Abstract
Introduction and Aims: Accumulating evidence suggests that bacterially derived endotoxins may contribute to aseptic loosening. This study determined whether lipopolysaccharide (LPS), the classical endotoxin from Gram-negative bacteria, can be detected in periprosthetic tissue from patients with aseptic loosening. We utilised an assay that detects all forms of LPS and is unaffected by beta-glucan-like molecules.
Method: Periprosthetic tissue from revision total hip arthroplasty and synovia from primary total joint arthroplasty were homogenised in PBS in endotoxin-free conditions. Non-specific amidases in the homogenates were inactivated at 100 degrees C. LPS was measured using the Endospecy assay (Associate of Cap Cod). Multiple dilutions of the homogenates were assayed to maximise sensitivity, while avoiding assay inhibition assessed by spike recovery determinations. Results were corrected for colour and spike recovery. Assay results were considered positive if the absorbances were higher than the lowest standard and the LPS level was significantly greater (p< 0.05) than the PBS control. Statistical analysis was by ANOVA with Bonferroni-Dunn (Control) post-hoc tests.
Results: Samples from 13 patients have been studied to date. Multiple assays of four of these samples showed no detectable LPS while nine of these samples resulted in both positive and negative assays. This inter-assay variability prevents measurement of the concentration of LPS in the samples. Nonetheless, many of the samples contain detectable amount of LPS. Thus, six out of eight samples from revision THA patients with aseptic loosening had positive assays, as did two of four primary TJA patients. LPS was also detected in a sample from a revision control. These results demonstrate that samples from THA patients with aseptic loosening and from primary TJA contain detectable amounts of LPS derived from Gram-negative bacteria.
Conclusion: This conclusion is consistent with numerous studies, showing that human serum contains LPS derived from minor infections, gut flora, or dental procedures. It is likely that many of these samples also contain molecules derived from Gram-positive bacteria that have very similar biological effects as LPS. However, detection of these Gram-positive molecules await further improvements in assay specificity and sensitivity.
These abstracts were prepared by Editorial Secretary, George Sikorski. Correspondence should be addressed to Australian Orthopaedic Association, Ground Floor, The William Bland Centre, 229 Macquarie Street, Sydney, NSW 2000, Australia.
None of the authors is receiving any financial benefit or support from any source.
