One of the most important factors that seems to be involved in total hip replacement is periprosthetic osteolysis. As it is well documented that several interleukins (ILs) are triggered in periprosthetic osteolysis, this article investigates the role of five ILs in primary and replacement total hip arthroplasty, understanding if one of them can also predict hip implant loosening, secondary surgery, and prosthesis breakage. The levels of IL-1alpha, 1beta, 6, 8, and 10 in synovial fluid were examined, using a high sensitivity enzyme-linked immunosorbent assay (ELISA) test kit (Pierce Biotechnology, Inc., Rockford, IL, USA) to determine whether these cytokines could be used as markers of enhanced periprosthetic osteolysis, leading to aseptic loosening of total/partial hip arthroplasty or revision surgery. Synovial fluid was harvested from 23 patients undergoing primary total hip arthroplasty and 35 patients undergoing total/partial hip revision due to aseptic loosening. In the revision group, four cases had suffered a prosthesis fracture and five were second revisions. ILs 6 and 8 were significantly higher in the revisions (305 and 817 pg/mL) compared with the primary arthroplasties (151 and 151 pg/mL), including cases with prosthesis fracture and those requiring a second revision. IL-10 levels were lower (not significantly) in second revision samples compared with those of revision samples. IL-1beta levels were significantly higher in prosthesis fracture samples compared with those of all the other revision samples. No statistically significant differences in IL levels were found between osteoarthritis samples and those of other diseases. These results are a step forward to elucidating the complex network of events that are involved in loosening of hip implants.
"The joint fluid samples harvested by arthrocentesis at revision surgery were centrifuged at 2000x g for 10 minutes at room temperature in order to remove cells and other debris. The supernatants aliquoted can be stored until 2 month at - 20ºC . For electrophoresis of proteins and lipoproteins, the viscosity of joint fluid was reduced by using Fluidil SEBIA (25μl Fluidil and 75μl joint fluid), a solution frequently used for dilution of viscous or turbid sera. "
[Show abstract][Hide abstract] ABSTRACT: Rationale:
Prevention of wear-mediated osteolysis, the most common complication in total joint arthroplasty, is a great challenge for orthopedic surgery. Despite the diversity of current biomarkers of periprosthetic osteolysis (products of wear, bone turnover and inflammatory biomarkers), the major interferences and the great amount of sample necessary for analysis limit their use in clinical practice.
The aim of this paper is to present three new electrophoretic methods using Hyrys-Hydrasys SEBIA system that have been used for the first time in Electrophoresis Laboratory of our hospital in the analysis of joint fluid for the prevention of periprosthetic osteolysis in revision arthroplasty.
Methods and results:
Analytical aspects of agarose gel electrophoresis of joint fluid proteins and lipoproteins as well as SDS-agarose gel electrophoresis of joint fluid proteins, their performances and clinical value are presented. The decreased level of albumin and increased level of alpha1 and alpha2 globulins were frequent changes detected on SEBIA electropherograms and good indicator for the presence of an inflammatory reaction generated by particle debris. In addition, a slightly increase of LDL mobility could provide good information about a high oxidative stress. Moreover, the Ig G assessed by using SDS-agarose gel electrophoresis could be a potential biomarker for an immunological reaction towards orthopedic implants.
Electrophoresis of joint fluid using Hyrys-Hydrasys SEBIA France system is a new analytical technique able to remove the most of current biomarkers disadvantages due to the determination of particular proteins (acute phase proteins, albumin, lipoproteins, and immunoglobulins) by using minimal amounts of joint fluid with minor interferences, minimal cost and rapid results.
Journal of medicine and life 09/2013; 6(3):249-253.
[Show abstract][Hide abstract] ABSTRACT: Titanium particles generated from orthopedic and dental implants are suspected to play an important role in peri-implant osteolysis. Elucidation of the mechanisms involved in the effects of titanium particles on osteolysis may provide methods for preventing osteolysis. Therefore, in the present study, we investigated the effect of titanium particles on osteoclast activity in vitro. We evaluated bone resorption pits on dentin slices and osteoclast F-actin rings after treatment with titanium particles. The influence of titanium particles on the expression of TRAP, Cat K and CA II mRNA was also evaluated. We found that osteoclast bone resorption activity was enhanced at a lower concentration of titanium particles, but inhibited by a higher concentration of titanium particles. Titanium particles can be phagocytosed by osteoclasts, but the F-actin ring was not destroyed after phagocytosis. In addition, a lower concentration of titanium particles enhanced the mRNA expression of TRAP and Cat K, while higher concentrations of titanium particles decreased their expression. By contrast, the mRNA expression of CA II was decreased at all concentrations of titanium particles. These results suggest that direct exposure to titanium particles at a low concentration enhances osteoclast activity, while exposure to a high concentration inhibits their activity. This understanding of the direct effect of titanium particles on osteoclast activity suggests that aseptic loosening is caused not only by the generation of osteoclasts, but also by enhanced osteoclast activity.
Molecular Medicine Reports 11/2010; 3(6):1065-9. DOI:10.3892/mmr.2010.368 · 1.55 Impact Factor
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