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Emerging electron microscopy and 3D methodologies to interrogate Staphylococcus aureus osteomyelitis in murine
Karen L de Mesy Bentley1,2,3, Chad A Galloway2, Gowrishankar Muthukrishnan1,3
1Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USA.
Summary
Advanced transmission electron microscopy reveals Staphylococcus aureus colonization within bone's microscopic networks. Surprisingly, some networks resist infection, suggesting osteocyte defense mechanisms against chronic osteomyelitis.
Area of Science:
- Orthopaedic research
- Microbial pathogenesis
- Advanced imaging techniques
Background:
- Chronic osteomyelitis is a persistent bone infection.
- Staphylococcus aureus invades osteocyte-lacuno canalicular networks.
- Understanding microbial invasion mechanisms is crucial for developing new treatments.
Purpose of the Study:
- To detail protocols for preparing bone tissue for transmission electron microscopy (TEM).
- To apply immunoelectron microscopy and 3D reconstruction to study Staphylococcus aureus in bone.
- To investigate Staphylococcus aureus colonization within the osteocyte-lacuno canalicular network.
Main Methods:
- Detailed protocols for bone tissue fixation, decalcification, and epoxy embedment for TEM.
- Immunoelectron microscopy to confirm bacterial presence in canaliculi.
- Automated-Tape-UltraMicrotome system for 3D reconstruction and volumetric analysis.
Main Results:
- Established protocols for high-resolution TEM imaging of bone infections.
- Confirmed Staphylococcus aureus presence within sub-micron canaliculi.
- 3D reconstruction revealed Staphylococcus aureus colonization patterns and adjacent uninfected canaliculi.
Conclusions:
- Osteocyte-lacuno canalicular networks are sites of Staphylococcus aureus colonization in chronic osteomyelitis.
- The presence of uninfected canaliculi suggests potential osteocyte resistance mechanisms.
- Further research is needed to understand osteocyte responses and the persistence of bone infections.
