Longitudinal intravital imaging of the bone marrow for analysis of the race for the surface in a murine osteomyelitis model.

Autor: Xie C; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA., Ren Y; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA., Weeks J; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA., Rainbolt J; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Kenney HM; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Xue T; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Allen F; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Shu Y; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Tay AJH; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Lekkala S; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA., Yeh SA; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA., Muthukrishnan G; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA., Gill AL; Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USA., Gill SR; Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USA., Kim M; Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USA., Kates SL; Department of Orthopaedic Surgery, Virginia Commonwealth University, Richmond, Virginia, USA., Schwarz EM; Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, USA.
Jazyk: angličtina
Zdroj: Journal of orthopaedic research : official publication of the Orthopaedic Research Society [J Orthop Res] 2024 Mar; Vol. 42 (3), pp. 531-538. Date of Electronic Publication: 2023 Oct 17.
DOI: 10.1002/jor.25716
Abstrakt: Critical knowledge gaps of orthopedic infections pertain to bacterial colonization. The established dogma termed the Race for the Surface posits that contaminating bacteria compete with host cells for the implant post-op, which remains unproven without real-time in vivo evidence. Thus, we modified the murine longitudinal intravital imaging of the bone marrow (LIMB) system to allow real-time quantification of green fluorescent protein (GFP+) host cells and enhanced cyan fluorescent protein (ECFP+) or red fluorescent protein (RFP+) methicillin-resistant Staphylococcus aureus (MRSA) proximal to a transfemoral implant. Following inoculation with ~10 5 CFU, an L-shaped metal implant was press-fit through the lateral cortex at a 90° angle ~0.150 mm below a gradient refractive index (GRIN) lens. We empirically derived a volume of interest (VOI) = 0.0161 ± 0.000675 mm 3 during each imaging session by aggregating the Z-stacks between the first (superior) and last (inferior) in-focus LIMB slice. LIMB postimplantation revealed very limited bacteria detection at 1 h, but by 3 h, 56.8% of the implant surface was covered by ECFP+ bacteria, and the rest were covered by GFP+ host cells. 3D volumetric rendering of the GFP+ and ECFP+ or RFP+ voxels demonstrated exponential MRSA growth between 3 and 6 h in the Z-plane, which was validated with cross-sectional ex vivo bacterial burden analyses demonstrating significant growth by ~2 × 10 4 CFU/h on the implant from 2 to 12 h post-op (p < 0.05; r 2  > 0.98). Collectively, these results show the competition at the surface is completed by 3 h in this model and demonstrate the potential of LIMB to elucidate mechanisms of bacterial colonization, the host immune response, and the efficacy of antimicrobials.
(© 2023 Orthopaedic Research Society.)
Databáze: MEDLINE
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