RESUMO
Most animal models of infection utilize planktonic bacteria as initial inocula. However, this may not accurately mimic scenarios where bacteria in the biofilm phenotype contaminate a site at the point of injury. We developed a modified CDC biofilm reactor in which biofilms can be grown on the surface of simulated fracture fixation plates. Multiple reactor runs were performed and demonstrated that monomicrobial biofilms of a clinical strain of methicillin-resistant Staphylococcus aureus, S. aureus ATCC 6538, and Pseudomonas aeruginosa ATCC 27853 consistently developed on fixation plates. We also identified a method by which to successfully grow polymicrobial biofilms of S. aureus ATCC 6538 and P. aeruginosa ATCC 27853 on fixation plates. This customized reactor can be used to grow biofilms on simulated fracture fixation plates that can be inoculated in animal models of biofilm implant-related infection that, for example, mimic open fracture scenarios. The reactor provides a method for growing biofilms that can be used as initial inocula and potentially improve the testing and development of antibiofilm technologies.
RESUMO
Aim: Treatment of infected orthopedic implants remains a major medical challenge, involving prolonged antibiotic therapy and revision surgery, and adding a >$1 billion annual burden to the health care system in the US alone. Exposure of metallic implants to alternating magnetic fields (AMF) generates heat that can provide a noninvasive means to target biofilm adhered to the surface. In this study, an AMF system with a solenoid coil was constructed for targeting a metal plate surgically implanted in a sheep model.Methods: A tissue-mimicking phantom of the sheep leg was developed along with simulation model of phantom and the live sheep leg. This was used evaluate heating with the AMF system and to compare experimental results with numerical simulations. Comparative AMF exposures were performed/simulated in these model for feasibility of design, verification, and validation of simulations.Results: The system produced magnetic field strengths up to 12mT and achieved plate temperatures of 65-80 °C within 10-14 s. Single and intermittent AMF exposures of a tissue-mimicking phantom agreed with numerical simulations within 5 °C. Similar agreement between experimental measurements and simulations was also observed in the live sheep metal implant model. The simulations also predicted 2-3 mm of tissue damage using a CEM43 thermal dose model for 1-h AMF exposures targeting 65 °C for pulse delays of 2.5 and 5 mins.Conclusion: This study confirmed that AMF technology can be scaled up to treat implants in a large animal model with the same rates of heating and peak temperatures achieved in prior in vitro studies. Further, numerical simulations provided accurate predictions of the heating produced by AMF on metal implants and surrounding tissues, and can be used to design AMF coils for treating human prosthetic joint implants with more complex geometrical shapes.
