Establishment of a human osteocyte model to reveal mechanisms of chronic Staphylococcus epidermidis bone and joint infections
Abstract
Aims: Chronic bone and joint infections caused by Staphylococcus epidermidis can be difficult to treat and are prone to recurrence. In this study, established human osteoblast- and osteocyte-like cell line models were used in vitro to investigate the intracellular persistence of S. epidermidis as a potential mechanism for infection persistence.
Methods: The SaOS2 human osteosarcoma cell line was differentiated into osteoblast-like (SaOS2-OB) and osteocyte-like (SaOS2-OY) stages and infected with four S. epidermidis strains at varying multiplicities of infection. Susceptibilities to lysostaphin, a known bacteriolytic enzyme, and a panel of antibiotics were determined to identify an antimicrobial capable of clearing extracellular bacteria while preserving intracellular bacterial viability. Bacterial survival and number were quantified by colony-forming unit and droplet digital polymerase chain reaction (PCR), while host gene expression was analyzed by real-time PCR.
Results: Lysostaphin was ineffective against all four S. epidermidis strains, while 10 × minimal bactericidal concentration levofloxacin consistently eliminated extracellular but not intracellular bacteria. Intracellular S. epidermidis at multiplicities of infection of 1,000 persisted in SaOS2-OY for 14 days at 10 × minimal bactericidal concentration levofloxacin, while it was cleared within five days in SaOS2-OB cells. Droplet digital PCR genome copy analysis identified a higher number of bacteria than…
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