Physicochemical and Inflammatory Analysis of Unconjugated and Conjugated Bone-Binding Carbon Dots

Carbon nanodots (CDs) have drawn significant attention for their potential uses in diagnostic and therapeutic applications due to their small size, tissue biocompatibility, stable photoluminescence, and modifiable surface groups. However, the effect of cargo molecules on CD photoluminescence and their ability to interact with tissues are not fully understood. Our previous work has shown that CDs produced from the acidic oxidation of carbon nanopowder can bind to mineralized bone with high affinity and specificity in a zebrafish animal model system. Using this model, we investigated the impact of loading Cy5 and biotin cargo on CDs' photoluminescence and bone-binding properties. We report that CD cargo loading alters CD photoluminescence in a pH- and cargo-dependent manner without interfering with the CDs' bone binding properties. In a reciprocal analysis, we show that cargo loading of CDs does not affect the cargo's fluorescence. Significantly, CDs do not trigger nitric oxide production in a mouse macrophage assay, suggesting that they are noninflammatory. Together, these results further support the development of carbon nanopowder-derived CDs for the precise delivery of therapeutic agents to bone tissue.