Abstract
Biofilm-associated tissue and device infection is a major threat to therapy. The present work aims to potentiate β-lactam antibiotics with biologically synthesized copper oxide nanoparticles. The synergistic combination of amoxyclav with copper oxide nanoparticles was investigated by checkerboard assay and time-kill assay against bacteria isolated from a burn wound and a urinary catheter. The control of biofilm formation and extracellular polymeric substance production by the synergistic combination was quantified in well plate assay. The effect of copper oxide nanoparticles on the viability of human dermal fibroblasts was evaluated. The minimum inhibitory concentration and minimum bactericidal concentration of amoxyclav were 70 μg/mL and 140 μg/mL, respectively, against Proteus mirabilis and 50 μg/mL and 100 μg/mL, respectively, against Staphylococcus aureus. The synergistic combination of amoxyclav with copper oxide nanoparticles reduced the minimum inhibitory concentration of amoxyclav by 16-fold against P. mirabilis and 32-fold against S. aureus. Above 17.5 μg/mL, amoxyclav exhibited additive activity with copper oxide nanoparticles against P. mirabilis. The time-kill assay showed the efficacy of the synergistic combination on the complete inhibition of P. mirabilis and S. aureus within 20 h and 24 h, respectively, whereas amoxyclav and copper oxide nanoparticles did not inhibit P. mirabilis and S. aureus until 48 h. The synergistic combination of amoxyclav with copper oxide nanoparticles significantly reduced the biofilm formed by P. mirabilis and S. aureus by 85% and 93%, respectively. The concentration of proteins, carbohydrates, and DNA in extracellular polymeric substances of the biofilm was significantly reduced by the synergistic combination of amoxyclav and copper oxide nanoparticles. The fibroblast cells cultured in the presence of copper oxide nanoparticles showed normal morphology with 99.47% viability. No cytopathic effect was observed. Thus, the study demonstrated the re-potentiation of amoxyclav by copper oxide nanoparticles.
Highlights
Biofilm is a community of bacteria that survives like multicellular organisms
The main goal of the present study is to evaluate the synergistic activity of CuO NPs with
Patient bacteriain bacteria isolated from a burn wound and a urinary catheterinofclinical a hospitalized were isolated from a burn wound and a urinary catheter of a hospitalized patient in Sivakasi, Sivakasi, India’s fireworks headquarters
Summary
Biofilm is a community of bacteria that survives like multicellular organisms. The biofilm phenotype is different from planktonic cells [1]. Biofilm forms through a complex cascade of events that encapsulate bacteria within self-assembled extracellular polymeric substances (EPS) [2]. 65% to 95% of biofilm is composed of water. The EPS of biofilm are made up of proteins (≥2%), carbohydrates (1–2%), and DNA (≤1%) [3]. Bacteria inside the biofilm are resistant to external stress and evade the host immune system [5]. The therapeutic failure of antibiotics in the treatment of tissue and medical device-associated infections is mainly due to persistent biofilm formation. About 80% of recalcitrant infections are due to biofilm [6]
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