Microbial nanocellulose as an effective lactonase immobilization matrix for enhanced wound healing

Chronic wounds pose a significant challenge due to bacterial biofilm infections and antibiotic resistance. Quorum-quenching enzymes like lactonases offer a promising therapeutic alternative. In this study, bacterial nanocellulose (BNC) is investigated as a biocompatible matrix for immobilization of YtnP lactonase, aiming to increase its efficacy against Pseudomonas aeruginosa. Two methods were employed: covalent cross-linking with glutaraldehyde (BNCL) and coordination with zinc ions (BNCML). Fourier transform infrared spectroscopy confirmed enzyme-matrix interactions, while differential scanning calorimetry and rheological analyses confirmed the stability of the immobilized enzyme. Both BNCL and BNCML preparations retained their activity when reused ten times and showed no significant loss of function after three months at +4 °C, while the free enzyme lost over 40 % of its initial activity. In a mouse wound model infected with P. aeruginosa, treatment with BNCL reduced the number of neutrophils, NK cells, and pro-inflammatory M1 macrophages, while increasing the number of anti-inflammatory M2 macrophages, which accelerated wound closure compared to untreated controls. In contrast, BNCML had a negative effect on healing. These results highlight BNC as a stable and biocompatible matrix for the immobilization of lactonase, making BNCL a promising infection-inhibiting alternative to conventional antimicrobial therapies for the treatment of chronic wounds.