ZnFe Layered Double Hydroxide Nanosheets Loaded with Cu Single-Atom Nanozymes with Multi-Enzyme-Like Catalytic Activities as an Effective Treatment for Bacterial Keratitis

Abstract

Bacterial keratitis (BK) is a type of corneal inflammation resulting from bacterial infection in the eye. Although nanozymes have been explored as promising materials in corneal wound healing, currently available nanozymes lack sufficient catalytic activity and the ability to penetrate bacterial biofilms, limiting their efficacy against the treatment of BK. To remedy this, ZnFe layered double hydroxide (ZnFe-LDH) nanosheets are loaded with Cu single-atom nanozymes (Cu-SAzymes) and aminated dextran (Dex-NH₂), resulting in the formation of the nanozyme DT-ZnFe-LDH@Cu, which possesses peroxidase (POD)-, oxidase (OXD)-, and catalase (CAT)-like catalytic activities. This enables the nanozyme to generate reactive oxygen species (ROS), such as hydroxyl radicals (^•OH), superoxide anion radical (O₂^•−), and singlet oxygen (¹O₂) from hydrogen peroxide (H₂O₂), thereby killing the bacteria causing the infections. The surface Dex-NH₂ enabled the DT-ZnFe-LDH@Cu to penetrate the biofilm and adsorb onto extracellular polymeric substances (EPS) produced by bacteria in the biofilm. Additionally, the DT-ZnFe-LDH@Cu successfully repaired P. aeruginosa-infected corneal injury in a BK rabbit model more effectively than commercially available tobramycin eye drops. This was enabled, in part, by the ability of DT-ZnFe-LDH@Cu to reduce inflammation by promoting the polarization of pro-inflammatory macrophages (M1) to anti-inflammatory macrophages (M2) and decrease the expression of α-smooth muscle actin (α-SMA) to promote wound healing without scar formation. This study provides an innovative concept for the treatment of BK and holds great scientific value and clinical application potential.