In vitro Bioluminescence Used as a Method for Real-Time Inhibition Zone Testing for Antibiotic-Releasing Composites
Noora M. Mannisto
Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 10, 33720 Tampere, Finland.
Niina Ahola *
Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 10, 33720 Tampere, Finland and Department of Electronics and Communications Engineering, Tampere University of Technology, Korkeakoulunkatu 3, 33720 Tampere, Finland.
Matti T. Karp
Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 10, 33720 Tampere, Finland.
Minna Veiranto
Bioretec Ltd., Hermiankatu 22, 33720 Tampere, Finland.
Minna Kellomaki
Department of Electronics and Communications Engineering, Tampere University of Technology, Korkeakoulunkatu 3, 33720 Tampere, Finland and BioMediTech, Tampere, Finland.
*Author to whom correspondence should be addressed.
Abstract
Aims: This study describes the potential of real-time bioluminescence imaging in evaluating the antibiotic efficiency of two cylinder-shaped bioabsorbable antibiotic-releasing composites by in vitro inhibition zone tests. The bacterial infections of bone tissue can cause extensive hard and soft tissue damage and decrease the efficiency of oral antibiotic therapy due to the poor blood circulation in the infected area. To overcome this problem, new, locally antibiotic-releasing biodegradable composites have been developed.
Study Design & Methodology: The two composites evaluated in this study were composed of poly(L-lactide-co-ε-caprolactone) matrix, β-tricalcium phosphate ceramic and either ciprofloxacin or rifampicin antibiotic. The composites were tested with genetically modified model pathogens of osteomyelitis (Pseudomonas aeruginosa and Staphylococcus epidermidis) in vitro in inhibition zone tests using a method of real-time bioluminescence.
Results: The first signs of the effect of the released ciprofloxacin or rifampicin became visible after four hours of incubation and were seen as changed bioluminescence around the composite pellet on a culture dish. Both of the composite types showed excellent effects against the sensor bacteria within the diffusion area. Bioluminescence measurements suggested that no survivor bacteria capable of evolving resistant strains were left inside the inhibition zones. The S. epidermidis bacterial strain was an inhibition sensor and P. aeruginosa was a stress sensor.
Conclusion: These results highlight the potential of the composite materials against the pathogens of osteomyelitis. The approach allows continuous visual inspection of the efficacy of the antibiotics against the bacteria.
Keywords: Bioluminescence, ciprofloxacin, controlled drug delivery, inhibition zone, rifampicin