Competitive Binding of Bilirubin and Fatty Acid on Serum Albumin Affects Wear of UHMWPE
Total Joint Replacement (TJR) devices undergo standardized wear testing in mechanical simulators while submerged in a proteinaceous testing solution to mimic the environmental conditions of artificial joints in the human body. Typically, bovine calf serum is used to provide the required protein content. However, due to lot-to-lot variability, an undesirable variance in testing outcome is observed. Based on an earlier finding that yellowish-orange serum color saturation is associated with wear rate, we examined potential sources of this variability, by running a comparative wear test with bilirubin; hemin; and a fatty acid, oleic acid, in the lubricant. All these compounds readily bind to albumin, the most abundant protein in bovine serum. Ultrahigh molecular weight polyethylene (UHMWPE) pins were articulated against CoCrMo discs in a pin-on-disc tribometer, and the UHMWPE wear rates were compared between lubricants. We found that the addition of bilirubin increased wear by 121%, while hemin had a much weaker, insignificant effect. When added at the same molar ratio as bilirubin, the fatty acid tended to reduce wear. Additionally, there was a significant interaction with respect to bilirubin and hemin in that UHMWPE wear rate decreased with increasing fatty acid concentration. We believe the conformational change in albumin by binding bilirubin makes it more likely to form molecular bridges between UHMWPE and the metal counterface, thus increasing adhesive wear. However, fatty acids compete for binding sites on albumin, and can prevent this conformational change. Hence, the protein is stabilized, and the chance for albumin to form bridges is lowered. Ultimately, UHMWPE wear rate is driven by the competitive binding of bilirubin and fatty acid to albumin.
Albumin Protein Cleavage Affects the Wear and Friction of Ultra-High Molecular Weight Polyethylene
It is well established that the total protein concentration and albumin-to-globulin ratio influence the wear of ultra-high molecular polyethylene (UHMWPE, "polyethylene") in joint prostheses. A factor on wear not yet studied, but of possible clinical relevance, is protein cleavage. Such cleavage is expected in the presence of an inflammatory response and as a result of wear processes at the articular interface. The aim of this study was to compare the tribological behavior of polyethylene articulated against an orthopedic wrought CoCrMo alloy for three lubricants: cleaved albumin, uncleaved albumin, and newborn calf serum (control). We hypothesized that the cleavage of albumin will increase the friction and wear rate of polyethylene, with a concomitant roughening of the polymer surface and the generation of larger wear debris particles. Cleavage of the bovine albumin into five fragments was performed by digestion with cyanogen bromide. In pin-on-flat (POF) wear tests of polyethylene pins made of Ticona GUR 1020/1050 against CoCrMo alloy discs, the cleaved albumin led to the lowest polyethylene wear and highest friction coefficients, whereas albumin led to the highest wear rates. In knee simulator tests, the albumin lubricant also led to a 2.7-fold increase in the tibial insert wear rate compared to the regular bovine serum lubricant (a wear rate for the cleaved albumin could not be obtained). The generated polyethylene wear particles were of increasing size and fibrillar shape in going from serum to albumin to cleaved albumin, although only the shape achieved statistical significance. Unlike bovine serum, cleaved albumin led to wear scars for both the POF and simulator wear tests that closely emulated the morphological features observed on explanted polyethylene tibial inserts from total knee replacements. We posit that the smaller protein fragments can more efficiently adsorb on the surfaces of both the polyethylene and the metal, thus offering protection against wear, while at the same time leading to an increase in friction, particle size, and particle elongation, as the protein fragment films interact adhesively during sliding. The results of this study have implications for pre-clinical wear testing methodology as they suggest that albumin concentration may be more pertinent than total protein concentration for wear testing polyethylene.
Phospholipid Vesicles in Media for Tribological Studies against Live Cartilage
Pre-clinical testing of hemiarthroplasty devices requires that the tribological conditions present in vivo with live cartilage be closely duplicated. A current limitation in the tribological testing of live cartilage involves the use of cell-culture media as lubricant.