A stimuli-responsive separating microneedle (MN) system composed of photo-crosslinked GelMA and 5-FuA-Pep-MA prodrug was developed to treat hypertrophic scars (HS) by remodeling the pathological microenvironment. The MNs respond to high reactive oxygen species (ROS) levels and overexpression of matrix metalloproteinases (MMPs) in HS tissues. In vivo experiments in female mice showed that the MN tips remained in the tissue, providing sustained drug release. The drug-loaded MNs reduced fibroblast proliferation and collagen deposition by scavenging ROS and consuming MMPs, while regulating inflammatory responses and keratinocyte differentiation. Bulk and single-cell RNA sequencing revealed that the MNs reversed skin fibrosis by down-regulating BAD and IGF1R pathways, and up-regulating TOLL, IL1R, and keratinocyte pathways. The MNs also promoted interactions between fibroblasts and keratinocytes via HSPG2-DAG1 signaling. The MNs demonstrated effective ROS scavenging and MMP consumption, leading to remodeling of the HS microenvironment. The study highlights the potential of drug-loaded MNs for HS treatment and clinical application. The MNs showed minimal skin irritation and long-term retention, with efficient drug release in response to endogenous stimuli. The results suggest that drug-loaded MNs could inhibit fibroblast over-proliferation and collagen synthesis, offering a safer and more effective treatment for HS compared to traditional methods. The study also emphasizes the role of fibroblasts and keratinocytes in HS pathogenesis and the importance of their interactions in therapeutic mechanisms. The MNs provide a minimally invasive, painless, and efficient treatment option for HS, with potential for long-term clinical use.A stimuli-responsive separating microneedle (MN) system composed of photo-crosslinked GelMA and 5-FuA-Pep-MA prodrug was developed to treat hypertrophic scars (HS) by remodeling the pathological microenvironment. The MNs respond to high reactive oxygen species (ROS) levels and overexpression of matrix metalloproteinases (MMPs) in HS tissues. In vivo experiments in female mice showed that the MN tips remained in the tissue, providing sustained drug release. The drug-loaded MNs reduced fibroblast proliferation and collagen deposition by scavenging ROS and consuming MMPs, while regulating inflammatory responses and keratinocyte differentiation. Bulk and single-cell RNA sequencing revealed that the MNs reversed skin fibrosis by down-regulating BAD and IGF1R pathways, and up-regulating TOLL, IL1R, and keratinocyte pathways. The MNs also promoted interactions between fibroblasts and keratinocytes via HSPG2-DAG1 signaling. The MNs demonstrated effective ROS scavenging and MMP consumption, leading to remodeling of the HS microenvironment. The study highlights the potential of drug-loaded MNs for HS treatment and clinical application. The MNs showed minimal skin irritation and long-term retention, with efficient drug release in response to endogenous stimuli. The results suggest that drug-loaded MNs could inhibit fibroblast over-proliferation and collagen synthesis, offering a safer and more effective treatment for HS compared to traditional methods. The study also emphasizes the role of fibroblasts and keratinocytes in HS pathogenesis and the importance of their interactions in therapeutic mechanisms. The MNs provide a minimally invasive, painless, and efficient treatment option for HS, with potential for long-term clinical use.