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Dandelion herb-derived carbon dots (DH-CDs) have been developed as a biocompatible antimicrobial agent with potential for wound healing. These nanoparticles exhibit broad-spectrum antibacterial activity against S. aureus and E. coli, with minimum inhibitory concentrations of 62.5 μg/mL at pH 7.4 and 31.25 μg/mL at pH 5.5. They disrupt bacterial membranes, inhibit energy metabolism, generate intracellular ROS, and inactivate antioxidant enzymes. DH-CDs can completely disrupt mature S. aureus biofilms at 300 μg/mL and show excellent biocompatibility, with a 10% hemolysis concentration exceeding 4000 μg/mL and over 95% viability of NIH 3T3 fibroblasts at 2000 μg/mL. They uniquely modulate ROS, eradicating bacteria while preserving mammalian cell redox homeostasis. In vivo, DH-CDs significantly accelerate wound healing in a murine model by promoting re-epithelialization, granulation tissue maturation, collagen deposition, and hair follicle regeneration, outperforming commercial wound healing sprays. These findings suggest DH-CDs as a promising candidate for treating bacteria-infected wounds.