Development of a novel chitosan-based two-photon fluorescent nanoprobe with enhanced stability for the specific detection of endogenous H2O2

Abstract

Hydrogen peroxide (H₂O₂) acts as an important reactive oxygen species (ROS) and maintains the redox equilibrium in organisms. Imbalance of H₂O₂ concentration is associated with the development of many diseases. Traditional small molecular based fluorescent probes often show drawbacks of cytotoxicity and easily metabolic clearance. Herein, a chitosan-based two-photon fluorescent nanoprobe (DC-BI) was constructed and applied for H₂O₂ detection in live organisms. DC-BI was composed by chitosan nanoparticles and a two-photon fluorophore of naphthalimide analogues (BI) with H₂O₂-responsive property. The structure of DC-BI was characterized by NMR, FTIR, XPS, XRD, DLS and MLS analyses. As study shown, the nanoprobe DC-BI exhibited improved distribution stability and smaller cytotoxicity. In the presence of H₂O₂, both the absorption and emission spectra show dramatic changes, the fluorescence intensity at 580 nm obviously enhanced. Furthermore, fluorescence imaging results indicate that DC-BI is capable of imaging endogenous H₂O₂ in cells and zebrafish. The design and development of chitosan-based nanoprobe DC-BI has provided a general example of nanoprobe construction with excellent distribution stability, two-photon property, and biocompatibility.

Keywords: Chitosan; Hydrogen peroxide; Nanoprobe; Two-photon; Zebrafish.