Spatially Separated Dual-Probiotic Hydrogel Enables Synergistic Infection Control and Tissue Healing

Abstract

Bacterial infection and oxidative stress are major barriers to wound healing, while conventional antibiotic therapies are constrained by cytotoxicity and antimicrobial resistance. Probiotic-based strategies present safer alternatives, yet most current efforts largely rely on single-strain systems with limited functionality, as multi-strain combinations risk antagonism in complex wound environments. Herein, a dual-probiotic microsphere hydrogel is reported fabricated via parallel microfluidic technology, where Lactobacillus reuteri and Lactobacillus fermentum are individually encapsulated into distinct microspheres and integrated within a hydrogel matrix. By leveraging spatial separation and functional complementarity, the hydrogel achieved outstanding antibacterial efficacy of approximate to 97.8% against Gram-negative E. coli and approximate to 98.5% against Gram-positive S. aureus, together with strong antioxidant activity that reduces intracellular ROS by up to approximate to 83.9%. Notably, these multifunctional effects are realized without the use of any conventional biochemical antibiotics or antioxidants. In a full-thickness infected wound model, the dual-probiotic hydrogel markedly accelerated wound closure and enhanced tissue regeneration, outperforming single-strain controls, with complete closure achieved as early as day 8. This work demonstrates, for the first time, the full exploitation of dual probiotic synergy to achieve potent antibacterial, antioxidant, and wound-healing performances, highlighting a promising therapeutic platform for managing complex wound environments.