Validation of input parameters and the resulting polyethylene damage is essential for knee joint wear simulators to be useful in prospective evaluations. The purpose of this study was to compare damage patterns on polyethylene inserts wear tested on a knee simulator with inserts retrieved after well-functioning total knee arthroplasty (TKA). Five polyethylene tibial inserts from a PCL-retaining knee prosthesis (Natural Knee) were wear tested on an Instron/Stanmore simulator in 50% bovine serum. The input consisted of ISO 14243 force-controlled testing standard to simulate human gait for 5 million cycles. Nine polyethylene tibial inserts (Natural Knee) were retrieved from patients after 52+45 months (13-124) of successful function. The inserts were retrieved post-mortem (n=7) and for pain (n=2). Articular damage was assessed and the circumference of each damage region digitized. The damage size, location and linear surface deformation were measured and the deformation rate (mm/106 cycles or mm/year) was calculated. The linear deformation rate for all inserts decreased considerably with time. Lateral damage was located significantly more posterior than the medial damage on both the simulated and retrieved inserts, corresponding to femoral external rotation. Retrieved inserts had larger lateral damage, whereas simulator inserts had larger medial damage. The AP extent of damage on the retrieved inserts was significantly greater than the extent on the simulator inserts. Three retrieved inserts had substantial delamination, whereas none of the simulator inserts had delamination. There was good agreement in the deformation rates for the simulator and retrieved inserts. However, retrieved inserts with delamination show an increased deformation rate and this type of damage did not occur on simulator inserts. The greater AP extent of damage and larger lateral damage on the retrieved inserts suggest that in vitro wear simulation should perhaps include a more complete range of patient activity dynamics to better predict in vivo damage.