Ultrasound-assisted preparation of shikonin-loaded emulsions for the treatment of bacterial infections

Bacteria can encapsulate themselves in a self-generated matrix of hydrated extracellular polymeric substances such as polysaccharides, proteins, and nucleic acids, thereby forming bacterial biofilm infections. These biofilms are drug resistant and will diminish the efficacy of antimicrobial agents, rendering treatment of such infections challenging. Herein, an innovative strategy is proposed to synergistically degrade bacterial biofilms and eradicate the entrapped bacteria through integrating alpha-amylase (alpha-Amy), shikonin (SK) and epigallocatechin gallate (EGCG) within an emulsion. The natural protein alpha-Amy is deployed to enzymatically hydrolyze the polysaccharide of biofilms. Due to the amphipilic properties of alpha-Amy and the cross-linking capability of EGCG, the formed alpha-Amy/SK@EGCG emulsion possess high stability. SK was encapsulated within the emulsion through ultrasound-assisted assembly, targeting to treat bacterial infection after biofilm degradation. In vitro and in vivo experiments demonstrate that the polyphenol-protein stabilized emulsion loaded with antibacterial SK achieves profound penetration into the biofilms due to the extracellular polysaccharide hydrolysis mediated by alpha-Amy. As a result, the alpha-Amy/SK@EGCG emulsion can significantly alleviate inflammation symptoms and accelerate the healing process of biofilm-infected wounds. This study provides a promising therapeutic strategy for the development of novel materials aimed for the enhanced treatment of bacterial biofilm infections.