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Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds

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Antimicrobial activity against intraosteoblastic Staphylococcus aureus.

Florent Valour¹, Sophie Trouillet-Assant², Natacha Riffard²

¹Infectious Diseases Department, Hospices Civils de Lyon, Lyon, France INSERM U1111, International Centre for Research in Infectiology, Université Claude Bernard Lyon 1, Lyon, France florent.valour@chu-lyon.fr.

Antimicrobial Agents and Chemotherapy

|January 22, 2015

Summary

Ofloxacin demonstrated superior intracellular activity against Staphylococcus aureus in osteoblasts, effectively limiting small-colony variant (SCV) emergence. This finding supports using fluoroquinolones for bone and joint infection (BJI) treatment.

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Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

Published on: January 17, 2025

Area of Science:

Microbiology
Pharmacology
Orthopedics

Background:

Staphylococcus aureus intracellular persistence in osteoblasts, particularly as small-colony variants (SCVs), contributes to relapsing bone and joint infections (BJIs).
Current BJI treatment strategies do not adequately consider the intracellular activity of antimicrobials against bacteria within osteoblasts.

Purpose of the Study:

To evaluate the intraosteoblastic activity of antistaphylococcal antimicrobials.
To assess the impact of these antimicrobials on the intracellular emergence of SCVs in an ex vivo osteoblast infection model.

Main Methods:

Osteoblastic MG63 cells were infected with S. aureus.
Following extracellular bacterial killing, infected cells were treated with various antimicrobials at therapeutic intraosseous concentrations.
Intracellular bacteria and SCVs were quantified via cell lysate plating.

Main Results:

Several antimicrobials, including fosfomycin, linezolid, tigecycline, oxacillin, rifampin, ofloxacin, and clindamycin, exhibited significant bactericidal effects intracellularly.
Ofloxacin, daptomycin, and vancomycin were effective in limiting intracellular SCV emergence.
Ofloxacin uniquely combined potent intracellular bactericidal activity with significant SCV emergence limitation.

Conclusions:

Ofloxacin is a promising agent for BJI management due to its strong intracellular activity and ability to prevent SCV formation.
These findings suggest refining antibiotic choices for BJIs, potentially combining intracellularly active agents like fluoroquinolones with anti-biofilm drugs such as rifampin.