Conventional UHMWPE particles were generated in total knee replacement simulator and isolated from the test supernatants following previously established protocol.16 (link),17 (link),19 (link) Particles were then washed twice in ethanol (96 and 70%), resuspended in phosphate buffered saline (PBS), and stored in −80°C until used. Electron microscopy showed that most of the particles were spherical with a mean diameter of 0.48 ± 0.10 μm (range 0.26–0.81 μm). Particles did not contain a significant endotoxin contamination as tested with Limulus Amebocyte Lysate Kit (BioWhittaker, Walkersville, MD). Alzet osmotic pumps (Model 2006, Durect Corporation, Cupertino, CA) were loaded either with (a) carrier solution (1% BSA-PBS); (b) carrier solution with 15 mg/ml UHMWPE particles; or (c) carrier solution with UHMWPE particles and 10 μg/ml mouse recombinant IL-4 (R&D Systems, Minneapolis, MN). Six-centimeter long vinyl tubing (Durect Corporation) was prefilled with the appropriate solution, connected to the pumps, and subsequently used to connect the pumps to hollow A-40 titanium rods (6 mm long, 21 g, New England Small Tube, Litchfield, NH).
Protocol detail
Generating and Characterizing UHMWPE Particles for Implant Debris Studies
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Amebocyte lysate
Electron microscopy
Endotoxin
Ethanol
Mouse
Osmotic
Phosphate
Rods
Saline
Titanium
Total knee replacement
Uhmwpe
Vinyl
Corresponding Organization : Orthopaedic Research Laboratories
Other organizations : Helsinki University Hospital, University of Helsinki
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Variable analysis
independent variables
- Presence of UHMWPE particles in the carrier solution
- Presence of mouse recombinant IL-4 in the carrier solution
- Not explicitly mentioned
- Carrier solution (1% BSA-PBS)
- Particle size and morphology (spherical with a mean diameter of 0.48 ± 0.10 μm, range 0.26–0.81 μm)
- Endotoxin contamination (tested with Limulus Amebocyte Lysate Kit)
- Positive control: Carrier solution (1% BSA-PBS) with UHMWPE particles
- Negative control: Carrier solution (1% BSA-PBS) without UHMWPE particles
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