Therapeutic innovation in Prosthetic Joint Infections: A biofilm-disrupting and drug-delivering hydrogel incorporating an enzymatic cocktail

Prosthetic joint infections pose a major challenge in orthopedic surgery, affecting 0.5–2% of patients following hip or knee arthroplasty. Their management is complicated by the rapid formation of a bacterial biofilm, a protective matrix that limits antibiotic diffusion and undermines treatment efficacy. In this context, novel approaches specifically targeting the biofilm are essential to improve therapeutic outcomes. Building on prior in vitro work demonstrating the benefit of combining a tri‐enzymatic cocktail with an antibiotic, but lacking in vivo validation, this doctoral research set out to explore a strategy that integrates enzymes, antibiotic, and a thermosensitive hydrogel to enhance in situ retention. We coformulated these components in a poloxamer 407 hydrogel, which undergoes sol–gel transition at 37 °C to allow minimally invasive administration and prolonged residence at the implant site. Two formulation strategies were developed: the first, yielded promising results but produced only a transient effect in vivo; the second, employing a double‐injection regimen, achieved undetectable bacterial loads both in vitro and in vivo. In a preclinical model, local injection maintained high antibiotic concentrations while minimizing systemic exposure and local complications. These findings validate a dual‐action concept, enzymatic biofilm disruption coupled with controlled antibiotic delivery and pave the way for optimizing new combinations and designing relevant protocols for a pilot clinical trial.