomaterial degradation affected the balance of bone formation and resorption. However, there is a lack of techniques for conveniently detecting the pH of the extracellular environment. In light of the development of fluorescent pH-sensing probes, herein, we fabricated a novel ratiometric fluorescent microgel (F-MG) for real-time and spatiotemporal monitoring of microenvironment pH. F-MGs were prepared from polyurethane with a size of around 75 μm by loading with pH-sensitive bovine serum albumin nanoparticles (BNPs) and pH-insensitive Nile red as a reference. The pH probes exhibited reversible fluorescence response to pH change and worked in a linear range of 6–10. F-MGs were biocompatible and could be used for long-term pH detection. It could be used to map interfacial pH on biomaterials during their degradation through pseudocolored images formed by the fluorescence intensity ratio between the green fluorescence of BNPs and the red fluorescence of Nile red. This study provided a useful tool for studying the influence of biomaterial microenvironment on biological functions of surrounding cells.