Tracking Wound Healing in Real Time: A Multi-Parameter Assessment Using mRNA-Driven DNA Nanospheres with Signal Amplification

Abstract Effective wound management requires accurate assessment of wound status through the whole healing process to support clinical decision-making. However, current methods are limited by weak signals output resulting from insufficient biomarkers and by their inability to monitor multiple indicators of healing throughout the recovery process. In this study, we developed mRNA-responsive DNA nanospheres (DNS) capable of dynamically visualizing and tracking the sequential phases of wound healing. Functional elements were precisely organized into “nanosphere-shaped” delivery vectors using a programmable self-assembly of three Y-shaped modules and hairpin-locked DNAzymes via sequence-specific hybridization. These DNSs are selectively activated by mRNA biomarkers in skin cells, where sensitivity is significantly enhanced by DNAzyme-catalyzed signal amplification. The enhanced biostability and sensing performance of the proposed nanoplatform facilitate real-time monitoring of mRNA signal dynamics during wound progression. In vivo studies demonstrated that the nanoplatform could support real-time staging and differentiation of the healing stages, while also enabling comparative evaluation of normal and diabetic wound healing processes.