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Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_23 | Pages 48 - 48
1 Dec 2016
Sophie T Dupieux C Camus C Chidiac C Lustig S Ferry T Laurent F Valour F
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Aim

Intracellular persistence of S. aureus is believed to be one of the major mechanisms leading to bone and joint infection (BJI) chronicity and relapses. Despite its poor intracellular activity, daptomycin (DAP) is increasingly used in the treatment of staphylococcal BJI. The well-known in vitro synergy of daptomycin with various betalactam antibiotics consequently led us to investigate whether these combinations enhance the activity of daptomycin against the intracellular reservoir of methicillin-susceptible (MSSA) and -resistant (MRSA) S. aureus in an ex-vivo model of human osteoblast infection.

Method

Osteoblastic MG63 cells were infected for 2h with MSSA strain or its isogenic MRSA. After killing the remaining extracellular bacteria with lysostaphin, infected cells were then incubated for 24h with DAP, oxacillin (OXA) or ceftaroline (CPT) alone or in combination, at the intraosseous concentrations reached with standard human therapeutic doses. Intracellular bacteria were then quantified by plating cell lysates. Minimum inhibitory concentrations (MICs) of these molecules alone and in combination were determined using the checkerboard method at pH7, but also at pH5 to mimic intracellular conditions.