Objectives: Osteomyelitis, marked by bone inflammation due to microbial infection, presents significant healthcare challenges. While the protective role of biofilm in bacterial immunity and persistence is well-documented, its direct impact on bone degradation in osteomyelitis remains inadequately characterized. This study aims to comprehensively examine the direct effects of biofilm-forming pathogens on human bone, providing new insights into the mechanisms of bone destruction in osteomyelitis. Methods: Bone sections were collected from patients undergoing total knee replacement surgery between January 2021 and December 2022 for this study at Sultan Qaboos University Hospital. Then samples were inoculated with Staphylococcus aureus to simulate in-vitro bone infection. We then used Raman spectroscopy and Scanning Electron Microscopy (SEM) for detailed analysis of the bacterial interaction with bone tissue. Results: SEM images depicted trabecular destruction caused by biofilm. Biofilm-forming pathogens contribute directly to bone exhibited by SEM images with marked trabecular destruction. Raman analysis showed a significant increase in the carbonate-phosphate ratio in inoculated samples (619%) compared to controls (47%). Mineral content decreased in inoculated samples, and the carbonate-to-amide I ratio reduced by 47% in inoculated and 80% in controls. The inoculated samples exhibited an 82% shift in collagen crosslinking compared to 72% in controls. Conclusion: This research enhances our comprehension of the mechanisms behind bone destruction in osteomyelitis and underscores the intricate role of biofilm in the disease's development. These findings highlight the importance of biofilm in bone degradation and its potential implications for managing infections.
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