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In vitro evaluation of Pseudomonas aeruginosa chronic lung infection models: Are agar and calcium-alginate beads interchangeable?

Abstract : Animal models of chronic lung infection with P. aeruginosa (PA) are useful tools to improve antibiotic (ATB) treatment. Two main models based on the pulmonary instillation of PA embedded in agar or calcium-alginate beads are currently used. However, these two polymers used to prepare the beads have different properties; for example, agar is a neutral polysaccharide while alginate is anionic. We hypothesized that the effect of an ATB on PA entrapped in agar or calcium-alginate beads depends on its physicochemical properties, including charge, and concentration. To test this hypothesis, PAs were entrapped in agar or calcium-alginate beads dispersed in a growth medium containing either tobramycin (TOB), selected as a cationic ATB, or ciprofloxacin (CIP) selected as a neutral zwitterionic ATB. In vitro, time-kill curves evaluating the efficacy of ATBs over time were performed by measuring the light emitted by a bioluminescent PA for 42 h in the presence of ATB concentrations ranging from 0 to 100 times the MIC. In the presence of CIP, time-kill curves obtained with PA trapped in agar or calcium-alginate beads were comparable, whatever the CIP concentration used. In the presence of TOB, a clear difference was observed between the kill curves obtained with PA embedded in agar or calcium-alginate beads. While PA trapped within agar displayed the same susceptibility than the planktonic one, it was unresponsive to TOB for concentrations up to 1-fold MIC when trapped in calcium-alginate. At 10-fold the TOB's MIC, the luminescence emitted by PA01 in the agar beads was reduced by 95% after 40 h, whereas it returned to the same initial value for PA01 trapped in alginate-based beads. The reduction in TOB efficiency was even greater when alginate-based beads were dispersed in a mucus-simulating medium. These results show that the agar and alginate beads models can be interchangeable only for uncharged ATB, such as CIP, but not for cationic ATB, like TOB. In vitro experiments performed in this study could be a quick way to evaluate the effect of each model on a given ATB before performing animal experiments.
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Submitted on : Wednesday, July 20, 2022 - 2:12:16 PM
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Bruna Gaelzer Silva Torres, Rana Awad, Sandrine Marchand, William Couet, Frédéric Tewes. In vitro evaluation of Pseudomonas aeruginosa chronic lung infection models: Are agar and calcium-alginate beads interchangeable?. European Journal of Pharmaceutics and Biopharmaceutics, Elsevier, 2019, 143, pp.35-43. ⟨10.1016/j.ejpb.2019.08.006⟩. ⟨hal-02477575⟩



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